Prevalence of pelvic floor dysfunction in women attending obstetrics and gynaecology OPD at PES Institute of Medical Sciences asnd Research, Kuppam

Background: Millions of women are affected with pelvic floor dysfunction globally. But when the literature was reviewed, studies assessing the prevalence of PFD (pelvic floor dysfunction) and related factors were limited in India. Hence the present study was undertaken to assess the prevalence of PFD.Methods: 300 women aged 18-70years attending Obstetrics and Gynaecology OPD at Kuppam were interviewed and details like age, number of children, mode of delivery, BMI, education, occupation as primary outcome variables and type of pelvic floor dysfunction as explanatory variable were collected using a semi structured questionnaire. Frequency and proportions were calculated for quantitative variables and Chi-square test was used for comparison of categorical variables. p-value of 45 years old, 38 (65.52%) women had 2 or 3 children, and 12 (20.69%) women had ≥4 children. 47 (81.03%) had vaginal delivery. 37 (63.79%) subjects had no schooling and 13 (22.41%) were unskilled workers. 36 (62.07%) participants BMI ranged between 25 to 34.99. Age, education, occupation, number of children, mode of delivery, BMI were found to be associated with increased incidence of pelvic floor dysfunction.Conclusions: In conclusion the study assessed high prevalence of pelvic floor dysfunction associated with vaginal delivery, increased age, number of children and BMI

Evaluation of TNF-α, IL-10 and IL-6 Cytokine Production and Their Correlation with Genotype Variants amongst Tuberculosis Patients and Their Household Contacts.

BackgroundHousehold contacts of diagnostically established tuberculosis (TB) patients are highly susceptible to disease development. It is surmised that cytokines perhaps play a synergistic and a prognostic role in the activation of the otherwise latent infection in these house hold contacts. Evaluation of the cytokines and any of their inherent polymorphisms might provide a useful diagnostic tool in evaluating the immune regulation and the progression of the disease. The cytokines thus released in a paracrine manner in serum may also provide an indirect measure of the cytokine function.ObjectiveThe present study was aimed to evaluate the levels of TNF-α, IL-10 & IL-6 cytokines and their correlation with genotype variants amongst tuberculosis patients and their household contacts.MethodsThe cytokine levels were estimated in serum by enzyme-linked immunosorbent assay (ELISA) and their polymorphisms were studied by amplification refractory mutation system polymerase chain reaction (ARMs PCR) in active pulmonary tuberculosis patients (APTB = 150), household contacts (HHC = 190), and healthy controls (HC = 150).ResultsThe median values of TNF-α cytokine were significantly high among APTB and HHC compared to HCs (PConclusionLarge sample size with follow-up at different time points may further illuminate the role of IL-10 and IL-6 cytokines as a prognostic marker in house hold contacts

Mycobacterial and HIV Infections Up-Regulated Human Zinc Finger Protein 134, a Novel Positive Regulator of HIV-1 LTR Activity and Viral Propagation

<div><p>Background</p><p>Concurrent occurrence of HIV and Tuberculosis (TB) infections influence the cellular environment of the host for synergistic existence. An elementary approach to understand such coalition at the molecular level is to understand the interactions of the host and the viral factors that subsequently effect viral replication. Long terminal repeats (LTR) of HIV genome serve as a template for binding trans-acting viral and cellular factors that regulate its transcriptional activity, thereby, deciding the fate of HIV pathogenesis, making it an ideal system to explore the interplay between HIV and the host.</p><p>Methodology/Principal Findings</p><p>In this study, using biotinylated full length HIV-1 LTR sequence as bait followed by MALDI analyses, we identified and further characterized human-Zinc-finger-protein-134 (hZNF-134) as a novel positive regulator of HIV-1 that promoted LTR-driven transcription and viral production. Over-expression of hZNF-134 promoted LTR driven luciferase activity and viral transcripts, resulting in increased virus production while siRNA mediated knockdown reduced both the viral transcripts and the viral titers, establishing hZNF-134 as a positive effector of HIV-1. HIV, <i>Mycobacteria</i> and HIV-TB co-infections increased hZNF-134 expressions in PBMCs, the impact being highest by mycobacteria. Corroborating these observations, primary TB patients (n = 22) recorded extraordinarily high transcript levels of hZNF-134 as compared to healthy controls (n = 16).</p><p>Conclusions/Significance</p><p>With these observations, it was concluded that hZNF-134, which promoted HIV-1 LTR activity acted as a positive regulator of HIV propagation in human host. High titers of hZNF-134 transcripts in TB patients suggest that up-regulation of such positive effectors of HIV-1 upon mycobacterial infection can be yet another mechanism by which mycobacteria assists HIV-1 propagation during HIV-TB co-infections. hZNF-134, an uncharacterized host protein, thus assumes a novel regulatory role during HIV-host interactions. Our study provides new insights into the emerging role of zinc finger proteins in HIV-1 pathogenesis.</p></div

HIV-1 and <i>Mycobacterium bovis</i>BCG infections increased the transcript levels of hZNF-134.

