Down Regulation of Genes Involved in T Cell Polarity and Motility during the Induction of Heart Allograft Tolerance by Allochimeric MHC I

BACKGROUND:The allochimeric MHC class I molecule [alpha1h1/u]-RT1.Aa that contains donor-type (Wistar Furth, WF; RT1u) epitopes displayed on recipient-type (ACI, RT1a) administered in conjunction with sub-therapeutic dose of cyclosporine (CsA) induces indefinite survival of heterotopic cardiac allografts in rat model. In vascularized transplantation models, the spleen contributes to graft rejection by generating alloantigen reactive T cells. The immune response in allograft rejection involves a cascade of molecular events leading to the formation of immunological synapses between T cells and the antigen-presenting cells. METHODOLOGY/PRINCIPAL FINDINGS:To elucidate the molecular pathways involved in the immunosuppressive function of allochimeric molecule we performed microarray and quantitative RTPCR analyses of gene expression profile of splenic T cells from untreated, CsA treated, and allochimeric molecule + subtherapeutic dose of CsA treated animals at day 1, 3 and 7 of post transplantation. Allochimeric molecule treatment caused down regulation of genes involved in actin filament polymerization (RhoA and Rac1), cell adhesion (Catna1, Vcam and CD9), vacuolar transport (RhoB, Cln8 and ATP6v1b2), and MAPK pathway (Spred1 and Dusp6) involved in tubulin cytoskeleton reorganization and interaction between actin and microtubule cytoskeleton. All these genes are involved in T cell polarity and motility, i.e., their ability to move, scan and to form functional immunological synapse with antigen presenting cells (APCs). CONCLUSIONS:These results indicate that the immunosuppressive function of allochimeric molecule may depend on the impairment of T cells' movement and scanning ability, and possibly also the formation of immunological synapse. We believe that these novel findings may have important clinical implications for organ transplantation

Hsp72 (HSPA1A) Prevents Human Islet Amyloid Polypeptide Aggregation and Toxicity: A New Approach for Type 2 Diabetes Treatment

Type 2 diabetes is a growing public health concern and accounts for approximately 90% of all the cases of diabetes. Besides insulin resistance, type 2 diabetes is characterized by a deficit in β-cell mass as a result of misfolded human islet amyloid polypeptide (h-IAPP) which forms toxic aggregates that destroy pancreatic β-cells. Heat shock proteins (HSP) play an important role in combating the unwanted self-association of unfolded proteins. We hypothesized that Hsp72 (HSPA1A) prevents h-IAPP aggregation and toxicity. In this study, we demonstrated that thermal stress significantly up-regulates the intracellular expression of Hsp72, and prevents h-IAPP toxicity against pancreatic β-cells. Moreover, Hsp72 (HSPA1A) overexpression in pancreatic β-cells ameliorates h-IAPP toxicity. To test the hypothesis that Hsp72 (HSPA1A) prevents aggregation and fibril formation, we established a novel C. elegans model that expresses the highly amyloidogenic human pro-IAPP (h-proIAPP) that is implicated in amyloid formation and β-cell toxicity. We demonstrated that h-proIAPP expression in body-wall muscles, pharynx and neurons adversely affects C. elegans development. In addition, we demonstrated that h-proIAPP forms insoluble aggregates and that the co-expression of h-Hsp72 in our h-proIAPP C. elegans model, increases h-proIAPP solubility. Furthermore, treatment of transgenic h-proIAPP C. elegans with ADAPT-232, known to induce the expression and release of Hsp72 (HSPA1A), significantly improved the growth retardation phenotype of transgenic worms. Taken together, this study identifies Hsp72 (HSPA1A) as a potential treatment to prevent β-cell mass decline in type 2 diabetic patients and establishes for the first time a novel in vivo model that can be used to select compounds that attenuate h-proIAPP aggregation and toxicity

A mouse model for triple-negative breast cancer tumor-initiating cells (TNBC-TICs) exhibits similar aggressive phenotype to the human disease

<p>Abstract</p> <p>Background</p> <p>Triple-negative breast cancer (TNBC) exhibit characteristics quite distinct from other kinds of breast cancer, presenting as an aggressive disease--recurring and metastasizing more often than other kinds of breast cancer, without tumor-specific treatment options and accounts for 15% of all types of breast cancer with higher percentages in premenopausal African-American and Hispanic women. The reason for this aggressive phenotype is currently the focus of intensive research. However, progress is hampered by the lack of suitable TNBC cell model systems.</p> <p>Methods</p> <p>To understand the mechanistic basis for the aggressiveness of TNBC, we produced a stable TNBC cell line by sorting for 4T1 cells that do not express the estrogen receptor (ER), progesterone receptor (PgR) or the gene for human epidermal growth factor receptor 2 (HER2). As a control, we produced a stable triple-positive breast cancer (TPBC) cell line by transfecting 4T1 cells with rat HER2, ER and PgR genes and sorted for cells with high expression of ER and PgR by flow cytometry and high expression of the HER2 gene by Western blot analysis.</p> <p>Results</p> <p>We isolated tumor-initiating cells (TICs) by sorting for CD24⁺/CD44^high/ALDH1⁺cells from TNBC (TNBC-TICs) and TPBC (TPBC-TICs) stable cell lines. Limiting dilution transplantation experiments revealed that CD24⁺/CD44^high/ALDH1⁺cells derived from TNBC (TNBC-TICs) and TPBC (TPBC-TICs) were significantly more effective at repopulating the mammary glands of naïve female BALB/c mice than CD24^-/CD44^-/ALDH1^-cells. Implantation of the TNBC-TICs resulted in significantly larger tumors, which metastasized to the lungs to a significantly greater extent than TNBC, TPBC-TICs, TPBC or parental 4T1 cells. We further demonstrated that the increased aggressiveness of TNBC-TICs correlates with the presence of high levels of mouse twenty-five kDa heat shock protein (Hsp25/mouse HspB1) and seventy-two kDa heat shock protein (Hsp72/HspA1A).</p> <p>Conclusions</p> <p>Taken together, we have developed a TNBC-TICs model system based on the 4T1 cells which is a very useful metastasis model with the advantage of being able to be transplanted into immune competent recipients. Our data demonstrates that the TNBC-TICs model system could be a useful tool for studies on the pathogenesis and therapeutic treatment for TNBC.</p

