Dead End Homologue (DND1) Protein and Its Target mRNA is Influenced By Age and Estradiol in Normal and Systemic Lupus Erythematosus (SLE) T Cells

SLE is gender biased autoimmune disease (9:1 female to male). The immune system is a complex biological system that fights with foreign antigens including bacteria and viruses. The immune system works by producing antibodies against antigens. In SLE, the immune system produces antibodies against its own tissues. These antibodies are known as auto-antibodies. The female sex steroid, estradiol, alters T cell signaling pathways in SLE T cells and contributes to SLE onset and progression. Estradiol inappropriately regulates genes involved in T cell signaling pathways only in SLE T cells and not in control T cells. This suggest that the changes in the gene expression patterns in SLE T cells could lead to impaired peripheral tolerance in women and SLE onset. Peripheral tolerance is immunological tolerance developed after T and B cells mature and enter the periphery. Dead end homolog (Dnd1), an RNA-binding protein, inhibits miRNA by binding to target mRNAs. DND1 targets, such as p27 –Kip1, contribute to the maintenance of peripheral tolerance. Microarray analysis suggested estradiol suppresses DND1 expression in SLE T cells but no in normal T cells. The present study measured DND1 protein in freshly isolated normal T cells and tested the effect of estradiol (10-8M) on DND1 expression. T cells were purified from normal blood samples (n = 16) by Histopaque gradient and negative selection. Proteins were extracted and DND1 immunoprecipitated. The immunoprecipitates were size fractionated by SDS-PAGE and transferred to nitrocellulose membranes. The membranes were reacted with DND1 antibody. The protein amount was determined in triplicate using chemiluminesence. T cells from women \u3c35 years contained significantly higher amounts (p = 0.37, Pearson’s rho test, resulting R value was used to calculate significance p-value) of DND1 compared with T cells from donors \u3e35 years. DND1 was 63% higher in younger age group than older age donors. DND1 increased in response to estradiol with maximum expression (3.5-fold) at 3 h post estradiol stimulation. p27 expression was measured by the real-time (Step-one, Applied Bio systems) polymerase chain reaction (PCR). p27 expression was suppressed (p = 27, two tail T test was used to calculate the significance P value) in SLE T cells (n=12) compared with normal T cells (n=7). Together, the results confirm the microarray data indicating that estradiol does not suppress DND1 nor p27 mRNA expression in normal T cells. p27 has an established role in the development/maintenance of tolerance via complex regulation of CD4+ T cell proliferation and effector function. Future experiments will test the hypothesis that the inappropriate suppression of Dnd1 allows miRNA inhibition of p27 (Kip 1) and thereby alters CD4+ T cell effector subtypes in SLE. Changes in CD4+ T cells subtypes lead to a reduction in suppressor function and impaired peripheral tolerance. Also look at the aspect in detail, weather the estradiol effect is age dependent.https://digitalcommons.pittstate.edu/posters_2015/1007/thumbnail.jp

Molecular evidence for genetic distinctions between Chlamydiaceae detected in Siamese crocodiles (Crocodylus siamensis) and known Chlamydiaceae species

Chlamydiosis, caused by Chlamydiaceae, is a zoonotic disease found in humans and several species of animals, including reptiles and amphibians. Although chlamydiosis in saltwater crocodiles has been previously reported in South Africa and Papua New Guinea, the reported strains have not been identified or confirmed. Therefore, the main aim of this study was to sequence and characterize Chamydiaceae isolated from Siamese crocodiles. Results showed the 16S ribosomal (r) RNA and the 16S/23S rRNA gene of the crocodile isolates were closely related to the genus Chlamydophila with matched identity greater than 98%. The phylogenetic tree constructed from the 16S/23S rRNA gene showed the crocodile cluster diverges far from Cp. caviae with a 100% bootstrap value. The tree based on the ompA gene loci distinguished the crocodile strains into genotypes I, II, and III. The present study is the first report on Chlamydophila detected in Siamese crocodiles that is genetically distinct from the known species of Chlamydiaceae

Takotsubo Cardiomyopathy as a Sequela of Elective Direct-Current Cardioversion for Atrial Fibrillation

In takotsubo cardiomyopathy, the clinical appearance is that of an acute myocardial infarction in the absence of obstructive coronary artery disease, with apical ballooning of the left ventricle. The condition is usually precipitated by a stressful physical or psychological experience. The mechanism is unknown but is thought to be related to catecholamine excess. We present the case of a 67-year-old woman who experienced cardiogenic shock caused by takotsubo cardiomyopathy, immediately after undergoing elective direct-current cardioversion for atrial fibrillation. After a course complicated by left ventricular failure, cardiogenic shock, and ventricular tachycardia, she made a complete clinical and echocardiographic recovery In addition to this case, we discuss the possible direct effect of cardioversion in takotsubo cardiomyopathy

The occurrence of elephant endotheliotropic herpesvirus infection in wild and captive Asian elephants in Thailand: Investigation based on viral DNA and host antibody

Background and Aim: Elephant endotheliotropic herpesvirus (EEHV) is a serious disease, threatening the life of young elephants. Many elephants have been infected with no clinical signs and may serve as carriers spreading this disease. It is important to monitor the disease through clinical signs and molecular diagnosis. In this study we investigated the occurrence of EEHV and the efficiency of different techniques used to monitor EEHV infection in various samples and populations of Asian elephants. Materials and Methods: Blood and trunk swabs were collected from live elephants, while visceral organs (lung, digestive tract, spleen, lymph nodes, and kidney) were collected from dead elephants. EEHV was detected by polymerase chain reaction (PCR) in whole blood, trunk swabs, and visceral organs as samples, while elephant anti-EEHV immunoglobulin G (IgG) in serum was detected by enzyme-linked immunosorbent assay (ELISA). A total of 162 samples were analyzed in this study: 129 from healthy, 26 from dead, and 7 from sick elephants. Results: The present study showed that the overall incidence of EEHV was 40.1% (n=65/162). Approximately 46.2% (n=12/26) and 85.7% (n=6/7) of dead and sick elephants were positive for EEHV by PCR, respectively. All sick elephants that were young and affected by EEHV clinical disease tested negative for the IgG antibody ELISA, suggesting primary EEHV infection in this group. In addition, 2.3% (n=3/129) of subclinical infections were detected using PCR, and trunk swab samples showed slightly higher sensitivity (5.3%, n=2/38) to detect EEHV than whole blood (1.2%, n=1/84). As many as, 48.4% (n=44/91) of healthy elephants were EEHV seropositive (ELISA-positive), suggesting that many elephants in Thailand had previously been infected. Overall, 30% of dead wild elephants had been infected with EEHV (n=3/10). Moreover, statistical analysis revealed no significant differences in the EEHV detection rate between different age groups or sexes (p>0.05). Conclusion: PCR is better than ELISA to detect EEHV active infection in dead/sick elephants and to monitor EEHV in young elephants. ELISA is suitable for detecting previous EEHV infection and carriers, particularly adults. Theoretically, we could use both PCR and ELISA to increase the sensitivity of testing, along with observing abnormal behavior to efficiently monitor this disease. Identification of EEHV carriers within elephant populations is important to prevent transmission to healthy individuals, especially young elephants with high mortality from EEHV. This is the first report from Thailand regarding EEHV infection in wild elephants, showing the importance of preventing disease transmission between captive and wild elephants