S. Sharma

Plants respond to heavy metal stress by metal complexation process like production of phytochelations or by other metal chelating peptides. In this paper we report the synthesis of silver nanoparticles (AgNPs) from the room dried stem and root of Ocimum sanctum. The broth of the plant is used as a reducing agent for the synthesis of Ag nanoparticles at room temperature. The reaction process was simple and was monitored by ultraviolet-visible spectroscopy (UV-vis). There was formation of highly stable silver nanoparticles in the solution. The morphology and crystalline phase of the NPs were determined from transmission electron microscopy (TEM), selected area electron diffraction (SAED) and X-ray diffraction (XRD) spectra. Transmission Electron Microscopy studies showed that the silver nanoparticles obtained from roots and stem were of sizes 10+/-2 and 5+/-1.5 nm, respectively. The various phytochemicals present within the ocimum plant result in effective reduction of silver salts to nanoparticles but their chemical framework is also effective at wrapping around the nanoparticles to provide excellent robustness against agglomeration.

A single-step environmental friendly approach is employed to synthesize silver nanoparticles. The biomolecules found in plants induce the reduction of Ag + ions from silver nitrate to silver nanoparticles (AgNPs). UV-visible spectrum of the aqueous medium containing silver ions demonstrated a peak at 425 nm corresponding to the plasmon absorbance of silver nanoparticles. Transmission electron microscopy (TEM) showed the formation of well-dispersed silver nanoparticles in the range of 5–20 nm. X-ray diffraction (XRD) spectrum of the AgNPs exhibited 2θ values corresponding to the silver nanocrystal. The process of reduction is extracellular and fast which may lead to the development of easy biosynthesis of silver nanoparticles. Plants during glycolysis produce a large amount of H + ions along with NAD which acts as a strong redoxing agent; this seems to be responsible for the formation of AgNPs. Water-soluble antioxidative agents like ascorbic acids further seem to be responsible for the reduction of AgNPs. These AgNPs produced show good antimicrobial activity against common pathogens.

The biosynthesis of nanoparticles has been proposed as a cost effective and environmental friendly alternative to chemical and physical methods.Plant mediated synthesis of nanoparticles is a green chemistry approach that intercomnects nanotechnology and plant biotechnology.In the present study, synthesis of silver nanoparticles (AgNPs) or (Green-Silver) has been demonstrated using extracts of Ananas comosus reducing aqueous silver nitrate.The AgNPs were characterized by Ultraviolet-Visible (UV-vis) Spectrometer, Energy Dispersive X-ray Analysis (EDAX), Selected Area Diffraction Pattern (SAED) and High Resolution Transmission Electron Microscopy (HRTEM).TEM micrographs showed spherical particles with an average size of 12 nm.The XRD pattern showed the characteristic Bragg peaks of ( 111), ( 200), ( 220) and ( 311) facets of the face center cubic (fcc) silver nanoparticles and confirmed that these nanoparticles are crystalline in nature.The different types of antioxidants presented in the pineapple juice synergistically reduce the Ag metal ions, as each antioxidant is unique in terms of its structure and antioxidant function.The reaction process was simple for formation of silver nanoparticles and AgNPs presented in the aqueous medium were quite stable, even up to 4 months of incubation.This work proved the capability of using biomaterial towards the synthesis of silver nanoparticle, by adopting the principles of green chemistry.

Purpose To evaluate the toxicity and clinical outcome in patients with locally advanced cervical cancer (LACC) treated with whole pelvic conventional radiation therapy (WP-CRT) versus intensity modulated radiation therapy (WP-IMRT). Methods and Materials Between January 2010 and January 2012, 44 patients with International Federation of Gynecology and Obstetrics (FIGO 2009) stage IIB-IIIB squamous cell carcinoma of the cervix were randomized to receive 50.4 Gy in 28 fractions delivered via either WP-CRT or WP-IMRT with concurrent weekly cisplatin 40 mg/m2. Acute toxicity was graded according to the Common Terminology Criteria for Adverse Events, version 3.0, and late toxicity was graded according to the Radiation Therapy Oncology Group system. The primary and secondary endpoints were acute gastrointestinal toxicity and disease-free survival, respectively. Results Of 44 patients, 22 patients received WP-CRT and 22 received WP-IMRT. In the WP-CRT arm, 13 patients had stage IIB disease and 9 had stage IIIB disease; in the IMRT arm, 12 patients had stage IIB disease and 10 had stage IIIB disease. The median follow-up time in the WP-CRT arm was 21.7 months (range, 10.7-37.4 months), and in the WP-IMRT arm it was 21.6 months (range, 7.7-34.4 months). At 27 months, disease-free survival was 79.4% in the WP-CRT group versus 60% in the WP-IMRT group (P=.651), and overall survival was 76% in the WP-CRT group versus 85.7% in the WP-IMRT group (P=.645). Patients in the WP-IMRT arm experienced significantly fewer grade ≥2 acute gastrointestinal toxicities (31.8% vs 63.6%, P=.034) and grade ≥3 gastrointestinal toxicities (4.5% vs 27.3%, P=.047) than did patients receiving WP-CRT and had less chronic gastrointestinal toxicity (13.6% vs 50%, P=.011). Conclusion WP-IMRT is associated with significantly less toxicity compared with WP-CRT and has a comparable clinical outcome. Further studies with larger sample sizes and longer follow-up times are warranted to justify its use in routine clinical practice. To evaluate the toxicity and clinical outcome in patients with locally advanced cervical cancer (LACC) treated with whole pelvic conventional radiation therapy (WP-CRT) versus intensity modulated radiation therapy (WP-IMRT). Between January 2010 and January 2012, 44 patients with International Federation of Gynecology and Obstetrics (FIGO 2009) stage IIB-IIIB squamous cell carcinoma of the cervix were randomized to receive 50.4 Gy in 28 fractions delivered via either WP-CRT or WP-IMRT with concurrent weekly cisplatin 40 mg/m2. Acute toxicity was graded according to the Common Terminology Criteria for Adverse Events, version 3.0, and late toxicity was graded according to the Radiation Therapy Oncology Group system. The primary and secondary endpoints were acute gastrointestinal toxicity and disease-free survival, respectively. Of 44 patients, 22 patients received WP-CRT and 22 received WP-IMRT. In the WP-CRT arm, 13 patients had stage IIB disease and 9 had stage IIIB disease; in the IMRT arm, 12 patients had stage IIB disease and 10 had stage IIIB disease. The median follow-up time in the WP-CRT arm was 21.7 months (range, 10.7-37.4 months), and in the WP-IMRT arm it was 21.6 months (range, 7.7-34.4 months). At 27 months, disease-free survival was 79.4% in the WP-CRT group versus 60% in the WP-IMRT group (P=.651), and overall survival was 76% in the WP-CRT group versus 85.7% in the WP-IMRT group (P=.645). Patients in the WP-IMRT arm experienced significantly fewer grade ≥2 acute gastrointestinal toxicities (31.8% vs 63.6%, P=.034) and grade ≥3 gastrointestinal toxicities (4.5% vs 27.3%, P=.047) than did patients receiving WP-CRT and had less chronic gastrointestinal toxicity (13.6% vs 50%, P=.011). WP-IMRT is associated with significantly less toxicity compared with WP-CRT and has a comparable clinical outcome. Further studies with larger sample sizes and longer follow-up times are warranted to justify its use in routine clinical practice.

We present a simple and eco-friendly biosynthesis of silver nanoparticles using Pomegranate peel extract as the reducing agent.Peel extract of Pomegranate was challenged with silver nitrate (AgNO 3 ) and chloroauric acid (HAuCl 4 ) solution for the production of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs), respectively.The reaction process was simple for the formation of highly stable silver and gold nanoparticles at room temperature by using the biowaste of the fruit.The morphology and crystalline phase of the NPs were determined from UV-Vis spectroscopy, transmission electron microscopy (TEM), selected area electron diffraction (SAED) and X-ray diffraction (XRD) spectra.TEM studies showed that the average particle size of silver nanoparticles were 5 ±1.5 nm whereas the gold nanoparticles were found to be 10 ±1.5 nm.An effort has been also been made to understand the possible involved mechanism for the biosynthesis of the NPs.Presumably biosynthetic products or reduced cofactors play an important role in the reduction of respective salts to nanoparticles.

Targeted top-down (TD) and middle-down (MD) mass spectrometry (MS) offer reduced sample manipulation during protein analysis, limiting the risk of introducing artifactual modifications to better capture sequence information on the proteoforms present. This provides some advantages when characterizing biotherapeutic molecules such as monoclonal antibodies, particularly for the class of biosimilars. Here, we describe the results obtained analyzing a monoclonal IgG1, either in its ∼150 kDa intact form or after highly specific digestions yielding ∼25 and ∼50 kDa subunits, using an Orbitrap mass spectrometer on a liquid chromatography (LC) time scale with fragmentation from ion–photon, ion–ion, and ion–neutral interactions. Ultraviolet photodissociation (UVPD) used a new 213 nm solid-state laser. Alternatively, we applied high-capacity electron-transfer dissociation (ETD HD), alone or in combination with higher energy collisional dissociation (EThcD). Notably, we verify the degree of complementarity of these ion activation methods, with the combination of 213 nm UVPD and ETD HD producing a new record sequence coverage of ∼40% for TD MS experiments. The addition of EThcD for the >25 kDa products from MD strategies generated up to 90% of complete sequence information in six LC runs. Importantly, we determined an optimal signal-to-noise threshold for fragment ion deconvolution to suppress false positives yet maximize sequence coverage and implemented a systematic validation of this process using the new software TDValidator. This rigorous data analysis should elevate confidence for assignment of dense MS2 spectra and represents a purposeful step toward the application of TD and MD MS for deep sequencing of monoclonal antibodies.

In the present work silver nanoparticles (AgNPs) were synthesized extracellularly by bacteria Bacillus cereus collected from the riverine belt of Gangetic Plain of India.The microbes were isolated, screened and characterized by morphological and biochemical analyses.The silver resistant strain was exposed to different concentrations of silver salt (Ag-NO 3 ).UV-visible spectrum of the supernatant of cell culture showed absorbance peak of AgNPs at ~ 435 nm.The shape and size of AgNPs were ascertained by High Resolution Transmission Electron Micrography (HRTEM), X-ray diffraction (XRD) and Energy Dispersive spectroscopy (EDS).Average size of the synthesized AgNPs was found to be in the range of 10 -30 nm with spherical shape.AgNPs were tested against antibacterial potential of some common human pathogens.

