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Find video protocols related to scientific articles indexed in Pubmed.
Polymorphisms in nucleotide excision repair genes and susceptibility to colorectal cancer in the Polish population.
Mol. Biol. Rep.
PUBLISHED: 11-08-2014
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Xeroderma pigmentosum (XP) is a rare autosomal recessive disease that is associated with a severe deficiency in nucleotide excision repair. Genetic polymorphisms in XP genes may be associated with a change in DNA repair capacity, which could be associated with colorectal cancer development. We assessed the association between 94 single nucleotide polymorphisms (SNPs) within seven XP genes (XPA-XPG) and the colorectal cancer risk in the Polish population. We genotyped 758 unselected patients with colorectal cancer and 1,841 healthy adults. We found that a significantly decreased risk of colorectal cancer was associated with XPC polymorphism rs2228000_CT genotype (OR 0.59; p < 0.0001) and the rs2228000_TT genotype (OR 0.29; p < 0.0001) compared to the reference genotype (CC). And an increased disease risk was associated with the XPD SNP, rs1799793_AG genotype (OR 1.44, p = 0.018) and rs1799793_AA genotype (OR 3.31, p < 0.0001) compared to the reference genotype. Haplotype analysis within XPC, XPD and XPG revealed haplotypes associated with an altered colorectal cancer risk. Stratified analysis by gender showed differences between the association of three SNPs: XPC rs2228000, XPD rs1799793 and XPD rs238406 in females and males. Association analysis between age of disease onset and polymorphisms in XPD (rs1799793) and XPC (rs2228000) revealed differences in the prevalence of these variants in patients under and over 50 years of age. Our results confirmed that polymorphisms in XPC and XPD may be associated with the risk of colorectal cancer.
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Methylation of the BRCA1 promoter in peripheral blood DNA is associated with triple-negative and medullary breast cancer.
Breast Cancer Res. Treat.
PUBLISHED: 08-25-2014
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It has been proposed that methylation signatures in blood-derived DNA may correlate with cancer risk. In this study, we evaluated whether methylation of the promoter region of the BRCA1 gene detectable in DNA from peripheral blood cells is a risk factor for breast cancer, in particular for tumors with pathologic features characteristic for cancers with BRCA1 gene mutations. We conducted a case-control study of 66 breast cancer cases and 36 unaffected controls. Cases were triple-negative or of medullary histology, or both; 30 carried a constitutional BRCA1 mutation and 36 did not carry a mutation. Blood for DNA methylation analysis was taken within three months of diagnosis. Methylation of the promoter of the BRCA1 gene was measured in cases and controls using methylation-sensitive high-resolution melting (MS-HRM). A sample with any detectable level of methylation was considered to be positive. Methylation of the BRCA1 promoter was detected in 15 of 66 cases and in 2 of 36 controls (OR 5.0, p = 0.03). Methylation was present in 15 of 36 women with breast cancer and without germline BRCA1 mutation, but in none of 30 women with breast cancer and a germline mutation (p < 0.01). The association between methylation and breast cancer was restricted to women with no constitutional BRCA1 mutation (OR 12.1, p = 0.0006). Methylation of the promoter of the BRCA1 gene detectable in peripheral blood DNA may be a marker of increased susceptibility to triple-negative or medullary breast cancer.
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Unraveling the intricacies of mammalian fertilization.
Asian J. Androl.
PUBLISHED: 07-20-2014
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It is imperative to understand the molecular basis of various steps involved during fertilization. In the manuscript by Bianchi et al. [1] a novel protein, Juno on egg membrane (oolemma) has been characterized that binds to sperm specific protein, Izumo-1. Monoclonal antibodies against Juno inhibited in vitro fertilization. Juno knock-out female mice failed to deliver litters on mating. It is rapidly shed from oolemma after fertilization, suggesting its role in preventing polyspermy. Taken together these studies will help in our understanding of sperm-egg recognition mechanisms and also facilitate development of new fertility treatment regimens and novel contraceptives.
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Influence of iodothyronine conjugates of bovine serum albumin and horseradish peroxidase on enzyme immunosorbent assay of thyroid hormones.
J Immunoassay Immunochem
PUBLISHED: 05-28-2014
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Enzyme-linked immunosorbent assays (ELISA's) reported for thyroxine (T?) and 3,5,3'-triiodothyronine (T?), involved coupling of the haptens through (i) carboxylic group to carrier protein for producing antibodies and (ii) amino group to detection labels. To improve the titer and specificity of antibodies, immunogens were prepared by coupling of carboxyl group to bovine serum albumin (BSA) either directly or through adipic acid dihydrazide (ADH), after protecting amino group through acetylation of T? and T?. Direct coupling resulted in the incorporation of 40-50 moles of T? and T? per BSA molecule and helped in improving immunogenic response and specificity, especially of T?. High epitope density of immunogens evoked better antibody response, since attachement of ADH as spacer, introduced 18-27 moles of haptens into carrier protein and had less effect on antibody development, with T? being exception. Detection labels were prepared by coupling horseradish peroxidase (HRP) to amino group of thyroid hormones directly and after preparing their methyl esters, which provided sensitive displacement curves in combination with the antibodies developed against N-acetylated-T? and T?. Unlike methyl esters, T?-HRP and T?-HRP showed higher sensitivity and seemed to be related to the affinity of the labels for binding the antibody.
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Diagnostic utility of adenosine deaminase in exudative pleural effusions.
Lung India
PUBLISHED: 04-30-2014
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To determine the diagnostic utility of adenosine deaminase (ADA) in exudative pleural effusions of different etiologies.
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Hyponatremia in stroke.
Ann Indian Acad Neurol
PUBLISHED: 04-23-2014
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Hyponatremia is a common electrolyte disorder encountered in patients of neurological disorders which is usually either due to inappropriate secretion of Antidiuretic hormone (SIADH) or cerebral salt wasting syndrome (CSWS). We conducted this study in a tertiary care hospital to determine the incidence and etiology of hyponatremia in patients of stroke admitted in the hospital.
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Humanized antibody neutralizing 2009 pandemic H1N1 virus.
Biotechnol J
PUBLISHED: 03-24-2014
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The 2009 pandemic H1N1 S-OIV (swine origin influenza A virus) caused noticeable morbidity and mortality worldwide. In addition to vaccine and antiviral drug therapy, the use of influenza virus neutralizing monoclonal antibodies (MAbs) for treatment purposes is a viable alternative. We previously reported the isolation of a high affinity, potently neutralizing murine MAb MA2077 against 2009 pandemic H1N1 virus. We describe here the humanization of MA2077 and its expression in a mammalian cell line. Six complementarity-determining regions (CDRs) of MA2077 were grafted onto the human germline variable regions; along with six and eight back mutations in the framework of heavy and light chains, respectively, pertaining to the vernier zone and interchain packing residues to promote favorable CDR conformation and facilitate antigen binding. The full length humanized antibody, 2077Hu2, expressed in CHO-K1 cells, showed high affinity to hemagglutinin protein (KD = 0.75 ± 0.32 nM) and potent neutralization of pandemic H1N1 virus (IC50 = 0.17 ?g/mL), with marginally higher IC50 as compared to MA2077 (0.08 ?g/mL). In addition, 2077Hu2 also retained the epitope specificity for the "Sa" antigenic site on pandemic HA. To the best of our knowledge, this is the first report of a humanized neutralizing antibody against pandemic H1N1 virus.
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Dupuytren's disease and the risk of malignant neoplasms.
Hered Cancer Clin Pract
PUBLISHED: 02-06-2014
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The object of this study was the investigation of the risk of occurrence of malignant neoplasms in 508 patients with Dupuytren's disease (DD) and in 2157 of their 1st degree relatives. In the first stage of the study, we evaluated the tumour spectrum as well as the age of the patient at diagnosis of cancers in DD families along with the observed and expected frequencies of malignancies. In the second stage of the study, we examined the distribution of 20 common mutations/polymorphisms in 12 known cancer susceptibility genes among DD patients and 508 matched healthy controls. No such study has been published to date. Results. No significant differences were noted between malignancies diagnosed among members of DD families and the general population. Molecular examination of 20 mutations/polymorphisms in 12 cancer susceptibility genes in Dupuytren's patients and controls showed a statistically significant association of one mutation with Dupuytren disease: D312M in XPD (OR?=?1.75, p?=?0.004). We observed a tendency toward changed frequencies of occurrence of central nervous system tumors, laryngeal cancer and non-melanoma skin cancers in DD families. The results of our study indicate a lack of a strong association between Dupuytren disease and familial cancer risk.
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STAT3 and ERK1/2 cross-talk in leukaemia inhibitory factor mediated trophoblastic JEG-3 cell invasion and expression of mucin 1 and Fos.
Am. J. Reprod. Immunol.
PUBLISHED: 01-14-2014
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The aim of this study was to investigate the relative importance of STAT3 and ERK1/2 activation in leukaemia inhibitory factor (LIF)-mediated invasion of JEG-3 cells.
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Common variants of xeroderma pigmentosum genes and prostate cancer risk.
Gene
PUBLISHED: 01-09-2014
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The genetic basis of prostate cancer (PC) is complex and appears to involve multiple susceptibility genes. A number of studies have evaluated a possible correlation between several NER gene polymorphisms and PC risk, but most of them evaluated only single SNPs among XP genes and the results remain inconsistent. Out of 94 SNPs located in seven XP genes (XPA-XPG) a total of 15 SNPs were assayed in 720 unselected patients with PC and compared to 1121 healthy adults. An increased risk of disease was associated with the XPD SNP, rs1799793 (Asp312Asn) AG genotype (OR=2.60; p<0.001) and with the AA genotype (OR=531; p<0.0001) compared to the control population. Haplotype analysis of XPD revealed one protective haplotype and four associated with an increased disease risk, which showed that the A allele (XPD rs1799793) appeared to drive the main effect on promoting prostate cancer risk. Polymorphism in XPD gene appears to be associated with the risk of prostate cancer.
