Anti-galectin-3 peptides increase apoptosis in galectin-3 expressing human breast cancer cells

Abstract only availableA critical factor in the proliferation and the metastatic nature of carcinoma cells appears to be their resistance to natural programmed cell death (apoptosis). However, the molecular mechanisms that enable carcinoma cells to become resistant to cell death are unclear. Galectin-3 (Gal-3) is a protein that is found at elevated levels in a variety of primary and metastatic tumor cells that may play a key role in chemo-resistance and proliferation of carcinoma cells. Gal-3 has also been found to play a key role in the regulation of common apoptosis commitment pathways. Therefore, we hypothesize that peptides, which bind to and inhibit Gal-3 functions, could be used to reduce the anti-apoptotic activity of Gal-3 thus increasing the occurrence of cell death in carcinoma cells. Two cell lines were cultured, the human breast cancer cell line BT549 and a Gal-3-transfected derivative of BT549 (BT549/V). After undergoing apoptosis induction with 0.5 M staurosporine, apoptosis markers were detected fluorescently using flow cytometry. Our preliminary data suggests that, in the absence of anti-galectin-3 peptides, the parent BT549 cell line exhibits mitochondrial damage (decrease in mitochondrial membrane potential as detected by using MitoTracker Red fluorescence) by 6 hours of staurosporine treatment, whereas the BT549/V cell line shows little change in MitoTracker Red fluorescence even after 8 hours of apoptosis induction. A similar pattern is observed when changes in MitoTracker Red fluorescence are correlated with changes in phosphatidylserine translocation from the inner to outer surface of the plasma membrane. The current data suggest that cells transfected with Gal-3 have an increased rate of survival after apoptosis induction. In the next phase of this ongoing project, flow cytometric studies of changes in membrane permeability and DNA damage in parent and galectin-3 transfected BT549 cells will be conducted to further define the time-dependent apoptotic response of the BT549 parent versus BT549/V cells. Finally, we will observe and compare the effect of anti-Gal-3 peptides on induction of apoptosis in these two cell lines in order to determine if Gal-3 plays a key role in the anti-apoptotic nature of carcinoma cells and to test if anti-Gal-3 peptides are efficacious in inhibiting the anti-apoptotic functions of Gal-3.Molecular Imaging Progra

Preventing human immunodeficiency virus infection among sexual assault survivors in Cape Town, South Africa: an observational study.

We describe 131 South African sexual assault survivors offered HIV post-exposure prophylaxis (PEP). While the median days completed was 27 (IQR 27, 28), 34% stopped PEP or missed doses. Controlling for baseline symptoms, PEP was not associated with symptoms (OR = 1.30, 95% CI = 0.66, 2.64). Factors associated with unprotected sex included prior unprotected sex (OR = 6.46, 95% CI = 3.04, 13.74), time since the assault (OR = 1.33, 95% CI = 1.12, 1.57) and age (OR = 1.30, 95% CI = 1.08, 1.57). Trauma counseling was protective (OR = 0.18, 95% CI = 0.05, 0.58). Four instances of seroconversion were observed by 6 months (risk = 3.7%, 95% CI = 1.0, 9.1). Proactive follow-up is necessary to increase the likelihood of PEP completion and address the mental health and HIV risk needs of survivors. Adherence interventions and targeted risk reduction counseling should be provided to minimize HIV acquisition

The Impact of Provider Accessibility and Usability of Antibiotic Stewardship Guidelines

Aims for Improvement We worked with the antimicrobial stewardship team on the rollout of a new digital platform encompassing antibiotic prescribing guidelines and institutional antibiograms with userfriendly features such as mobile accessibility and clear selection menus. Our goal was to increase provider awareness and use of institutional guidelines by at least 25% with the ultimate hope of improving antimicrobial stewardship (measured by frequency of broad-spectrum antibiotic ordering) for the hospital within the calendar year

Constitutive fusion of ubiquitin to PCNA provides DNA damage tolerance independent of translesion polymerase activities

In response to replication-blocking DNA lesions, proliferating cell nuclear antigen (PCNA) can be conjugated with a single ubiquitin (Ub) or Lys63-linked Ub chains at the Lys164 residue, leading to two modes of DNA damage tolerance (DDT), namely translesion synthesis (TLS) and error-free DDT, respectively. Several reports suggest a model whereby monoubiquitylated PCNA recruits TLS polymerases through an enhanced physical association. We sought to examine this model in Saccharomyces cerevisiae through artificial fusions of Ub to PCNA in vivo. We created N- and C- terminal gene fusions of Ub to PCNA-K164R (collectively called PCNA·Ub) and found that both conferred tolerance to DNA damage. The creation of viable PCNA·Ub strains lacking endogenous PCNA enabled a thorough analysis of roles for PCNA mono-Ub in DDT. As expected, the DNA damage resistance provided by PCNA·Ub is not dependent on RAD18 or UBC13. Surprisingly, inactivation of TLS polymerases did not abolish PCNA·Ub resistance to DNA damage, nor did PCNA·Ub cause elevated spontaneous mutagenesis, which is a defining characteristic of REV3-dependent TLS activity. Taken together, our data suggest that either the monoubiquitylation of PCNA does not promote TLS activity in all cases or PCNA·Ub reveals a currently undiscovered role for monoubiquitylated PCNA in DNA damage tolerance