Histones, and their modifications, are critical components of cellular programming and epigenetic inheritance. Recently, cancer genome sequencing has uncovered driver mutations in chromatin modifying enzymes spurring high interest how such mutations change histone modification patterns. Here, we applied Top-Down mass spectrometry for the characterization of combinatorial modifications (i.e. methylation and acetylation) on full length histone H3 from human cell lines derived from multiple myeloma patients with overexpression of the histone methyltransferase MMSET as the result of a t(4;14) chromosomal translocation. Using the latest in Orbitrap-based technology for clean isolation of isobaric proteoforms containing up to 10 methylations and/or up to two acetylations, we provide extensive characterization of histone H3.1 and H3.3 proteoforms. Differential analysis of modifications by electron-based dissociation recapitulated antagonistic crosstalk between K27 and K36 methylation in H3.1, validating that full-length histone H3 (15 kDa) can be analyzed with site-specific assignments for multiple modifications. It also revealed K36 methylation in H3.3 was affected less by the overexpression of MMSET because of its higher methylation levels in control cells. The co-occurrence of acetylation with a minimum of three methyl groups in H3K9 and H3K27 suggested a hierarchy in the addition of certain modifications. Comparative analysis showed that high levels of MMSET in the myeloma-like cells drove the formation of hypermethyled proteoforms containing H3K36me2 co-existent with the repressive marks H3K9me2/3 and H3K27me2/3. Unique histone proteoforms with such “trivalent hypermethylation” (K9me2/3-K27me2/3-K36me2) were not discovered when H3.1 peptides were analyzed by Bottom-Up. Such disease-correlated proteoforms could link tightly to aberrant transcription programs driving cellular proliferation, and their precise description demonstrates that Top-Down mass spectrometry can now decode crosstalk involving up to three modified sites. Histones, and their modifications, are critical components of cellular programming and epigenetic inheritance. Recently, cancer genome sequencing has uncovered driver mutations in chromatin modifying enzymes spurring high interest how such mutations change histone modification patterns. Here, we applied Top-Down mass spectrometry for the characterization of combinatorial modifications (i.e. methylation and acetylation) on full length histone H3 from human cell lines derived from multiple myeloma patients with overexpression of the histone methyltransferase MMSET as the result of a t(4;14) chromosomal translocation. Using the latest in Orbitrap-based technology for clean isolation of isobaric proteoforms containing up to 10 methylations and/or up to two acetylations, we provide extensive characterization of histone H3.1 and H3.3 proteoforms. Differential analysis of modifications by electron-based dissociation recapitulated antagonistic crosstalk between K27 and K36 methylation in H3.1, validating that full-length histone H3 (15 kDa) can be analyzed with site-specific assignments for multiple modifications. It also revealed K36 methylation in H3.3 was affected less by the overexpression of MMSET because of its higher methylation levels in control cells. The co-occurrence of acetylation with a minimum of three methyl groups in H3K9 and H3K27 suggested a hierarchy in the addition of certain modifications. Comparative analysis showed that high levels of MMSET in the myeloma-like cells drove the formation of hypermethyled proteoforms containing H3K36me2 co-existent with the repressive marks H3K9me2/3 and H3K27me2/3. Unique histone proteoforms with such “trivalent hypermethylation” (K9me2/3-K27me2/3-K36me2) were not discovered when H3.1 peptides were analyzed by Bottom-Up. Such disease-correlated proteoforms could link tightly to aberrant transcription programs driving cellular proliferation, and their precise description demonstrates that Top-Down mass spectrometry can now decode crosstalk involving up to three modified sites. The field of epigenetics has seen an explosion of research in the past decade as scientists from different fields discovered its critical roles in many aspects related to human health, ranging from stem cell pluripotency to aging (1.Ladewig J. Koch P. Brustle O. Leveling Waddington: the emergence of direct programming and the loss of cell fate hierarchies.Nat. Rev. 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Driver mutations in histone H3.3 and chromatin remodelling genes in paediatric glioblastoma.Nature. 2012; 482: 226-231Crossref PubMed Scopus (1707) Google Scholar, 8.Jaffe J.D. Wang Y. Chan H.M. Zhang J. Huether R. Kryukov G.V. Bhang H.E. Taylor J.E. Hu M. Englund N.P. Yan F. Wang Z. Robert McDonald 3rd, E. Wei L. Ma J. Easton J. Yu Z. deBeaumount R. Gibaja V. Venkatesan K. Schlegel R. Sellers W.R. Keen N. Liu J. Caponigro G. Barretina J. Cooke V.G. Mullighan C. Carr S.A. Downing J.R. Garraway L.A. Stegmeier F. Global chromatin profiling reveals NSD2 mutations in pediatric acute lymphoblastic leukemia.Nat. Genet. 2013; 45: 1386-1391Crossref PubMed Scopus (185) Google Scholar), from memory processing to drug addiction (9.Gräff J. Tsai L.H. Histone acetylation: molecular mnemonics on the chromatin.Nat. Rev. Neurosci. 2013; 14: 97-111Crossref PubMed Scopus (426) Google Scholar, 10.Schmidt H.D. McGinty J.F. West A.E. Sadri-Vakili G. Epigenetics and psychostimulant addiction.Perspectives Med. 2013; 3: a012047Google Scholar). Histone modifications, including methylation (me), acetylation (ac), monoubiquitylation (ub1), etc., are related to the study of epigenetics (11.Badeaux A.I. Shi Y. Emerging roles for chromatin as a signal integration and storage platform.Nat. Rev. Mol. Cell Biol. 2013; 14: 211-224Crossref Scopus (214) Google Scholar, 12.Berger S.L. Kouzarides T. Shiekhattar R. Shilatifard A. An operational definition of epigenetics.Genes Develop. 2009; 23: 781-783Crossref PubMed Scopus (1239) Google Scholar). These modifications, as well as their different states in case of methylation (i.e. mono-, di-, and trimethylation) and positions on the histone, play important and distinct roles in almost every activity operative on the chromatin template. The significance of these modifications are further underscored by the unexpected identification of many driver mutations underlying cancer biology within histone modifying enzymes (13.Zaidi S. Choi M. Wakimoto H. Ma L. Jiang J. Overton J.D. Romano-Adesman A. Bjornson R.D. Breitbart R.E. Brown K.K. Carriero N.J. Cheung Y.H. Deanfield J. DePalma S. Fakhro K.A. Glessner J. Hakonarson H. Italia M.J. Kaltman J.R. Kaski J. Kim R. Kline J.K. Lee T. Leipzig J. Lopez A. Mane S.M. Mitchell L.E. Newburger J.W. Parfenov M. Pe'er I. Porter G. Roberts A.E. Sachidanandam R. Sanders S.J. Seiden H.S. State M.W. Subramanian S. Tikhonova I.R. Wang W. Warburton D. White P.S. Williams I.A. Zhao H. Seidman J.G. Brueckner M. Chung W.K. Gelb B.D. Goldmuntz E. Seidman C.E. Lifton R.P. De novo mutations in histone-modifying genes in congenital heart disease.Nature. 2013; 498: 220-223Crossref PubMed Scopus (630) Google Scholar, 14.Suvà M.L. Riggi N. Bernstein B.E. Epigenetic reprogramming in cancer.Science. 2013; 339: 1567-1570Crossref PubMed Scopus (529) Google Scholar) and somatic mutations in histone H3.3 (7.Schwartzentruber J. Korshunov A. Liu X.-Y. Jones D.T.W. Pfaff E. Jacob K. Sturm D. Fontebasso A.M. Quang D.-A. K. Tonjes M. Hovestadt V. Albrecht S. Kool M. Nantel A. Konermann C. Lindroth A. Jäger N. Rausch T. Ryzhova M. Korbel J.O. Hielscher T. Hauser P. Garami M. Klekner A. Bognar L. Ebinger M. Schuhmann M.U. Scheurlen W. Pekrun A. Fruhwald M.C. Roggendorf W. Kramm C. Durken M. Atkinson J. Lepage P. Montpetit A. Zakrzewska M. Zakrzewski K. Liberski P.P. Dong Z. Siegel P. Kulozik A.E. Zapatka M. Guha A. Malkin D. Felsberg J. Reifenberger G. von Deimling A. Ichimura K. Collins V.P. Witt H. Milde T. Witt O. Zhang C. Castelo-Branco P. Lichter P. Faury D. Tabori U. Plass C. Majewski J. Pfister S.M. Jabado N. Driver mutations in histone H3.3 and chromatin remodelling genes in paediatric glioblastoma.Nature. 2012; 482: 226-231Crossref PubMed Scopus (1707) Google Scholar, 15.Behjati S. Tarpey P.S. Presneau N. Scheipl S. Pillay N. Van Loo P. Wedge D.C. Cooke S.L. Gundem G. Davies H. Nik-Zainal S. Martin S. McLaren S. Goodie V. Robinson B. Butler A. Teague J.W. Halai D. Khatri B. Myklebost O. Baumhoer D. Jundt G. Hamoudi R. Tirabosco R. Amary M.F. Futreal P.A. Stratton M.R. Campbell P.J. Flanagan A.M. Distinct H3F3A and H3F3B driver mutations define chondroblastoma and giant cell tumor of bone.Nat. Genet. 2013; 45: 1479-1482Crossref PubMed Scopus (525) Google Scholar, 16.Herz H.M. Morgan M. Gao X. Jackson J. Rickels R. Swanson S.K. Florens L. Washburn M.P. Eissenberg J.C. Shilatifard A. Histone H3 lysine-to-methionine mutants as a paradigm to study chromatin signaling.Science. 2014; 345: 1065-1070Crossref PubMed Scopus (130) Google Scholar). Widely used antibody-based measurements of histone modifications face two analytical challenges: (1) similar chemical structure of modification (e.g. three distinct methylation states of mono-, di-, and tri-methylation) and closely related flanking sequence can lead to cross-reactivity (17.Peach S.E. Rudomin E.L. Udeshi N.D. Carr S.A. Jaffe J.D. Quantitative assessment of chromatin immunoprecipitation grade antibodies directed against histone modifications reveals patterns of co-occurring marks on histone protein molecules.Mol. Cell. Proteomics. 2012; 11: 128-137Abstract Full Text Full Text PDF PubMed Scopus (61) Google Scholar); (2) close proximity of many modifications can have unexpected effects in antibody recognition (18.Fuchs S.M. Krajewski K. Baker R.W. Miller V.L. Strahl B.D. Influence of combinatorial histone modifications on antibody and effector protein recognition.Curr. Biol. 2011; 21: 53-58Abstract Full Text Full Text PDF PubMed Scopus (134) Google Scholar). For example, H4K20me2 antibody can lose its recognition when acetylation is present in the neighboring H4K16 (19.Hsiao K.Y. Mizzen C.A. Histone H4 deacetylation facilitates 53BP1 DNA damage signaling and double-strand break repair.J. Mol. Cell Biol. 2013; 5: 157-165Crossref PubMed Scopus (124) Google Scholar). Therefore, analyzing histone modifications by MS can provide a highly valuable orthogonal measurement. There are two general modes of interrogation by MS: Bottom-Up analysis of tryptic peptides, and Top-Down or Middle-Down measurement of full-length histones or large tail peptides, respectively (20.Yuan Z.F. Arnaudo A.M. Garcia B.A. Mass spectrometric analysis of histone proteoforms.Annu. Rev. Anal. Chem. 2014; 7: 113-128Crossref PubMed Scopus (46) Google Scholar). The value of mass spectrometric analysis of histone modifications in the field of cancer epigenetics was further demonstrated by the recent identification of a recurrent point mutation of E1099K in MMSET in lymphoid malignancies (8.Jaffe J.D. Wang Y. Chan H.M. Zhang J. Huether R. Kryukov G.V. Bhang H.E. Taylor J.E. Hu M. Englund N.P. Yan F. Wang Z. Robert McDonald 3rd, E. Wei L. Ma J. Easton J. Yu Z. deBeaumount R. Gibaja V. Venkatesan K. Schlegel R. Sellers W.R. Keen N. Liu J. Caponigro G. Barretina J. Cooke V.G. Mullighan C. Carr S.A. Downing J.R. Garraway L.A. Stegmeier F. Global chromatin profiling reveals NSD2 mutations in pediatric acute lymphoblastic leukemia.Nat. Genet. 2013; 45: 1386-1391Crossref PubMed Scopus (185) Google Scholar, 21.Oyer J.A. Huang X. Zheng Y. Shim J. Ezponda T. Carpenter Z. Allegretta M. Okot-Kotber C.I. Patel J.P. Melnick A. Levine R.L. Ferrando A. Mackerell Jr., A.D. Kelleher N.L. Licht J.D. Popovic R. Point mutation E1099K in MMSET/NSD2 enhances its methyltranferase activity and leads to altered global chromatin methylation in lymphoid malignancies.Leukemia. 2014; 28: 198-201Crossref PubMed Scopus (94) Google Scholar). The “histone code” hypothesis posits that the combinatorial nature of histone modifications can serve as the binding platform to elicit specific cellular processes (22.Jenuwein T. Allis C.D. Translating the histone code.Science. 2001; 293: 1074-1080Crossref PubMed Scopus (7632) Google Scholar). Recently, the combination of modifications has been used to define chromatin states, which are generated from meta-analysis of multiple ChIP-Seq data sets and found to be highly dynamic among different cell lines (23.Ernst J. Kheradpour P. Mikkelsen T.S. Shoresh N. Ward L.D. Epstein C.B. Zhang X. Wang L. Issner R. Coyne M. Ku M. Durham T. Kellis M. Bernstein B.E. Mapping and analysis of chromatin state dynamics in nine human cell types.Nature. 2011; 473: 43-49Crossref PubMed Scopus (2063) Google Scholar). However, this type of antibody-based technique relies on the associated DNA sequence to infer PTM co-occurrence from an average of histone modifications in the same locus but not necessarily on the same molecule. MMSET (also known as NSD2 or WHSC1) is one of the eight known histone methyltransferases targeting H3K36 (24.Wagner E.J. Carpenter P.B. Understanding the language of Lys36 methylation at histone H3.Nat. Rev. Mol. Cell Biol. 2012; 13: 115-126Crossref PubMed Scopus (632) Google Scholar) with specificity toward dimethylation (25.Li Y. Trojer P. Xu C.F. Cheung P. Kuo A. Drury 3rd, W.J. Qiao Q. Neubert T.A. Xu R.M. Gozani O. Reinberg D. The target of the NSD family of histone lysine methyltransferases depends on the nature of the substrate.J. Biol. Chem. 2009; 284: 34283-34295Abstract Full Text Full Text PDF PubMed Scopus (211) Google Scholar). Overexpression of MMSET has been documented in ∼20% of multiple myeloma cases as the result of chromosomal translocation t(4;14) (26.Keats J.J. Reiman T. Belch A.R. Pilarski L.M. Ten years and counting: so what do we know about t(4;14)(p16;q32) multiple myeloma.Leuk. Lymphoma. 2006; 47: 2289-2300Crossref PubMed Scopus (75) Google Scholar), which places the MMSET gene under the strong immunoglobulin enhancers (27.Stec I. Wright T.J. van Ommen G.J. de Boer P.A. van Haeringen A. Moorman A.F. Altherr M.R. den Dunnen J.T. WHSC1, a 90 kb SET domain-containing gene, expressed in early development and homologous to a Drosophila dysmorphy gene maps in the Wolf-Hirschhorn syndrome critical region and is fused to IgH in t(4;14) multiple myeloma.Human Mol. Genet. 1998; 7: 1071-1082Crossref PubMed Scopus (251) Google Scholar). A pair of cell lines, TKO and NTKO, were engineered from a t(4;14)+ multiple myeloma patient-derived cell line, KMS11. In the targeted knockout (TKO) 1The abbreviations used are:TKOtargeted knockoutHILIChydrophilic interaction chromatographyLCliquid chromatographyMSmass spectrometryETDelectron transfer dissociationEThcDelectron transfer dissociation – higher-energy collision dissociation. cell line, the translocated copy of MMSET was knocked out by homologous recombination, which leads to close to normal expression level of MMSET. In the nontargeted knockout (NTKO) cell line, the non-translocated gene was knocked out and the expression level of MMSET remains high (28.Lauring J. Abukhdeir A.M. Konishi H. Garay J.P. Gustin J.P. Wang Q. Arceci R.J. Matsui W. Park B.H. The multiple myeloma associated MMSET gene contributes to cellular adhesion, clonogenic growth, and tumorigenicity.Blood. 2008; 111: 856-864Crossref PubMed Scopus (129) Google Scholar). Our quantitative Bottom-Up MS assay using selective reaction monitoring revealed how overexpression of a HMT targeting H3K36 led to the global changes in both H3K27 and H3K36 methylation (29.Zheng Y. Sweet S.M. Popovic R. Martinez-Garcia E. Tipton J.D. Thomas P.M. Licht J.D. Kelleher N.L. Total kinetic analysis reveals how combinatorial methylation patterns are established on lysines 27 and 36 of histone H3.Proc. Natl. Acad. Sci. U.S.A. 2012; 109: 13549-13554Crossref PubMed Scopus (93) Google Scholar, 30.Popovic R. Martinez-Garcia E. Giannopoulou E.G. Zhang Q. Ezponda T. Shah M.Y. Zheng Y. Will C.M. Small E.C. Hua Y. Bulic M. Jiang Y. Carrara M. Calogero R.A. Kath W.L. Kelleher N.L. Wang J.P. Elemento O. Licht J.D. Histone methyltransferase MMSET/NSD2 alters EZH2 binding and reprograms the myeloma epigenome through global and focal changes in H3K36 and H3K27 methylation.PLoS Genet. 2014; 10: e1004566Crossref PubMed Scopus (148) Google Scholar, 31.Kuo A.J. Cheung P. Chen K. Zee B.M. Kioi M. Lauring J. Xi Y. Park B.H. Shi X. Garcia B.A. Li W. Gozani O. NSD2 links dimethylation of histone H3 at lysine 36 to oncogenic programming.Mol. Cell. 2011; 44: 609-620Abstract Full Text Full Text PDF PubMed Scopus (298) Google Scholar). Here, our goal was to make differential Top-Down measurement of two histone H3 variants, whose synthesis is (H3.1) and is not (H3.3) dependent on replication during S phase (32.Filipescu D. Muller S. Almouzni G. Histone H3 variants and their chaperones during development and disease: contributing to epigenetic control.Ann. Rev. Cell Develop. Biol. 2014; 30: 615-646Crossref PubMed Scopus (77) Google Scholar). targeted knockout hydrophilic interaction chromatography liquid chromatography mass spectrometry electron transfer dissociation electron transfer dissociation – higher-energy collision dissociation. To directly catalogue modifications co-occurring on the same histone (i.e. combinatorial modifications), we have reported Top-Down MS analysis of all histones (33.Pesavento J.J. Mizzen C.A. Kelleher N.L. Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry: human histone H4.Anal. Chem. 2006; 78: 4271-4280Crossref PubMed Scopus (201) Google Scholar, 34.Boyne 2nd, M.T. Pesavento J.J. Mizzen C.A. Kelleher N.L. Precise characterization of human histones in the H2A gene family by top down mass spectrometry.J. Proteome Res. 2006; 5: 248-253Crossref PubMed Scopus (142) Google Scholar, 35.Siuti N. Roth M.J. Mizzen C.A. Kelleher N.L. Pesavento J.J. Gene-specific characterization of human histone H2B by electron capture dissociation.J. Proteome Res. 2006; 5: 233-239Crossref PubMed Scopus (94) Google Scholar, 36.Thomas C.E. Kelleher N.L. Mizzen C.A. Mass spectrometric characterization of human histone H3: a bird's eye view.J. 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Mass spectrometry identifies and quantifies 74 unique histone H4 isoforms in differentiating human embryonic stem cells.Proc. Natl. Acad. Sci. U.S.A. 2008; 105: 4093-4098Crossref PubMed Scopus (142) Google Scholar, 40.Tian Z. Tolic N. Zhao R. Moore R.J. Hengel S.M. Robinson E.W. Stenoien D.L. Wu S. Smith R.D. Pasa-Tolic L. Enhanced top-down characterization of histone post-translational modifications.Genome Biol. 2012; 13: R86Crossref PubMed Google Scholar, 41.Moradian A. Kalli A. Sweredoski M.J. Hess S. The top-down, middle-down, and bottom-up mass spectrometry approaches for characterization of histone variants and their post-translational modifications.Proteomics. 2014; 14: 489-497Crossref PubMed Scopus (113) Google Scholar, 42.Sidoli S. Lin S. Karch K.R. Garcia B.A. Bottom-up and middle-down proteomics have comparable accuracies in defining histone post-translational modification relative abundance and stoichiometry.Anal. Chem. 2015; 87: 3129-3133Crossref PubMed Scopus (44) Google Scholar, 43.Harshman S.W. Hoover M.E. Huang C. Branson O.E. Chaney S.B. Cheney C.M. Rosol T.J. Shapiro C.L. Wysocki V.H. Huebner K. Freitas M.A. Histone H1 phosphorylation in breast cancer.J.Proteome Res. 2014; 13: 2453-2467Crossref PubMed Scopus (31) Google Scholar, 44.Young N.L. DiMaggio P.A. Plazas-Mayorca M.D. Baliban R.C. Floudas C.A. Garcia B.A. High throughput characterization of combinatorial histone codes.Mol. Cell. Proteomics. 2009; 8: 2266-2284Abstract Full Text Full Text PDF PubMed Scopus (245) Google Scholar). However, it is still very challenging to apply Top-Down approach for the routine analysis of histone proteoforms (45.Smith L.M. Kelleher N.L. Proteoform: a single term describing protein complexity.Nat. Methods. 2013; 10: 186-187Crossref PubMed Scopus (884) Google Scholar). This was demonstrated in the first pilot project from the Consortium for Top-Down Proteomics to assess intra-laboratory variation in the characterization of histone H4 (46.Dang X. Scotcher J. Wu S. Chu R.K. Tolic N. Ntai I. Thomas P.M. Fellers R.T. Early B.P. Zheng Y. Durbin K.R. Leduc R.D. Wolff J.J. Thompson C.J. Pan J. Han J. Shaw J.B. Salisbury J.P. Easterling M. Borchers C.H. Brodbelt J.S. Agar J.N. Pasa-Tolic L. Kelleher N.L. Young N.L. The first pilot project of the consortium for top-down proteomics: a status report.Proteomics. 2014; 14: 1130-1140Crossref PubMed Scopus (77) Google Scholar). One of the key limitations identified in that study was a need for continued improvement in high-resolution isolation and high-efficiency fragmentation. These two critical aspects for high quality proteoform characterization align with the development of a new Orbitrap-based tribrid mass spectrometer, whose architecture includes a segmented quadrupole for narrow precursor isolation with high transmission efficiency, improved vacuum conditions, and the optimization of multiple ion dissociation techniques including electron transfer dissociation (ETD) (47.Syka J.E. Coon J.J. Schroeder M.J. Shabanowitz J. Hunt D.F. Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry.Proc. Natl. Acad. Sci. U.S.A. 2004; 101: 9528-9533Crossref PubMed Scopus (1999) Google Scholar), higher-energy collisional dissociation (HCD) (48.Olsen J.V. Macek B. Lange O. Makarov A. Horning S. Mann M. Higher-energy C-trap dissociation for peptide modification analysis.Nat. Methods. 2007; 4: 709-712Crossref PubMed Scopus (719) Google Scholar) and their combination (EThcD) (49.Frese C.K. Altelaar A.F. van den Toorn H. Nolting D. Griep-Raming J. Heck A.J. Mohammed S. Toward full peptide sequence coverage by dual fragmentation combining electron-transfer and higher-energy collision dissociation tandem mass spectrometry.Anal. Chem. 2012; 84: 9668-9673Crossref PubMed Scopus (216) Google Scholar, 50.Brunner A.M. Lossl P. Liu F. Huguet R. Mullen C. Yamashita M. Zabrouskov V. Makarov A. Altelaar A.F. Heck A.J. Benchmarking multiple fragmentation methods on an orbitrap fusion for top-down phospho-proteoform characterization.Anal. Chem. 2015; 87: 4152-4158Crossref PubMed Scopus (85) Google Scholar). Therefore, we aimed to develop proper workflow and informatic tools to enable Top-Down comparative interrogation of the most highly modified core histone, H3, upon cellular perturbation. KMS11 TKO and NTKO cells were cultured in RPMI media supplemented with 10% FBS (Sigma, St. Louis, MI). Harvested cells were flash frozen in liquid nitrogen and stored at −80 °C before sample preparation. Nuclei were isolated with NIB buffer (15 mm Tris, 60 mm KCl, 15 mm NaCl, 5 mm MgCl2, 1 mm CaCl2, 250 mm Sucrose, 0.5 mm 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), 10 mm sodium butyrate, and 1 mm DTT, pH 7.5) containing 0.3% Nonidet P-40 and subsequently washed twice with NIB buffer without detergent. Isolated nuclei were recovered by centrifugation at 600 rcf. Crude histones were extracted from isolated nuclei with 0.4 N H2SO4 and recovered by precipitation with 20% (w/v, final) trichloroacetic acid (TCA). The precipitate was washed first with 0.1% HCl in acetone and twice with pure acetone. Crude histones were then resuspended in water and subjected to fractionation using reverse phase high pressure liquid chromatography (RP-HPLC) as described previously (51.Zheng Y. Tipton J.D. Thomas P.M. Kelleher N.L. Sweet S.M. Site-specific human histone H3 methylation stability: fast K4me3 turnover.Proteomics. 2014; 14: 2190-2199Crossref PubMed Scopus (17) Google Scholar). Briefly, histones were separated using a Jupiter C18 analytical column (Phenomenex, Torrance, CA), 15 cm × 4.6 mm, 5 μm diam., 300 Å pores, using a gradient of 30–57% B in 90 min (Buffer A: 5% ACN, 0.1% TFA; Buffer B: 90% ACN, 0.094% TFA) at a flow rate of 0.8 ml/min using an Agilent 1100 HPLC system (Agilent, Santa Clara, CA) monitored by UV absorbance at 214 nm. Fractionated histone H3.1 and H3.3 were collected and dried by speed-vacuum. Dried histone pellets were resuspended in 49.95:49.95:0.1 (v:v:v) water/acetonitrile/formic acid (LC-MS grade) at ∼1 μm final concentration, and were sprayed using a NanoFlex ion source (Thermo Fisher Scientific, San Jose, CA) equipped with a nanoelectrospray static probe and coated glass emitters, applying a 1.7–1.9 kV potential at the emitter. All mass spectrometry measurements were performed on an fETD-enabled (52.Earley L. Anderson L.C. Bai D.L. Mullen C. Syka J.E. English A.M. Dunyach J.J. Stafford Jr., G.C. Shabanowitz J. Hunt D.F. Compton P.D. Front-end electron transfer dissociation: a new ionization source.Anal. Chem. 2013; 85: 8385-8390Crossref PubMed Scopus (50) Google Scholar) Orbitrap Fusion Lumos mass spectrometer (Thermo Fisher Scientific) operating in Intact Protein Mode (N2 pressure at the ion routing multipole of 1 mTorr), using a resolving power of 60,000 (at m/z 200) and averaging five microscans for every scan, with transfer capillary temperature set at 275 °C, the RF of the source ion funnel operating at 20% and a source offset of 15 V to favor adduct removal. For each histone fraction, broadband MS1 spectra were recorded over a 500–2000 m/z window using an AGC target value of 2e5. MS1 spectra were used to define a list of histone proteoform peaks differing in mass for ∼14 Da, corresponding to the mass of one methylation. The list included m/z values corresponding to the most abundant isotopic distribution for each isobaric proteoform cluster. MS2 experiments were based on the isolation and subsequent fragmentation of each of these clusters for the 18+ precursor. Each proteoform cluster was quadrupole isolated using a 0.6 Th isolation window, and subjected sequentially first to high capacity ETD (ETD HD) performed with increasing duration, and then to EThcD, performed as previously described (50.Brunner A.M. Lossl P. Liu F. Huguet R. Mullen C. Yamashita M. Zabrouskov V. Makarov A. Altelaar A.F. Heck A.J. Benchmarking multiple fragmentation methods on an orbitrap fusion for top-down phospho-proteoform characterization.Anal. Chem. 2015; 87: 4152-4158Crossref PubMed Scop