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Evaluation of effectiveness of connective tissue graft as a barrier with bioresorbable collagen membrane in the treatment of mandibular Class II furcation defects in humans: 4-year clinical results.
Quintessence Int
PUBLISHED: 01-07-2014
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Connective tissue grafts (CTGs) have been used as a barrier for closing mandibular Class II furcation defects. The present study compared long-term (48 months) effectiveness of CTG as a barrier with bioresorbable collagen membrane in the treatment of mandibular Class II furcation defects in humans.
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Milestones in contraceptive vaccines development and hurdles in their application.
Hum Vaccin Immunother
PUBLISHED: 11-23-2013
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Contraceptive vaccines have been proposed for controlling the growing human population and wildlife population management. Multiple targets such as gonadotropin releasing hormone (GnRH), luteinizing hormone, follicle stimulating hormone, gonadotropin receptors, sperm-specific proteins and zona pellucida glycoproteins have been exploited to develop contraceptive vaccine and their efficacy investigated and shown in various experimental animal models. Vaccines based on GnRH have found application in immuno-castration of male pigs for prevention of boar-taint. Vaccines based on zona pellucida glycoproteins have shown promising results for population management of wild horses and white-tailed deer. Phase II clinical trials in women with ?-human chorionic gonadotropin (?-hCG)-based contraceptive vaccine established proof of principle that these can be developed for human application. Block in fertility by ?-hCG contraceptive vaccine was reversible. Further research inputs are required to establish the safety of contraceptive vaccines, improve their immunogenicity and to develop novel vaccine delivery platforms for providing long lasting immunity.
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Production of tag-free recombinant fusion protein encompassing promiscuous T cell epitope of tetanus toxoid and dog zona pellucida glycoprotein-3 for contraceptive vaccine development.
Mol. Biotechnol.
PUBLISHED: 11-05-2013
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Affinity tags can interfere in various physicochemical properties and immunogenicity of the recombinant proteins. In the present study, tag-free recombinant fusion protein encompassing promiscuous T cell epitope of tetanus toxoid [TT; amino acid (aa) residues 830-844] followed by dilysine linker and dog zona pellucida glycoprotein-3 (ZP3; aa residues 23-348) (TT-KK-ZP3) was expressed in Escherichia coli. The recombinant protein, expressed as inclusion bodies (IBs), was purified by isolation of IBs, processed to remove host cell proteins, followed by solubilization and refolding. A specific 39 kDa protein including ZP3 was identified by SDS-PAGE. CD spectra showed the presence of ?-helices and ?-sheets, and fluorescent spectroscopy revealed emission maxima of 265 A.U. at 339 nm for refolded protein and showed red shift in the presence of 6 M guanidine hydrochloride. Immunization of inbred FvB/J female mice with purified recombinant TT-KK-ZP3 (25 ?g/animal) led to generation of high antibody titers against the recombinant protein. The antibodies reacted specifically with ZP matrix surrounding mouse oocytes. Immunized mice showed significant reduction in fertility as compared to the control group. The studies described herein provide a simple method to produce and purify tag-free recombinant protein for the development of a contraceptive vaccine.
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Evaluation of recombinant fusion protein comprising dog zona pellucida glycoprotein-3 and Izumo and individual fragments as immunogens for contraception.
Vaccine
PUBLISHED: 10-14-2013
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Vaccines based on gamete specific proteins have been proposed for fertility inhibition. In the present study, immunogenicity and contraceptive potential of E. coli-expressed recombinant fusion protein TT-KK-ZP3-GGG-Iz, comprising promiscuous T cell epitope of tetanus toxoid (TT) followed by dilysin linker (KK), a fragment of dog zona pellucida glycoprotein-3 (ZP3), triglycine spacer (GGG) and a fragment of dog Izumo (Iz) without any affinity tag has been evaluated in female FvB/J mice. In addition, recombinant TT-KK-ZP3 and Izumo linked to the promiscuous T cell epitope of bovine RNase (bRNase-KK-Iz) and their physical mixture were also used. SDS-PAGE and immunoblot studies revealed ?32kDa band corresponding to TT-KK-ZP3-GGG-Iz, ?22kDa band of TT-KK-ZP3 and ?11kDa band of bRNase-KK-Iz. Groups of mice immunized with the above recombinant proteins led to the generation of high antibody titres against the respective proteins. Immunization with recombinant TT-KK-ZP3-GGG-Iz generated higher antibody titre as compared to mice immunized with physical mixture of TT-KK-ZP3 and bRNase-KK-Iz. Antibodies against TT-KK-ZP3-GGG-Iz and TT-KK-ZP3 recognized mouse and dog ZP and those against TT-KK-ZP3-GGG-Iz and bRNase-KK-Iz recognized mouse and dog acrosome-reacted sperm in an indirect immunofluorescence assay. Immune sera from groups of mice immunized with the above recombinant proteins led to a significant reduction in mouse in vitro fertilization. Mating studies revealed significant reduction in fertility as compared to adjuvant control group. Highest infertility was observed in group of mice immunized with TT-KK-ZP3 followed by TT-KK-ZP3-GGG-Iz. Infertility was associated with the antibody titres against ZP3, whereas no association in the inhibition of fertility and antibody titres against Izumo was observed. In conclusion, these studies revealed the contraceptive potential of ZP3, which could not be further enhanced by the inclusion of Izumo.
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Antibiotic uptake by vegetable crops from manure-applied soils.
J. Agric. Food Chem.
PUBLISHED: 10-09-2013
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This study quantified the uptake of five antibiotics (chlortetracycline, monensin, sulfamethazine, tylosin, and virginiamycin) by 11 vegetable crops in two different soils that were fertilized with raw versus composted turkey and hog manures or inorganic fertilizer. Almost all vegetables showed some uptake of antibiotics from manure treatments. However, statistical testing showed that except for a few isolated treatments the concentrations of all antibiotics in vegetable tissues were generally less than the limits of quantification. Further testing of the significant treatments showed that antibiotic concentrations in vegetables from many of these treatments were not significantly different than the corresponding concentrations from the fertilizer treatment (matrix effect). All five antibiotic concentrations in the studied vegetables were <10 ?g kg(-1). On the basis of the standards for maximum residue levels in animal tissues and suggested maximum daily intake based on body weight, this concentration would not pose any health risk unless one is allergic to that particular antibiotic.
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Can selenium be a modifier of cancer risk in CHEK2 mutation carriers?
Mutagenesis
PUBLISHED: 10-08-2013
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Selenium is an essential trace element for humans, playing an important role in various major metabolic pathways. Selenium helps to protect the body from the poisonous effects of heavy metals and other harmful substances. Medical studies have provided evidence of selenium supplementation in preventing certain cancers. Low and too high selenium (Se) status correlates with increased risk of e.g. lung, larynx, colorectal and prostate cancers. A higher level of selenium and supplementation with selenium has been shown to be associated with substantially reduced cancer mortality. Selenium exerts its biological roles through selenoproteins, which are involved in oxidoreductions, redox signalling, antioxidant defence, thyroid hormone metabolism and immune responses. Checkpoint kinase 2 (CHEK2) is an important signal transducer of cellular responses to DNA damage and acts as a tumour suppressor gene. Mutations in the CHEK2 gene have been shown to be associated with increased risks of several cancers. Four common mutations in CHEK2 gene (1100delC, IVS2+1G>A, del5395 and I157T) have been identified in the Polish population. Studies have provided evidence that CHEK2-truncating and/or missense mutations are associated with increased risk of breast, prostate, thyroid, colon and kidney cancers. The variability in penetrance and cancer expression in CHEK2 mutation carriers can probably be explained by the influence of other genetic or environmental factors. One of the possible candidates is Se, which together with genetic variations in selenoprotein genes may influence susceptibility to cancer risk.
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Synthesis, anti-HIV activity, integrase enzyme inhibition and molecular modeling of catechol, hydroquinone and quinol labdane analogs.
Bioorg. Med. Chem. Lett.
PUBLISHED: 07-24-2013
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Labdane analogs with o-quinol, catechol and hydroquinone moiety have been synthesized using Diels-Alder reaction of methyl 3,4-dioxocyclohexa-1,5-diene-carboxylate, 3,4-dioxocyclohexa-1,5-diene-carboxylic acid and 3,6-dioxocyclohexa-1,4-dienecarboxylic acid with mono terpene 1,3-dienes, namely ocimene and myrcene. The resulting molecules and their derivatives were evaluated for their anti-HIV-1 activity using TZM-bl cell based virus infectivity assay. Two molecules 13 and 18 showed anti-HIV activity with IC50 values 5.0 (TI=11) and 4.6 (TI=46)?M, respectively. The compounds 17, 18 and 20 showed efficacy against HIV-1 integrase activity and showed inhibition with IC50 13.4, 11.1 and 11.5?M, respectively. The HIV-1 integrase inhibition activity of these synthetic molecules was comparable with integric acid, the natural fungal metabolite. Molecular modeling studies for the HIV-1 integrase inhibition of these active synthetic molecules indicated the binding to the active site residues of the enzyme.
