Impaired spermatogenesis associated with changes in spatial arrangement of Sertoli and spermatogonial cells following induced diabetes

The current study was conducted to assess the relationship between testicular cells in spermatogenesis, through which the production of healthy and mature sperm is essential. However, it seems necessary to obtain more information about the three-dimensional pattern of the testis cells arrangement, which is directly related to the function of the testis after induction of diabetes. Twelve adult mice (28-30 g) were assigned into two experimental groups: (1) control and (2) diabetic (40 mg/kg STZ). The epididymal sperm collected from the tail of the epididymis and testes samples were taken for stereology, immunocytochemistry and RNA extraction. Our data showed that diabetes could notably decrease the number of testicular cells, together with a reduction of total sperm count. In addition, the results from the second-order stereology indicated the significant changes in the spatial arrangement of Sertoli cells and spermatogonial cells in the diabetic groups, in comparison with the control (P < .05). Moreover, the immunohistochemistry results showed a significant reduction in Sex-determining Region Y (SRY) box 9 gene (SOX9), vimentin, occludin, and connexin-43 positive cells in the diabetic groups compared with the control (P < .05). Furthermore, our data showed that the expression of steroidogenic acute regulatory protein steroidogenic acute regulatory protein (StAR) and peripheral benzodiazepine receptor peripheral benzodiazepine receptor (PBR) was significantly reduced in the diabetic groups, in comparison with the control (P < .05). These findings suggest that structural and functional changes of testis cells after induction of diabetes cause the alterations in the spatial arrangement of Sertoli and spermatogonial cells, ultimately influencing the normal spermatogenesis in mice.     

Hepatocyte differentiation of human induced pluripotent stem cells is modulated by stearoyl‐CoA desaturase 1 activity

Stearoyl‐CoA desaturase 1 (SCD1) plays important roles in organ development, glucose tolerance, insulin sensitivity, and cancer. Here, we examined the role of SCD1 for the differentiation of human induced pluripotent stem (hiPS) cells to liver cells by using drug inhibition and biochemical experiments. hiPS cells cultured in a pro‐hepatic medium were exposed to an SCD1 inhibitor at various stages throughout differentiation. Liver‐specific markers, specifically α‐fetoprotein, albumin and urea in conditioned medium, and hepatocyte nuclear factor 4α (HNF4α) and cytochrome P450 7A1 (CYP7A1) gene expressions and triglyceride in cellular extracts were analyzed at various development stages. Measures of hepatocyte‐specific function and triglyceride accumulation in later stages were strongly inhibited a minimum of −29% (P < 0.05) by SCD1 inhibitor in the early stage of hepatic differentiation and effectively reversed (>30%, P < 0.01) by the addition of oleate. The results were also reproducible with human primary mononuclear cells (hPMN). SCD1 inhibitor had no significant effect on liver‐specific markers when it was added in the hepatic maturation stage. However, it strikingly led to higher albumin (1.6‐fold, P = 0.03) and urea (1.9‐fold, P = 0.02) production, and HNF4α (1.9‐fold, P = 0.02) and CYP7A1 (1.3‐fold, P = 0.03) expression upon incubation during the lineage‐commitment stage. Hepatic differentiation from cultured hiPS cells is sensitive to SCD1 inhibition and this sensitivity is affected by the stage of cellular differentiation. Notably, findings also indicate that this notion can be extended to hPMN. The requirement for SCD1 activity in functional differentiation of hepatocytes may have relevance for human liver disease and metabolic dysregulation.     

Is urine a reliable clinical sample for the diagnosis of human visceral leishmaniasis? A systematic review and meta-analysis

Visualization of amastigotes in lymph nodes, bone marrow, and other tissues samples remains the gold standard method for the diagnosis of visceral leishmaniasis (VL) in humans. This gold standard diagnostic method uses a technically challenging microscopy procedure that is often not accessible in many places in the world where VL is endemic. Here, we report the current systematic review and meta-analysis to evaluate whether urine is a reliable clinical sample for diagnosis of human VL. Data were extracted from ten available databases during the period from 2002 to 2017. Overall, 29 articles fulfilled the inclusion criteria and were used for data extraction in this systematic review. Most studies (72.4%) using urine specimens were reported from five countries: India 6 (20.7%), Iran 5 (17.2%), Bangladesh 4 (13.8%), Japan 3 (10.3%) and Spain 3 (10.3%), respectively. The most common diagnostic tests performed on urine were Katex (62.1%), ELISA (24.1%), and the rK39 (17.2%) assays. In meta-analysis the sensitivity and specificity of the three most commonly used diagnostic assays were rK39 (97%; CI: 91–99; 98%;76–100), ELISA (91%; 82–95; 99%; CI: 94–100), and Katex (83%; 73–90; 98%; 98–100), suggesting that the rK39 assay provided the highest sensitivity and the ELISA assay provided the highest specificity for diagnosis of VL from urine samples. Our findings suggest that urine is a valuable clinical sample for the diagnosis of human VL, particularly in areas where the gold standard test for VL is not available.     

Effects of rye and triticale seed proteins on Leptinotarsa decemlineata (Say) digestive α-amylase and protease and some biological parameters

The current study aimed to investigate the effect of rye and triticale seed proteins on the Leptinotarsa decemlineata, gut enzymes. Results showed that ammonium sulphate precipitation fractions; 0–30, 30–50, 50–70 and 70–100% had no inhibition on the fourth instar larval (L₄) protease activity, while first two fractions of triticale and all fractions of rye had inhibitory effects on the all larval stages and adult’s α-amylase activity. Mode of inhibition in rye and triticale was partial mixed and uncompetitive, respectively. Zymograms approved the results. Feeding assays were conducted using four cultivars of potato leaves treated with extracts. Weight of L₄ on Marx in both trials and the L₄ evolution in all cultivars in rye and just on Picasso in triticale were reduced, the developmental durations were increased on Marx and Picasso in triticale trial significantly. Also, rye extract caused inhibition in amylase activity of survived individuals that feed from treated Burren leaves.     