<p>PBMCs were isolated from three healthy donors and infected either with HIV, <i>Mycobacterium bovis</i>BCG or both. The levels of hZNF-134 was measured by qRT-PCR and compared to uninfected controls. (A) hZNF-134 transcript levels upon HIV infection. (B) hZNF-134 transcript levels upon <i>Mycobacterium bovis</i>BCG infection. (C) Comparison of hZNF-134 transcript levels upon HIV and HIV-<i>Mycobacterium bovis</i>BCG co-infections. Uninfected cells were used as controls. The transcript levels were normalized to transcript levels of β-Actin. (D) The relative expression of 9 kb viral transcript was measured in HIV and HIV-<i>Mycobacterium bovis</i>BCG co-infections. The transcript levels were normalized to transcript levels of β-Actin. All experiments were done in triplicate and error bars represent mean ± SD.</p

hZNF-134 over expression enhanced the transcriptional potential of LTR and increased viral titers.

<p>(A) Luciferase activity was measured in the presence or absence of Tat following transfection with pEGFP-C3 or ZNF-134-GFP-C3 vector into HEK293T cells along with the reporter construct, pLTR-luc. Luciferase activity was measured in terms of relative luminescence units (RLU) from HEK293T cell lysates after 48 hours and plotted as fold increase in the luciferase activity. Cells over-expressing hZNF-134-GFP showed increased luciferase activity when compared to GFP alone. (B) pNL4-3 was transfected in HEK293T cells with either GFP or hZNF-134-GFP. After 48 hours, the viral transcription was measured by qRT-PCR. The plot shows that viral transcripts increased in the presence of over-expressed hZNF-134. (C) HEK293T cells over-expressing either GFP or hZNF-134-GFP were transfected with pNL4-3 vector and after 48 hours scored for viral p24 by ELISA. Inset shows the Western blot confirming the over-expression of either GFP or hZNF-134-GFP in HEK293T cells. For loading control, expression of GAPDH was detected by Western blots in both the cases. Over-expression of hZNF-134 increased viral titers. All the transfection efficiencies were normalized to pRL-TK transfections. All experiments were done more than three times and error bars represents mean ± SD. Student's <i>t</i>-test was performed and *p<0.05 was considered significant.</p

hZNF-134 could bind to HIV-1 LTR.

<p>(A) GFP or hZNF-134-GFP fusion proteins were transiently expressed in HEK293T cells. After 48 hours post transfection, cell lysates were prepared and Western blot with anti-GFP antibody was performed to check for the expression of GFP or hZNF-134-GFP. GAPDH was used as a loading control. (B) Transiently expressed GFP or hZNF-134-GFP in HEK293T cells were used for pull-down assays with biotinylated LTR. Western blot analyses of the pull-down samples using anti-GFP antibody showed that biotinylated LTR could capture hZNF-134-GFP successfully but GFP alone was not captured by LTR. Pull-down experiments were performed more than three times.</p

hZNF-134-GFP is localized in the nucleus of HEK293T and Astrocytoma 1321N1 cells.

<p>HEK293T and 1321N1 cells were transfected with expression plasmids pEGFP-C3 or ZNF-134-GFP-C3. The cells were fixed after 48 hours and visualized under confocal microscopy. Panels show Trans, DAPI, GFP, and Merged images. hZNF-134-GFP protein was localized in the nucleus of HEK293T and 1321N1 cells whereas GFP control showed both nuclear and cytoplasmic localization. Localization was confirmed by 3 independent experiments.</p

siRNA mediated knockdown of hZNF-134 decreased the viral transcript levels and the final viral titer output measured by p24 ELISA.

<p>(A) qRT-PCR showing reduced transcript levels of hZNF-134 after siRNA mediated knockdown. Scrambled siRNA was used as a control. 50picomoles of each siRNA were used. The transcript levels were normalized to the β-actin levels. (B) pNL4-3 was transfected in HEK293T cells treated with 50 picomoles of scrambled siRNA or hZNF-134 siRNA. After 48 hours, the viral transcription was measured by qRT-PCR. The plot shows that viral transcripts decreased following the knockdown of hZNF-134. (C) Viral p24 levels were measured in pNL4-3 transfected HEK293T cells after treatment with 50picomoles of hZNF-134 specific or scrambled siRNA. All experiments were done more than three times and error bars represents mean ± SD. Student's <i>t</i>-test was performed and *p<0.05 was considered significant.</p

Comparison of cytokines in APTB, HHC and HC categories.

<p>(a) Ratio of TNF-α/IL-10 serum levels (b) Ratio of IL-6/IL-10 serum levels. Results are represented as box plot. The threshold for significance was set at p ≤ 0.05. Bars above the plots represent the statistical differences between the groups.</p

Serum levels of TNF-α, IL-10, IL-6.

<p>(2a, 2b, 2c) Data are shown as box plots, where the boxes represent the first through third quartiles, the lines within the boxes represent the median, and the lines outside the boxes represent the minimum and maximum values (excluding outliers). APTB- active pulmonary Tuberculosis patients; HHC-household contacts; HC-healthy controls.</p