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Palm top plasma focus device as a portable pulsed neutron source.
Rev Sci Instrum
PUBLISHED: 07-05-2013
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Development of a palm top plasma focus device generating (5.2 ± 0.8) × 10(4) neutrons?pulse into 4? steradians with a pulse width of 15 ± 3 ns is reported for the first time. The weight of the system is less than 1.5 kg. The system comprises a compact capacitor bank, a triggered open air spark gap switch, and a sealed type miniature plasma focus tube. The setup is around 14 cm in diameter and 12.5 cm in length. The energy driver for the unit is a capacitor bank of four cylindrical commercially available electrolytic capacitors. Each capacitor is of 2 ?F capacity, 4.5 cm in diameter, and 9.8 cm in length. The cost of each capacitor is less than US$ 10. The internal diameter and the effective length of the plasma focus unit are 2.9 cm and 5 cm, respectively. A DC to DC converter power supply powered by two rechargeable batteries charges the capacitor bank to the desired voltage and also provides a trigger pulse of -15 kV to the spark gap. The maximum energy of operation of the device is 100 J (8 ?F, 5 kV, 59 kA) with deuterium gas filling pressure of 3 mbar. The neutrons have also been produced at energy as low as 36 J (3 kV) of operation. The neutron diagnostics are carried out with a bank of (3)He detectors and with a plastic scintillator detector. The device is portable, reusable, and can be operated for multiple shots with a single gas filling.
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Rosin: a naturally derived excipient in drug delivery systems.
Polim Med
PUBLISHED: 07-02-2013
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Natural polymers are primarily attractive because they are biodegradable, inexpensive, and readily available. The most important benefit of natural polymers is that they are capable for chemical modifications. One such biopolymer, rosin, and its derivatives have been pharmaceutically evaluated as microencapsulating materials, film forming agent and as binding agent in formulation of tablets. They are also employed in formulation of chewing gum bases and cosmetics. This review article provides an overview of pharmaceutical use of rosin and its derivatives as excipient in dosage forms as well as novel drug delivery systems.
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Anti-HIV-1 activity, protease inhibition and safety profile of extracts prepared from Rhus parviflora.
BMC Complement Altern Med
PUBLISHED: 06-27-2013
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In the present study, extracts prepared from the leaves of Rhus parviflora Roxb.(Anacardiaceae) were evaluated for their anti-HIV activity, which have been traditionally used for the treatment of neurological disorders such as anxiety, insomnia and epilepsy.
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Ellagic acid & gallic acid from Lagerstroemia speciosa L. inhibit HIV-1 infection through inhibition of HIV-1 protease & reverse transcriptase activity.
Indian J. Med. Res.
PUBLISHED: 05-04-2013
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Banaba (Lagerstroemia speciosa L.) extracts have been used as traditional medicines and are effective in controlling diabetes and obesity. The aim of this study was to evaluate the anti-HIV property of the extracts prepared from the leaves and stems of banaba, and further purification and characterization of the active components.
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Durability and restoring of superhydrophobic properties in silica-based coatings.
J Colloid Interface Sci
PUBLISHED: 02-14-2013
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The durability and restorable properties of superhydrophobicity are a most important issue essential for utilization of superhydrophobic materials in industrial and domestic fields. In this work, we have focused on the sol-gel dip coating synthesis, durability, and restoring properties of a superhydrophobic surface composed of spherical silica particles with low energy free surface successfully achieved on the surface of glass substrates. The water contact angle (WCA) of the as-prepared superhydrophobic coatings reached as high as 170±1° (WCA=170±1°) and the sliding angle was 3±1° (WSA=3±1°). The behavior of wettability on superhydrophobic coating surface under various external disturbances including outdoor environments, heat, and mild acidic condition is investigated. It is shown in particular that degradation and repairing of surface methyl groups between two such surfaces leads to a fully restorable coating surface by the surface modification with trimethylchlorosilane (TMCS) at room temperature. The coating also holds good thermal stability; it holds superhydrophobicity up to 550 °C and gets degrade into superhydrophilicity above 600 °C without deforming surface morphology. The results confirm the durability of the silica coating under different external disturbances and sol-gel dip coating method and restoring property provide the best solution to fabricate superhydrophobic silica coating surface with extended durability at low cost.
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A low selenium level is associated with lung and laryngeal cancers.
PLoS ONE
PUBLISHED: 02-11-2013
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It has been suggested that selenium deficiency is a risk factor for several cancer types. We conducted a case-control study in Szczecin, a region of northwestern Poland, on 95 cases of lung cancer, 113 cases of laryngeal cancer and corresponding healthy controls.
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Quantitative analysis of conjugated and free estrogens in swine manure: solutions to overcome analytical problems due to matrix effects.
J Chromatogr A
PUBLISHED: 02-01-2013
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Although animal manure is an important source for environmental estrogens, quantitative analysis of estrogens in manure is complicated due to matrix interference. In the present study, chromatographic methods have been developed for quantification of conjugated and free estrogens in manure samples collected from pig farms. The whole manure samples, immediately after collection, were stored at 4°C, acidified (pH?2.0) and spiked with (i) (13)C-labeled internal standards to account for possible storage related degradation and (ii) deuterium labeled internal standards for calibration and quantitative analysis. The liquid samples were extracted with ethyl acetate for separating conjugated and free estrogens. The solid samples were eluted with water for desorbing conjugated hormones followed by methanol for desorbing free hormones. The water and extracts were further purified using hydrophilic-lipophilic balance and/or aminopropyl cartridges. The conjugated estrogens were analyzed using high-performance liquid chromatograph-mass spectrometer, while the free estrogens were analyzed using gas chromatograph-mass spectrometer. The extraction and calibration methods used in the present study yielded excellent sensitivity, reproducibility and >85% recovery of both free and conjugated estrogens that was independent of the manure matrix. In general, the total estrogen loads in liquid and solid samples were 5.1mg/l and 4.93mg/kg, respectively. This may represent the hormonal load of approximately 2.3tons estrogen per day.
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Isolation of a high affinity neutralizing monoclonal antibody against 2009 pandemic H1N1 virus that binds at the Sa antigenic site.
PLoS ONE
PUBLISHED: 01-31-2013
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Influenza virus evades host immunity through antigenic drift and shift, and continues to circulate in the human population causing periodic outbreaks including the recent 2009 pandemic. A large segment of the population was potentially susceptible to this novel strain of virus. Historically, monoclonal antibodies (MAbs) have been fundamental tools for diagnosis and epitope mapping of influenza viruses and their importance as an alternate treatment option is also being realized. The current study describes isolation of a high affinity (K(D)?=?2.1±0.4 pM) murine MAb, MA2077 that binds specifically to the hemagglutinin (HA) surface glycoprotein of the pandemic virus. The antibody neutralized the 2009 pandemic H1N1 virus in an in vitro microneutralization assay (IC(50)?=?0.08 µg/ml). MA2077 also showed hemagglutination inhibition activity (HI titre of 0.50 µg/ml) against the pandemic virus. In a competition ELISA, MA2077 competed with the binding site of the human MAb, 2D1 (isolated from a survivor of the 1918 Spanish flu pandemic) on pandemic H1N1 HA. Epitope mapping studies using yeast cell-surface display of a stable HA1 fragment, wherein Sa and Sb sites were independently mutated, localized the binding site of MA2077 within the Sa antigenic site. These studies will facilitate our understanding of antigen antibody interaction in the context of neutralization of the pandemic influenza virus.
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Can selenium levels act as a marker of colorectal cancer risk?
BMC Cancer
PUBLISHED: 01-23-2013
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Selenium has attracted attention because of its antioxidant properties. Antioxidants protects cells from damage. Certain breakdown products of selenium are believed to prevent tumor growth by enhancing the immune cell activity and suppressing the development of tumor blood vessels. In this observational study, selenium level was measured in a series of patients from Poland and Estonia to determine a correlation between levels of this microelement and colorectal cancer risk.
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Immunogenicity and contraceptive efficacy of Escherichia coli-expressed recombinant porcine zona pellucida proteins.
Am. J. Reprod. Immunol.
PUBLISHED: 01-19-2013
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To overcome availability of the purified native zona pellucida (ZP) glycoproteins for immunocontraception, porcine ZP3, and ZP4 were expressed in E. coli.
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Effect of excipients on dissolution enhancement of aceclofenac solid dispersions studied using response surface methodology: a technical note.
Arch. Pharm. Res.
PUBLISHED: 01-18-2013
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The aim of present study was to enhance the dissolution rate of poorly water-soluble drug aceclofenac by solid dispersion technique using corn starch, dicalcium phosphate, lactose, and microcrystalline cellulose as carriers. Solid dispersions were prepared by solvent wetting method using 3(2) full factorial design for each of the carrier. The prepared solid dispersions were evaluated for differential scanning calorimetry, X-ray diffraction, scanning electron microscopy, Fourier-transform infrared spectroscopy (FTIR), and angle of repose. In vitro dissolution studies were carried out in phosphate buffer (pH 7.5) and 0.1 N HCl (pH 1.2). The results of solid state characterization bring to view that in solid dispersions the crystalline drug gets converted to its amorphous form. FTIR study results indicated the absence of interaction between aceclofenac and carriers. For prepared solid dispersions, angle of repose was found to be in the range of 26.19° to 35.29°, which indicates good flowability. Enhanced drug dissolution was obtained with carrier in order lactose > corn starch > microcrystalline cellulose > dicalcium phosphate. Hence, these carriers could be used to enhance the dissolution rate of poorly water-soluble drug.
