Curcumin abrogates bile-induced NF-κB activity and DNA damage <Emphasis Type="Italic">in vitro</Emphasis> and suppresses NF-κB activity whilst promoting apoptosis <Emphasis Type="Italic">in vivo</Emphasis>, suggesting chemopreventative potential in Barrett’s oesophagus

Introduction

Curcumin has been suggested to possess anti-neoplastic properties. As oesophageal adenocarcinoma (OA) and Barrett’s oesophagus (BO) represent a neoplastic series, we postulated that curcumin supplementation may slow neoplastic progression at this site. Our aim was to investigate the effects of curcumin in vitro and in vivo on markers of oesophageal cancer progression.

Methods

We investigated the in vitro ability of curcumin to prevent bile acid-induced DNA damage using micronucleus assay and nuclear factor-kappaB (NF-κB) activity in the oesophageal cell lines (OE33) using real-time PCR of the extracted RNA. We also analysed NF-κB p65 activation in curcumin-pre-treated OE33 cells exposed to deoxycholic acid (DCA) using ELISA. In another pilot study, BO patients took a daily 500 mg curcumin tablet for 7 days prior to their endoscopy. In biopsies collected from these patients (n=33, 16 curcumin, 17 control), we examined NF-κB-driven gene expression (interleukin (IL)-8, inhibitor-kappaB (I-κB)) using real-time PCR of the extracted RNA from the biopsy sample. The apoptotic frequency was assessed by counting the number of apoptotic bodies in the epithelial cells from the Barrett’s tissue with and without curcumin.

Results

In vitro, curcumin (50 μM) significantly abrogated DNA damage and NF-κB activity induced by bile. Pretreating OE33 cells with curcumin (50 μM) completely abolished the ability of DCA (300 μM) to activate NF-κB. In vivo, IL-8 expression was non-significantly suppressed in the curcumin-supplemented patients compared to the squamous control tissue, whilst also showing a doubling in the apoptotic frequency compared to non-supplemented control patients.

Conclusions

Curcumin abrogated bile-driven effects in vitro. The in vivo data also suggests that curcumin supplementation had beneficial effects (increased apoptosis, potentially reduced NF-κB activity) in the Barrett’s tissues themselves, despite poor delivery of the curcumin to the oesophagus.

     

Hsp 70/Hsp 90 organizing protein as a nitrosylation target in cystic fibrosis therapy

The endogenous signaling molecule S-nitrosoglutathione (GSNO) and other S-nitrosylating agents can cause full maturation of the abnormal gene product ΔF508 cystic fibrosis (CF) transmembrane conductance regulator (CFTR). However, the molecular mechanism of action is not known. Here we show that Hsp70/Hsp90 organizing protein (Hop) is a critical target of GSNO, and its S-nitrosylation results in ΔF508 CFTR maturation and cell surface expression. S-nitrosylation by GSNO inhibited the association of Hop with CFTR in the endoplasmic reticulum. This effect was necessary and sufficient to mediate GSNO-induced cell-surface expression of ΔF508 CFTR. Hop knockdown using siRNA recapitulated the effect of GSNO on ΔF508 CFTR maturation and expression. Moreover, GSNO acted additively with decreased temperature, which promoted mutant CFTR maturation through a Hop-independent mechanism. We conclude that GSNO corrects ΔF508 CFTR trafficking by inhibiting Hop expression, and that combination therapies—using differing mechanisms of action—may have additive benefits in treating CF.     

Reduction Mammoplasty Approach to Oncoplasty—Zone-Wise Planning in Indian Patients

Introduction This article is an attempt to formulate certain guidelines for planning of zone-wise reconstruction after breast conservation surgery. The planning involves applying reduction mammoplasty principles with certain modifications to address the defect. Patients and Methods This is a retrospective study of 61 patients with breast cancer who underwent breast conservation surgery and reconstruction of partial breast defects with oncoplastic techniques between January 2014 to March 2019. Patients having low tumor to breast ratio and thus good candidates for volume displacement techniques were included in the study. Results A total of 61 breast cancer cases were included; 22 cases were located in zone 1, nine in zone 2, seven in zone 3, three in zone 4, four in zone 5, one in zone 6, 12 in zone 7, two in zone 8, and three in zone 9. The most common pedicle design used was superomedial in 38 cases, followed by inferior in 19 and medial in 6 cases. Vertical short scar technique was used in 33 cases and Wise pattern skin incision in 30 cases. Follow-up period ranged from 4 months to 65 months, with a mean of 31 months. Four patients had partial skin necrosis, three had suture line dehiscence, two had wound infection, one had seroma, and eight patients had fat necrosis. All patients were satisfied with the cosmetic outcome. Conclusion Breast oncoplastic techniques are effective, reliable, oncologically safe, and conducted with minimal complications in patients with moderately large ptotic breasts, thereby making planning easier and more reproducible by following the reconstruction procedures described in the article. We believe that these techniques should be incorporated in the armamentarium of every plastic surgeon to manage the defects created after breast conservation surgery, in order to achieve the best cosmetic outcomes.     