We investigated the combination of weak anion exchange (WAX) fractionation and on-line reversed-phase liquid chromatography (RPLC) separation using a 12 T FTICR mass spectrometer for the detection of intact proteins from a Shewanella oneidensis MR-1 cell lysate. This work aimed at optimizing intact protein detection for profiling proteins at a level that incorporates their modification state. A total of 715 intact proteins were detected, and the combined results from the WAX fractions and the unfractionated cell lysate were aligned using LC-MS features to facilitate protein abundance measurements. Protein identifications and post-translational modifications were assigned for ∼10% of the detected proteins by comparing intact protein mass measurements to proteins identified in peptide MS/MS analysis of an aliquot of the same fraction. Intact proteins were also detected for S. oneidensis lysates obtained from cells grown on 13C-, 15N-depleted media under aerobic and sub-oxic conditions. The strategy can be readily applied for measuring differential protein abundances and provides a platform for high-throughput selection of biologically relevant targets for further characterization. Keywords: Comparative proteomics • FTICR MS • intact proteins • post-translational modifications

Electron transfer dissociation (ETD) is a valuable tool for protein sequence analysis, especially for the fragmentation of intact proteins. However, low product ion signal-to-noise often requires some degree of signal averaging to achieve high quality MS/MS spectra of intact proteins. Here we describe a new implementation of ETD on the newest generation of quadrupole-Orbitrap-linear ion trap Tribrid, the Orbitrap Fusion Lumos, for improved product ion signal-to-noise via ETD reactions on larger precursor populations. In this new high precursor capacity ETD implementation, precursor cations are accumulated in the center section of the high pressure cell in the dual pressure linear ion trap prior to charge-sign independent trapping, rather than precursor ion sequestration in only the back section as is done for standard ETD. This new scheme increases the charge capacity of the precursor accumulation event, enabling storage of approximately 3-fold more precursor charges. High capacity ETD boosts the number of matching fragments identified in a single MS/MS event, reducing the need for spectral averaging. These improvements in intra-scan dynamic range via reaction of larger precursor populations, which have been previously demonstrated through custom modified hardware, are now available on a commercial platform, offering considerable benefits for intact protein analysis and top down proteomics. In this work, we characterize the advantages of high precursor capacity ETD through studies with myoglobin and carbonic anhydrase.

Under changing climate scenarios, developing sustainable adaptation strategies in agriculture will be obligatory. To understand the effect of carbohydrate-based superabsorbent polymer (SAP) and crop-residue mulch (CRM) on pearl millet productivity and water-use efficiency (WUE), field experiments were undertaken for three consecutive years at 11 different locations (representing ⁓30 % of the global pearl millet growing area). Eight treatments, namely, Control; CRM 5.0 t/ha; SAP 2.5 kg/ha; SAP 5.0 kg/ha; SAP 7.5 kg/ha; SAP 2.5 kg/ha + CRM 5.0 t/ha; SAP 5.0 kg/ha + CRM 5.0 t/ha and SAP 7.5 kg/ha + CRM 5.0 t/ha were evaluated. Co-application of CRM and SAP increased pearl millet grain and stover yield by up to ⁓45 % and ⁓36 %, respectively. Pearl millet responded significantly up to 2.5 kg/ha SAP application (with or without CRM) only. Further, soil microbial biomass carbon improved significantly with CRM (20 %) and SAP (10.9–12.1 %) individually and with simultaneous application of CRM and SAP (⁓30 %). Likewise, dehydrogenase, alkaline phosphatase, acid phosphatase, and urease activities also improved significantly due to the co-use of CRM and SAP. Positive effects of CRM, SAP, and their co-application were also witnessed on soil microbial (bacterial, fungal, actinobacteria) populations and water-use efficiency (WUE) across environments. Among the locations, New Delhi and Aurangabad were the most desirable and stable ecologies, whereas Bikaner and Vijayapur remained the least consistent. Hence, to tackle the moisture-stress problem under pearl millet production systems and to achieve stable productivity, greater WUE and better soil microbial activity, CRM 5 t/ha in conjunction with SAP 2.5 kg/ha may be recommended across diverse ecologies.

Solid solution of 32 mol% of Lead Titanate in PMN–PT system has been prepared by columbite precursor method. Room temperature X-ray diffraction study reveals the formation of perovskite phase with tetragonal structure. Dielectric measurements have been carried out at different frequencies (0.1 kHz–1 MHz) as a function of temperature (RT to 235 °C). The phase transition was found to be of diffused type. The polarization studies show ferroelectric nature of the material with a high value of remnant polarization (Pr) ∼21 μC/cm2 and spontaneous polarization (Ps) ∼29 μC/cm2. Strain versus electric field (S–E) behavior shows piezoelectric nature of the material with high value of maximum strain 0.14% at 60 kV/cm. Pyroelectric coefficient at room temperature has been found to be 3×10−2 μC/cm2 K.

Thin films of Pb1−xCaxTiO3 [x=0.20, 0.24 and 0.28] have been prepared on ITO coated Corning glass substrates by sol gel technique. The perovskite phase of PCT films is formed at 650 °C with a polycrystalline tetragonal structure. The tetragonal factor (c/a) decreases with increasing Ca concentration. Dielectric, pyroelectric and ferroelectric studies have been carried out on these films. The effects of introduction of Ca ion in PbTiO3 have also been discussed.