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Pharmaceutical solid dispersion technology: a strategy to improve dissolution of poorly water-soluble drugs.
Recent Pat Drug Deliv Formul
PUBLISHED: 01-08-2013
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Oral bioavailability is the major problem when a poorly water-soluble active agent is delivered via oral route. To overcome such problems, solid dispersion systems have been demonstrated in literature to enhance the dissolution property of poorly water-soluble drugs. In the present review, the important aspects to be considered during preparation of solid dispersion systems viz., properties of polymer and preparation techniques of solid dispersion which affect the dissolution rate are discussed. Formulation and evaluation techniques for solid dispersions have been described. The final section of article highlights the recent patents and studies related to solid dispersion systems.
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Clinical use of vaginal or rectally applied microbicides in patients suffering from HIV/AIDS.
HIV AIDS (Auckl)
PUBLISHED: 01-01-2013
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Microbicides, primarily used as topical pre-exposure prophylaxis, have been proposed to prevent sexual transmission of HIV. This review covers the trends and challenges in the development of safe and effective microbicides to prevent sexual transmission of HIV Initial phases of microbicide development used such surfactants as nonoxynol-9 (N-9), C13G, and sodium lauryl sulfate, aiming to inactivate the virus. Clinical trials of microbicides based on N-9 and C31G failed to inhibit sexual transmission of HIV. On the contrary, N-9 enhanced susceptibility to sexual transmission of HIV-1. Subsequently, microbicides based on polyanions and a variety of other compounds that inhibit the binding, fusion, or entry of virus to the host cells were evaluated for their efficacy in different clinical setups. Most of these trials failed to show either safety or efficacy for prevention of HIV transmission. The next phase of microbicide development involved antiretroviral drugs. Microbicide in the form of 1% tenofovir vaginal gel when tested in a Phase IIb trial (CAPRISA 004) in a coitally dependent manner revealed that tenofovir gel users were 39% less likely to become HIV-infected compared to placebo control. However, in another trial (VOICE MTN 003), tenofovir gel used once daily in a coitally independent mode failed to show any efficacy to prevent HIV infection. Tenofovir gel is currently in a Phase III safety and efficacy trial in South Africa (FACTS 001) employing a coitally dependent dosing regimen. Further, long-acting microbicide-delivery systems (vaginal ring) for slow release of such antiretroviral drugs as dapivirine are also undergoing clinical trials. Discovering new markers as correlates of protective efficacy, novel long-acting delivery systems with improved adherence in the use of microbicides, discovering new compounds effective against a broad spectrum of HIV strains, developing multipurpose technologies incorporating additional features of efficacy against other sexually transmitted infections, and contraception will help in moving the field of microbicide development forward.
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Expanding and improving urban outreach immunization in Patna, India.
Trop. Med. Int. Health
PUBLISHED: 12-14-2011
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We conducted a case study of an urban immunization outreach strategy to determine the feasibility of the intervention and to measure administrative immunization coverage outcomes.
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Microbicides: a new hope for HIV prevention.
Indian J. Med. Res.
PUBLISHED: 09-01-2011
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Human immunodeficiency virus (HIV), causative agent of acquired immunodeficiency syndrome (AIDS), is a global health concern. To control its transmission, safe sex has been proposed as one of the strategies. Microbicides- intravaginal/intrarectal topical formulations of anti-HIV agents have also been proposed to prevent HIV transmission. Microbicides would provide protection by directly inactivating HIV or preventing the attachment, entry or replication of HIV in susceptible target cells as well as their dissemination from target cells present in semen or the host cells lining the vaginal/rectal wall to other migratory cells. Microbicides must be safe, effective following vaginal or rectal administration, and should cause minimal or no genital symptoms or inflammations following long-term repeated usage. However, a safe and efficacious anti-HIV microbicide is not yet available despite the fact that more than 60 candidate agents have been identified to have in vitro activity against HIV, several of which have advanced to clinical testing. Nonetheless, proof-of-concept of microbicides has been established based on the results of recent CAPRISA 004 clinical trials. In this article, the trends and challenges in the development of effective and safe microbicides to combat HIV transmission are reviewed.
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Regression of coronary atherosclerosis through healthy lifestyle in coronary artery disease patients--Mount Abu Open Heart Trial.
Indian Heart J
PUBLISHED: 09-01-2011
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To evaluate the efficacy of a unique healthy and happy lifestyle (HLS) program in regression of coronary atherosclerosis and reduction in cardiac events in an open trial.
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Leukaemia inhibitory factor mediated proliferation of HTR-8/SVneo trophoblast cells is dependent on activation of extracellular signal-regulated kinase 1/2.
Reprod. Fertil. Dev.
PUBLISHED: 06-04-2011
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Leukaemia inhibitory factor (LIF) is one of the cytokines that is indispensable for embryo implantation. The aim of the present study was to investigate the role of activation of extracellular signal-regulated kinase (ERK) 1/2 in LIF-mediated proliferation of HTR-8/SVneo cells. Stimulation of HTR-8/SVneo cells with LIF (50 ng mL(-1)) resulted in an increase in cell proliferation (P < 0.05) via increased transition of cells to the G(2)/M phase of cell cycle. Stimulation with LIF resulted in the activation of both signal transducer and activator of transcription (STAT) 3 Tyr(705) and ERK1/2, but inhibition of ERK1/2 signalling by pretreatment of cells with U0126 (10 µM) for 2h resulted in abrogation of LIF-mediated increases in G(2)/M transition, with a significant decrease (P < 0.05) in absolute cell numbers compared with control. Although STAT3 silencing had no effect on LIF-dependent proliferation of HTR-8/SVneo cells, it did result in an increase in cell apoptosis, which increased further upon inhibition of ERK1/2 activation irrespective of LIF stimulation. Stimulation of cells with LIF increased the Bcl-2/Bax ratio, whereas ERK1/2 inhibition decreased the Bcl-2/Bax ratio, even after LIF stimulation. Hence, it can be inferred that ERK1/2 activation is essential for LIF-mediated increases in proliferation and that both STAT3 and ERK1/2 activation are important for the survival of HTR-8/SVneo cells.
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Relevance of syndecan-1 in the trophoblastic BeWo cell syncytialization.
Am. J. Reprod. Immunol.
PUBLISHED: 05-30-2011
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To investigate the role of syndecan-1 in the differentiation of the BeWo cells into syncytiotrophoblast.
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Contraceptive vaccines based on the zona pellucida glycoproteins for dogs and other wildlife population management.
Am. J. Reprod. Immunol.
PUBLISHED: 04-19-2011
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Zona pellucida (ZP) glycoproteins, by virtue of their critical role in fertilization, have been proposed as candidate antigens for the development of contraceptive vaccines. In this review, the potential of a ZP-based contraceptive vaccine for the management of wildlife population, with special reference to street dogs, is discussed. Immunization of various animal species, including female dogs, with native porcine ZP led to inhibition of fertility, which was associated with the ovarian dysfunction. Immunization of female dogs with Escherichia coli-expressed recombinant dog ZP glycoprotein-3 (ZP3) either coupled to diphtheria toxoid or expressed as fusion protein with promiscuous T non-B-cell epitope of tetanus toxoid also led to inhibition of fertility. To improve the contraceptive efficacy of ZP-based contraceptive vaccine, various groups are working on improving the immunogen, use of DNA vaccine as prime-boost strategy, and delivering the zona proteins/peptides presented on either virus-like particles or entrapped in microsphere. Host-specific live vectors such as ectromelia virus and cytomegalovirus have also been used to deliver mouse ZP3 in mice. Various studies show the enormous potential of the ZP-based vaccine for the management of wildlife population, where permanent sterilization may be desirable.
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Safety of food crops on land contaminated with trace elements.
J. Sci. Food Agric.
PUBLISHED: 03-28-2011
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Contamination of agricultural soils with trace elements (TEs) through municipal and industrial wastes, atmospheric deposition and fertilisers is a matter of great global concern. Since TE accumulation in edible plant parts depends on soil characteristics, plant genotype and agricultural practices, those soil- and plant-specific options that restrict the entry of harmful TEs into the food chain to protect human and animal health are reviewed. Soil options such as in situ stabilisation of TEs in soils, changes in physicochemical parameters, fertiliser management, element interactions and agronomic practices reduce TE uptake by food crops. Furthermore, phytoremediation and solubilisation as alternative techniques to reduce TE concentrations in soils are also discussed. Among plant options, selection of species and cultivars, metabolic processes and microbial transformations in the rhizosphere can potentially affect TE uptake and distribution in plants. For this purpose, genetic variations are exploited to select cultivars with low uptake potential, especially low-cadmium accumulator wheat and rice cultivars. The microbial reduction of elements and transformations in the rhizosphere are other key players in the cycling of TEs that may offer the basis for a wide range of innovative biotechnological processes. It is thus concluded that appropriate combination of soil- and plant-specific options can minimise TE transfer to the food chain.
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Effectiveness of Muskaan Ek Abhiyan (the smile campaign) for strengthening routine immunization in bihar, India.
Indian Pediatr
PUBLISHED: 03-15-2011
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In Bihar State, proportion of fully immunized children was only 19% ;in Coverage Evaluation Survey of 2005. In October 2007, a special campaign called Muskaan Ek Abhiyan (The Smile Campaign) was launched under National Rural Health Mission to give a fillip to the immunization program.
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Phenotype, function, and gene expression profiles of programmed death-1(hi) CD8 T cells in healthy human adults.