A cytogenetic study of couples with recurrent spontaneous abortions and infertile patients with recurrent IVF/ICSI failure

PURPOSE:

This study was conducted to determine the frequency and contribution of chromosomal abnormalities in miscarriages and in couples with recurrent in vitro fertilization/intra cytoplasmic sperm injection (IVF/ICSI) failure.

MATERIALS and METHODS:

A total of 221 individuals; 79 with three or more recurrent spontaneous abortions and 142 with at least three IVF/ICSI failures. Chromosomal analysis from peripheral blood lymphocytes was performed according to standard cytogenetic methods using G-banding technique.

RESULTS:

Abnormal karyotype was found in 21 (9.50%) individuals. Of these 21 subjects, 4 (19.04%) exhibited sex chromosomal abnormalities and 17 (80.96%) had autosomal abnormalities. Male partners had significantly higher chromosomal abnormalities (5.88%) than of females (3.61%). These abnormalities were also higher in patients with recurrent spontaneous abortions than with IVF/ICSI failure (P < 0.05).

CONCLUSIONS:

These data may be indicative that chromosomal abnormalities are involved more in spontaneous abortions than in recurrent IVF/ICSI failure. Cytogenetic analysis could be valuable for these couples when clinical data fail to clarify the cause.     

Titanium Dioxide Nanoparticles As Guardian against Environmental Carcinogen Benzo[alpha]Pyrene

Polycyclic aromatic hydrocarbons (PAH), like Benzo[alpha]Pyrene (BaP) are known to cause a number of toxic manifestations including lung cancer. As Titanium dioxide Nanoparticles (TiO₂ NPs) have recently been shown to adsorb a number of PAHs from soil and water, we investigated whether TiO₂ NPs could provide protection against the BaP induced toxicity in biological system. A549 cells when co-exposed with BaP (25 µM, 50 µM and 75 µM) along with 0.1 µg/ml,0.5 µg/ml and 1 µg/ml of TiO₂ NPs, showed significant reduction in the toxic effects of BaP, as measured by Micronucleus Assay, MTT Assay and ROS Assay. In order to explore the mechanism of protection by TiO₂ NP against BaP, we performed in silico studies. BaP and other PAHs are known to enter the cell via aromatic hydrocarbon receptor (AHR). TiO₂ NP showed a much higher docking score with AHR (12074) as compared to the docking score of BaP with AHR (4600). This indicates a preferential binding of TiO₂ NP with the AHR, in case if both the TiO₂ NP and BaP are present. Further, we have done the docking of BaP with the TiO₂ NP bound AHR-complex (score 4710), and observed that BaP showed strong adsorption on TiO₂ NP itself, and not at its original binding site (at AHR). TiO₂ NPs thereby prevent the entry of BaP in to the cell via AHR and hence protect cells against the deleterious effects induced by BaP.     

A quantitative study of detection mechanism of a label-free impedance biosensor using ultrananocrystalline diamond microelectrode array

It is well recognized that label-free biosensors are the only class of sensors that can rapidly detect antigens in real-time and provide remote environmental monitoring and point-of-care diagnosis that is low-cost, specific, and sensitive. Electrical impedance spectroscopy (EIS) based label-free biosensors have been used to detect a wide variety of antigens including bacteria, viruses, DNA, and proteins due to the simplicity of their detection technique. However, their commercial development has been hindered due to difficulty in interpreting the change in impedance upon antigen binding and poor signal reproducibility as a result of surface fouling and non-specific binding. In this study, we develop a circuit model to adequately describe the physical changes at bio functionalized surface and provide an understanding of the detection mechanism based on electron exchange between electrolyte and surface through pores surrounding antibody-antigen. The model was successfully applied to extract quantitative information about the bio surface at different stages of surface functionalization. Further, we demonstrate boron-doped ultrananocrystalline diamond (UNCD) microelectrode array (3 × 3 format, 200 μm diameter) improves signal reproducibility significantly and increases sensitivity by four orders of magnitude. This study marks the first demonstration of UNCD array based biosensor that can reliably detect a model Escherichia coli K12 bacterium using EIS, positioning this technology for rapid adoption in point-of-use applications.     

Identification and characterization of protein composition in Withania somnifera—an Indian ginseng

To have better understanding of the processes that occur in Withania somnifera L. Dunal, proteome analyses were initiated on two tissues (seeds & leaves) of this plant. Protein extracts were separated by two-dimensional gel electrophoresis (2-DE) across a broad 3.0?C10.0 immobilized pH gradient (IPG) strip that yielded 434 protein spots. A total of 167 individual spots (82 from seeds and 85 from leaves) were excised from the gel and were characterized by peptide mass fingerprinting. From these analyses, 70 individual proteins from seeds and 74 from leaves were identified by protein sequence database interrogation and were catalogued accordingly to different protein functions. A comparative analysis of the two tissues indicated that some enzymes/proteins involved in housekeeping pathways were common to both, whereas some were exclusively tissue specific with specialized metabolic complement. The knowledge gained by this study towards the tissue specific protein expression in W. somnifera would form the basis for our future endeavor of characterization of proteins to understand the physiology and the associated complex metabolic network during its ontogenetic development.