Trypsin dominates bottom-up proteomics, but there are reasons to consider alternative enzymes. Improving sequence coverage, exposing proteomic “dark matter,” and clustering post-translational modifications in different ways and with higher-order drive the pursuit of reagents complementary to trypsin. Additionally, enzymes that are easy to use and generate larger peptides that capitalize upon newer fragmentation technologies should have a place in proteomics. We expressed and characterized recombinant neprosin, a novel prolyl endoprotease of the DUF239 family, which preferentially cleaves C-terminal to proline residues under highly acidic conditions. Cleavage also occurs C-terminal to alanine with some frequency, but with an intriguingly high “skipping rate.” Digestion proceeds to a stable end point, resulting in an average peptide mass of 2521 units and a higher dependence upon electron-transfer dissociation for peptide-spectrum matches. In contrast to most proline-cleaving enzymes, neprosin effectively degrades proteins of any size. For 1251 HeLa cell proteins identified in common using trypsin, Lys-C, and neprosin, almost 50% of the neprosin sequence contribution is unique. The high average peptide mass coupled with cleavage at residues not usually modified provide new opportunities for profiling clusters of post-translational modifications. We show that neprosin is a useful reagent for reading epigenetic marks on histones. It generates peptide 1–38 of histone H3 and peptide 1–32 of histone H4 in a single digest, permitting the analysis of co-occurring post-translational modifications in these important N-terminal tails. Trypsin dominates bottom-up proteomics, but there are reasons to consider alternative enzymes. Improving sequence coverage, exposing proteomic “dark matter,” and clustering post-translational modifications in different ways and with higher-order drive the pursuit of reagents complementary to trypsin. Additionally, enzymes that are easy to use and generate larger peptides that capitalize upon newer fragmentation technologies should have a place in proteomics. We expressed and characterized recombinant neprosin, a novel prolyl endoprotease of the DUF239 family, which preferentially cleaves C-terminal to proline residues under highly acidic conditions. Cleavage also occurs C-terminal to alanine with some frequency, but with an intriguingly high “skipping rate.” Digestion proceeds to a stable end point, resulting in an average peptide mass of 2521 units and a higher dependence upon electron-transfer dissociation for peptide-spectrum matches. In contrast to most proline-cleaving enzymes, neprosin effectively degrades proteins of any size. For 1251 HeLa cell proteins identified in common using trypsin, Lys-C, and neprosin, almost 50% of the neprosin sequence contribution is unique. The high average peptide mass coupled with cleavage at residues not usually modified provide new opportunities for profiling clusters of post-translational modifications. We show that neprosin is a useful reagent for reading epigenetic marks on histones. It generates peptide 1–38 of histone H3 and peptide 1–32 of histone H4 in a single digest, permitting the analysis of co-occurring post-translational modifications in these important N-terminal tails. The most widely used “bottom-up” proteomics approach requires the proteolytic digestion of proteins into peptides. These peptides are detected by MS and identified through database searching (1.Aebersold R. Mann M. Mass-spectrometric exploration of proteome structure and function.Nature. 2016; 537: 347-355Crossref PubMed Scopus (1113) Google Scholar). Trypsin is the standard protease and cleaves almost exclusively after the basic amino acids lysine (Lys) and arginine (Arg) (2.Olsen J.V. Ong S.E. Mann M. Trypsin cleaves exclusively C-terminal to arginine and lysine residues.Mol. Cell. Proteomics. 2004; 3: 608-614Abstract Full Text Full Text PDF PubMed Scopus (872) Google Scholar). However, in practice, due to the natural distribution of these amino acids in proteomes, a large segment of the proteome is not identifiable. Most peptides are too small for unambiguous identification (56% ≤6 residues) (3.Swaney D.L. Wenger C.D. Coon J.J. Value of using multiple proteases for large-scale mass spectrometry-based proteomics.J. Proteome Res. 2010; 9: 1323-1329Crossref PubMed Scopus (331) Google Scholar). Improvements in proteome representation and the characterization of post-translational modifications (PTMs) 1The abbreviations used are: PTM, post-translational modification; CID, collision-induced dissociation; DUF239, domain of unknown function 239; E:S ratio, enzyme-to-substrate ratio; ETD, electron-transfer dissociation; EThcD, electron-transfer/higher-energy collision dissociation; HCD, higher-energy collisional dissociation; PEP, prolyl endoprotease; POP, prolyl oligopeptidase; PSM, peptide spectrum match; MBP, maltose-binding protein; FA, folic acid; ACN, acetonitrile; NCE, normalized collision energy. 1The abbreviations used are: PTM, post-translational modification; CID, collision-induced dissociation; DUF239, domain of unknown function 239; E:S ratio, enzyme-to-substrate ratio; ETD, electron-transfer dissociation; EThcD, electron-transfer/higher-energy collision dissociation; HCD, higher-energy collisional dissociation; PEP, prolyl endoprotease; POP, prolyl oligopeptidase; PSM, peptide spectrum match; MBP, maltose-binding protein; FA, folic acid; ACN, acetonitrile; NCE, normalized collision energy. and proteoforms are possible with proteolytic enzymes such as LysC, ArgC, AspN, and GluC (3.Swaney D.L. Wenger C.D. Coon J.J. Value of using multiple proteases for large-scale mass spectrometry-based proteomics.J. Proteome Res. 2010; 9: 1323-1329Crossref PubMed Scopus (331) Google Scholar, 4.Huesgen P.F. Lange P.F. Rogers L.D. Solis N. Eckhard U. Kleifeld O. Goulas T. Gomis-Rüth F.X. Overall C.M. LysargiNase mirrors trypsin for protein C-terminal and methylation-site identification.Nat. Methods. 2015; 12: 55-58Crossref PubMed Scopus (111) Google Scholar, 5.Tsiatsiani L. Heck A.J. Proteomics beyond trypsin.FEBS J. 2015; 282: 2612-2626Crossref PubMed Scopus (204) Google Scholar). They all share the property of cleaving either the C or N terminus to charged residues. An enzyme with specificity for neutral amino acids would be a welcome addition. For example, an advantage in membrane protein representation was demonstrated recently using WaLP and MaLP (6.Meyer J.G. Kim S. Maltby D.A. Ghassemian M. Bandeira N. Komives E.A. Expanding proteome coverage with orthogonal-specificity α-lytic proteases.Mol. Cell. Proteomics. 2014; 13: 823-835Abstract Full Text Full Text PDF PubMed Scopus (38) Google Scholar). These are lower-specificity enzymes that cleave after Thr, Val, Ala, Ser, and Met (WaLP) and Met, Leu, Phe, Tyr, Thr, and Val (MaLP). It would be useful to have enzymes in this general class with even higher specificity to generate longer sequence “read lengths” and possibly improved representation of heterogeneity in PTMs. Enzymes recognizing dibasic motifs such as Sap9 (7.Laskay Ü.A. Srzentić K. Monod M. Tsybin Y.O. Extended bottom-up proteomics with secreted aspartic protease Sap9.J. Proteomics. 2014; 110: 20-31Crossref PubMed Scopus (20) Google Scholar) and OmpT (8.Wu C. Tran J.C. Zamdborg L. Durbin K.R. Li M. Ahlf D.R. Early B.P. Thomas P.M. Sweedler J.V. Kelleher N.L. A protease for ‘middle-down’ proteomics.Nat. 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We considered proline-cleaving enzymes as an attractive alternative for number of reasons. Proline (Pro) is less abundant than Lys + Arg (14.Morgan A.A. Rubenstein E. Proline: the distribution, frequency, positioning, and common functional roles of proline and polyproline sequences in the human proteome.PLoS ONE. 2013; 8: e53785Crossref PubMed Scopus (148) Google Scholar). It often delineates structural transitions in proteins and is generally unmodified in cellular proteins; thus digestion products could provide a better grouping of PTMs. Unfortunately, there are very few validated prolyl endoproteases (PEPs) currently available. Numerous prolyl oligopeptidases (POPs) are known to cleave after Pro and alanine (Ala), but their activity is restricted to small substrates (15.Gass J. Khosla C. Prolyl endopeptidases.Cell. Mol. Life Sci. 2007; 64: 345-355Crossref PubMed Scopus (115) Google Scholar). 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Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics.J. Mass Spectrom. 2009; 44: 1587-1595Crossref PubMed Scopus (17) Google Scholar). An effective PEP would be particularly useful for characterizing histone modifications. Histones are Lys- and Arg-rich proteins that form the core components of nucleosomes, around which the DNA is wound in the chromatin of eukaryotes (19.Biterge B. Schneider R. Histone variants: key players of chromatin.Cell Tissue Res. 2014; 356: 457-466Crossref PubMed Scopus (99) Google Scholar). Histones are tightly packed into octamers, but they maintain highly modified N-terminal tails that are flexible and integrate a variety of post-translational modifications. Many of these modifications are dynamically installed and removed. They play a key role in epigenetically regulating cell fate (20.Probst A.V. Dunleavy E. Almouzni G. Epigenetic inheritance during the cell cycle.Nat. Rev. Mol. Cell Biol. 2009; 10: 192-206Crossref PubMed Scopus (580) Google Scholar), and their deregulation is often associated with cancer (21.Audia J.E. Campbell R.M. Histone modifications and cancer.Cold Spring Harb. Perspect. Biol. 2016; 8: a019521Crossref PubMed Scopus (418) Google Scholar). The high frequency of positively charged residues in the histone tails complicates tryptic analysis, as the resulting peptides are small, and the connectivity between modifications is lost (22.Garcia B.A. Mollah S. Ueberheide B.M. Busby S.A. Muratore T.L. Shabanowitz J. Hunt D.F. Chemical derivatization of histones for facilitated analysis by mass spectrometry.Nat. Protoc. 2007; 2: 933-938Crossref PubMed Scopus (279) Google Scholar). GluC and AspN enzymes show more promise, especially for presenting the combinatorial modifications of histones H3 and H4, but they need to be used separately to cluster the most important PTM sites (23.Phanstiel D. Brumbaugh J. Berggren W.T. Conard K. Feng X. Levenstein M.E. McAlister G.C. Thomson J.A. Coon J.J. Mass spectrometry identifies and quantifies 74 unique histone H4 isoforms in differentiating human embryonic stem cells.Proc. Natl. Acad. Sci. U.S.A. 2008; 105: 4093-4098Crossref PubMed Scopus (142) Google Scholar, 24.Young N.L. DiMaggio P.A. Plazas-Mayorca M.D. Baliban R.C. Floudas C.A. Garcia B.A. High throughput characterization of combinatorial histone codes.Mol. Cell. Proteomics. 2009; 8: 2266-2284Abstract Full Text Full Text PDF PubMed Scopus (245) Google Scholar). A PEP may provide a more universal approach to mapping the “histone code” and complement the existing methods for analysis. We recently discovered neprosin from the secretions of the carnivorous pitcher plant (25.Lee L. Zhang Y. Ozar B. Sensen C.W. Schriemer D.C. Carnivorous nutrition in pitcher plants (Nepenthes spp.) via an unusual complement of endogenous enzymes.J. Proteome Res. 2016; 15: 3108-3117Crossref PubMed Scopus (38) Google Scholar). It appears to be a proline-cleaving enzyme, based on our initial characterizations (26.Rey M. Yang M. Lee L. Zhang Y. Sheff J.G. Sensen C.W. Mrazek H. Halada P. Man P. McCarville J.L. Verdu E.F. Schriemer D.C. Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease.Sci. Rep. 2016; 6: 30980Crossref PubMed Scopus (40) Google Scholar). In this study, we produced neprosin recombinantly and characterized the enzyme for use in proteomics. We show that neprosin is a legitimate low-molecular-weight PEP, active at low concentrations and low pH. We demonstrate strong complementarity with conventional enzymes for whole-proteome analysis and histone mapping. The gene corresponding to pro-neprosin (residues 25–380) from Nepenthes ventrata was synthesized by Genscript with codon optimization for expression in Escherichia coli and yeast (26.Rey M. Yang M. Lee L. Zhang Y. Sheff J.G. Sensen C.W. Mrazek H. Halada P. Man P. McCarville J.L. Verdu E.F. Schriemer D.C. Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease.Sci. Rep. 2016; 6: 30980Crossref PubMed Scopus (40) Google Scholar). The optimized neprosin gene was then inserted into pET28a(+) at the EcoRI and SalI restriction sites in-frame and downstream of an N-terminal His tag, yielding the plasmid pDS36. The MBP gene was amplified from e3884 (generous gift from Dr. A. Schryvers, University of Calgary) with forward primer LSO13F (5′GGC TCG CATATG AAA ACT GAA GAA GGT AAA CTG GTA ATC TG3′, NdeI site underlined) and reverse primer LSO14R (5′CAC TCA GCTAGC CCT TCC CTC GAT GTT GTT GTT ATT GTT ATT GTT GTT GTT GTT CG3′, the NheI site is underlined, and the factor Xa cleavage site is italicized) and inserted into pDS36 at the NdeI and NheI restriction sites in-frame and between the His tag and neprosin gene, to form the His-MBP-neprosin expression plasmid termed pDS42. The constructs were verified by DNA sequencing. The His-MBP-pro-neprosin expression plasmid (pDS42) was transformed into E. coli Artic Express (generous gift from Dr. Peter Facchini, University of Calgary) competent cells. The pDS42-transformed cells (DSB90) were grown in 2YT with kanamycin 50 μg ml−1 at 37 °C until an OD of ∼0.8 was reached, and then expression was induced with 0.3 mm isopropyl β-d-thiogalactopyranoside at 16 °C overnight with shaking. Cells were harvested by centrifugation and frozen at −80 °C. The cells were then thawed and resuspended in lysis buffer (50 mm Tris-HCl, pH 8.0, 0.5 m NaCl, 10% glycerol, 1 mm DTT, 1 mm PMSF, 0.5% Triton X-100, 0.025% sodium azide, 1 μg ml−1 lysozyme, RNase A) supplemented with a mixture of protease inhibitors (Roche Applied Science) at a ratio of 10 ml of buffer per 1 g of wet cells. The cells were lysed by sonication (Qsonica Sonicators) on ice at 30% amplitude using six 30-s pulses with a 30-s rest between pulses. Following centrifugation at 15,000 rpm at 4 °C for 30 min, the supernatant was applied to a 5-ml HisTrap HP (GE Healthcare) pre-equilibrated with 50 mm Tris-HCl, pH 8.0, 0.5 m NaCl (buffer A). After washing with 20 ml of buffer A, His-MBP-pro-neprosin was eluted with 50 ml of a linear gradient of 0–100% buffer B (50 mm Tris-HCl, pH 8.0, 0.5 m NaCl, 0.5 m imidazole) at a flow rate of 1 ml min−1. Fractions containing His-MBP-pro-neprosin were pooled and dialyzed against 50 mm Tris-HCl, pH 7.5, 150 mm NaCl. Multiple rounds of dialysis against 100 mm Gly-HCl, pH 2.5, at 37 °C for 7 days allowed for acid auto-activation to the mature, active neprosin. Neprosin was then concentrated in a 10-kDa molecular mass cutoff filter (Millipore) and analyzed by SDS-PAGE. The concentration of active recombinant neprosin was estimated by comparing its activity to native neprosin purified from Nepenthes digestive fluid, using the GFP activity assay (below). The green fluorescent protein (GFP) proteolysis assay was performed as reported (27.Malik A. Rudolph R. Söhling B. Use of enhanced green fluorescent protein to determine pepsin at high sensitivity.Anal. Biochem. 2005; 340: 252-258Crossref PubMed Scopus (14) Google Scholar) with minor modifications. Briefly, 0.1 mg ml−1 of GFP S65T (in 10 mm Tris-HCl, pH 8) was denatured with 0.1× volume of 1 m Gly-HCl, pH 2.4 (final pH of solution was 2.5), in the absence or presence of neprosin (1 nm) at 37 °C (or the indicated temperature) for 1 h. Reaction was quenched, and the GFP was renatured by addition of 0.25× volume of 1 m Tris-HCl, pH 8.4, and 0.1× volume of 1 m DTT (final pH of solution was 8). Fluorescence of renatured GFP S65T was determined using a Molecular Devices Filter Max F5 microplate spectrophotometer with excitation at 485 nm, and emission was monitored at 535 nm. For the analysis of the effect of protease inhibitors on proteolytic activity, neprosin was pre-incubated with the protease inhibitor in the reaction volume at room temperature for 30 min prior to the addition of GFP S65T and acid denaturation. For the analysis of the effect of pH on neprosin activity, GFP S65T and neprosin were incubated in 100 mm solution of various buffers (pH 2–9: Gly-HCl, ammonium formate, sodium phosphate, Tris-HCl). Neprosin activity was expressed as percentage of fluorescence lost (100% − % fluorescence recovery) relative to the untreated GFP S65T under the same reaction conditions. For gel-based analysis using recombinant neprosin, 1 mg ml−1 of BSA was reduced with 10 mm DTT at 50 °C for 30 min. The reduced protein substrate (0.05 mg ml−1) was incubated with recombinant Npr1 (∼1 nm) and different concentrations of urea in 100 mm Gly-HCl, pH 2.5, at 37 °C for 1 h and analyzed by SDS-PAGE. HeLa S3 cells were grown at the National Cell Culture Centre in Joklik-modified minimum Eagle's medium supplemented with 5% newborn calf serum to high cell density (1 × 106 cells/ml). Cells were collected by centrifugation at 2500 × g, followed by two washes in warm 37 °C PBS (Ca/Mg free) and stored at −80 °C. After thawing, the cells were resuspended in 50 mm HEPES buffer (pH 8, 150 mm KCl, 1 mm MgCl2), 10% glycerol, 0.5% Nonidet P-40, 5 units/ml benzonase supplemented with a mixture of protease inhibitors (Roche Applied Science). The cells were lysed by sonication on ice at 30% amplitude for six rounds of 30-s pulses with a 30-s rest between pulses. Following centrifugation at 37,000 rpm at 4 °C for 30 min, the supernatant lysate was aliquoted, frozen in liquid nitrogen, and stored at −80 °C. Protein concentration was determined by Bradford assay, using BSA as standards. HeLa whole-cell lysate was digested with recombinant and endogenous neprosin, AN-PEP, LysC, and trypsin using the FASP protocol (28.Wiśniewski J.R. Zougman A. Nagaraj N. Mann M. Universal sample preparation method for proteome analysis.Nat. Methods. 2009; 6: 359-362Crossref PubMed Scopus (5097) Google Scholar). Briefly, 100 μg of precipitated lysate was loaded onto a 10-kDa filter device and subsequently denatured, reduced, and alkylated at pH 8.5. Depending on the protease, buffer exchange was conducted using a volume of 120 μl of 100 mm Gly-HCl, pH 2.5, for both neprosin preparations and AN-PEP or 50 mm ammonium bicarbonate, pH 8.5, for LysC and trypsin. Addition of buffer was followed by a centrifugation step for 30 min at 14,000 × g. This was repeated three times for a complete buffer exchange. The enzyme was then added to an estimated enzyme-to-substrate ratio of 1:50 (w/w) in case of trypsin and LysC, 1:100 for AN-PEP, and 1:500 for both neprosin preparations. Samples were subsequently incubated overnight at 37 °C. Released peptides were eluted from the filter device in three steps. First, the filter unit was centrifuged for 30 min at 14,000 rpm. Second, 50 μl of the corresponding buffer solution was added to the filter unit, followed by a second centrifugation step for 30 min at 14,000 rpm. Final elution was performed by adding 50 μl of 0.5 m NaCl to the filter device and subsequent centrifugation for 15 min at 14,000 rpm, and the eluates were combined. Prior to mass spectrometric data acquisition, all samples were desalted and concentrated using Stage tips (29.Rappsilber J. Mann M. Ishihama Y. Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips.Nat. Protoc. 2007; 2: 1896-1906Crossref PubMed Scopus (2589) Google Scholar). Peptides were eluted with 50% ACN in 0.1% TFA and evaporated to dryness using a SpeedVac. Finally, peptides were reconstituted in 0.1% FA for mass spectrometric analysis. Cleavage specificity experiments using recombinant and endogenous neprosin, AN-PEP, and trypsin were conducted in biological duplicates. Experiments for the evaluation of proteome coverage with recombinant neprosin, trypsin, and LysC were done in single experiments. To evaluate whether acidic solution induced unspecific cleavages under long incubation times at pH 2.5, tryptic digests were reconstituted in 20 μl of H2O and split in 2 aliquots of 10 μl. One aliquot was kept at −20 °C until analysis. The other aliquot was adjusted to pH 2.5 with 100 mm Gly-HCl buffer and incubated at 37 °C overnight. For whole histone analysis, unfractionated whole histone from calf thymus (Sigma-Aldrich) was dissolved in Gly-HCl, pH 2.5, to a final concentration of 1 μg μl−1. Recombinant neprosin was added to an estimated enzyme-to-substrate ratio of 1:500. Samples were subsequently incubated overnight at 37 °C. For the determination of cleavage specificity, HeLa digests were analyzed using an EASY-nLC 1000 nano-LC coupled to an Orbitrap Velos mass spectrometer (Thermo Fisher Scientific, San Jose CA) equipped with a Nanospray Flex Ion Source. Peptides were chromatographically separated using a 15-cm PicoTip fused silica emitter with an inner diameter of 75 μm (New Objective Inc., Woburn MA) packed in-house with reversed-phase Reprosil-Pur C18-AQ 3-μm resin (New Objective). The flow rate was 300 nl min−1, and peptides were eluted using a 140-min gradient running linearly from 5 to 40% B (97% ACN in 0.1% FA). Data were acquired using data-dependent MS/MS mode. Each high-resolution product ion scan in the Orbitrap (m/z 300 to 2000, R = 60,000) was followed by high-resolution product ion scans (isolation window 3 Th) in the Orbitrap after HCD fragmentation at 35% NCE. Resolution was set to 7500. Top 10 most abundant signals with a charge state greater than 1 were selected for fragmentation, followed by dynamic exclusion for 60 s. Data acquisition was controlled with Xcalibur software (version 3.0.63). For the comparative proteomic analysis of HeLa cells digested with LysC, trypsin, or recombinant neprosin, samples were measured using an EASY-nLC 1000 nano-LC coupled to an Orbitrap Fusion Lumos Tribrid mass spectrometer (Thermo Fisher Scientific) equipped with an EASY-spray source. Peptides were separated using a 25-cm Easy-Spray PepMap analytical column (Thermo Fisher Scientific) packed with reversed-phase C18 beads (2-μm particle diameter, 100-Å pores). The injection volume was adjusted to achieve similar TIC levels for all samples. The flow rate was set to 300 nl min−1. Peptides were eluted using a 110-min gradient running linearly from 5 to 25% B (100% ACN in 0.1% FA) at a spray voltage of 2.4 kV. Each high-resolution precursor ion scan in the Orbitrap (m/z 350 to 1500, R = 120,000) was followed by product ion scans (isolation window 3 Th) in the linear ion trap with a fixed scan cycle time of 3 s. Dissociation in MS2 mode was carried out via CID and ETD using a data-dependent decision tree (30.Swaney D.L. McAlister G.C. Coon J.J. Decision tree-driven tandem mass spectrometry for shotgun proteomics.Nat. Methods. 2008; 5: 959-964Crossref PubMed Scopus (268) Google Scholar). The fragmentation mode was chosen depending on the nature of the selected ions. Doubly charged ions, triply charged ions with m/z >650, quadruply charged ions with m/z >900, and quintuply charged ions with m/z >950 were triggered with CID. Triply charged ions with m/z ≤650, quadruply charged ions with m/z ≤900, quintuply charged ions with m/z ≤950, and ions with a charge state of six and higher were fragmented using ETD. CID was performed at 30% normalized collision energy (NCE), and ETD was carried out with a maximum injection time of 100 ms. A target value of 20,000 was selected for MS2 automatic gain control, and precursor ions were dynamically excluded for 30 s. Whole histone digests were measured on an Orbitrap Fusion Lumos Tribrid mass spectrometer coupled to an EASY-nLC 1200 nano-LC system. Peptides were separated using a 15-cm Easy-Spray PepMap analytical column (Thermo Fisher Scientific) packed with reversed-phase C18 beads (2-μm particle diameter, 100-Å pores) using first a 30-min gradient running linearly from 5 to 15% B (80% ACN in 0.015% FA), followed by increasing B to 60% within the next 10 min. Each high-resolution precursor ion scan in the Orbitrap (m/z 350 to 1200, R = 120,000) was followed by product ion scans (isolation window 2 Th) in the Orbitrap (R = 15,000) with a fixed scan cycle time of 3 s. Charge states of three and less were excluded. Peptides were fragmented using EThcD (maximum injection time of 70 ms and HCD at 25% NCE) at a spray voltage of 1.9 kV. A target value of 50,000 was selected for MS2 automatic gain control, and precursor ions were dynamically excluded for 15 s. Data acquisition was controlled using Xcalibur software (version 4.0). All unprocessed data files (RAW-format) were directly loaded into PEAKS studio (31.Zhang J. Xin L. Shan B. Chen W. Xie M. Yuen D. Zhang W. Zhang Z. Lajoie G.A. Ma B. PEAKS DB: de novo sequencing assisted database search for sensitive and accurate peptide identification.Mol. Cell. Proteomics. 2012; 11Abstract Full Text Full Text PDF Scopus (105) Google Scholar) (version 7.5; Bioinformatics Solutions), and precursor masses were subsequently corrected in the software. Peptides were identified by de novo sequencing and database search, with matching to the human Swiss-Prot database (downloaded from uniprot.org in May 2016; containing 20,201 entries). For whole proteome analysis of HeLa digests, cleavage sites were restricted in a number of ways. For trypsin (P1: Lys and Arg, with P1′: any amino acid, according to Rodriguez et al. (32.Rodriguez J. Gupta N. Smith R.D. Pevzner P.A. Does trypsin cut before proline?.J. Proteome Res. 2008; 7: 300-305Crossref PubMed Scopus (185) Google Scholar)), we allowed up to two missed cleavages for a conventional search. We also performed a semi-tryptic search (unspecific cleavage at one end) and a fully nonspecific search. For LysC (P1: Lys, with P1′: any amino acid), we also allowed up to two missed cleavages for a conventional search, and both semi-LysC and fully nonspecific searches. For neprosin (P1: Pro and Ala, with P1′: any amino acid except Pro; and P1: Asp with P1′: Pro), we allowed a maximum number of seven missed cleavages, and we also allowed for both semi-neprosin and fully nonspecific cleavage in separate searches. Met oxidation was selected as a variable modification with a maximum number of two variable modifications per peptide. Carbamidomethylation of cysteine residues was set as a fixed modification. To decrease FDR while searching for multiple post-translationally modified peptides, cleavages were restricted to the same rules as stated above for the three proteases, but applying strict cleavage rules for both ends. Acetylation of the protein's N terminus and methylation and dimethylation of Lys and Arg were chosen as variable modifications with a maximum of three variable modifications per peptide. For a minimally biased cleavage site analysis in the proteomics experiments, the enzyme was set to “none,” and a modified database file was used, in which initiator methionine residues and signal peptides (35.Choo K.H. Tan T.W. Ranganatha

The current study highlights rapid, sustainable, and cost-effective biosynthesis of silver (Ag), gold (Au) nanoparticles (NPs), and bimetallic Au-AgNPs composites using bio-waste extract of Trapa natans. Growth of the NPs was monitored spectrophotometrically and peak was observed at ∼525 nm, ∼450 nm, and ∼495 nm corresponding to Plasmon absorbance of AuNPs, AgNPs, and Au-AgNPs, respectively. Transmission electron microscopy (TEM) revealed the size of AgNPs (∼15 nm), AuNPs (∼25 nm), and Au-AgNPs (∼26–90 nm). Synthesized NPs follow the Gaussian bell curve and its crystalline nature was identified by X-ray diffraction (XRD). Furthermore, Au-AgNPs induced cytotoxicity in various cancer cells (HCT116, MDA-MB-231, and HeLa) effectively at 200 μg/mL. Au-AgNPs-exposed cancer cells exhibited apoptotic features such as nuclear condensation, mitochondrial membrane potential loss, and cleavage of casp-3 and poly (ADP-ribose) polymerase-1 (PARP). Au-AgNPs exposure enhanced reactive oxygen species (ROS) and upon inhibition of ROS, apoptosis was reduced effectively. NPs treatment killed HCT116 WT and p53 knockout cells without any significant difference. Mechanistically, Au-AgNPs derived with Trapa peel extract significantly enhance ROS which trigger p53-independent apoptosis in various cancer cells effectively. Our study explores the use of bio-waste for the green synthesis of NPs, which can be attractive candidates for cancer therapy.

Nanotechnology is one of the rapidly growing scientific disciplines due to its enormous potential in creating novel materials that have advanced applications. Electrospinning has been found to be a viable technique to produce materials in nanofiber form. Ferroelectric and/or piezoelectric materials in nanofiber and/or nanowire form have been utilized for producing energy harvesting devices, high frequency transducers, implanted biosensors, vibration absorbers and composite force sensors, etc. An in-depth review of research activities on the development of ferroelectric nanofibers, fundamental understanding of the electrospinning process, and properties of nanostructured fibrous materials and their applications is provided in this article. A detailed account on the type of fibers that have been electrospun and their characteristics is also elaborated. It is hoped that the overview article will serve as a good reference tool for nanoscience researchers in ferroelectric materials.

Online news platforms greatly influence our society and culture in both positive and negative ways. As online media becomes more dependent for source of information, a lot of fake news is posted online, that widespread with people following it without any prior or complete information of event authenticity. Such misinformation has the potential to manipulate public opinions. The exponential growth of fake news propagation have become a great threat to public for news trustworthiness. It has become a compelling issue for which discovering, examining and dealing with fake news has increased in demand. However, with the limited availability of literature on the issue of uncovering fake news, a number of potential methodologies and techniques remains unexplored. The primary aim of this paper is to review existing methodologies, to propose and implement a method for automated deception detection. The proposed methodology uses deep learning in discourse-level structure analysis to formulate the structure that differentiates fake and real news. The baseline model achieved 74% accuracy.

Tuberculosis is predicted to be a major undocumented cause of mortality in children. This systematic review with meta-analysis assessed the diagnostic accuracy of Lipoarabinomannan antigen testing (FujiLAM) in urine in HIV-negative children with TB-like signs and symptoms. PubMed, EMBASE, Scopus, Cochrane database and Google Scholar search engine were searched to identify relevant studies from earliest records to June 2022 without any language restriction. Three studies were finalized, patients were recruited from Africa and Haiti. Among microbiologically confirmed pediatric TB patients, pooled sensitivity and specificity of FujiLAM (with 95% CI) was 52% (35%-69%) and 90% (85%-93%) respectively. In both clinical (unconfirmed) and microbiological confirmed TB cases, sensitivity reduced to 24% (16%-34%) while specificity was 91%(80%-97%). We concluded that due to ease in obtaining urine sample, FujiLAM can be used as point-of-care TB test in HIV negative children, however more data from different population is needed.

22Apr 2017 IMRT AND VMAT COMMISSIONING FOR VERSA HD LINEAR ACCELERATOR USING AAPM TG-119. Seema Sharma MSc Dip , RP Subhash Chander , MD Velliyan Subramani , PhD Pratik Kumar , PhD Suman Bhaskar , MD Sushmita Pathy , MD Sanjay Thulkar , MD N.Gopishankar , PhD Megha Sairem , MSc , Dip. RP Ashish Binjola MSc Dip , RP Dhayanethi N , MSc Pradeep Kumar S. , MSc Priyanka Agarwal , MSc Dip. RP. All India Institute of Medical Sciences, New Delhi, India.