Comparison of F(ab') versus Fab antivenom for pit viper envenomation: A prospective, blinded, multicenter, randomized clinical trial

BACKGROUND: Crotalidae Polyvalent Immune Fab (Ovine) has been the only antivenom commercially available in the US since 2007 for treatment of Crotalinae envenomation. Late coagulopathy can occur or recur after clearance of Fab antivenom, often after hospital discharge, lasting in some cases more than 2 weeks. There have been serious, even fatal, bleeding complications associated with recurrence phenomena. Frequent follow-up is required, and additional intervention or hospitalization is often necessary. F(ab')2 immunoglobulin derivatives have longer plasma half life than do Fab. We hypothesized that F(ab')2 antivenom would be superior to Fab in the prevention of late coagulopathy following treatment of patients with Crotalinae envenomation. METHODS: We conducted a prospective, double-blind, randomized clinical trial, comparing late coagulopathy in snakebitten patients treated with F(ab')2 with maintenance doses [F(ab')2/F(ab')2], or F(ab')2 with placebo maintenance doses [F(ab')2/placebo], versus Fab with maintenance doses [Fab/Fab]. The primary efficacy endpoint was coagulopathy (platelet count < 150 K/mm(3), fibrinogen level < 150 mg/dL) between end of maintenance dosing and day 8. RESULTS: 121 patients were randomized at 18 clinical sites and received at least one dose of study drug. 114 completed the study. Of these, 11/37 (29.7%) in the Fab/Fab cohort experienced late coagulopathy versus 4/39 (10.3%, p < 0.05) in the F(ab')2/F(ab')2 cohort and 2/38 (5.3%, p < 0.05) in the F(ab')2/placebo cohort. The lowest heterologous protein exposure was with F(ab')2/placebo. No serious adverse events were related to study drug. In each study arm, one patient experienced an acute serum reaction and one experienced serum sickness. CONCLUSIONS: In this study, management of coagulopathic Crotalinae envenomation with longer-half-life F(ab')2 antivenom, with or without maintenance dosing, reduced the risk of subacute coagulopathy and bleeding following treatment of envenomation

Male Use of Female Sex Work in India: A Nationally Representative Behavioural Survey

Heterosexual transmission of HIV in India is driven by the male use of female sex workers (FSW), but few studies have examined the factors associated with using FSW. This nationally representative study examined the prevalence and correlates of FSW use among 31,040 men aged 15–49 years in India in 2006. Nationally, about 4% of men used FSW in the previous year, representing about 8.5 million FSW clients. Unmarried men were far more likely than married men to use FSW overall (PR = 8.0), but less likely than married men to use FSW among those reporting at least one non-regular partner (PR = 0.8). More than half of all FSW clients were married. FSW use was higher among men in the high-HIV states than in the low-HIV states (PR = 2.7), and half of all FSW clients lived in the high-HIV states. The risk of FSW use rose sharply with increasing number of non-regular partners in the past year. Given the large number of men using FSW, interventions for the much smaller number of FSW remains the most efficient strategy for curbing heterosexual HIV transmission in India

Guidelines to support HIV-affected individuals and couples to achieve pregnancy safely: Update 2018

No abstract available

Integrating Prevention of Mother-to-Child HIV Transmission Programs to Improve Uptake: A Systematic Review

BACKGROUND: We performed a systematic review to assess the effect of integrated perinatal prevention of mother-to-child transmission of HIV interventions compared to non- or partially integrated services on the uptake in low- and middle-income countries. METHODS: We searched for experimental, quasi-experimental and controlled observational studies in any language from 21 databases and grey literature sources. RESULTS: Out of 28 654 citations retrieved, five studies met our inclusion criteria. A cluster randomized controlled trial reported higher probability of nevirapine uptake at the labor wards implementing HIV testing and structured nevirapine adherence assessment (RRR 1.37, bootstrapped 95% CI, 1.04-1.77). A stepped wedge design study showed marked improvement in antiretroviral therapy (ART) enrolment (44.4% versus 25.3%, p<0.001) and initiation (32.9% versus 14.4%, p<0.001) in integrated care, but the median gestational age of ART initiation (27.1 versus 27.7 weeks, p = 0.4), ART duration (10.8 versus 10.0 weeks, p = 0.3) or 90 days ART retention (87.8% versus 91.3%, p = 0.3) did not differ significantly. A cohort study reported no significant difference either in the ART coverage (55% versus 48% versus 47%, p = 0.29) or eight weeks of ART duration before the delivery (50% versus 42% versus 52%; p = 0.96) between integrated, proximal and distal partially integrated care. Two before and after studies assessed the impact of integration on HIV testing uptake in antenatal care. The first study reported that significantly more women received information on PMTCT (92% versus 77%, p<0.001), were tested (76% versus 62%, p<0.001) and learned their HIV status (66% versus 55%, p<0.001) after integration. The second study also reported significant increase in HIV testing uptake after integration (98.8% versus 52.6%, p<0.001). CONCLUSION: Limited, non-generalizable evidence supports the effectiveness of integrated PMTCT programs. More research measuring coverage and other relevant outcomes is urgently needed to inform the design of services delivering PMTCT programs