The progestin receptor interactome in the female mouse hypothalamus: Interactions with synaptic proteins are isoform specific and ligand dependent

Progestins bind to the progestin receptor (PR) isoforms, PR-A and PR-B, in brain to influence development, female reproduction, anxiety, and stress. Hormone-activated PRs associate with multiple proteins to form functional complexes. In the present study, proteins from female mouse hypothalamus that associate with PR were isolated using affinity pull-down assays with glutathione S-transferase–tagged mouse PR-A and PR-B. Using complementary proteomics approaches, reverse phase protein array (RPPA) and mass spectrometry, we identified hypothalamic proteins that interact with PR in a ligand-dependent and isoform-specific manner and were confirmed by Western blot. Synaptic proteins, including synapsin-I and synapsin-II, interacted with agonist-bound PR isoforms, suggesting that both isoforms function in synaptic plasticity. In further support, synaptogyrin-III and synapsin-III associated with PR-A and PR-B, respectively. PR also interacted with kinases, including c-Src, mTOR, and MAPK1, confirming phosphorylation as an integral process in rapid effects of PR in the brain. Consistent with a role in transcriptional regulation, PR associated with transcription factors and coactivators in a ligand-specific and isoform-dependent manner. Interestingly, both PR isoforms associated with a key regulator of energy homeostasis, FoxO1, suggesting a novel role for PR in energy metabolism. Because many identified proteins in this PR interactome are synaptic proteins, we tested the hypothesis that progestins function in synaptic plasticity. Indeed, progesterone enhanced synaptic density, by increasing synapsin-I–positive synapses, in rat primary cortical neuronal cultures. This novel combination of RPPA and mass spectrometry allowed identification of PR action in synaptic remodeling and energy homeostasis and reveals unique roles for progestins in brain function and disease

Gain and loss of function of P2X7 receptors: Mechanisms, pharmacology and relevance to diabetic neuropathic pain

Background: Genetic causes of exaggerated or reduced pain sensitivity in humans are well known. Recently, single nucleotide polymorphisms (SNPs) in the gene P2RX7, coding for the ATP-gated ion channel P2X7, have been described that cause gain-of-function (GOF) and loss-of-function (LOF), respectively of this channel. Importantly, P2RX7 SNPs have been associated with more or less severe pain scores in patient suffering of post-mastectomy pain and osteoarthritis. Results: The functional consequences of some P2RX7 SNPs (rs208294 (His155Tyr), rs1718119 (Ala348Thr) and rs3751143 (Glu496Ala)) were studied in recombinant cells in vitro. Our findings suggest a correlation between GOF and LOF of P2X7 and actual channel protein expression. Both channel and pore function for these mutant P2X7 receptors changed in parallel to protein levels. On the other hand, the mutant receptors did not differ in their sensitivity to known P2X7 agonists and antagonists. We further demonstrated that in patients with diabetic peripheral neuropathic pain (DPNP), the presence of the GOF SNPs rs208294 (His155Tyr) and rs1718119 (Ala348Thr) is associated, in females, with higher pain intensity scores. Conclusions: Our present results confirm the physiological relevance of some of the SNPs in the P2RX7 gene and show that the presence of these genetic variants correlates with pain sensitivity also in a diabetic neuropathic pain patient population

The purification and properties of a ribonuclease from Salmonella typhimurium extract

A ribonuclease has been purified about 140-fold from extracts of Salmonella typhimurium through ammonium sulfate fractionation, gel filtration, and DEAE-cellulose chromatography. The optimum pH for the hydrolytic breakdown of RNA is 7. The enzyme hydrolyzes polyadenylic acid (poly A), polycytidylic acid (poly C), and polyuridylic acid (poly U) at much faster rates than transfer RNA. Polyinosinic acid (poly I) was not hydrolyzed at all. Higher concentrations of poly A and poly U (above 200 μg per ml) were inhibitory. A mixture of poly A and poly U in the proportion 1:2, which is known to produce maximum secondary interaction, is also inhibitory. These results indicate that the secondary structures of nucleic acids interfere with the action of the nuclease. The first product of hydrolysis is a 2',3'-cyclic nucleotide which is poorly hydrolyzed to the 3'-nucleotide. The enzyme behaves as an endonuclease. The properties of the S. typhimurium nuclease have been compared with those of RNase I of Escherichia coli. Although there are some differences, the S. typhimurium nuclease is like RNase I in its mode of action

REPRESENTATIONS OF EDGE REGULAR BIPOLAR FUZZY GRAPHS

V. Ramadass & D. Kalpana, “Representations of Edge Regular Bipolar Fuzzy Graphs”, International Journal of Applied and Advanced Scientific Research, Volume 2, Issue 2, Page Number 267-272, 2017