J. Immunol.
PUBLISHED: 03-07-2011
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T cell dysfunction is an important feature of many chronic viral infections. In particular, it was shown that programmed death-1 (PD-1) regulates T cell dysfunction during chronic lymphocytic choriomeningitis virus infection in mice, and PD-1(hi) cells exhibit an intense exhausted gene signature. These findings were extended to human chronic infections such as HIV, hepatitis C virus, and hepatitis B virus. However, it is not known if PD-1(hi) cells of healthy humans have the traits of exhausted cells. In this study, we provide a comprehensive description of phenotype, function, and gene expression profiles of PD-1(hi) versus PD-1(lo) CD8 T cells in the peripheral blood of healthy human adults as follows: 1) the percentage of naive and memory CD8 T cells varied widely in the peripheral blood cells of healthy humans, and PD-1 was expressed by the memory CD8 T cells; 2) PD-1(hi) CD8 T cells in healthy humans did not significantly correlate with the PD-1(hi) exhausted gene signature of HIV-specific human CD8 T cells or chronic lymphocytic choriomeningitis virus-specific CD8 T cells from mice; 3) PD-1 expression did not directly affect the ability of CD8 T cells to secrete cytokines in healthy adults; 4) PD-1 was expressed by the effector memory compared with terminally differentiated effector CD8 T cells; and 5) finally, an interesting inverse relationship between CD45RA and PD-1 expression was observed. In conclusion, our study shows that most PD-1(hi) CD8 T cells in healthy adult humans are effector memory cells rather than exhausted cells.
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Note: A portable pulsed neutron source based on the smallest sealed-type plasma focus device.
Rev Sci Instrum
PUBLISHED: 03-03-2011
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Development and operation of a portable and compact pulsed neutron source based on sealed-type plasma focus (PF) device are reported. The unit is the smallest sealed-type neutron producing PF device. The effective volume of the PF unit is 33 cm(3) only. A compact size single capacitor (4 ?F) is used as the energy driver. A battery based power supply unit is used for charging the capacitor and triggering the spark gap. The PF unit is operated at 10 kV (200 J) and at a deuterium gas filling pressure of 8 mb. The device is operated over a time span of 200 days and the neutron emissions have been observed for 200 shots without changing the gas in between the shots. The maximum yield of this device is 7.8 × 10(4) neutrons/pulse. Beyond 200 shots the yield is below the threshold (1050 neutrons/pulse) of our (3)He detector. The neutron energy is evaluated using time of flight technique and the value is (2.49 ± 0.27) MeV. The measured neutron pulse width is (24 ± 5) ns. Multishot and long duration operations envisage the potentiality of such portable device for repetitive mode of operation.
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Sperm proteasomes degrade sperm receptor on the egg zona pellucida during mammalian fertilization.
PLoS ONE
PUBLISHED: 01-27-2011
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Despite decades of research, the mechanism by which the fertilizing spermatozoon penetrates the mammalian vitelline membrane, the zona pellucida (ZP) remains one of the unexplained fundamental events of human/mammalian development. Evidence has been accumulating in support of the 26S proteasome as a candidate for echinoderm, ascidian and mammalian egg coat lysin. Monitoring ZP protein degradation by sperm during fertilization is nearly impossible because those few spermatozoa that penetrate the ZP leave behind a virtually untraceable residue of degraded proteins. We have overcome this hurdle by designing an experimentally consistent in vitro system in which live boar spermatozoa are co-incubated with ZP-proteins (ZPP) solubilized from porcine oocytes. Using this assay, mimicking sperm-egg interactions, we demonstrate that the sperm-borne proteasomes can degrade the sperm receptor protein ZPC. Upon coincubation with motile spermatozoa, the solubilized ZPP, which appear to be ubiquitinated, adhered to sperm acrosomal caps and induced acrosomal exocytosis/formation of the acrosomal shroud. The degradation of the sperm receptor protein ZPC was assessed by Western blotting band-densitometry and proteomics. A nearly identical pattern of sperm receptor degradation, evident already within the first 5 min of coincubation, was observed when the spermatozoa were replaced with the isolated, enzymatically active, sperm-derived proteasomes. ZPC degradation was blocked by proteasomal inhibitors and accelerated by ubiquitin-aldehyde(UBAL), a modified ubiquitin protein that stimulates proteasomal proteolysis. Such a degradation pattern of ZPC is consistent with in vitro fertilization studies, in which proteasomal inhibitors completely blocked fertilization, and UBAL increased fertilization and polyspermy rates. Preincubation of intact zona-enclosed ova with isolated active sperm proteasomes caused digestion, abrasions and loosening of the exposed zonae, and significantly reduced the fertilization/polyspermy rates after IVF, accompanied by en-mass detachment of zona bound sperm. Thus, the sperm borne 26S proteasome is a candidate zona lysin in mammals. This new paradigm has implications for contraception and assisted reproductive technologies in humans, as well as animals.
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Rhodium(II) acetate-catalyzed stereoselective synthesis, SAR and anti-HIV activity of novel oxindoles bearing cyclopropane ring.
Eur J Med Chem
PUBLISHED: 01-24-2011
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Novel oxindole derivatives bearing substituted cyclopropane ring have been designed on the basis of docking studies with HIV-1 RT using the software DS 2.5 and synthesized as probable NNRTIs against HIV-1 using rhodium(II) acetate-catalyzed stereoselective cyclopropanation reaction. The cyclopropane isomer, having trans relationship with respect to carbonyl of lactam moiety and functional group on the cyclopropane ring, was the major product in all cases along with a small amount of cis and methylene products. The trans isomers interacted well with HIV-1 RT through H-bonding with amino acids, like Lys101, Lys103, His235, Tyr318, constituting the non-nucleoside inhibitor binding pocket (NNIBP) during docking experiments. However, the compounds showed very little activity when subjected to in vitro anti-HIV-1 screening using ?-galactosidase assay (TZM-bl cells) and GFP quantification (CEM-GFP cells). The very low level of in vitro HIV inhibition, in comparison to predicted EC(50) values on the basis of computational studies, during CEM-GFP screening using AZT as positive control indicated that probably the HIV RT is not the viral target and the molecules work through some different mechanism.
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Implementation and evaluation of the Reaching Every District (RED) strategy in Assam, India, 2005-2008.
Vaccine
PUBLISHED: 01-19-2011
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In 2005, UNICEF and the Centers for Disease Control and Prevention implemented and evaluated the Reaching Every District (RED) approach, an intervention designed to improve key components of immunization services including planning, outreach, community mobilization, supervision, and monitoring, in select districts of Assam, India. Two intervention and 3 comparison districts were selected for a 2-year evaluation trial. In intervention districts, immunization staff received comprehensive training and ongoing supervision by a fulltime consultant, and regular monitoring of progress was conducted. Population-based vaccination coverage surveys were conducted at baseline and 2 years after the start of implementation in the 5 districts. Post-intervention process indicators were systematically collected and focus group discussions were held. At follow-up, children in both the intervention and comparison districts were twice as likely to be fully vaccinated as they were at baseline. However, sites that received intervention training were better performing than those that did not, as measured by process indicators, including a higher number of outreach visits planned and held (p=0.02), having a monitoring chart (p<0.01), and correctly calculating dropout (p<0.01). The number of supervisory visits was significantly and positively associated with other key process indicators. Although coverage did not differ significantly between intervention and comparison districts, among individual districts, process data indicate significant improvements in program quality in the intervention districts. Further studies are needed to determine if the improved process indicators have sustainable impact on maintaining improvements in coverage.
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Zona pellucida-based contraceptive vaccines for human and animal utility.
J. Reprod. Immunol.
PUBLISHED: 01-16-2011
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Contraceptive vaccines can be designed to inhibit (i) production of the gametes (sperm and oocyte), (ii) functions of gametes leading to block in fertilization, and (iii) the gamete outcome (pregnancy). The zona pellucida (ZP) glycoproteins have been proposed as candidates for developing contraceptive vaccines by virtue of their critical role in fertilization. Immunization of non-human primates with either native or recombinant ZP proteins leads to curtailment of fertility, which however is invariably associated with ovarian pathology. To avoid oophoritis, immunogens corresponding to mapped B cell epitopes of ZP proteins that are devoid of oophoritogenic T cell epitopes have been proposed. However, ways to overcome the observed oophoritis associated with the ZP-based contraceptive vaccines are yet to be fully defined. This is essential if their use for control of human fertility is to be considered. Nonetheless, contraceptive vaccines based on ZP proteins have shown very promising results in controlling wildlife population such as wild horses, white-tailed deers, elephants, marsupials, grey seals and dogs, where long term infertility or even permanent sterility is desirable.
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Ethyl 3-oxo-2-(2-phenyl-hydrazinyl-idene)butano-ate: a re-determination.
Acta Crystallogr Sect E Struct Rep Online
PUBLISHED: 01-06-2011
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The previous crystallographic studies [Wang et al. (2005 ?). Huaxue Yanjiu16, 29-32; Wang et al. (2007 ?). Youji Huaxue, 27, 524] of the title compound, C(12)H(14)N(2)O(3), gave only the unit-cell dimensions and an R factor with no other details available: the full structure is presented here. The eth-oxy group is disordered over two orientations with refined occupancies of 0.642?(15):0.358?(15). The nine C atoms and two N atoms of the 1-phenyl-2-(propan-2-yl-idene)hydrazine segment of the mol-ecule are close to being coplanar, with a maximum deviation of 0.0779?(14)?Å for the phenyl-amino N atom and an intra-molecular N-H?O hydrogen bond generates an S(6) ring. In the crystal, pairs of C-H?O hydrogen bonds link mol-ecules into inverson dimers, generating R(2) (2)(16) loops.