Arrest of cell differentiation is one of the leading causes of leukemia and other cancers. Induction of cell differentiation using pharmaceutical agents has been clinically attempted for the treatment of these cancers. Epigenetic regulation may be one of the underlying molecular mechanisms controlling cell proliferation or differentiation. Here, we report on the use of proteomics-based differential protein expression analysis in conjunction with quantification of histone modifications to decipher the interconnections among epigenetic modifications, their modifying enzymes or mediators, and changes in the associated pathways/networks that occur during cell differentiation. During phorbol-12-myristate 13-acetate-induced differentiation of U937 cells, fatty acid synthesis and its metabolic processing, the clathrin-coated pit endocytosis pathway, and the ubiquitin/26 S proteasome degradation pathways were up-regulated. In addition, global histone H3/H4 acetylation and H2B ubiquitination were down-regulated concomitantly with impaired chromatin remodeling machinery, RNA polymerase II complexes, and DNA replication. Differential protein expression analysis established the networks linking histone hypoacetylation to the down-regulated expression/activity of p300 and linking histone H2B ubiquitination to the RNA polymerase II-associated FACT-RTF1-PAF1 complex. Collectively, our approach has provided an unprecedentedly systemic set of insights into the role of epigenetic regulation in leukemia cell differentiation.

Identifying protein post-translational modifications (PTMs) from tandem mass spectrometry data of complex proteome mixtures is a highly challenging task. Here we present a new strategy, named iterative search for identifying PTMs (ISPTM), for tackling this challenge. The ISPTM approach consists of a basic search with no variable modification, followed by iterative searches of many PTMs using a small number of them (usually two) in each search. The performance of the ISPTM approach was evaluated on mixtures of 70 synthetic peptides with known modifications, on an 18-protein standard mixture with unknown modifications and on real, complex biological samples of mouse nuclear matrix proteins with unknown modifications. ISPTM revealed that many chemical PTMs were introduced by urea and iodoacetamide during sample preparation and many biological PTMs, including dimethylation of arginine and lysine, were significantly activated by Adriamycin treatment in nuclear matrix associated proteins. ISPTM increased the MS/MS spectral identification rate substantially, displayed significantly better sensitivity for systematic PTM identification compared with that of the conventional all-in-one search approach, and offered PTM identification results that were complementary to InsPecT and MODa, both of which are established PTM identification algorithms. In summary, ISPTM is a new and powerful tool for unbiased identification of many different PTMs with high confidence from complex proteome mixtures.

We present measurements of the reduction of light output by plastic scintillators irradiated in the CMS detector during the 8 TeV run of the Large Hadron Collider and show that they indicate a strong dose rate effect. The damage for a given dose is larger for lower dose rate exposures. The results agree with previous measurements of dose rate effects, but are stronger due to the very low dose rates probed. We show that the scaling with dose rate is consistent with that expected from diffusion effects.

The importance of natural resources in sustaining productivity and environmental protection is now relatively more realized than the past. Over the past few decades or so, more and more attention is being paid all over the world to conserve the Natural Resources. Natural resources are important material basis for a stable economy and social development too With Industrialization and Urbanization, mankind’s great demand for natural resources and their large scale exploitation and consumption has resulted in the weakening, deterioration and exhaustion of these resources. Human existence depends on the natural resources and the environment and the maintenance of which is now increasingly being considered as essential for mankind. As human populations increase and natural resources become more limited, there is a critical need for trained conservation professionals in natural resources conservation.
 Natural Resources are those environmental gifts which satisfy the human wants. They are the means of attaining social objectives. Conservation of natural resources is the wise use of the earth's resources by humanity to achieve its benefits for the longest possible period of time and ensure availability of these resources for the further generation. One difficult task faced by all countries is to guarantee the lasting utilization of natural resources at the lowest possible environmental cost while still assuring economical and social development.

In mass spectrometry-based lipidomics, complex lipid mixtures undergo chromatographic separation, are ionized, and are detected using tandem MS (MSn) to simultaneously quantify and structurally characterize eluting species. The reported structural granularity of these identified lipids is strongly reliant on the analytical techniques leveraged in a study. For example, lipid identifications from traditional collisionally activated data-dependent acquisition experiments are often reported at either species level or molecular species level. Structural resolution of reported lipid identifications is routinely enhanced by integrating both positive and negative mode analyses, requiring two separate runs or polarity switching during a single analysis. MS3+ can further elucidate lipid structure, but the lengthened MS duty cycle can negatively impact analysis depth. Recently, functionality has been introduced on several Orbitrap Tribrid mass spectrometry platforms to identify eluting molecular species on-the-fly. These real-time identifications can be leveraged to trigger downstream MSn to improve structural characterization with lessened impacts on analysis depth. Here, we describe a novel lipidomics real-time library search (RTLS) approach, which utilizes the lipid class of real-time identifications to trigger class-targeted MSn and to improve the structural characterization of phosphotidylcholines, phosphotidylethanolamines, phosphotidylinositols, phosphotidylglycerols, phosphotidylserine, and sphingomyelins in the positive ion mode. Our class-based RTLS method demonstrates improved selectivity compared to the current methodology of triggering MSn in the presence of characteristic ions or neutral losses.

In perovskite oxides, the multiple charge states of cations influence their local structure and correlated physical properties. In this report, we studied the role of multivalence cations on the local structure and magnetic behavior of the oxygen-deficient system Sr2FeCoO6-δ (SFCO), prepared by the solid-state synthesis technique. A detailed structural and geometrical analysis confirmed the coexistence of orthorhombic (Brownmillerite) and cubic crystal symmetries and a gesture of the Jahn-Teller (JT) distortion. The intense Raman modes are allowed to be indexed for BO-polyhedra and JT-distortion. Experimental and theoretical analysis of the X-ray photoelectron spectroscopy (XPS) spectra supported the mixed oxidation state of Fe (Fe3+/Fe4+) and Co (Co2+/Co3+), further confirmed by theoretically simulated X-ray absorption spectroscopy (XAS) spectra. The presence of the multivalence states leads to AFM/FM exchange interaction in the sample which further leads to glassy magnetic behavior.

Abstract In the present work, we report a systematic theoretical study of the log ft values for the forbidden β − decay transitions in the 208 Pb region. For this, we have considered 206 Hg → 206 Tl, 208 Hg → 208 Tl, 206 Tl → 206 Pb and 208 Tl → 208 Pb transitions. We have performed shell model calculations using KHH7B interaction in valence shell 58-114 for protons and 100-164 for neutrons by considering 1p − 1h excitations for both protons and neutrons simultaneously for daughter nuclei. This study presents the first shell model results of β − -decay corresponding to the recent experimental data.

When it comes to its use in various food and feed applications, wheat is the cereal that is farmed most frequently worldwide. It is utilized for feedstock and grain production in both developed and developing nations. In 2020–21, India is predicted to produce a record 109.24 MT of wheat, up from 107.86 MT in 2019–20. The illness has been continuously controlled with the use of chemical treatments, resistant cultivars, and cultural practices. Fungicides significantly improved seed germination in one year, with Tilt 250 EC and Folicur being most effective, with other fungicides also showing consistent efficacy. The study examined wheat seed that had been administered with different fungicides to prevent disease. The results showed that all fungicides significantly boosted seed germination in a single year, with Folicur and Tilt 250 EC showing the highest efficacy. The trend of efficacy remained unchanged.

We use a thermodynamically consistent form of Tsallis distribution to study the dependence of various thermodynamic quantities on the system size in high-energy collisions. The charged hadron spectra obtained in $p$+$p$, $p$+Pb, Xe+Xe, and Pb+Pb collisions at LHC are used to determine the energy density, pressure, particle density, entropy density, mean free path, Knudsen number, heat capacity, isothermal compressibility, expansion coefficient, and speed of sound at the kinetic freeze-out surface. These quantities are studied as a function of the system size. Notably, the rate of increase (or decrease) in these thermodynamic variables is found to be more rapid in small systems such as $p$+$p$ and $p$+Pb collisions than in large systems such as Xe+Xe and Pb+Pb collisions. This may be due to the small volume of the hadronic system in small collision systems at kinetic freeze-out. It is observed that high-multiplicity $p$+$p$ collisions produce similar thermodynamic conditions as peripheral heavy-ion collisions at kinetic freeze-out.

Ideally, a solid electrolyte which is a central component of SOFC should exhibit high anionic or cationic ionic conductivity at the proposed operating temperatures. In most case, the performance is compromised when operating above 1000 °C due to poor mechanical, thermal and chemical stability of selected functional and non-functional materials. In this context, pyrochlores are one of the potential candidates due to their high conductivity, flexibility to accommodate large cations, and high mechanical, thermal and chemical stability. In this study, we report the synthesis of nano-powders of Ho2Hf2O7 (HH) and Ho2Zr2O7 (HZ) pyrochlore ceramics by eco-friendly alginate mediated ion-exchange process also known as Leeds Alginate Process (LAP) and provide further insight into the structure conductivity relationship of HH and HZ compounds by EXAFS studies. Both the compositions were sintered at temperatures, ranging from 1100 ºC-1500 °C at 2 h dwell time to achieve the desired high-density ceramic with stable pyrochlore structure. X-ray diffraction and Extended X-ray absorption fine structural (EXAFS) analysis showed superlattice reflections and complex coordination geometry of these oxides. The coordination number and disorder factor in the case of HZ were found to be more stable than the HH sample, as evident from the EXAFS. Impedance spectroscopy and dc-conductivity analysis showed a better charge transport behavior in HZ ceramics than in HH making HZ as a preferred solid electrolyte for SOFC. The conductivity of Ho2Zr2O7 is comparable with the best-known fluorite oxide-ion conductor such as Sc2O3-stabilized ZrO2 at 500 °C.

The 5 s/6 s (A-site)-3d (B-site) based double perovskites enounce multiple aspects to study the interactive effects on structural (global and local), electronic, and other physical properties of synthesized compounds. The characteristic feature of this family is the B-site cations, which have comparative ionic radii and multivalency. Here, we report a detailed analysis of the structural, electronic, and magnetic properties of Sr2FeMnO6 (SFMO) and La-substituted SrLaFeMnO6 (SLFMO). The relatively smaller radii but large valance La3+ (1.36 Å) substitution at Sr2+ (1.44 Å) sites led to adequate changes in the structural and physical properties. SLFMO crystallized in a lower crystal symmetry than SFMO, further confirmed by Raman analysis. Spectroscopic analyses showed that the Mn-valency compensates for the cationic disorder introduced by the La-substitution. These analyses also revealed that Mn and Fe hold high spin states of their 3+ and 4+ valence states, and thus, the studied compounds also have Jahn-Teller (JT) distortion. These structural and electronic distortions affect the magnetic properties of SFMO and SLFMO. SFMO showed antiferromagnetic behavior, while SLFMO showed spin-glass-type characteristics.

Ayurveda is a holistic science that emphasizes the maintenance and promotion of health through the prevention and curing aspects. According to Sushruta Samhita, term swasthya is the equilibrium state of Dosha, Agni, Dhatu, and Mala. Three doshas (vata, pitta, kapha), Agni (dhatvagni, bhutagni and jatharagni), Sapta Dhatu (Ras, rakta, mansa, meda, asthi, majja, shukra) and Trimala are responsible for maintaining good health. Rasayan is one of the eight clinical specialties of Ayurveda. The Rasayan therapy improves the qualities of Rasa and enriches it with nutrients so one can attain longevity, memory, intelligence, freedom from disorder, youthfulness, excellence of luster, complexion, and voice. The word “Rasayana” means the way to attain excellent Rasadi Dhatus. In Ayurveda, one of the major methods of presentation of positive health has been described i.e. Rasayana. This resistance power of the body, which prevents the development of diseases, is called Immunity or Vyadhikshamatva. The ultimate aim of Rasayana therapy is to correct Dosha imbalance and improve Agni and Dhatu function which overall improves strength and immunity. The application of Rasayan therapy comes in perspective of premature ageing and death. Rasayanas are used for preventive, curative, and health-promotive purposes.

Improving maternal & child health has been an essential tool for achieving health for all. The discourse on the Accredited Social Health Activist’s (ASHA’s) role centers around three typologies -as an activist, as a link worker or facilitator, and as a community level health care provider. To assess knowledge of ASHA workers regarding antenatal and neonatal care in a block of Haryana. The present cross-sectional, community-based study was conducted in block Barwala, district Hisar of Haryana. Universal sampling technique was used. A predesigned, pretested, semi-structured schedule was used to collect the information, which included details of socio-demographic profile of ASHA workers and questions about knowledge of antenatal & postnatal care. Appropriate statistical tests like frequencies and percentages test were applied to analyze the collected data. Only 6.9% ASHA workers told about benefits of early registration of pregnancy. Majority of ASHA workers had inadequate knowledge about identification of danger signs during pregnancy and infancy, duration of trimester. Majority of ASHA workers had adequate knowledge about breast-feeding practices however about half of ASHA workers had inadequate knowledge about duration of colostrum. Only about one third ASHA workers had adequate knowledge regarding weighing machine & colour zone used for identify malnutrition. : It is important to reorient ASHA workers at field level and supportive supervision also plays an important role.

We report the measurement of first-order event plane-correlated directed flow (v1) and triangular flow (v3) for identified hadrons (π±, K±, and p), net particle (net-K, net-p), and light nuclei (d and t) in Au + Au collisions at sNN = 3.2, 3.5, and 3.9 GeV in the fixed-target mode from the second phase of the beam energy scan (BES-II) program at RHIC-STAR. The v1 slopes at mid-rapidity for identified hadrons and net particles except π+ are found to be positive, implying the effect of dominant repulsive baryonic interactions. The slope of v1 for net-kaon undergoes a sign change from negative to positive at a lower collision energy compared to net-proton. An approximate atomic mass number scaling is observed in the measured v1 slopes of light nuclei at mid-rapidity, which favors the nucleon coalescence mechanism for the production of light nuclei. The v3 slope for all particles decreases in magnitude with increasing collision energy, suggesting a notable integrated impact of the mean-field, baryon stopping, and collision geometry at lower collision energies.

To study the safety and efficacy of high-dose-rate interstitial brachytherapy (HDRIBT) in patients with liver metastases (LM).Between 2009 and 2011, 10 patients with 12 metastatic lesions in the liver were enrolled in this prospective trial. All patients had either refused surgery or found ineligible for surgery due to various factors. Under CT guidance, 16 gauze blind end stainless steel or rigid plastic brachytherapy needle was inserted in the center of lesion through the percutaneous route. Generally, a single interstitial brachytherapy (IBT) needle for lesions up to 3 cm and multiple needles for lesions more than 3 cm in diameter were inserted. Treatment was delivered with a single high-dose-rate (HDR) dose of 20 Gy prescribed to the target. The needles were removed immediately after the treatment. The endpoints of study were acute complications and local control of the disease.The median size of the lesion was 3.8 cm (2.7-7.0 cm). The average time for the entire IBT procedure was 65 minutes (50-105 minutes). Median follow up was 9 months (3-17 months). None of the patients had fatal complications. Minor complications like pain, nausea/vomiting, and asymptomatic pleural effusion were observed in 3, 2 and 1 patients, respectively. Local control rate at 12 months was 75%. The 1-year local progression free survival (LPFS) was 33%.Although limited by small sample size, the results of our first study from India suggest that HDRIBT is a safe and effective non surgical option for LM.

Interstitial brachytherapy (IBT) is the standard alternative treatment for patients with cervical carcinoma not suitable for intracavitary radiotherapy. There is an emerging belief that intensity-modulated radiotherapy (IMRT) has the potential to replace IBT. We aimed to compare the dosimetry achieved by IBT and IMRT in such patients.The CT imaging data, previously used for IBT planning of 12 patients with cervical carcinoma, were transferred to IMRT planning system to generate parallel IMRT plans. Prescribed dose to the planning target volume (PTV) was 20Gy delivered in 2-weekly high-dose-rate fractions of 10Gy each with IBT (biologically equivalent dose [BED10] 40Gy) and 33Gy/13 fractions/2.5 wk with IMRT (BED10 41Gy). For comparison, dose-volume parameters for target and organs at risk were recorded and expressed in terms of BED10 and BED3, respectively.For PTV, the mean D95 (dose received by 95% of PTV) was better with IBT (57.16Gy vs. 41.47Gy, p=0.003). The mean conformity index was 0.94 and 0.90 with IBT and IMRT, respectively (p=0.034). IBT delivered significantly reduced doses to 1.0cc (Dmax), 5.0cc (D5 cc), 50% (D50), and 75% (D75) of bladder volume as compared with IMRT. The mean rectal Dmax was significantly better with IBT as compared with IMRT (54.64Gy vs. 62.63Gy, p=0.02).IBT provides superior PTV coverage and organs at risk sparing to IMRT. Thus, IBT remains the standard treatment for patients with cervical carcinoma unsuitable for intracavitary radiotherapy.

Bacteria obtained from the isolates of the biodiversity of riverine coast of Ganga identified as Bacillus Koriensis, when challenged with silver nitrate solution accumulated silver nanoparticles on the surface of its cell wall.These nanoparticles showed an absorption peak at 438 nm in UV-visible spectrum corresponding to the plasmon resonance of AgNPs.The transmission electron micrographs of nanoparticles in aqueous solution showed the production of reasonably monodisperse AgNPs (average particle size: 9.92 ± 1.311 nm) by the bacteria.X-ray diffraction spectrum of the nanoparticles confirmed the formation of metallic silver.

NWe present a simple and eco-friendly biosynthesis of silver nanoparticles using Pomegranate peel extract as the reducing agent. The extract was challenged with AgNO3 solution for the production of silver nanoparticles (AgNPs). The reaction process was simple for the formation of highly stable silver nanoparticles at room temperature by using the biowaste of the fruit. The morphology and crystalline phase of the NPs was determined from UV-Vis spectroscopy, transmission electron microscopy (TEM), selected area electron diffraction (SAED), X-ray diffraction (XRD) spectra and Fourier-Transform IR spectroscopy (FTIR). TEM studies showed that the silver nanoparticles obtained were of sizes 5 ± 1.5 nm. An effort has also been made to understand the possible involved mechanism for the biosynthesis of AgNPs. Presumably biosynthetic products or reduced cofactors play an important role in the reduction of respective salts to nanoparticles.

Tin oxide (SnO 2 ) nanofibers are successfully prepared by electrospinning homogeneous viscous solutions of tin acetate in polyvinylpyrrolidone (PVP).The electrospinning is carried out by applying a DC voltage to the tip of a syringe and maintaining the tip to collector distance (TCD), i.e. at DC electric field of 1.25 kV•cm -1 .The electrospun nanofibers are calcined between 550˚C and 650˚C for 4 h.Both spun and heat treated nanofibers are characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infra red spectroscopy (FTIR) etc. XRD analysis of calcined nanofibers confirms the formation of pure tin oxide.TEM study showed that fibers have a polycrystalline structure with multiple nano-grains.