Genome fingerprinting of the silkworm, Bombyx mori, using random arbitrary primers

The random amplified polymorphic DNA (RAPD) technique was used to study DNA profiling of thirteen silkworm genotypes. The genotypes included six diapausing and seven nondiapausing varieties that represent a high degree of divergence with respect to geographic origin, and morphological, qualitative, quantitative and biochemical characters. Two hundred sixteen amplified products were generated using 40 random primers. Genotype-specific amplification products were identified. Amplification products specific to diapausing genotypes were also identified. Segregation of the RAPD marker was analyzed in a backcross population and found to be inherited as dominant Mendelian traits. Based on pairwise comparison of amplified products, the genetic similarity coefficient was calculated using the NeiLi similarity coefficient, and cluster analysis was performed by a hierarchical clustering technique. Silkworm genotypes were clustered into two groups, one consisting of six diapausing and the other of seven nondiapausing genotypes. The results of our study suggest that the RAPD technique could be used as a powerful tool to generate genetic markers that are linked to traits of interest in the silkworm

Awareness of gestational diabetes mellitus among pregnant women attending a tertiary health center

BACKGROUND AND OBJECTIVES: Awareness of gestational diabetes mellitus (GDM) among pregnant women is poor. However, increasing awareness may help in diagnosis and prevention of maternal and fetal complications. Hence, this study was aimed at evaluating the knowledge in diagnosed GDM pregnant women. METHODOLOGY: This cross-sectional study was conducted from October 2014 to March 2017. A total of 500 registered pregnant women diagnosed to have GDM residing within 5 km of the study area aged more than 18 years who were willing to participate in the study were enrolled in the study. The participants were provided with the questionnaire which was designed to assess their knowledge about GDM. RESULTS: The age of the women ranged from 22 to 44 years with mean age as 27.53 ± 2.42 years and median age as 27 years. The most common age group was 26–30 years which comprised 67.6% of the women. Maximum women had primary education (61%) and were Hindus (54.8%). Most of the women were working (54.8%), resided in slum areas (43.2%), and had body mass index (BMI) between 19.8 and 26 kg/m2 (67%). The mean BMI level was 28.07 ± 4.11 kg/m2. The mean blood sugar levels at diagnosis ranged between 88 and 300 mg/dL and the mean blood sugar level was 201.36 ± 38.67 mg/dL and the median blood sugar level was 190 mg/dL. Majority of the women, that is, 57.6% of the women, had an average knowledge about GDM while 21.8% of the women had good knowledge, 1.6% had excellent, and 19% had poor knowledge. The mean knowledge score was 6.51 ± 3.41. The mean percentage of the knowledge was 36.14% ± 18.94%. Statistically significant association was noted between knowledge about GDM with maternal age and educational status, religion, and occupation (P < 0.050), but the GDM knowledge was independent of that found between place of residence (P = 0.715) and family history of DM (P = 0.661). CONCLUSION: There is poor knowledge about GDM in the study area. Hence, there is need to create awareness of this condition through counseling and use of mass media

Prospective Study of Medication Adherence Pattern in Chronic Obstructive Pulmonary Disease and Asthma Patient&apos;s in Tertiary Care teaching Hospital

ABSTRACT Lack of patient compliance with prescribed regimen is an important fascinating problem in medical care, especially in patients suffering from chronic illnesses. Asthma and Chronic Obstructive Pulmonary Disease (COPD) are the two main chronic lung disorders which are particularly vulnerable for medication non-adherence problem. Aim of the study was to assess medication adherence by self-report method to understand various determinants of medication non-adherence, and to enhance adherence using the strategies of counseling, education, and interviewing the patient. The prospective study was conducted for period of six months, assessed medication and inhaler adherence by administering Morisky self-reported questionnaires and inhaler technique were assessed by the standard checklist. Also reported the reasons for non-adherence.The study shows there was statistically significant increase in medication (P&lt;0.001) and inhaler adherence score (P&lt;0.001) and inhaler technique score (P&lt;0.001) from baseline after pharmacist counseling regarding disease and medication. Comparison between various demographic factors like age, gender, diagnosis, duration of disease, and education status showed equal impact of pharmacist counseling on improvement of overall medication adherence score. Overall medication adherence and inhaler technique improvement was found to be statistically significant in asthma and COPD patients after counseling by pharmacist. The major reasons for medication non-compliance were felt better and stopped, high cost of medication, forgetfulness and lack of access to drug store/hospital