Distinct genomic profiles in hereditary breast tumors identified by array-based comparative genomic hybridization

Mutations in BRCA1 and BRCA2 account for a significant proportion of hereditary breast cancers. Earlier studies have shown that inherited and sporadic tumors progress along different somatic genetic pathways and that global gene expression profiles distinguish between these groups. To determine whether genomic profiles similarly discriminate among BRCA1, BRCA2, and sporadic tumors, we established DNA copy number profiles using comparative genomic hybridization to BAC-clone microarrays providing <1 Mb resolution. Tumor DNA was obtained from BRCA1 (n = 14) and BRCA2 (n = 12) mutation carriers, as well as sporadic cases (n = 26). Overall, BRCA1 tumors had a higher frequency of copy number alterations than sporadic breast cancers (P = 0.00078). In particular, frequent losses on 4p, 4q, and 5q in BRCA1 tumors and frequent gains on 7p and 17q24 in BRCA2 tumors distinguish these from sporadic tumors. Distinct amplicons at 3q27.1-q27.3 were identified in BRCA1 tumors and at 17q23.3-q24.2 in BRCA2 tumors. A homozygous deletion on 5q12.1 was found in a BRCA1 tumor. Using a set of 169 BAC clones that detect significantly (P < 0.001) different frequencies of copy number changes in inherited and sporadic tumors, these could be discriminated into separate groups using hierarchical clustering. By comparing DNA copy number and RNA expression for genes in these regions, several candidate genes affected by up- or down-regulation were identified. Moreover, using support vector machines, we correctly classified BRCA1 and BRCA2 tumors (P < 0.0000004 and 0.00005, respectively). Further validation may prove this tumor classifier to be useful for selecting familial breast cancer cases for further mutation screening, particularly, as these data can be obtained using archival tissue.     

Assessing public-private reproductive health efforts to reach young married couples in rural Bangladesh

Young married couples (YMC) in Bangladesh receive insufficient attention from service providers for reproductive health and family planning needs. The ACQUIRE Project, undertaken by EngenderHealth, Bangladesh, provides intervention for service providers, social and local leaders, and mothers-in-law as effective agents of channeling information to YMCs. EngenderHealth, in collaboration with the public sector, examined the extent to which an intervention program enhances overall quality of services, respondents' knowledge and attitude, and service-seeking behavior related to reproductive health issues. A quasi-experimental design with two matching groups, one watching the intervention, was used. The endline survey was carried out 10 months after the Baseline survey. Key informants interviews and FGDs were conducted. The findings were mixed. Importantly, young married men and women need friendly services and service providers with positive attitudes.     

Public and Professional Educational Needs for Downstaging Breast Cancer in Egypt

We conducted focus groups with women from urban and rural areas in the Nile Delta region to investigate their attitudes regarding breast cancer diagnosis, treatment, and screening. Six 60-min focus groups, each group comprised of 6–10 women with ages between 20–69 years, were conducted. Discussions included breast health, breast cancer diagnosis, treatment, early detection and screening, and communication for breast health. Almost all urban and rural women reported that women do not see physicians until they are seriously ill or have advanced cancer. They reported that oncologists or gynecologists were important to be seen first if a woman suspected breast cancer and primary care physician are not the primary line of cancer diagnosis. Other deterring factors besides distrust in primary care physicians included attitude that breast cancer equals death and lack of knowledge of early detection and screening techniques. Women felt that public education campaigns must be implemented to improve early detection and screening methods for breast cancer. The majority of beliefs regarding breast cancer and screening were common among urban and rural women. Culture-specific and tailored professional and public education programs in developing countries are essential for achieving downstaging cancer.     

Hepatitis C virus induces oxidative stress,DNA damage and modulates the DNA repair enzyme NEIL1

Background and Aims: Hepatitis C virus (HCV)‐induced chronic inflammation may induce oxidative stress which could compromise the repair of damaged DNA, rendering cells more susceptible to spontaneous or mutagen‐induced alterations, the underlying cause of liver cirrhosis and hepatocellular carcinoma. In the current study we examined the induction of reactive oxygen species (ROS) resulting from HCV infection and evaluated its effect on the host DNA damage and repair machinery. Methods: HCV infected human hepatoma cells were analyzed to determine (i) ROS, (ii) 8‐oxoG and (iii) DNA glycosylases NEIL1, NEIL2, OGG1. Liver biopsies were analyzed for NEIL1. Results: Human hepatoma cells infected with HCV JFH‐1 showed 30–60‐fold increases in ROS levels compared to uninfected cells. Levels of the oxidatively modified guanosine base 8‐oxoguanine (8‐oxoG) were significantly increased sixfold in the HCV‐infected cells. Because DNA glycosylases are the enzymes that remove oxidized nucleotides, their expression in HCV‐infected cells was analyzed. NEIL1 but not OGG1 or NEIL2 gene expression was impaired in HCV‐infected cells. In accordance, we found reduced glycosylase (NEIL1‐specific) activity in HCV‐infected cells. The antioxidant N‐acetyl cystein (NAC) efficiently reversed the NEIL1 repression by inhibiting ROS induction by HCV. NEIL1 expression was also partly restored when virus‐infected cells were treated with interferon (IFN). HCV core and to a lesser extent NS3‐4a and NS5A induced ROS, and downregulated NEIL1 expression. Liver biopsy specimens showed significant impairment of NEIL1 levels in HCV‐infected patients with advanced liver disease compared to patients with no disease. Conclusion: Collectively, the data indicate that HCV induction of ROS and perturbation of NEIL1 expression may be mechanistically involved in progression of liver disease and suggest that antioxidant and antiviral therapies can reverse these deleterious effects of HCV in part by restoring function of the DNA repair enzyme/s.     