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Effect of concentration of trimethylchlorosilane (TMCS) and hexamethyldisilazane (HMDZ) silylating agents on surface free energy of silica aerogels.
J Colloid Interface Sci
PUBLISHED: 01-03-2011
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The surface free energy of a solid determines its surface and interfacial behavior in processes like wetting and adhesion which is crucial for silica aerogels in case of organic liquid absorption and transportation of chemicals at nano-scale for biotechnological applications. Here, we have demonstrated that the surface free energy of aerogels can be tuned in wide range from 5.5892 to 0.3073 mJ/m(2) by modifying their surface using TMCS and HMDZ silylating reagents. The alcogels were prepared by two step acid-base catalyzed process where the molar ratio of precursors Tetraethoxysilane (TEOS):Methanol (MeOH):Oxalic acid:NH(4)OH:NH(4)F was kept at optimal value of 1:2.7:0.18×10(-4):0.02:0.22×10(-3), respectively. To modify gel surfaces, TMCS and HMDZ concentration have been varied from 3% to 12% and such alcogels were dried at ambient pressure. It is observed from FTIR for aerogels that increase in concentration of silylating reagent resulted increase in hydrophobicity. This leads to increase in contact angle for water from 123° to 155° but leads to decrease in surface free energy from 5.5892 to 0.3073 mJ/m(2). As there is not direct method, we have used Neumanns equation of state to estimate surface energy of aerogels.
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Acrosome reaction: relevance of zona pellucida glycoproteins.
Asian J. Androl.
PUBLISHED: 11-01-2010
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During mammalian fertilisation, the zona pellucida (ZP) matrix surrounding the oocyte is responsible for the binding of the spermatozoa to the oocyte and induction of the acrosome reaction (AR) in the ZP-bound spermatozoon. The AR is crucial for the penetration of the ZP matrix by spermatozoa. The ZP matrix in mice is composed of three glycoproteins designated ZP1, ZP2 and ZP3, whereas in humans, it is composed of four (ZP1, ZP2, ZP3 and ZP4). ZP3 acts as the putative primary sperm receptor and is responsible for AR induction in mice, whereas in humans (in addition to ZP3), ZP1 and ZP4 also induce the AR. The ability of ZP3 to induce the AR resides in its C-terminal fragment. O-linked glycans are critical for the murine ZP3-mediated AR. However, N-linked glycans of human ZP1, ZP3 and ZP4 have important roles in the induction of the AR. Studies with pharmacological inhibitors showed that the ZP3-induced AR involves the activation of the G(i)-coupled receptor pathway, whereas ZP1- and ZP4-mediated ARs are independent of this pathway. The ZP3-induced AR involves the activation of T-type voltage-operated calcium channels (VOCCs), whereas ZP1- and ZP4-induced ARs involve both T- and L-type VOCCs. To conclude, in mice, ZP3 is primarily responsible for the binding of capacitated spermatozoa to the ZP matrix and induction of the AR, whereas in humans (in addition to ZP3), ZP1 and ZP4 also participate in these stages of fertilisation.
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Zona pellucida-induced acrosome reaction in human spermatozoa is potentiated by glycodelin-A via down-regulation of extracellular signal-regulated kinases and up-regulation of zona pellucida-induced calcium influx.
Hum. Reprod.
PUBLISHED: 09-14-2010
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Glycodelin-A interacts with spermatozoa before fertilization, but its role in modulating sperm functions is not known. Zona pellucida-induced acrosome reaction is crucial to fertilization and its dysfunction is a cause of male infertility. We hypothesized that glycodelin-A, a glycoprotein found in the female reproductive tract, potentiates human spermatozoa for zona pellucida-induced acrosome reaction.
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Decidualized endometrial stromal cell derived factors promote trophoblast invasion.
Fertil. Steril.
PUBLISHED: 06-21-2010
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To evaluate the effects of decidua-derived factors on trophoblast invasion.
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Note: Printed circuit board based electrically triggered compact rail gap switch.
Rev Sci Instrum
PUBLISHED: 06-03-2010
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An electrically triggered rail gap switch has been designed over a commercially available copper clad fiberglass sheet commonly used in making printed circuit boards for applications requiring compact design and direct integration to parallel plate transmission lines. Switch performance has been investigated in terms of its inductance, jitter, and gap closing time. With an electrode separation of 9.0 mm, it has been found to have an inductance of 6 nH, gap closing time of 5 ns, and jitter of about 4-10 ns measured at 95% of self-breakdown voltage. An application of this switch has been demonstrated as an electrically exploding foil accelerator developed over the same board and velocities up to 1.6 km/s have been achieved on Kapton flyers with diameter of 3.0 mm and thickness of 125 microm using a compact 1 microF capacitor bank.
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ZP domain of human zona pellucida glycoprotein-1 binds to human spermatozoa and induces acrosomal exocytosis.
Reprod. Biol. Endocrinol.
PUBLISHED: 05-28-2010
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The human egg coat, zona pellucida (ZP), is composed of four glycoproteins designated as zona pellucida glycoprotein-1 (ZP1), -2 (ZP2), -3 (ZP3) and -4 (ZP4) respectively. The zona proteins possess the archetypal ZP domain, a signature domain comprised of approximately 260 amino acid (aa) residues. In the present manuscript, attempts have been made to delineate the functional significance of the ZP domain module of human ZP1, corresponding to 273-551 aa fragment of human ZP1.
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Water repellent porous silica films by sol-gel dip coating method.
J Colloid Interface Sci
PUBLISHED: 05-27-2010
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The wetting of solid surfaces by water droplets is ubiquitous in our daily lives as well as in industrial processes. In the present research work, water repellent porous silica films are prepared on glass substrate at room temperature by sol-gel process. The coating sol was prepared by keeping the molar ratio of methyltriethoxysilane (MTES), methanol (MeOH), water (H(2)O) constant at 1:12.90:4.74, respectively, with 2M NH(4)OH throughout the experiments and the molar ratio (M) of MTES/Ph-TMS was varied from 0 to 0.22. A simple dip coating technique is adopted to coat silica films on the glass substrates. The static water contact angle as high as 164° and water sliding angle as low as 4° was obtained for silica film prepared from M=0.22. The surface morphological studies of the prepared silica film showed the porous structure with pore sizes typically ranging from 200nm to 1.3?m. The superhydrophobic silica films prepared from M=0.22 retained their superhydrophobicity up to a temperature of 285°C and above this temperature the films became superhydrophilic. The porous and water repellent silica films are prepared by proper alteration of the Ph-TMS in the coating solution. The prepared silica films were characterized by surface profilometer, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier Transform Infrared (FT-IR) spectroscopy, humidity tests, chemical aging tests, static and dynamic water contact angle measurements.
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In humans, zona pellucida glycoprotein-1 binds to spermatozoa and induces acrosomal exocytosis.
Hum. Reprod.
PUBLISHED: 05-26-2010
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It has been suggested that the zona pellucida (ZP) may mediate species-specific fertilization. In human the ZP is composed of four glycoproteins: ZP1, ZP2, ZP3 and ZP4. In the present study, the expression profile of ZP1 in human oocytes and ovaries, and its role during fertilization, is presented.
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Evaluation of subantimicrobial dose doxycycline as an adjunct to scaling and root planing in chronic periodontitis patients with diabetes: a randomized, placebo-controlled clinical trial.
J Contemp Dent Pract
PUBLISHED: 05-13-2010
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Diabetic patients have more severe periodontal destruction, but periodontal therapy can improve metabolic control. Recently, interest has focused on the use of subantimicrobial dose doxycycline (SDD) as a treatment paradigm. Therefore, this study was undertaken to evaluate clinical efficacy of SDD with scaling and root planning (SRP) in chronic periodontitis patients with diabetes.
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Shock experiments and numerical simulations on low energy portable electrically exploding foil accelerators.
Rev Sci Instrum
PUBLISHED: 04-08-2010
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Two low energy (1.6 and 8 kJ) portable electrically exploding foil accelerators are developed for moderately high pressure shock studies at small laboratory scale. Projectile velocities up to 4.0 km/s have been measured on Kapton flyers of thickness 125 microm and diameter 8 mm, using an in-house developed Fabry-Perot velocimeter. An asymmetric tilt of typically few milliradians has been measured in flyers using fiber optic technique. High pressure impact experiments have been carried out on tantalum, and aluminum targets up to pressures of 27 and 18 GPa, respectively. Peak particle velocities at the target-glass interface as measured by Fabry-Perot velocimeter have been found in good agreement with the reported equation of state data. A one-dimensional hydrodynamic code based on realistic models of equation of state and electrical resistivity has been developed to numerically simulate the flyer velocity profiles. The developed numerical scheme is validated against experimental and simulation data reported in literature on such systems. Numerically computed flyer velocity profiles and final flyer velocities have been found in close agreement with the previously reported experimental results with a significant improvement over reported magnetohydrodynamic simulations. Numerical modeling of low energy systems reported here predicts flyer velocity profiles higher than experimental values, indicating possibility of further improvement to achieve higher shock pressures.
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Delineation of downstream signalling components during acrosome reaction mediated by heat solubilized human zona pellucida.
Reprod. Biol. Endocrinol.
PUBLISHED: 01-23-2010
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Human egg is enveloped by a glycoproteinaceous matrix, zona pellucida (ZP), responsible for binding of the human spermatozoa to the egg and induction of acrosomal exocytosis in the spermatozoon bound to ZP. In the present manuscript, attempts have been made to delineate the downstream signalling components employed by human ZP to induce acrosome reaction.