Motion-compensated noninvasive periodontal health monitoring using handheld and motor-based photoacoustic-ultrasound imaging systems

Simultaneous visualization of the teeth and periodontium is of significant clinical interest for image-based monitoring of periodontal health. We recently reported the application of a dual-modality photoacoustic-ultrasound (PA-US) imaging system for resolving periodontal anatomy and periodontal pocket depths in humans. This work utilized a linear array transducer attached to a stepper motor to generate 3D images via maximum intensity projection. This prior work also used a medical head immobilizer to reduce artifacts during volume rendering caused by motion from the subject (e.g., breathing, minor head movements). However, this solution does not completely eliminate motion artifacts while also complicating the imaging procedure and causing patient discomfort. To address this issue, we report the implementation of an image registration technique to correctly align B-mode PA-US images and generate artifact-free 2D cross-sections. Application of the deshaking technique to PA phantoms revealed 80% similarity to the ground truth when shaking was intentionally applied during stepper motor scans. Images from handheld sweeps could also be deshaken using an LED PA-US scanner. In ex vivo porcine mandibles, pigmentation of the enamel was well-estimated within 0.1 mm error. The pocket depth measured in a healthy human subject was also in good agreement with our prior study. This report demonstrates that a modality-independent registration technique can be applied to clinically relevant PA-US scans of the periodontium to reduce operator burden of skill and subject discomfort while showing potential for handheld clinical periodontal imaging.     

Maternal Immunity and Vaccination Influence Disease Severity in Progeny in a Novel Mast Cell-Deficient Mouse Model of Severe Dengue

Sub-neutralizing concentrations of antibodies in dengue infected patients is a major risk factor for the development of dengue hemorrhagic fever and dengue shock syndrome. Here, we describe a mouse model with a deficiency in mast cells (MCs) in addition to a deficiency in Type-I and II IFN receptors for studying dengue virus (DENV) infection. We used this model to understand the influence of MCs in a maternal antibody-dependent model of severe dengue, where offspring born to DENV-immune mothers are challenged with a heterologous DENV serotype. Mice lacking both MCs and IFN receptors were found susceptible to primary DENV infection and showed morbidity and mortality. When these mice were immunized, pups born to DENV-immune mothers were found to be protected for a longer duration from a heterologous DENV challenge. In the absence of MCs and type-I interferon signaling, IFN-γ was found to protect pups born to naïve mothers but had the opposite effect on pups born to DENV-immune mothers. Our results highlight the complex interactions between MCs and IFN-signaling in influencing the role of maternal antibodies in DENV-induced disease severity.     

Cocaine Enhances HIV-1–Induced CD4+ T-Cell Apoptosis: Implications in Disease Progression in Cocaine-Abusing HIV-1 Patients

Substance abuse is a major barrier in eradication of the HIV epidemic because it serves as a powerful cofactor for viral transmission, disease progression, and AIDS-related mortality. Cocaine, one of the commonly abused drugs among HIV-1 patients, has been suggested to accelerate HIV disease progression. However, the underlying mechanism remains largely unknown. Therefore, we tested whether cocaine augments HIV-1–associated CD4⁺ T-cell decline, a predictor of HIV disease progression. We examined apoptosis of resting CD4⁺ T cells from HIV-1–negative and HIV-1–positive donors in our study, because decline of uninfected cells plays a major role in HIV-1 disease progression. Treatment of resting CD4⁺ T cells with cocaine (up to 100 μmol/L concentrations) did not induce apoptosis, but 200 to 1000 μmol/L cocaine induced apoptosis in a dose-dependent manner. Notably, treatment of CD4⁺ T cells isolated from healthy donors with both HIV-1 virions and cocaine significantly increased apoptosis compared with the apoptosis induced by cocaine or virions alone. Most important, our biochemical data suggest that cocaine induces CD4⁺ T-cell apoptosis by increasing intracellular reactive oxygen species levels and inducing mitochondrial depolarization. Collectively, our results provide evidence of a synergy between cocaine and HIV-1 on CD4⁺ T-cell apoptosis that may, in part, explain the accelerated disease observed in HIV-1–infected drug abusers.The HIV/AIDS pandemic has claimed the lives of an estimated 35 million people (http://www.who.int/mediacentre/factsheets/fs360/en/index.html, last updated October 2013). Although anti-retroviral therapy (ART) has dramatically reduced HIV/AIDS-related mortality,¹ substance use is a major barrier for combating the HIV/AIDS pandemic because it is associated with transmission, delayed diagnosis, delayed initiation of therapy, and poor adherence to therapy.² Cocaine is a commonly abused drug among HIV-1 patients,^3–5 and studies suggest that cocaine abuse may accelerate HIV-1 disease progression. For example, Vittinghoff et al⁶ documented an increased risk of HIV-1 disease progression among frequent cocaine users. Arnsten et al^7,8 have reported that active cocaine use strongly predicts failure to viral suppression. Similarly, Webber et al⁹ suggested that use of cocaine, along with alcohol, might accelerate HIV-1 disease progression. In addition, Lucas and colleagues^10–12 found that cocaine users have inferior virological and immunological responses to ART. The effects of cocaine on disease progression in these studies can be attributed, in part, to nonadherence to ART, because substance use is often associated with reduced adherence and/or access to ART.¹³ There are also reports that did not find significant association between cocaine abuse and HIV-1 disease progression.^14,15 However, Baum et al³ found that cocaine users have higher viral load and were twice as likely to progress to AIDS when controlled for ART use. Notably, Palepu et al¹⁶ reported that HIV-1–positive drug users, while taking ART, were less likely to suppress the viral load. Recently, Rasbach et al¹⁷ have suggested that active cocaine use among HIV-1 patients is associated with lack of virological suppression, independent of ART adherence. Although in vitro studies suggest that increased HIV-1 replication by cocaine^18–21 may play a role, the mechanism by which cocaine accelerates HIV-1 disease progression remains unclear. Therefore, we evaluated whether cocaine could potentiate HIV-1–induced CD4⁺ T-cell apoptosis because CD4⁺ T-cell decline is an important predictor of HIV-1 disease progression. Our data suggest a synergy between cocaine and HIV-1 on CD4⁺ T-cell apoptosis and highlight the molecular interplay between cocaine abuse and HIV-1 disease progression.