Imaging apoptosis with (99m)Tc-annexin-V in experimental subacute myocarditis.

99mTc-Annexin-V (ANX), which allows in vivo detection of apoptotic cells, is potentially a promising noninvasive tool to diagnose myocarditis. To test this assumption, we compared the myocardial uptake of ANX (imaging and quantitative autoradiography) in experimental subacute myocarditis (Wistar Bonn/Kobori rats [WBN/Kob]) and in normal Wistar rats. WBN/Kob is an inbred strain of Wistar rat in which myocardial injury mimicking subacute catecholamine-induced myocarditis spontaneously develops (course duration, 18 mo). The apoptotic myocardial rates were determined by immunohistochemical studies. METHODS: Fourteen WBN/Kob rats (8-10 mo old) and 12 control rats were injected with ANX (7.4 MBq/100 g). Ten-minute anterior planar thoracic images (matrix, 128 x 128) were obtained using a pinhole collimator, 1 and 4 h after injection. Heart-to-lung activity ratios were calculated on the scintigrams. Four hours after ANX injection, quantitative autoradiography of myocardial slices was performed, as well as histologic studies with hematoxylin-eosin and with a staining assay specific for apoptotic cells. RESULTS: Heart-to-lung activity ratios were higher in WBN/Kob rats than in control rats on 4-h images (2.07 +/- 0.07 vs. 1.66 +/- 0.06, P = 0.0007). Autoradiographic studies showed moderate diffuse, homogeneous myocardial ANX uptake significantly higher in WBN/Kob rats than in control rats: 54 +/- 4 versus 37 +/- 3 counts/mm(2) (P < 0.007). The apoptotic rate, evaluated with an apoptotic cell-staining assay, was 0.51% +/- 0.14% of cells in WBN/Kob rats versus 0.0042% +/- 0.0008% in control rats (P < 0.008). CONCLUSION: Compared with control rats, rats with subacute myocarditis mimicking catecholamine-induced myocarditis showed increased ANX myocardial uptake. This suggests a potential role for ANX imaging in the diagnosis of myocarditis.     

Antidiarrheal and Antispasmodic Activities of Buddleja polystachya are Mediated Through Dual Inhibition of Ca++ Influx and Phosphodiesterase Enzyme

This study describes the antidiarrheal and antispasmodic activities of the hydro‐alcoholic extract of Buddleja polystachya (Bp.Cr) with possible mode of action explored along with activity‐directed fractionation. Bp.Cr and its aqueous (Bp.Aq) and organic fractions, petroleum ether (Bp.Pet), dichloromethane (Bp.DCM), ethylacetate (Bp.EtAc) and butanol (Bp.But), were tested using the in‐vivo and in‐vitro assays. The crude extract (100–300 mg/kg) showed 20 and 60% protection of castor oil‐induced diarrhea in mice. In isolated rabbit jejunum, Bp.Cr like papaverine inhibited spontaneous and high K⁺ (80 mM)‐induced contractions equi‐potently. In guinea‐pig ileum, Bp.Cr showed a moderate spasmogenic effect. The activity‐directed fractionation revealed that the spasmolytic activity was concentrated in the organic fractions and spasmogenic component in the aqueous fraction. Amongst the organic fractions, BP.DCM and Bp.Pet inhibited spontaneous and high K⁺‐induced contractions equi‐potently, while Bp.But, like verapamil was more potent against high K⁺. The crude extract and its organic fractions caused rightward shift in the Ca⁺⁺‐concentration response curves (CRCs), similar to verapamil, and all except Bp.But potentiated the isoprenaline‐inhibitory CRCs to the left, similar to papaverine. The results of this study indicate that the crude extract of B. polystachya possesses antidiarrheal and antispasmodic activities, mediated possibly through dual inhibition of Ca⁺⁺ influx and phospodiesterase enzyme. Copyright © 2015 John Wiley & Sons, Ltd.     