A comparative study to assess the awareness of antibiotic resistance amongst first and second year medical undergraduate students in a medical college

Background: With the rising instances of antibiotic resistance and decline in the discovery of newer antibiotics, it is high time to prevent the emergence of antibiotic resistance. One of the best measures regarding the above problem is education and creating awareness regarding antibiotic resistance, and that too primarily among healthcare professionals. The initiation should start at the basic level and that too in the early days of medical profession, so that there is greater awareness and strong foundation for antibiotic prescribing pattern. The present study is taken up to compare awareness among 1st year and 2nd year medical students in a private medical college with 2nd year medical students having prior education regarding antibiotics resistance and first year students representing general public without any education regarding antibiotics resistance.Methods: This study was a comparative, cross-sectional, questionnaire based study. The questionnaire was validated for face validity and content validity from subject experts. The two groups selected were 1st year and 2nd year medical undergraduates. Data was analysed using simple descriptive statistics.Results: 136 students from 1st year and 88 students from 2nd year participated in the study. In both the groups studied, majority of the respondents completed the full course of antibiotics. 77% of 1st year and all 2nd year students were aware of the antibiotic resistance. 99% of 2nd year students and only 49% of the 1st year students were aware of the reasons for it (p<0.01). Awareness regarding prophylactic usage of antibiotics, usage for simple common cold and usage of higher antibiotics for mid infection leading to Antibiotic Resistance was significantly less among first year students. 2nd year students were well aware of consequences of antibiotic resistance like ineffective treatment, prolongation of illness, emergence of bacterial resistance and additional cost burden than 1st year students. Awareness regarding other aspects of antibiotic resistance was not significant between two groups.Conclusions: Stressing upon awareness and education regarding antibiotic resistance in the early days of medical curriculum will help in change of behaviour and habits of antibiotic usage and rational prescribing of antibiotics by the future doctors.

MATLAB based Brain Tumour Extraction using Artificial Neural Network

Brain tumour is the major cause of mortality among children and adults. The chance of survival can be greater when the tumour is detected correctly at its early stage. This paper presents a neural network techniques for the classification of magnetic resonance brain image. The proposed technique consists of three stages, features extraction from gray scale MR Image using gray level co-occurrence matrix, MR image segmentation using k-mean clustering method and classification of MR Image into normal and abnormal (tumourous) image using feed-forward neural network. This technique have been developed on MATLAB version 7.5.0 platform

Insight into Diabetes Management - A Kriyatmak Approach

Madhumeha is one of the leading lifestyle disorders which is posing a big challenge for the health care fraternity. Before planning the treatment it is very essential to know the Samprapti Ghatakas as well as the principles behind each Yoga which is recommended for treating Madhumeha. An insight into these aspects of Madhumeha Chikitsa has been reviewed in this article

An Anti-HIV-1 V3 Loop Antibody Fully Protects Cross-Clade and Elicits T-Cell Immunity in Macaques Mucosally Challenged with an R5 Clade C SHIV

Neutralizing antibodies have been shown to protect macaques against SHIV challenge. However, genetically diverse HIV-1 clades have evolved, and a key question left unanswered is whether neutralizing antibodies can confer cross-clade protection in vivo. The novel human monoclonal antibody HGN194 was isolated from an individual infected with an HIV-1 clade AG recombinant circulating recombinant form (CRF). HGN194 targets an epitope in the third hypervariable loop (V3) of HIV-1 gp120 and neutralizes a range of relatively neutralization-sensitive and resistant viruses. We evaluated the potential of HGN194 to protect infant rhesus monkeys against a SHIV encoding a primary CCR5-tropic HIV-1 clade C envelope. After high-dose mucosal challenge, all untreated controls became highly viremic while all HGN194-treated animals (50 mg/kg) were completely protected. When HGN194 was given at 1 mg/kg, one out of two monkeys remained aviremic, whereas the other had delayed, lower peak viremia. Interestingly, all protected monkeys given high-dose HGN194 developed Gag-specific proliferative responses of both CD4+ and CD8+ T cells. To test whether generation of the latter involved cryptic infection, we ablated CD8+ cells after HGN194 clearance. No viremia was detected in any protected monkeys, thus ruling out virus reservoirs. Thus, induction of CD8 T-cell immunity may have resulted from transient “Hit and Run” infection or cross priming via Ag-Ab-mediated cross-presentation. Together, our data identified the HGN194 epitope as protective and provide proof-of-concept that this anti-V3 loop mAb can prevent infection with sterilizing immunity after challenge with virus of a different clade, implying that V3 is a potential vaccine target