Andrographis paniculata (AP) (Burm.f.) Wall.ex Nees (Family: Acanthaceae), commonly known as kalmegh (in Trade), 'kirayat' (Hindi), 'kalamegha' (Sanskrit) and 'Indian Echinacea' in English (Kumar et al., 2012) is an annual herb, native of South India and Srilanka (Raina et al., 2013a) and grows in abundance in Asian countries like India, Pakistan, Java, Malaysia and Indonesia (Joseph, 2014).In India, found wild throughout the plains mainly in states of Madhya Pradesh, Chhattisgarh, Orissa, Maharashtra, Assam, West Bengal, Uttar Pradesh, Uttrakhand, Tamil Nadu, Karnataka and Kerala (Prajapati et al., 2003).Plant is erect, up to 30 cm -1m high, stem acutely quadrangular with profuse branching.Leaves are simple, opposite, lanceolate with acute apex and short petiole.Inflorescence is terminal, axillary and panicle.Flower violet to white in colour and fruit capsule with numerous seeds (Anonymous, 1999;Sareer et al., 2014).Kalmegh is a reputed herb, which is commonly used in Ayurveda, Siddha, Unani and Homoeopathy systems of medicine as well as tribal medicines.Ayurvedic properties such as Rasa-Tikta, Guna-Laghu, Ruksha, Veerya-Ushna, Vipaka-Katu, Doshaghnata-Kaphapittashamaka, etc., are found in plant

Structural characterization of novel metabolites in drug discovery or metabolomics is one of the most challenging tasks. Multilevel fragmentation (MSn) based approaches combined with various dissociation modes are frequently utilized for facilitating structure assignment of unknown compounds. As each of the MS precursors undergoes MSn, the instrument cycle time can limit the total number of precursors analyzed in a single LC run for complex samples. This necessitates splitting data acquisition into several analyses to target lower concentration analytes in successive experiments. Here we present a new LC/MS data acquisition strategy, termed Met-IQ, where the decision to perform an MSn acquisition is automatically made in real time based on the similarity between the experimental MS2 spectrum and a spectrum in a reference spectral library for the known compounds of interest. If similarity to a spectrum in the library is found, the instrument performs a decision-dependent event, such as an MS3 spectrum. Compared to an intensity-based, data-dependent MSn experiment, only a limited number of MS3 are triggered using Met-IQ, increasing the overall MS2 instrument sampling rate. We applied this strategy to an Amprenavir sample incubated with human liver microsomes. The number of MS2 spectra increased 2-fold compared to a data dependent experiment where MS3 was triggered for each precursor, resulting in identification of 14-34% more unique potential metabolites. Furthermore, the MS2 fragments were selected to focus likely sources of useful structural information, specifically higher mass fragments to maximize acquisition of MS3 data relevant for structure assignment. The described Met-IQ strategy is not limited to metabolism experiments and can be applied to analytical samples where the detection of unknown compounds structurally related to a known compound(s) is sought.

Objective: The incidence of oral cancer worldwide varies 2-18% and in India 0.1 to 13.5%.Early evaluation of oral precancerous lesions can have dramatic effect on oral cancer mortality rate.Among the premalignant stage, leukoplakia is the most commonly encountered clinical lesion and epithelial dysplasia is most important predictive factor.The objective of this paper is to compare the efficacy of exfoliative cytology, toluidine blue and chemiluminescent illumination (VIZILITE) to detect early dysplastic changes in leukoplakia.Study design: Study involved 50 patients of leukoplakia (homogeneous, speckled) in the age group of 20-72 years including males and females in the ratio of 7:3.All the diagnostic procedures were carried out on each patient followed by biopsy and data was subjected to statistical analysis.Results: Comparing the results of exfoliative cytology with toluidine blue in diagnosing dysplasia in leukoplakia, it showed 50% sensitivity and 83.3 % specificity.In comparison to chemiluminescent light examination, cytology showed 42.9% sensitivity and 79.3% specificity.Chemiluminescent light examination showed 60% sensitivity and 70% specificity compared to toluidine blue.Conclusions: Overall accuracy of exfoliative cytology was less than toluidine blue, whereas latter showed superior but comparable results to chemiluminescent illumination in detecting dysplasia.However role of chemiluminescent illumination should be further investigated in demarcating dysplatic lesions.

Electrocardiogram (ECG) is one of the most practiced methods to detect any abnormalities in heart function.ECG results are available as paper records.This leads to Archival and retrieval of patient's paper ECG records as a common practice for diagnosis of ailments.This process requires large storage space and extensive manual effort.This paper discusses method that involves processing of ECG paper records by an efficient and iterative set of digital image processing techniques for the conversion of ECG paper image data to time series digitized signal form, resulting in convenient storage and retrieval of ECG information.The method involves calculation of Heart rate, QRS Width and Stability (variation in R-R peaks) from the extracted signal.Comparison of the above calculated parameters with the manually calculated parameters shows an accuracy of 96.4%, hence proves the effectiveness of the process.This paper also proposes the development of fuzzy based ECG diagnosis system that assists the doctor in diagnosis.

This study compares the dosimetry of high-dose-rate intracavitary brachytherapy (HDR-ICBT) performed with and without general anesthesia/spinal anesthesia (GA/SA) in patients with cervical carcinoma. We retrospectively retrieved the records of 138 HDR-ICBT applicator insertions performed in 46 patients: 69 performed with GA/SA (anesthesia group known as AG) in 23 patients, and 69 performed without GA/SA (nonanesthesia group known as NAG) in 23 patients. The intracavitary brachytherapy (ICBT) application was done with central tandem and two vaginal ovoids. For each ICBT plan, a high-dose-rate (HDR) dose of 7 Gy was prescribed to point A. From each plan, the doses to Point B right (B(R)), Point B left (B(L)), bladder and rectal reference points (Bladder(ref) and Rectal(ref)) were recorded and compared in the two groups. Student's t-test was applied to find out the significance of difference. The two groups were comparable in terms of demography and clinical characteristics. Mean Point BL doses in AG and NAG were 1.89 Gy (27% of Point A dose) and 1.82 Gy (26% of Point A dose), respectively. Mean Point BR doses in AG and NAG were 1.91 Gy (27% of Point A dose) and 1.85 Gy (26% of point A), respectively (p-value 0.7). The mean dose to Bladder(ref) in AG and NAG was 5.03 Gy and 4.90 Gy, respectively (p-value 0.6). The mean dose to Rectal(ref) was significantly higher in AG than NAG (5.09 Gy vs. 4.49 Gy, p-value 0.01). Although based on conventional 2D dosimetry planning, our study has demonstrated that avoiding GA/SA does not result in inferior HDR-ICBT dosimetry.

We report bronchoalveolar lavage (BAL) cellular analysis in eight patients with miliary tuberculosis. The total cell count and lymphocytes count of BAL fluid were significantly increased and the alveolar macrophage count was significantly decreased in patient with miliary tuberculosis compared to normal subjects. Epithelioid cell granulomas were seen in BAL fluid in two patients.

Ferroelectric lead lanthanum titanate (Pb 1- x La x Ti 1- x /4 O 3 ) (PLT x ) thin films ( x =0.04, 0.08 and 0.12) have been prepared by sol–gel spin-coating process on ITO- coated 7059 Corning glass substrates. Investigations have been made on the crystal structure, surface morphology, dielectric and ferroelectric properties of the thin films. Characterization of these films by X-ray diffraction shows that the films annealed at 650°C exhibit tetragonal structure with perovskite phase. Replacement of lanthanum in lead titanate results in reduction of the tetragonal ratio ( c / a ), thereby resulting in better mechanical stability. Microstructural analyses of the films are carried out by obtaining atomic force microscope (AFM) images. AFM images are characterized by slight surface roughness with a uniform crack-free, densely packed structure. Dielectric, pyroelectric and ferroelectric studies carried out on these films have been reported. The dielectric constant increases with La concentration. The Curie temperature is found to decrease with the addition of lanthanum. The pyroelectric coefficient increases from 17 nC/cm 2 K to 31 nC/cm 2 K as La content increases from 4 to 12%. The pyroelectric figures of merit of the films have been calculated from these studies. These studies indicate that 8% lanthanum is the material best suited for infrared detectors owing to its high pyroelectric coefficient (∼29 nC/cm 2 K), high voltage responsivity (∼420 Vcm 2 /J), high detectivity (∼1.04×10 -5 Pa -1/2 ) and low variation of pyrocoefficient with temperature.

No AccessJournal of UrologyInvestigative Urology1 Oct 2010Origin and Characterization of Retrograde Labeled Neurons Supplying the Rat Urethra Using Fiberoptic Confocal Fluorescent Microscopy In Vivo and Immunohistochemistry Keon-Cheol Lee, Seema Sharma, Jeremy B. Tuttle, and William D. Steers Keon-Cheol LeeKeon-Cheol Lee Department of Urology, Inje University School of Medicine, Ilsanpaik Hospital, Goyang-si, Korea More articles by this author , Seema SharmaSeema Sharma Department of Urology, University of Virginia School of Medicine, Charlottesville, Virginia More articles by this author , Jeremy B. TuttleJeremy B. Tuttle Department of Neuroscience, University of Virginia School of Medicine, Charlottesville, Virginia More articles by this author , and William D. SteersWilliam D. Steers Department of Urology, University of Virginia School of Medicine, Charlottesville, Virginia More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.05.092AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract Purpose: Autonomic innervation of urethral smooth muscle may influence urinary continence after prostatectomy. It is unclear whether the cavernous nerves carry fibers that influence continence. Using a retrograde axonal tracer combined with real-time in vivo imaging and ex vivo immunohistochemistry we determined the course and type of neurons supplying urethral smooth muscle distal to the prostate in the rat. Materials and Methods: We injected the retrograde axonal tracers cholera toxin B fragment-Alexa Fluor® 488 and Fast Blue in the distal urethral smooth muscle in 10 rats each. Five days later the cavernous nerves and pelvic ganglion were imaged using fiberoptic confocal fluorescence microscopy (cholera toxin B fragment-Alexa Fluor 488) or harvested for immunohistochemistry (Fast Blue). Dual immunofluorescence of Fast Blue neurons with tyrosine hydroxylase or neuronal nitric oxide synthase was done to characterize neurons as noradrenergic or nitrergic. To ascertain whether the cavernous nerves contain fibers to the urethra that originate in the pelvic ganglia we cut the cavernous nerves with their ancillary branches in 3 rats and imaged them for Fast Blue. Results: Fluorescent neurons and axons were detected in cavernous nerves and the pelvic ganglion. Few neurons were seen in rats with cavernous nerve section. Of urethral neurons 53.1% showed neuronal nitric oxide synthase positivity while 40.6% were immunoreactive for tyrosine hydroxylase. About 6.2% of urethral neurons failed to show tyrosine hydroxylase or neuronal nitric oxide synthase immunoreactivity. Conclusions: Most of the autonomic innervation to the urethra beyond the prostatic apex travels in the cavernous nerves. Many nerves may be parasympathetic based on neuronal nitric oxide synthase immunoreactivity. Nerves supplying the urethra outside the cavernous nerves may course posterior to the prostate. Along with afferent fibers, tyrosine hydroxylase immunoreactivity expressing neuron fibers, ie noradrenergic nerves, traveling in the cavernous nerves may increase urethral resistance or regulate the reflex mechanisms controlling continence. References 1 : Improving the exposure of the prostate in radical retropubic prostatectomy: longitudinal bunching of the deep venous plexus. J Urol1989; 142: 1282. Link, Google Scholar 2 : Anatomic radical prostatectomy: evolution of the surgical technique. J Urol1998; 160: 2418. Link, Google Scholar 3 : Risk factors for urinary incontinence after radical prostatectomy. J Urol1996; 156: 1707. Link, Google Scholar 4 : Nerve sparing open radical retropubic prostatectomy—does it have an impact on urinary continence?. J Urol2006; 176: 189. Link, Google Scholar 5 : Complications of open radical retropubic prostatectomy in potential candidates for active monitoring. Urology2008; 72: 887. Google Scholar 6 : Urinary striated sphincter: what is its nerve supply?. Urology1982; 20: 41. Google Scholar 7 : The role of membranous urethral afferent autonomic innervation in the continence mechanism after nerve sparing radical prostatectomy: a clinical and prospective study. J Urol2008; 180: 2527. Link, Google Scholar 8 : Regional variations in the neural control of the female pig urethra. Br J Urol1995; 76: 730. Google Scholar 9 : Urethral pudendal afferent evoked bladder and sphincter reflexes in decerebrate and acute spinal cats. Neurosci Lett1998; 244: 137. Google Scholar 10 : Fiberoptic imaging of cavernous nerves in vivo. J Urol2007; 178: 2694. Link, Google Scholar 11 : The role of bladder-to-urethral reflexes in urinary continence mechanisms in rats. Am J Physiol Renal Physiol2004; 287: F434. Google Scholar 12 : Unusual autonomic ganglia: connections, chemistry, and plasticity of pelvic ganglia. Int Rev Cytol1999; 193: 1. Google Scholar 13 : The distribution of noradrenergic fibers in the human lower urinary tract. Eur Urol1999; 36: 23. Google Scholar 14 : Pharmacological properties of A-204176, a novel and selective alpha1A adrenergic agonist, in in vitro and in vivo models of urethral function. Life Sci2001; 70: 181. Google Scholar 15 : Urethral afferent nerve activity affects the micturition reflex; implications for stress incontinence and detrusor instability. J Urol1999; 162: 204. Link, Google Scholar 16 : Mediators and mechanisms of relaxation in rabbit urethral smooth muscle. Br J Pharmacol1998; 123: 617. Google Scholar 17 : Micturitional static urethral pressure profilometry in women. Neurourol Urodyn1982; 1: 149. 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Google Scholar © 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 184Issue 4October 2010Page: 1550-1554 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.Keywordsurinationsmoothmuscleprostateurethraautonomic pathwaysMetricsAuthor Information Keon-Cheol Lee Department of Urology, Inje University School of Medicine, Ilsanpaik Hospital, Goyang-si, Korea More articles by this author Seema Sharma Department of Urology, University of Virginia School of Medicine, Charlottesville, Virginia More articles by this author Jeremy B. Tuttle Department of Neuroscience, University of Virginia School of Medicine, Charlottesville, Virginia More articles by this author William D. Steers Department of Urology, University of Virginia School of Medicine, Charlottesville, Virginia More articles by this author Expand All Advertisement PDF downloadLoading ...

Dosimetric comparison of two dimensional (2D) radiography and three-dimensional computed tomography (3D-CT) based dose distributions with high-dose-rate (HDR) intracavitry radiotherapy (ICRT) for carcinoma cervix, in terms of target coverage and doses to bladder and rectum.Sixty four sessions of HDR ICRT were performed in 22 patients. External beam radiotherapy to pelvis at a dose of 50 Gray in 27 fractions followed by HDR ICRT, 21 Grays to point A in 3 sessions, one week apart was planned . All patients underwent 2D-orthogonal and 3D-CT simulation for each session. Treatment plans were generated using 2D-orthogonal images and dose prescription was made at point A. 3D plans were generated using 3D-CT images after delineating target volume and organs at risk. Comparative evaluation of 2D and 3D treatment planning was made for each session in terms of target coverage (dose received by 90%, 95% and 100% of the target volume: D90, D95 and D100 respectively) and doses to bladder and rectum: ICRU-38 bladder and rectum point dose in 2D planning and dose to 0.1cc, 1cc, 2cc, 5cc, and 10cc of bladder and rectum in 3D planning.Mean doses received by 100% and 90% of the target volume were 4.24 ± 0.63 and 4.9 ± 0.56 Gy respectively. Doses received by 0.1cc, 1cc and 2cc volume of bladder were 2.88 ± 0.72, 2.5 ± 0.65 and 2.2 ± 0.57 times more than the ICRU bladder reference point. Similarly, doses received by 0.1cc, 1cc and 2cc of rectum were 1.80 ± 0.5, 1.48 ± 0.41 and 1.35 ± 0.37 times higher than ICRU rectal reference point.Dosimetric comparative evaluation of 2D and 3D CT based treatment planning for the same brachytherapy session demonstrates underestimation of OAR doses and overestimation of target coverage in 2D treatment planning.

Study Objective To show laparoscopic management of disseminated peritoneal leiomyomatosis (DPL). Design Stepwise demonstration of the technique with narrated video footage. Setting DPL is characterized by dissemination and proliferation of peritoneal and subperitoneal lesions primarily originating from smooth muscle cells [ 1 Yang L Liu N Liu Y Leiomyomatosis peritonealis disseminata: three case reports. Medicine. 2020; 99: e22633 Crossref PubMed Scopus (3) Google Scholar ]. Generally considered benign, cases of malignant transformation to leiomyosarcoma have been reported [ 2 Sharma P Chaturvedi KU Gupta R Nigam S Leiomyomatosis peritonealis disseminata with malignant change in a post-menopausal woman. Gynecol Oncol. 2004; 95: 742-745 Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar , 3 Żyła MM Dzieniecka M Kostrzewa M et al. Leiomyomatosis peritonealis disseminata of unusual course with malignant transformation: case report. Acta Obstet Gynecol Scand. 2015; 94: 220-223 Crossref PubMed Scopus (13) Google Scholar ]. Iatrogenic DPL occurs because of unconfined morcellation resulting in small fragments of myoma that may implant on any organ and start deriving blood supply from it or may be pulled into port site while withdrawing laparoscopic cannulas [ 4 Nezhat C Kho K Iatrogenic myomas: new class of myomas?. J Minim Invasive Gynecol. 2010; 17: 544-550 Abstract Full Text Full Text PDF PubMed Scopus (66) Google Scholar ]. It is estimated that the overall incidence of DPL after laparoscopic uncontained morcellation was 0.12% to 0.95% [ 5 Cheng PJ Duan T China's new two-child policy: maternity care in the new multiparous era. BJOG. 2016; 123: 7-9 Crossref PubMed Scopus (51) Google Scholar ]. Mainstay of treatment is surgical resection of myomas and regular follow-up with imaging. A 28-year-old unmarried girl presented with complain of lump abdomen increasing in size for 1 year. She also complained of a 15 kg weight loss in the last 1 year; 4 years ago, patient had undergone laparoscopic myomectomy with unconfined morcellation for a 10 × 8 cm cervical myoma. Presently her menses were regular with a 28-day cycle and 3 to 4 days’ average flow. Magnetic resonance imaging showed multiple nodular lesions of varying sizes in relation to small bowel, colon, uterus, and anterior abdominal wall suggestive of DPL. Bilateral ovaries were normal. Tumor markers were as follows: CA 125 23.2 (<35) U/mL Carcinoembryonic antigen 1.67 (<8) ng/mL CA 19-9 47 (<37) U/mL Lactate dehydrogenase 809 (180–360) IU/L Alpha-fetoprotein 2.03 (<10) ng/mL Beta human chorionic gonadotropin 1.2(<2) mIU/mL Tru-cut biopsy was done elsewhere to rule out peritoneal carcinomatosis in view of raised CA 19-9 and lactate dehydrogenase, history of weight loss, and imaging showing multiple abdominal masses. Histopathological examination showed leiomyomatosis and immunohistochemistry for smooth muscle actin, desmin, and vimentin were positive. Interventions On laparoscopy the abdominal cavity was found studded with multiple leiomyomas of varying sizes deriving blood supply from ilium, transverse, descending and sigmoid colon, rectum, left tube, left ovary, pouch of Douglas, bilateral uterosacrals, uterovesical fold, and anterior abdominal wall. Large blood vessels were seen traversing between the descending and sigmoid colon and the myomas. Principles of surgery were as follows: 1. Complete removal of myomas 2. Cauterization of blood vessels feeding the parasitic myomas to minimize blood loss 3. Disscetion abutting the myoma to prevent injury to adjacent viscera. A total of 26 myomas were removed. All the myomas were retrieved by morcellation in a bag. Histopathology confirmed the diagnosis of diffuse peritoneal leiomyomatosis. Follow-up ultrasound at 6 months showed no recurrence of leiomyomatosis. Conclusion Proper mapping of lesions and surgery for complete removal of all masses is the mainstay of treatment. Contained morcellation in bag should be the norm to prevent iatrogenic DPL. Regular follow-up with imaging is required to rule out recurrence.