Protective effect of rutin on impairment of cognitive functions of due to antiepileptic drugs on zebrafish model

Aim:The severity of adverse reactions due to antiepileptics is observed during initiation and early treatment in which impairment of cognitive effects are common. Since long time, herbal medicine is a natural remedy to treat neural symptoms. Phytochemicals have been proven to be potent neuro-protective agents. Rutin, a bioflavonoid is established to be nootropic in many studies. In this study, we aimed to determine the protective effect of rutin in zebrafish against the side effects produced by AEDs.Results:Treatment with rutin reverted the locomotor behavior to normal. Treatment with AEDs caused fishes to move in all regions while, in case of treatment with rutin, the response reverted to normal. Treatment with AEDs altered swimming behavior of zebrafish, however, rutin showed a positive effect over this behavior. Treatment with AEDs resulted in restricted movement of zebrafish to the dark zone. Treatment with rutin caused increased latency of zebrafish to move in the light compartment. Similarly, time spent in the light compartment by zebrafish treated with rutin was significantly (P < 0.01) higher as compared to zebrafish treated with AEDs.Conclusion:The results suggest a protective role of rutin on cognition impaired by AEDs.KEY WORDS: Cognition, epilepsy, rutin, zebrafish     

Differential recruitment and activation of natural killer cell sub‐populations by Mycobacterium bovis‐infected dendritic cells

Through complex interplay with APCs, subsets of NK cells play an important role in shaping adaptive immune responses. Bovine tuberculosis, caused by Mycobacterium bovis, is increasing in incidence and detailed knowledge of host–pathogen interactions in the natural host is essential to facilitate disease control. We investigated the interactions of NK‐cell sub‐populations and M. bovis‐infected DCs to determine early innate mechanisms in the response to infection. A sub‐population of NK cells (NKp46⁺CD2^?) selectively expressing lymphoid homing and inflammatory chemokine receptors were induced to migrate towards M. bovis‐infected DCs. This migration was associated with increased expression of chemokines CCL3, 4, 5, 20 and CXCL8 by M. bovis‐infected DCs. Activation of NKp46⁺CD2^? NK cells and secretion of IFN‐γ was observed, a response reliant on localised IL‐12 release and direct cellular interaction. In a reciprocal manner, NKp46⁺CD2^? cells induced an increase in the intensity of cell surface MHC class II expression on DCs. In contrast, NKp46⁺CD2⁺ NK cells were unable to secrete IFN‐γ and did not reciprocally affect DCs. This study provides novel evidence to demonstrate distinct effector responses between bovine NK‐cell subsets during mycobacterial infection.     

Two cases of primary non-functioning kidney from the same donor

Abstract:  We report two cases of primary non-functioning kidney from the same non-heart beating donor (NHBD). A 42-yr-old man received a renal transplant from a NHBD. The donor was a 38-yr-old male who died of neck spinal injury. The warm ischemic time was 61 min. One h after reperfusion, the graft became edematous and congested in the presence of normal and adequate vascular anastomosis and lack of renal vein thrombosis. The one-h biopsy specimen showed acute tubular necrosis and severe congestion with occasional platelet aggregates in glomerular capillaries. The graft was resected on post-transplantation day 6. Graftectomy specimen revealed renal cortical necrosis. The contralateral kidney of the same donor was transplanted into a 54-yr-old male. The one-h biopsy specimen showed tubular degeneration and neutrophil infiltration in peritubular capillaries, but no glomerular congestion or thrombus. The recipient died on post-transplantation day 4. Although the cause of death was unknown, necropsy of the renal graft showed renal cortical necrosis, similar to case 1. Ischemia/reperfusion injury was considered the cause of non-functioning kidney in both cases, because of long warm ischemic time. The biopsy specimens at zero-h from both grafts showed focal tubular degeneration only without distinct necrosis, indicating it is difficult to predict viability based on the pathological findings at zero-h or one-h biopsies. There is a need for further understanding of clinical and histological patterns in order to define the criteria of viability of kidney transplant.     

Delivery of Amphotericin B for Effective Treatment of Candida Albicans Induced Dermal Mycosis in Rats via Emulgel System: Formulation and Evaluation

Background:

Amphotericin B (AmB) is among the gold standard antifungal agents used for the treatment of the wide range of fungal infections. However, the drug has various side- effects. Transdermal approach for the delivery of drug is one of the accepted and convenient modes of drug delivery. Aim: The current work was designed to formulate and to evaluate the AmB emulgel system.

Materials and Methods:

In the preparation of AmB emulgel, Carbopol 930 was used as a gel in this study. The formulation was evaluated for viscosity, spreadability, drug content, drug release and in vitro and in vivo antifungal testing.

Results:

AmB emulgel was found to penetrate skin effectively and without any irritation. Further, in vivo studies revealed effective therapeutic potential against Candida albicans induced dermal mycosis. Conclusions: The current work, for the first time, revealed effective delivery of AmB across the skin.