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Measles vaccination response during Kosi floods, Bihar, India 2008.
Indian Pediatr
PUBLISHED: 12-04-2009
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The Kosi floods of Bihar in 2008 led to initial rapid displacement followed by rehabilitation of the affected population. Strategically planned phase-wise activity of supplementary as well as primary measles vaccination combined with a variety of other interventions proved to be successful in preventing outbreaks and deaths due to measles. While 70% supplementary measles vaccination coverage was achieved in relief camps, the coverage of primary measles doses in the latter phases was dependant on accessibility of villages and previous vaccination status of eligible beneficiaries. The integrated diseases surveillance system, which became operational during the floods, also complemented the vaccination efforts by providing daily figures of cases with fever and rash. The overall response was not only successful in terms of preventing measles mortality, but also provided vital lessons that may be useful for planning future vaccination responses in emergency settings.
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Two doses of measles vaccine reduce measles dealths.
Indian Pediatr
PUBLISHED: 12-04-2009
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Two doses of measles vaccine to children reduce measles related deaths. The first dose is delivered through the routine immunization system to infants and the 2nd dose through campaigns or routine immunization system, whichever strategy reaches the highest coverage in the country. Experience in 46 out of 47 measles priority countries has shown that measles vaccination using mass vaccination campaigns can reduce measles related deaths, even in countries where routine immunization system fails to reach an important proportion of children. The gradual adoption of this strategy by countries has resulted in 74% reduction in measles related deaths between 2000 and 2007. The 2010 goal to reduce measles mortality by 90% compared with 2000 levels is achievable if India fully implements its plans to provide a second dose measles vaccine to all children either through campaigns in low coverage areas or through routine services in high coverage areas. Full implementation of measles mortality reduction strategies in all high burden countries will make an important contribution to achieving Millennium Development Goal 4 to reduce child mortality by two thirds in 2015 as compared to 1990.
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Shock Hugoniot of osmium up to 800 GPa from first principles calculations.
J Phys Condens Matter
PUBLISHED: 09-23-2009
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First principles total energy calculations on hcp, ? (a three atom simple hexagonal), ? (bcc) and fcc phases of osmium have been performed as a function of hydrostatic compression employing the FP-LAPW method. The comparison of total energies of these phases up to a maximum compression V/V(0) = 0.58 (pressure?700 GPa) shows that the hcp structure remains stable up to this compression. The 300 K isotherm is determined after adding finite temperature thermal contributions to the total energy calculated as a function of volume at 0 K. From the theoretically determined isotherm, we have derived the shock Hugoniot of this metal and determined the shock parameters C(0) and s to be 4.48 km s(-1) and 1.32, respectively. Employing the theoretically calculated Gruneisen parameter in the differential form of the Lindemann melting rule, we have determined the variation of melting point of the osmium with pressure. The theoretically derived melting curve and the temperature rise along the Hugoniot predict the shock melting of osmium at ?447 GPa with a corresponding temperature of ?9203 K.
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DNA vaccine encoding chimeric protein encompassing epitopes of human ZP3 and ZP4: immunogenicity and characterization of antibodies.
J. Reprod. Immunol.
PUBLISHED: 09-04-2009
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Immunization with zona pellucida (ZP) glycoproteins leads to curtailment of fertility often associated with ovarian dysfunction. To avoid ovarian dysfunction, synthetic peptides corresponding to ZP glycoproteins have been proposed as candidate immunogens. In the present study, plasmid DNA encoding a human ZP glycoprotein-3 (ZP3) epitope corresponding to amino acid (aa) residues 334-343 and a human ZP glycoprotein-4 (ZP4) epitope corresponding to aa residues 251-273 separated by a triglycine spacer was constructed using the mammalian expression vector, VR1020. The plasmid DNA construct expressed both human ZP3 and ZP4 epitopes, as revealed by transient transfection of COS-1 (African green monkey, kidney) mammalian cells. Active immunization of female BALB/cJ mice with the DNA vaccine led to generation of antibodies reactive with baculovirus-expressed recombinant human ZP3, ZP4 and ZP3((334-343aa))-GGG-ZP4((251-273aa)) synthetic peptide in an ELISA as well as T cell responses. Antibodies generated by the DNA vaccine also recognized native ZP. The immune sera significantly inhibited (p<0.005) the binding of FITC-labeled ZP3 to capacitated human sperm, whereas no inhibition in the binding of FITC-labeled ZP4 was observed. However, a significant decrease in acrosomal exocytosis mediated by both recombinant human ZP3 (p<0.005) and ZP4 (p<0.005) was observed in presence of the immune sera. These studies demonstrate that a DNA vaccine can be designed to elicit antibodies against small epitopes of ZP glycoproteins.
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Assessment of successful experiments and limitations of phytotechnologies: contaminant uptake, detoxification and sequestration, and consequences for food safety.
Environ Sci Pollut Res Int
PUBLISHED: 07-01-2009
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The term "phytotechnologies" refers to the application of science and engineering to provide solutions involving plants, including phytoremediation options using plants and associated microbes to remediate environmental compartments contaminated by trace elements (TE) and organic xenobiotics (OX). An extended knowledge of the uptake, translocation, storage, and detoxification mechanisms in plants, of the interactions with microorganisms, and of the use of "omic" technologies (functional genomics, proteomics, and metabolomics), combined with genetic analysis and plant improvement, is essential to understand the fate of contaminants in plants and food, nonfood and technical crops. The integration of physicochemical and biological understanding allows the optimization of these properties of plants, making phytotechnologies more economically and socially attractive, decreasing the level and transfer of contaminants along the food chain and augmenting the content of essential minerals in food crops. This review will disseminate experience gained between 2004 and 2009 by three working groups of COST Action 859 on the uptake, detoxification, and sequestration of pollutants by plants and consequences for food safety. Gaps between scientific approaches and lack of understanding are examined to suggest further research and to clarify the current state-of-the-art for potential end-users of such green options.
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Interleukin-11 increases invasiveness of JEG-3 choriocarcinoma cells by modulating STAT3 expression.
J. Reprod. Immunol.
PUBLISHED: 06-29-2009
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Amongst the interleukin-6 (IL-6) family of cytokines, leukemia inhibitory factor (LIF) has been shown to promote trophoblast invasion and proliferation. In the present study interleukin-11 (IL-11), another member of the IL-6 family, was investigated for its role in regulating invasion, migration and proliferation of JEG-3 choriocarcinoma cells. JEG-3 cells, like extra villous trophoblast (EVT), express mRNA transcripts encoding IL-11 and IL-11 receptor-alpha (IL-11Ralpha). Treatment of JEG-3 cells with IL-11 led to an increase in invasion across Matrigel extracellular matrix without an increase in proliferation. There was a dose-dependent increase in activation of STAT3 under the influence of IL-11 with maximum Tyr705 phosphorylation by 10min. In addition, treatment of JEG-3 cells with IL-11 for 24h led to an increase in expression of unphosphorylated STAT1 and STAT3. Analysis of the nuclear fraction showed an increased localization of STAT3 following IL-11 treatment while STAT1 was absent. Silencing the expression of STAT3 by siRNA caused a 25% reduction in invasion compared to control cells, however this was not significant. Furthermore, treatment of STAT3-silenced JEG-3 cells with IL-11 led to a significant increase in invasion compared to STAT3-silenced cells without cytokine, but this was not significant compared to non-transfected control cells. Silencing the expression of gp130 but not of IL-6R abrogated the increase in invasiveness of JEG-3 cells following IL-11 treatment. In conclusion, activation and upregulation of STAT3 appears to be critical for the IL-11-mediated increase in invasiveness of JEG-3 cells.
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Immune physiology and oogenesis in fetal and adult humans, ovarian infertility, and totipotency of adult ovarian stem cells.
Birth Defects Res. C Embryo Today
PUBLISHED: 03-24-2009
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It is still widely believed that while oocytes in invertebrates and lower vertebrates are periodically renewed throughout life, oocytes in humans and higher vertebrates are formed only during the fetal/perinatal period. However, this dogma is questioned, and clashes with Darwinian evolutionary theory. Studies of oogenesis and follicular renewal from ovarian stem cells (OSCs) in adult human ovaries, and of the role of third-party bone marrow-derived cells (monocyte-derived tissue macrophages and T lymphocytes) could help provide a better understanding of the causes of ovarian infertility, its prevention, and potential treatment. We have reported differentiation of distinct cell types from OSC and the production of new eggs in cultures derived from premenopausal and postmenopausal human ovaries. OSCs are also capable of producing neural/neuronal cells in vitro after sequential stimulation with sex steroid combinations. Hence, OSC represent a unique type of totipotent adult stem cells, which could be utilized for autologous treatment of premature ovarian failure and also for autologous stem cell therapy of neurodegenerative diseases without use of allogeneic embryonic stem cells or somatic cell nuclear transfer. The in vivo application of sex steroid combinations may augment the proliferation of existing neural stem cells and their differentiation into mature neuronal cells (systemic regenerative therapy). Such treatment may also stimulate the transdifferentiation of autologous neural stem cell precursors into neural stem cells useful for topical or systemic regenerative treatment.
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Human zona pellucida glycoproteins: functional relevance during fertilization.