In the present work, we report a systematic theoretical study of the $\log ft$ values for the forbidden $β^-$ decay transitions in the $^{208}$Pb region. For this, we have considered $^{206}$Hg $\rightarrow$ $^{206}$Tl, $^{208}$Hg $\rightarrow$ $^{208}$Tl, $^{206}$Tl $\rightarrow$ $^{206}$Pb and $^{208}$Tl $\rightarrow$ $^{208}$Pb transitions. We have performed shell model calculations using KHH7B interaction in valence shell 58-114 for protons and 100-164 for neutrons by considering ${\it 1p-1h}$ excitations for both protons and neutrons simultaneously for daughter nuclei. This study presents the first shell model results of $β^-$-decay corresponding to the recent experimental data.

Lanthanum modified lead titanate (Pb1−xLaxTi1−x/4O3) PLTx (x=0.08 i.e. PLT8) sol–gel derived thin films have been prepared on indium tin oxide (ITO) coated glass and quartz substrates using lead acetate trihydrate, lanthanum acetate hydrate and titanium isopropoxide as precursors along with 2-methoxyethanol as solvent and acetic acid as catalyst by spin coating method. The microstructure and surface morphology of the films annealed at 650 °C have been studied by X-ray diffraction technique and atomic force microscope (AFM). XRD has shown a single phase with tetragonal structure and AFM images have confirmed a smooth and crack-free surface with low surface roughness. The dependence of leakage current on applied voltage show ohmic behavior at low field region with a space charge conduction mechanism at high fields. The wavelength dispersion curve of thin films obtained from the transmission spectrum of thin films show that the films have high optical transparency in the visible region.

Induced pluripotent stem cells (iPSC) hold great promise for regenerative medicine as well as for investigations into the pathogenesis and treatment of various diseases. Understanding of key intracellular signaling pathways and protein targets that control development of iPSC from somatic cells is essential for designing new approaches to improve reprogramming efficiency. Here, we report the development and application of an integrated quantitative proteomics platform for investigating differences in protein expressions between mouse embryonic fibroblasts (MEF) and MEF-derived iPSC. This platform consists of 16O/18O labeling, multidimensional peptide separation coupled with tandem mass spectrometry, and data analysis with UNiquant software. With this platform, a total of 2481 proteins were identified and quantified from the 16O/18O-labeled MEF-iPSC proteome mixtures with a false discovery rate of 0.01. Among them, 218 proteins were significantly upregulated, while 247 proteins were significantly downregulated in iPSC compared to MEF. Many nuclear proteins, including Hdac1, Dnmt1, Pcna, Ccnd1, Smarcc1, and subunits in DNA replication and RNA polymerase II complex, were found to be enhanced in iPSC. Protein network analysis revealed that Pcna functions as a hub orchestrating complicated mechanisms including DNA replication, epigenetic inheritance (Dnmt1), and chromatin remodeling (Smarcc1) to reprogram MEF and maintain stemness of iPSC.

Abstract Advent of surface‐enhanced Raman spectroscopy (SERS) has engendered, the attention to exploit a facile, cost effective, 2D self‐assembly based architecture for sensing. Here, Au nanoparticles are decorated on controlled morphology of 2D polystyrene colloidal crystals (PSCCs), partially embedded into flexible polydimethylsiloxane elastomer. Self‐assembled flexible hexagonally close‐packed PSCCs form the 75 and 25 nm voids in the 500 and 200 nm, PSCCs monolayers and act as an effective plasmonic propagator for SERS sensors signal. Optical and sensing response investigates with UV‐Vis and Raman spectroscopy after the exposure of Au NP/2D PS CC/PDMS to NH 3 , H 2 O 2 , N 2 O and H 2 S gases environment. The increase in the SERS intensity at 491 cm −1 for the 200 nm, CCs based sensors as compared to 500 nm, indicates that the sensing response, strongly dependent on the CCs size. The red‐shift in the UV‐Vis spectra and enhancement in intensity of SERS designate that H 2 S forms the maximum Au NPs aggregates over the all tested gases. The chemisorptions on the phenyl ring, due to B 2 symmetric modes of the PSCCs with orthogonal polarizability of Au NPs, act as a hot spot for higher sensing response for the detection of gaseous precursors with higher SERS sensitivity and reliability.

In top quark production, the polarization of top quarks, decided by the chiral structure of couplings, is likely to be modified in the presence of any new physics contribution to the production. Hence, it is a good discriminator for those new physics models wherein the couplings have a chiral structure different than that in the Standard Model. In this paper, we construct probes of the polarization of a top quark decaying hadronically, using easily accessible kinematic variables such as the energy fraction or angular correlations of the decay products. Tagging the boosted top quark using the jet substructure technique, we study the robustness of these observables for a benchmark process, ${W}^{\ensuremath{'}}\ensuremath{\rightarrow}tb$. We demonstrate that the energy fraction of $b$ jet in the laboratory frame and a new angular variable, constructed by us in the top rest frame, are both very powerful tools to discriminate between the left and right polarized top quarks. Based on the polarization-sensitive angular variables, we construct asymmetries that reflect the polarization. We study the efficacy of these variables for two new physics processes that give rise to boosted top quarks: (i) the decay of the top squark in the context of supersymmetry searches and (ii) decays of the Kaluza-Klein (KK) graviton and KK gluon, in the Randall-Sundrum model. Remarkably, it is found that the asymmetry can vary over a wide range about $+20%$ to $\ensuremath{-}20%$. The dependence of asymmetry on top quark couplings of the new particles present in these models beyond the SM is also investigated in detail.

One dimensional nanofibers of organic and inorganic materials have been used in filters, optoelectronic devices, sensors etc.It is difficult to obtain ultra fine fibers of inorganic materials having lengths in the order of millimeter as they tend to break during formation due to thermal and other mechanical stresses.In this study, we have investigated the mechanism to prevent the defect formation and the breaking of CuO nanofibers by using optimized heat flow rates.CuO nanofibers were obtained by heat treating the poly(vinyl acetate) PVA composite fibers formed by electrospinning.The morphology and structural characteristics of prepared samples were investigated by Scanning electron microscopy, Transmission electron microscopy and X-ray diffraction.It was found that the morphology of the composite and annealed nanofibers could be influenced by the concentration of the polymer content.A lower concentration favors the formation of defects along the fiber and the number of defects reduces when the concentration is increased.

Spinal atypical teratoid/rhabdoid tumour (AT/RT) is exquisitely rare and constitutes 2% of all AT/RTs.A 6-year-old boy presented with low backache for the last 5 months. MRI of the spine showed a 1.5 × 1.5 × 4.7 cm intradural extramedullary mass extending from D10 to D12, causing compression of the conus medullaris. With a preoperative diagnosis of ependymoma, a gross total resection (GTR) of tumour was performed. Post-operative histopathology showed AT/RT. The tumour cells were immunopositive for cytokeratin, epithelial membrane antigen, smooth muscle actin, and p53 and immunonegative for MIC2, desmin, glial fibrillary acidic protein, and INI1. Post-operative neuraxis MRI revealed post-operative changes (D10-D12) with a 9 mm enhancing lesion at L5-S1 junction suggesting drop metastasis. There was no lesion in brain. Cerebrospinal fluid cytology did not show any malignant cell. The metastatic work-up was normal. He received 3 cycles of chemotherapy with ICE regimen (ifosfamide, carboplatin, and etoposide). Subsequently, he received craniospinal irradiation (CSI)-36 Gy/20 fractions/4 weeks followed by focal boost to primary tumour bed and spinal drop metastasis-14.4 Gy/8 fractions/1.5 weeks. Thereafter, he received 3 more cycles of ICE regimen. End-of-treatment MRI spine showed post-op changes (D10-D12) and 38.9% reduction of the L5-S1 lesion suggesting partial response. Six monthly spinal MRI showed serial reduction of the metastatic lesion leading to complete response (CR) 1 year after completion of treatment. On last follow-up (30 months from the initial diagnosis), he was neurologically intact and in CR.Multimodality management comprising GTR of tumour, CSI followed by focal boost, and multiagent chemotherapy (ICE) can lead to successful outcome in patients with this rare and aggressive spinal tumour.

Andrographis paniculata, a member of family Acanthaceae commonly found in plains of India, Pakistan and Sri Lanka and is an important medicinal plant, which is used in diseases like fevers, jaundice, colic dysentery and dyspepsia and also act as hepatoprotective, bitter tonic, stomachic, immunostimulant, antidote, antimalarial agent etc. The medicinal value of the plant is mainly due to active components as andrographolide, deoxy-andrographolide, neoandrographolide etc. present in the species. In the present study andrographolide, a major diterpenoid in Andrographis paniculata was quantified in different parts of plant using HPLC. The leaves were found to contain the highest content of andrographolide. The content ranged from 0.054 – 4.686%. The other parts studied were stem, flowering tops and roots. The andrographolide was detected in all four parts of the plant.

Polycrystalline samples of Bi(Mg 0.5 Ti 0.5 )O 3 -PbTiO 3 (BMT-PT) solid solutions exhibit high ferroelectric Curie temperatures and are promising materials for high temperature piezoelectric devices.A morphotropic phase boundary (MPB) between ferroelectric rhombohedral and tetragonal phases occurs between BMT-PT ratio of 70-30 and 65-35 compositions.In the present investigation, ceramics having BMT-PT ratios in the range from 70-30 to 50-50 were prepared by a conventional solid state reaction method.The change of crystal structure as a function of composition and temperature has been studied using high temperature X-ray diffraction.Polarisation-Electric field hysteresis curves at different temperatures show antiferroelectric behaviour for x = 0.30, 0.35 and 0.40 compositions while 0.45 and 0.50 show a ferroelectric behavior.

Non-target screening (NTS) based on the combination of liquid chromatography coupled to high-resolution mass spectrometry has become the key method to identify organic micro-pollutants (OMPs) in water samples. However, a large number of compounds remains unidentified with current NTS approaches due to poor quality fragmentation spectra generated by suboptimal fragmentation methods. Here, the potential of the alternative fragmentation technique ultraviolet photodissociation (UVPD) to improve identification of OMPs in water samples was investigated. A diverse set of water-relevant OMPs was selected based on k-means clustering and unsupervised artificial neural networks. The selected OMPs were analyzed using an Orbitrap Fusion Lumos equipped with UVPD. Therewith, information-rich MS2 fragmentation spectra of compounds that fragment poorly with higher-energy collisional dissociation (HCD) could be attained. Development of an R-based data analysis workflow and user interface facilitated the characterization and comparison of HCD and UVPD fragmentation patterns. UVPD and HCD generated both unique and common fragments, demonstrating that some fragmentation pathways are specific to the respective fragmentation method, while others seem more generic. Application of UVPD fragmentation to the analysis of surface water enabled OMP identification using existing HCD spectral libraries. However, high-throughput applications still require optimization of informatics workflows and spectral libraries tailored to UVPD.

The aim of present study was to assess the role of E-cadherin in oral carcinogenesis by comparing their expressions in normal oral mucosa, oral epithelial dysplasia (OED), and oral squamous cell carcinoma (OSCC).To elucidate the pattern of E-cadherin expression in oral carcinogenesis, 21 archival cases of formalin-fixed paraffin-embedded sections of OSCC, 21 OED, and 7 normal oral mucosa samples as control were used for the study.We observed reduction/loss of E-cadherin in membranous expression pattern and staining intensity with progression from dysplasia to oral cancer.A decrease in staining intensity and loss of E-cadherin membranous expression were noted from dysplasia to carcinoma, suggesting its role as a tumor suppressor gene.E-cadherin can be used as a biomarker to assess and evaluate the progression and prognosis of oral dysplastic lesions and OSCC.

Sodium potassium niobate (KNN) (K 0.5 Na 0.5 NbO 3 ) nanopowder with a mean particle size of about 20 -30 nm was synthesized by wet chemical route using Nb 2 O 5 as Nb source.A solution of K, Na and Nb cations was prepared, which resulted in a clear gel after the thermal treatment.Phase analysis, microstructure and morphology of the powder were determined by X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Field Emission Scanning Electron Microscopy (FESEM).The obtained gel was first analyzed by Thermo Gravimetric Analyzer (TGA) and Differential Scanning Calorimetry (DSC), and then calcined at different temperatures of 400˚C, 500˚C, 600˚C and 700˚C.The X-Ray Diffraction (XRD) patterns of the synthesized samples confirmed the formation of the orthorhombic crystal phase of K 0.5 Na 0.5 NbO 3 at 500˚C, a temperature significantly lower than that typically used in the conventional mixed oxide route.The process developed in this work is convenient to realize the mass production of KNN nanopowders at low cost and suitable for various industrial applications.

In the present paper, we synthesized the CdS hollow spheres by using PMMA sphere templates of 298-301 nm diameters and 20-51 nm of shell thickness. A CdS hollow sphere was characterized by X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), optical absorption and photoluminescence technique. CdS products are all cubic face-centered structure with the cell constant a = 5.815 Å. We also explore the morphology, structure and possible synthesis mechanism. A possible template mechanism has been proposed for the production of the hollow CdS nano-particles. The band gap of bulk CdS is about 2.45 eV, showing an absorption onset of bulk at about 513 nm. This shows a blue shift in the absorption spectra due to the quantum size effect, which is quite possible due to the small size of the CdS nano crystals as is evident in XRD pattern. The diameter of the beads is about 265-310 nm. The change in beads size due to the CdS over-layer is not so apparent in structures, due to its small thickness. The average diameter of the sphere is similar to that of the beads. Therefore, the spherical shells were obtained after the removal of PMMA core.

To describe our institutional experience with high dose rate (HDR) interstitial brachytherapy (IBT) compared with previously reported results on the low dose rate (LDR) practice for head and neck cancer.Eighty-four patients with oral cavity (n=70) or oropharyngeal cancer (n=14) were treated with 192Ir HDR-IBT. Seventy-eight patients had stage I or II tumour. The patients treated with IBT alone (n=42) received 39-42 Gy/10-14 fractions (median=40 Gy/10 fractions). With respect to the combination therapy group (n=42), prescription dose comprised of 12-18 Gy/3-6 fractions (median=15 Gy/5 fractions) for IBT and 40-50 Gy/20-25 fractions (median=50 Gy/25 fractions) for external radiotherapy. Brachytherapy was given as 2 fractions per day 6 hours apart with 4 Gy per fraction for monotherapy and 3 Gy per fraction for combination therapy.Four patients were not evaluable in the analysis of outcome. The primary site relapse rates were 23.8% (10/42) and 68.4% (26/38) in patients treated with IBT alone and combination therapy, respectively (p<0.001). Salvage surgery was performed in 19 patients. The 5-year local control rate was estimated at 62% and the disease-free survival (DFS) rate at 52% for all patients. Local control with respect to T1 and T2 tumours was 84% and 42%, respectively.Our present series on HDR-IBT and the previous report on LDR-IBT for head and neck cancer demonstrated similar DFS rates at 5 years (52%). The rate of regional failure in node-negative patients was <20% in both of our series. HDR-IBT offers similar results to LDR-IBT for head and neck cancer.

A ring chromosome 12 (p13; q24.33) was observed in all cells analyzed from peripheral blood lymphocytes of a 15-year-old female referred for academic difficulties and growth delay. In addition to clinical manifestations generally observed with ring chromosome 12 such as growth retardation, mental deficiency, microcephaly, the patient had bilateral pseudocamptodactyly of little fingers, mild hirsutism, exaggerated lumbar lordosis, and ostium secundum atrial septal defect (ASD). The clinical features of reported cases are analyzed. The only consistent features were growth retardation and mental deficiency. Breakpoint in all the cases has been at the telomeric region with minimal deletion of chromosomal material. An account of complex changes at mitosis and meiosis in ring chromosome has been given. Examination of 200 metaphases demonstrated 2% cell line was showing 45,XX, -12. Serum lactate dehydrogenase (LDH) level was normal ruling out overlapping monosomy 12 syndrome.

In recent years the Tsallis statistics is gaining popularity in describing charged particle produc- tion and their properties, in particular pT spectra and the multiplicities in high energy particle collisions. Motivated by its success, an analysis of the LHC data of proton-proton collisions at ener- gies ranging from 0.9 TeV to 7 TeV in different rapidity windows for charged particle multiplicities has been done. A comparative analysis is performed in terms of the Tsallis distribution, the Gamma distribution and the shifted-Gamma distribution. An interesting observation on the inapplicability of these distributions at sqrt{s}=7 TeV in the lower rapidity windows is intriguing. The non-extensive nature of the Tsallis statistics is studied by determining the entropic index and its energy depen- dence. The analysis is extrapolated to predict the multiplicity distribution at sqrt{s}=14 TeV for one rapidity window, |y| < 1.5 with the Tsallis function.

Coated bearings in high-performance turbine engines have the potential to provide considerable improvement in engine reliability and service-life extension. One coating performance indicator is the level of vibration generated by rolling elements and their cage separators at high-speed bearing operation. This article introduces a nonintrusive, cage-mounted sensor that detects cage vibration and transmits wirelessly to a nearby receiver. The sensitivity of the cage-mounted sensor was compared to a housing-mounted accelerometer in the detection of seeded faults in coated and uncoated bearings. The smallest spall on an uncoated bearing that the sensor could detect was 28 μm deep × 3.2 mm wide, with a reliability of ∼40%. In contrast, the accelerometer consistently detected all levels of flaws, the smallest being 0.4 μm deep × 3.2 mm wide on the coated bearing, with a reliability of 55%. Despite the sensor having a detection rate less than the accelerometer, this research successfully demonstrated that the sensor could detect inner-race-bearing flaws and established its sensitivity level.