J. Reprod. Immunol.
PUBLISHED: 03-12-2009
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The zona pellucida (ZP), a glycoproteinaceous matrix surrounding the mammalian oocyte plays an important role in species-specific sperm-egg binding, induction of acrosome reaction in the ZP-bound spermatozoa, avoidance of polyspermy and protection of the embryo prior to implantation. In contrast to mouse, human ZP matrix is composed of 4 glycoproteins designated as ZP1, ZP2, ZP3 and ZP4 (Zp4 pseudogene in mouse). Recent studies employing recombinant and immunoaffinity purified human zona proteins revealed that in addition to ZP3, capacitated acrosome-intact spermatozoa also bind ZP4. Human ZP2 primarily binds to the acrosome-reacted spermatozoa, supporting its role as secondary sperm receptor, as delineated in the murine model. For binding of human zona proteins to spermatozoa, glycosylation is not critical. Both human ZP3 and ZP4 induce dose-dependent acrosomal exocytosis in capacitated sperm. In contrast to the murine model, N-linked glycosylation is more critical for the human ZP3/ZP4 mediated induction of acrosomal exocytosis. Subtle differences in the downstream signaling events associated with ZP3 vs. ZP4 mediated induction of acrosomal exocytosis have been observed. To conclude, in humans, ZP3 and ZP4 are involved in binding of the spermatozoa to the egg and subsequent induction of acrosome reaction. The contribution, if any, of human ZP glycoprotein-1 (ZP1) during these stages of fertilization remains to be elucidated.
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Immunogenicity of zona pellucida glycoprotein-3 and spermatozoa YLP(12) peptides presented on Johnson grass mosaic virus-like particles.
Vaccine
PUBLISHED: 03-01-2009
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For safer and effective immunocontraception, zona (ZP3) and spermatozoa specific (YLP(12)) peptides have been presented on virus-like particles (VLPs) derived from Johnson grass mosaic virus coat protein. Immunization of FvB/cJ female mice with VLPs presenting YLP(12)-ZP3 fusion peptide and a physical mixture of VLPs presenting either YLP(12) or ZP3 epitope led to generation of specific antibody responses and a significant reduction in litters born per mice (p<0.005). Significant curtailment of fertility was also observed in animals immunized with adjuvnated ZP3 and YLP(12) synthetic peptides. These results suggest that VLPs can be used to present gamete epitopes for immunocontraception.
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Functional activity of human ZP3 primary sperm receptor resides toward its C-terminus.
Biol. Reprod.
PUBLISHED: 02-25-2009
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Zona pellucida glycoprotein 3 (ZP3) has been ascribed as a putative primary sperm receptor during fertilization in humans. Herein, attempts have been made to delineate the functional domain of human ZP3. ZP3 has been cloned and expressed in a baculovirus expression system as N-terminal fragments (amino acid [aa] residues 1-175 [pAc-ZP3(1-175 aa)] and 23-175 [pBg-ZP3(23-175 aa)]) and as C-terminal fragments (aa residues 214-305 [pBg-ZP3(214-305 aa)] and 214-348 [pBg-ZP3(214-348 aa)]). ZP3 encompassing both N- and C-terminal fragments corresponding to aa residues 1-370 (pAc-ZP3([1-370 aa])) has also been expressed. Lectin-binding analysis with these recombinant proteins revealed the presence of N- and O-linked glycosylation. Significant induction of acrosomal exocytosis was observed when capacitated sperm were incubated with pBg-ZP3(214-348 aa), pBg-ZP3(214-305 aa), and pAc-ZP3(1-370 aa) (P < 0.05), whereas incubation with pAc-ZP3(1-175 aa) and pBg-ZP3(23-175 aa) failed to do so under similar experimental conditions. However, N- and C-terminal fragments labeled with fluorescein isothiocyanate revealed binding to the anterior head of capacitated human spermatozoa. Escherichia coli-expressed ZP3 C-terminal fragments and chemically deglycosylated pBg-ZP3(214-348 aa) failed to induce a significant (P > 0.05) increase in acrosomal exocytosis, suggesting the relevance of glycosylation in imparting functional activity to ZP3 C-terminal fragments. pBg-ZP3(214-348 aa)-mediated induction of acrosomal exocytosis is regulated by G(i) protein, extracellular calcium, GABA(A) [gamma aminobutyric acid (A)] receptor-mediated Cl(-) channel, and T-type voltage-operated calcium channels. Taken together, the results of these studies suggest that the functional activity of human ZP3 resides in its C-terminal domain.
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Assessment of water quality trends in the Minnesota River using non-parametric and parametric methods.
J. Environ. Qual.
PUBLISHED: 01-01-2009
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Excessive loading of sediment and nutrients to rivers is a major problem in many parts of the United States. In this study, we tested the non-parametric Seasonal Kendall (SEAKEN) trend model and the parametric USGS Quality of Water trend program (QWTREND) to quantify trends in water quality of the Minnesota River at Fort Snelling from 1976 to 2003. Both methods indicated decreasing trends in flow-adjusted concentrations of total suspended solids (TSS), total phosphorus (TP), and orthophosphorus (OP) and a generally increasing trend in flow-adjusted nitrate plus nitrite-nitrogen (NO(3)-N) concentration. The SEAKEN results were strongly influenced by the length of the record as well as extreme years (dry or wet) earlier in the record. The QWTREND results, though influenced somewhat by the same factors, were more stable. The magnitudes of trends between the two methods were somewhat different and appeared to be associated with conceptual differences between the flow-adjustment processes used and with data processing methods. The decreasing trends in TSS, TP, and OP concentrations are likely related to conservation measures implemented in the basin. However, dilution effects from wet climate or additional tile drainage cannot be ruled out. The increasing trend in NO(3)-N concentrations was likely due to increased drainage in the basin. Since the Minnesota River is the main source of sediments to the Mississippi River, this study also addressed the rapid filling of Lake Pepin on the Mississippi River and found the likely cause to be increased flow due to recent wet climate in the region.
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Land application of sugar beet by-products: effects on runoff and percolating water quality.
J. Environ. Qual.
PUBLISHED: 01-01-2009
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Water quality concerns, including greater potential for nutrient transport to surface waters resulting in eutrophication and nutrient leaching to ground water, exist when agricultural or food processing industry wastes and by-products are land applied. Plot- and field-scale studies were conducted to evaluate the effects of sugar beet by-products on NO3-N and P losses and biochemical oxygen demand (BOD) in runoff and NO3-N concentrations in percolating waters. In the runoff plot study, treatments in the first year included two rates (224 and 448 Mg ha(-1) fresh weight) of pulp and spoiled beets and a nonfertilized control. In the second year, no by-products were applied on the treated plots, the control treatment was fertilized with N fertilizer, and an additional treatment was added as a nonfertilized control in buffer areas. Wheat (Triticum aestivum L.) was grown in the year of by-product application and sugar beet (Beta vulgaris L.) in the following year. In the percolation field study, the treatments were the control, pulp (224 Mg ha(-)(1)), and spoiled beets (224 Mg ha(-1)). Results from the runoff plot showed that both by-products caused immobilization of soil inorganic N and thus reduced NO3-N losses in runoff and soil waters during the first growing season. There was some risk of NO3-N exceeding the drinking water limit of 10 mg L(-1), especially between the period of wheat harvest and soil freezing in fall when pulp was applied at 448 Mg ha(-1). The field-scale study showed that by-product application at 224 Mg ha(-1) did not result in increased ground water NO3-N concentrations. Application of spoiled beets at both rates caused significantly higher BODs in runoff in the first year of application. The concentrations of total and soluble reactive P (SRP) were also higher from both rates of spoiled beet application and from the higher application rate of pulp during the 2-yr study period. These high BODs and total P and SRP concentrations in runoff waters from land application of sugar beet by-product suggest that application rates should not be higher than 224 Mg ha(-1). Best management practices that prevent runoff from entering surface waters directly from these fields are warranted.
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Land application of sugar beet by-products: effects on nitrogen mineralization and crop yields.
J. Environ. Qual.
PUBLISHED: 01-01-2009
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Land application of food processing wastes has become an acceptable practice because of the nutrient value of the wastes and potential cost savings in their disposal. Spoiled beets and pulp are among the main by-products generated by the sugar beet (Beta vulgaris L.) processing industry. Farmers commonly land apply these by-products at rates >224 Mg ha(-1) on a fresh weight basis. However, information on nutrient release in soils treated with these by-products and their subsequent impacts on crop yield is lacking. Field studies were conducted to determine the effects of sugar beet by-product application on N release and crop yields over two growing seasons. Treatments in the first year were two rates (224 and 448 Mg ha(-1) fresh weight) of pulp and spoiled beets and a nonfertilized control. In the second year after by-product application, the control treatment was fertilized with N fertilizer and an additional treatment was added as a nonfertilized control in buffer areas. Wheat (Triticum aestivum L.) was grown in the year of by-product application and sugar beet in the subsequent year. By-product treatments caused a significant reduction in wheat grain yield compared with the control. This was due to a decline in N availability as a result of immobilization. Based on microplots receiving 15N labeled beets, wheat took up <1% of spoiled beet-N (approximately 4.7 kg ha(-1)) during the year of by-product application. In the second cropping year, sugar beet root yields were significantly higher in the fertilized control and by-product treatments than the nonfertilized control. The lack of significant difference in sugar beet yield between the fertilized control and by-product treatments was likely due to the greater availability of N in the second year. Labeled 15N data also showed that the sugar beet crop recovered a 17% of sugar beet-N, an equivalent of 86 kg N ha(-1), during the second cropping year. There was no difference in sugar beet root yield, N uptake, or soil N mineralization during the sugar beet cropping season between the pulp and the spoiled beet treatments at comparable rates of application.
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