Polycrystalline samples of 1- x(Na0.5 K0.5)(Nb0.95 Ta0.05) ) -x(Bi Fe)O3 with x=0, 0.003, 0.005, 0.007) hereby denoted as NKNT-BF were prepared by the mixed oxide method. Preliminary structural studies carried out by X-ray diffraction technique showed the formation of perovskite structure with orthorhombic symmetry. Addition of BF in the NKNT system lowered the sintering temperature by 500C. The nature of the frequency dependence of ac conductivity of NKNT compounds follows Jonscher power law. Complex impedance and modulus spectra confirm the significant contribution of both grain and grain boundary to the electrical response of the materials. Above the ferroelectric–paraelectric phase transition temperature, the electrical conduction is governed by the thermal excitation of charge carriers from oxygen vacancies exhibiting Negative temperature coefficient (NTCR) behaviour. Detailed study on the multiferroic properties (where magnetism and ferroelectricity are strongly coupled together) of the system is under process which is likely to form key components in the development of future technology, for example, in memories and logic devices.

Currently the need for various developmental models in material sciences to develop green technologies has been reemphasized. For biodiversity rich countries like India, low investment and sustainability of biological synthesis adds to its attraction. Plants respond to heavy metal stress by metal complexation process like production of phyochelations or by other metal chelating peptides. In this paper we report the synthesis of AgNPS from the room dried stem and root of Ocimum sanctum at room temperature. The reaction process was simple for formation of highly stable silver nanoparticles. The morphology and crystalline phase of the NPs were determined from transmission electronmicroscopy (TEM), selected area electron diffraction (SAED) and X-ray diffraction(XRD) spectra. Transmission Electron Microscopy studies showed that the average silver nanoparticles obtained from roots and stem were of sizes 10 ± 2 nm and 5 ± 1.5 nm respectively.

Podophyllum hexandrum Royle, (Podophyllaceae) a critically endangered Himalayan herb is a rich source of a neurotoxin compound-podophyllotoxin, which has antitumour activity and after derivatization to etoposide, etoposide phosphate and teniposide is used for treatment of various malignant conditions. A simple, precise and accurate HPLC method with UV-detection was developed and validated for quantification of podophyllotoxin in Podophyllum hexandrum. The method was validated according to the ICH guidelines which includes the parameters like accuracy, precision, linearity, range, LOD, LOD and robustness. Seperation was achieved on a reverse-phase C18 column using methanol: water: 62: 38 (v/v) as mobile phase in isocratic mode with a flow rate of 0.9 ml/min and detection was done at 280 nm. The method showed high correlation coefficients (R > 0.998) for standard subjected to the entire procedure. Value of RSD % of area for Inter-day and Intra-day precision of the developed method was 0.68 % and 0.66 % respectively. Detection and quantitation limits of the developed method were 0.026µg/ml and 0.106µg/ml respectively.

The lightest neutralino, assumed to be the lightest supersymmetric particle, is proposed to be a dark matter (DM) candidate for the mass $\mathcal{O}(100)\text{ }\text{ }\mathrm{GeV}$. Constraints from various direct dark matter detection experiments and Planck measurements exclude a substantial region of parameter space of minimal supersymmetric standard model. However, a ``mild-tempered'' neutralino with dominant bino composition and a little admixture of Higgsino is found to be a viable candidate for DM. Within the minimal supersymmetric standard model framework, we revisit the allowed region of parameter space that is consistent with all existing constraints. Regions of parameters that are not sensitive to direct detection experiments, known as ``blind spots,'' are also revisited. Complimentary to the direct detection of DM particles, a mild-tempered neutralino scenario is explored at the LHC with the center of mass energy $\sqrt{\mathrm{s}}=13\text{ }\text{ }\mathrm{TeV}$ through the top-squark pair production, and its subsequent decays with the standard-model-like Higgs boson in the final state. Our considered channel is found to be very sensitive also to the blind spot scenario. Detectable signal sensitivities are achieved using the cut-based method for the high luminosity options 300 and $3000\text{ }\text{ }{\mathrm{fb}}^{\ensuremath{-}1}$, which are further improved by applying the multivariate analysis technique.

Purpose: The purpose of this study is to investigate the impact of patient position (supine and prone) on conventional bilateral field, three dimensional conformal radiotherapy (3DCRT) and intensity modulated radiotherapy (IMRT) treatment plans in patients of brainstem glioma with a view to exploring the possibility of avoiding beam entry through immobilization accessories. Methods: Five patients of brainstem glioma were immobilized and scanned in supine and prone positions with a combination of head rest and thermoplastic cast. Each patient was planned with three techniques: (i) 2-fields bilateral (ii) 3-fields 3DCRT, and (iii) 5-fields IMRT. Plan quality was analyzed in terms of planning target volume (PTV) coverage and dose to various critical organs at risk (OAR) for both the supine and prone treatment positions. Results: In case of bilateral fields (parallel opposed) planning, the PTV coverage and dose to the OAR were almost similar for both the supine and prone positions. In 3DCRT plan, although the PTV coverage and dose to critical structures were comparable for both the supine and prone position, dose to cochlea was lower for the prone position plan. A modest decrease in maximum dose to optic nerves and mean dose to temporal lobes were also observed for the prone position plan. In IMRT plans, the PTV coverage and homogeneity were comparable in both the supine and prone positions. Reduction in average maximum and mean doses to all OARs with functional subunit (FSU) in series and parallel respectively was observed in the IMRT plan for prone position when compared to the supine position. Conclusion: Supine and prone positions resulted in almost similar dose distribution in all the three techniques applied. At some instances, the prone position showed better normal tissues sparing when compared to supine. Moreover, prone position is more likely to avoid attenuation due to immobilization devices and uncertainty in dose calculation under large inhomogeneities.

We had earlier reported our results of early clinical outcome and acute toxicities in patients with locally advanced carcinoma of the cervix (LACC) treated with whole-pelvic conventional radiation therapy (WP-CRT) versus intensity modulated radiation therapy (WP-IMRT). This study aims to evaluate the long-term clinical outcome and late toxicities. Between January 2010 and January 2012, 44 patients with International Federation of Gynecology and Obstetrics (FIGO 2009) stage IIB-IIIB squamous cell carcinoma of the cervix were randomized to receive 50.4 Gy in 28 fractions delivered via either WP-CRT or WP-IMRT with concurrent weekly cisplatin 40 mg/m2 followed by high-dose-rate intracavitary brachytherapy (7 Gy in 3 fractions each 1 week apart). Late toxicity was graded as per the Radiation Therapy Oncology Group (RTOG) system. X2 test was used for comparison of toxicity between 2 arms. Survival was calculated from the date of initiation of treatment and was estimated by Kaplan-Meier method. A statistical software package was used for all statistical analysis and a P value <.05 was considered significant. Of 44 patients, 22 patients received WP-CRT and 22 received WP-IMRT. In the WP-CRT arm, 13 patients belonged to stage IIB and 9 to stage IIIB; in the IMRT arm 12 patients had stage IIB disease and 10 had stage IIIB disease. Median follow-up was 39 months (23-56 months). One patient each failed locally and 3 patients each failed at distant site in both the arms. One patient in the WP-CRT arm and 2 patients in the WP-IMRT arm failed at both local as well as distant sites. Four patients (2 in each arm) had persistent local disease detected on clinical examination/positron emission tomography scans after completion of the treatment but showed resolution of the disease on continued follow-up (median time of resolution: 3 months). Six patients in the WP-IMRT arm and 3 patients in the WP-CRT arm had died at the time of the last follow-up. Four-year disease-free survival and overall survival were 69.2% in the WP-CRT versus 72.4 % in the WP-IMRT (P=.667) and 85.2% in the WP-CRT versus 66.8% in the WP-IMRT (P=.326) arms. Patients in the WP-IMRT arm experienced significantly less chronic gastrointestinal (GI) toxicity (18.2% vs 50%, P=.027). In the IMRT arm, the percentage of patients having grade 2 and 3 chronic GI toxicities were 4.5% and 0%, respectively, while for patients in the conventional arm, the rates were 13.63% and 9.1%. One patient in each arm developed chronic grade 1 bladder toxicity. Grade 2 late bladder toxicity was seen more in the WP-CRT arm as compared to the WP-IMRT arm (13.6% vs 0%, P=.116). WP-IMRT continues to show noninferior long-term survival outcome and is associated with significantly less chronic GI toxicity compared to WP-CRT. WP-IMRT also leads to fewer late bladder toxicities. We suggest the use of WP-IMRT for LACC until further results from the ongoing multicentric trial are available.

To evaluate and compare chronic gastrointestinal (GI) toxicity in locally advanced carcinoma cervix treated with Conventional versus Intensity modulated radiation therapy (IMRT). Between March 2010 and January 2012, 44 untreated non-metastatic patients of squamous cell carcinoma cervix with stage IIB-IIIB, age 25-65 years, KPS ≥ 70, hemoglobin ≥10 gm/dl, WBC ≥ 3,000/μl, Absolute neutrophil count (ANC) ≥ 1500 cells/mm3; Platelets ≥ 100,000 cells/mm3; Creatinine clearance ≥ 50 mg/dl were randomized to receive either pelvic conventional or IMRT. Patients in both arms received external beam pelvic radiation therapy of 50.4 Gy in 28 fractions over 5.5 weeks with concurrent Cisplatin 40 mg/m2 weekly followed by high dose rate (HDR). Intracavitary radiation therapy (ICRT) 7 Gy in 3 fractions or Interstitial brachytherapy (ISBT) 10 Gy in 2 fractions, each one week apart. Patients were assessed for chronic GI toxicities and scored as grade 0 (no symptoms), 1 (mild symptoms, no medications required), 2 (moderate symptoms, medications required), and 3 (severe symptoms, hospitalization, surgery required) every three months after the completion of the treatment. Statistical analysis was done with SPSS (version 16.0). Chi-Square test/Fisher's Exact test was used for comparison of two arms. p value of ≤ 0.05 was taken as significant. Of 44 patients, 22 patients were treated with conventional and 22 with IMRT. Median age and KPS of patients in both the arms were 48 years and 90 respectively. In conventional arm 13 patients belonged to stage IIB and 9 to stage IIIB while in IMRT arm 12 patients were of stage IIB and 10 of stage IIIB. Median follow-up for patients in the IMRT arm was 21.6 months (Range, 7.7-34.4 months) and for the conventional arm was 21.7 months (10.7-37.4 months). Most of the grade 1 toxicities were in the form of increased number of stools, urgency and indigestion to certain kinds of food. Only 2 patients had Grade 3 toxicity, both in the conventional arm. One developed severe proctitis and bleeding requiring admission and blood transfusions while other developed rectovaginal fistula requiring surgery. Overall, patients in the IMRT arm had lesser chronic GI toxicity as compared with conventional arm (13.6% vs 50%, p = 0.011). In the IMRT arm, the percentage of patients having Grade 1, 2, and 3 chronic GI toxicities were 9%,4.5% and 0% respectively and for the patients in the conventional arm, the corresponding percentages were 27.3%,13.63% and 9.1% respectively. IMRT in locally advanced carcinoma cervix leads to significantly lesser chronic GI toxicities as compared to Conventional radiation therapy. We suggest IMRT should be used in favor of lesser radiation morbidities.

In the present work, a newly-developed MC-based treatment planning (MCTP) tool for proton therapy is proposed to support treatment planning studies and research applications.It allows for single-field and simultaneous multiple-fields optimization in realistic treatment scenarios and is based on the MC code FLUKA.Relative biological effectiveness (RBE)-weighted dose is optimized either with the common approach using a constant RBE of 1.1 or using a variable RBE according to radiobiological input tables.An example of treatment plan in a patient-CT geometry is presented for clinical treatment parameters as used at the Italian National Center for Oncological Hadron Therapy (CNAO).

HfO2 thin films with three different thickness (∼10 nm, ∼30 nm & ∼ 50 nm) were deposited on TiN/Si by Chemical Beam Vapor Deposition (CBVD) technique with combinatorial approach. The structural, morphological, and dielectric behavior of the films were examined in detail. XRD and Raman results revealed that the films have stable monoclinic phase irrespective of thickness. Atomic force microscopy (AFM) micrographs inferred homogeneity and crack free surfaces for all three thicknesses. Dielectric constant was found to be maximum (up to ∼ 67) for 2 kHz at room temperature which is three times higher than previously reported work. Furthermore, Impedance investigation explained the negative temperature coefficient of resistance (NTCR) behavior at the low-frequency region. CBVD grown HfO2 thin films represent promising properties in terms of structural, morphological, and dielectric in TiN/HfO2/Ag MIM structure thus could be a favorable candidate for the future capacitor applications.

National Rural Health Mission (NRHM) has created a cadre of trained female community health activists called Accredited Social Health Activist (ASHA) to strengthen the health care system and to mobilize community towards increased utilization of existing health services. The contribution of the Accredited Social Health ActivistASHA) to the achievement of the country’s targets for new-born &amp; child health services is critical. The present study was planned with objective to assess &amp; compare the level of knowledge of ASHA workers regarding child health services working in rural and urban areas of a block of Haryana. The present cross-sectional, community-based study was conducted in block Barwala, district Hisar of Haryana. The assessment of knowledge of ASHA workers was done on the basis of scoring. Appropriate statistical tests like percentages and chi-square (χ2) test were applied. Regarding child health services majority of ASHA workers had good knowledge, assessed by score gained by them and none of them was having poor knowledge about child health services. However, in rural area score gained was better than urban area &amp; the observed difference was found to be statistically significant. Majority of ASHA workers either working in rural or urban areas, were having inadequate knowledge about identification of danger signs during diarrhoea &amp; acute respiratory infection in a child and danger signs of new born. They had adequate knowledge regarding cord stump care; breast Feeding practices; routine Immunization; Vitamin A supplement and home based post natal care (HBPNC) visits. Inadequate knowledge of ASHA workers regarding new born care should be taken into consideration. Supportive supervision should be done in their working area.

Background Hypertension (HTN) is endemic in India and it is considered a public health challenge in both economically developed and developing nations. Unfortunately, despite its high prevalence, its awareness, treatment, and control status are low in urban as well as rural Indian populations. Objectives To determine the burden of hypertension and prehypertension in a Community Health Center (CHC) and to find the association of hypertension with the age group and sex of study subjects. Methodology A cross-sectional study was carried out among 713 patients of age 20 years and above attending the Out Patient Department (OPD) of the Community Health Centre (CHC), Barwala in the Hisar district of Haryana. JNC 7 classification of blood pressure was used to diagnose hypertension. The collected data was analysed using the Statistical Package for Social Sciences (SPSS) version 20.0. Appropriate statistical tests were used. Result Out of the total patients enrolled, 200 (28.1%) were found to be hypertensives. The burden of prehypertension was further observed to be 28.1%. About (61.6%) of OPD patients were female. Among hypertensive patients, nearly half (48.8%) were in the age group of 60-80 years whereas the majority of the hypertensives (56%) were females. Conclusion In our study, more than 50 percent of OPD patients were found to have hypertension and pre-hypertension. Health-seeking behavior was more among females.

In perovskite oxides, the multiple charge states of cations influence their local structure and correlated physical properties. In this report, we studied the role of multivalence cations on the local structure and magnetic behavior of the Sr2FeCoO6-δ, prepared by the solid-state synthesis technique. A detailed structural and geometrical analysis confirmed the co-existence of orthorhombic and cubic crystal symmetries and a gesture of the Jahn-Teller (JT) distortion. The intense Raman modes are allowed to be indexed for BO6- and JT-distortion. Experimental and theoretical analysis of the XPS spectra supported the mixed oxidation state of Fe (Fe3+/Fe4+) and Co (Co2+/Co3+), further confirmed by theoretically simulated XAS spectra. The presence of the multivalence states leads to AFM/FM exchange interaction in the sample which further leads to glassy magnetic behavior.

Negative health effects of synthetic and chemical ingredients have instilled in consumers an interest in natural bioactive food ingredients. The education and trainings have motivated consumers toward the regular use of bioactive food ingredients and to understand their health benefits for long-term survival. These natural bioactive food ingredients possess many types of bioactivities like antioxidative, antiobesity, and antimicrobial activities. The most common method for including all of these types of bioactivities in the food is fortification of these ingredients. However, the heat sensitivity of these ingredients is a limiting factor for their addition in food specifically during processing and less stability during storage leads to their decay with time. This chapter provides an in-depth discussion on the different techniques of encapsulation.

In the present study, variation in podophyllotoxin content was studied with growth and development in different parts of Podophyllum hexandrum, i.e., leaf lamina, petiole and rootstock using High Performance Liquid Chromatography (HPLC). It was found that podophyllotoxin content in rootstock and leaf lamina increased with increase in age of the plant while it was recorded vice-versa in petiole of the plant. The rootstock was found to contain the highest content of podophyllotoxin in comparison to other parts.

The social work profession lays a strong emphasis on internalizing and reinforcing core social work values and ethical standards in order to create a cadre of social workers who can withstand the criticalities of any crisis and commit themselves to firmly stand with those whose survival and well-being is fragile and “on the brink”. The performance of this role not only requires a dynamic and contextual knowledge base and evolving skills set, it mandates a deep rooted acquiescence with a fundamental set of values that shape the profession’s vision and mission, and establish the goals and priorities of the practitioners. The transition from an entry level neophyte to a competent social work professional entails a journey which is characterized by the assumption and bolstering of a professional persona that grounds the incumbent within the functional niche of social work. The present paper looks at the integration of a social work pledge as a viable means for social work incumbents to establish identification with social work values, don a professional identity, and assimilate with professional peers. It looks at the existing practice of pledge taking by highlighting the experience of a distinguished school of social work. It envisions the ‘pledge’ and the tradition of ‘taking the pledge’ evolving as a strong and more universal medium for creating an emotional connect of social work students with the value driven profession of social work. Keywords: Pledge, values, ethics, social work, profession

Ideally, a solid electrolyte which is a central component of SOFC should exhibit high anionic or cationic ionic conductivity at the proposed operating temperatures. In most case, the performance is compromised when operating above 1000 ºC due to poor mechanical, thermal and chemical stability of selected functional and non-functional materials. In this context, pyrochlores are one of the potential candidates due to their high conductivity, flexibility to accommodate large cations, and high mechanical, thermal and chemical stability. In this study, we report the synthesis of nano-powders of Ho2Hf2O7 (HH) and Ho2Zr2O7 (HZ) pyrochlore ceramics by eco-friendly alginate mediated ion-exchange process also known as Leeds Alginate Process (LAP) and provide further insight into the structure conductivity relationship of HH and HZ compounds by EXAFS studies. Both the compositions were sintered at temperatures, ranging from 1100 ºC-1500 ºC at 2h dwell time to achieve the desired high-density ceramic with stable pyrochlore structure. X-ray diffraction and Extended X-ray absorption fine structural (EXAFS) analysis showed superlattice reflections and complex coordination geometry of these oxides. The coordination number and disorder factor in the case of HZ were found to be more stable than the HH sample, as evident from the EXAFS. Impedance spectroscopy and dc-conductivity analysis showed a better charge transport behavior in HZ ceramics than in HH making HZ as a preferred solid electrolyte for SOFC.