A study on surgical management of undescended testis

Background: Cryptorchidism which is synonymous with Undescended testes. It is one of the most common congenital anomalies found in children. The aim is to study the surgical management of undescended testis in relation to age, position, side, symptoms, complications and management of undescended testis. This study will present an overview of the current understanding of this challenging problem, mainly concentrating on surgical management.Methods: About 30 cases of undescended testis were included in the study, which were diagnosed with regards to age of presentation, clinical features, inclusion and exclusion criteria and relevant investigations like ultrasonography, diagnostic laparoscopy, CT whereever required. Cases were managed accordingly in the course of study and sampling of cases was taken from Bapuji Hospital and C.G. Hospital, Davangere during the study period.Results: Undescended testis was found more common on right side and most of them presented with the age group of 1-15 years with absence of testis in the scrotum being the most common complaint and hernia was found to be most commonly associated with undescended testis. Most common position of undescended testis was found to be intra-canalicular with most of them being palpable. The only one case had torsion and few of the cases had post operative complications like, wound infection haematoma. Open and laparoscopic orchidopexy can be used as most common procedure in early age group.Conclusions: Undescended testes is eminently treatable in today’s practice with the surgical procedures, an early diagnosis, informed counselling of parents and optimal surgical management offer optimised opportunities for successful placement and function of the testes which lessen the burden of complications

Synthesis, spectral characterization and some biological studies of transition metal complexes with Schiff base ligand containing N,O and S as donor atoms

Transition metal complexes of Cu(II), Co(II), Ni(II), Zn(II), Cd(II) and Mn(II) have been synthesized with the Schiff base ligand 5-Sub-N-(2-mercaptophenyl)salicylideneimine. Elemental analysis of these complexes suggest that these metal ions forms complexes of type ML(H2O)stoichiometry for Cu(II), Co(II), Ni(II), Zn(II), Cd(II) and Mn(II). The ligand behaves as tridentate and forms coordinate bonds through O, S and N atoms. Magnetic susceptibility, IR, UV – Visible, Mass and ESR spectral studies suggest that Cu(II), Ni(II) complexes posses square planar geometry, whereas Co(II), Zn(II), Cd(II) and Mn(II) complexes posses tetrahedral geometry. The complexes were tested for their antimicrobial activity against the bacterial strains Staphylococcus aureus and Bacillus subtilis.The Schiff base metal complexes evaluated for their antifungal activity against the fungi A. niger and C. oxysporum. The DNA cleavage studies of Schiff base complexes werestudied using Calf – Thymus DNA by agarose gel electrophoresis method

Thermoanalytical and spectral characterization of cadmium(II) complexes of propantheline bromide

Synthesis, structure and properties of Cd(II) complexes of propantheline bromide (PPB) have been studied. The isolated complexes are characterised by various physico-chemical methods. Based on the data, tetrahedral structure has been suggested for the new metal complexes. Thermogravimetric studies of the complexes have Seen performed in order to establish the mode of their thermal stability. The thermal degradation process was found to proceed in two steps. Kinetic and thermodynamic parameters are evaluated on the basis of thermal degradation data. The values of activation energies are found to be in the range 8.5-33.5 kJ mol(-1)

Integrated Genomic Analysis of Diverse Induced Pluripotent Stem Cells from the Progenitor Cell Biology Consor tium

The rigorous characterization of distinct induced pluripotent stem cells (iPSC) derived from multiple reprogramming technologies, somatic sources, and donors is required to understand potential sources of variability and downstream potential. To achieve this goal, the Progenitor Cell Biology Consortium performed comprehensive experimental and genomic analyses of 58 iPSC from ten laboratories generated using a variety of reprogramming genes, vectors, and cells. Associated global molecular characterization studies identified functionally informative correlations in gene expression, DNA methylation, and/or copy-number variation among key developmental and oncogenic regulators as a result of donor, sex, line stability, reprogramming technology, and cell of origin. Furthermore, X-chromosome inactivation in PSC produced highly correlated differences in teratoma-lineage staining and regulator expression upon differentiation. All experimental results, and raw, processed, and metadata from these analyses, including powerful tools, are interactively accessible from a new online portal at https://www.synapse.org to serve as a reusable resource for the stem cell community

Lack of Guanylate Cyclase C results in increased mortality in mice following liver injury

<p>Abstract</p> <p>Background</p> <p>Guanylate Cyclase C (GC-C) expression in the intestine plays a role in the regulation of fluid and ion transport, as well as epithelial cell apoptosis and proliferation. In the adult rat liver, GC-C expression is increased in response to injury. We hypothesized that GC-C is required for repair/recovery from liver injury.</p> <p>Methods</p> <p>We subjected wild type (WT) and GC-C deficient mice to acute liver injury with a single injection of the hepatotoxin carbon tetrachloride. Changes in the level of expression of GC-C and its ligands uroguanylin and guanylin were quantified by real-time PCR. Liver morphology, and hepatocyte necrosis, apoptosis and proliferation, were examined at 1-3 days post-injury in mice on a mixed genetic background. Survival was followed for 14 days after carbon tetrachloride injection in wild type and GC-C deficient mice on both a mixed genetic background and on an inbred C57BL6/J background.</p> <p>Results</p> <p>GC-C deficient mice on the mixed genetic background nearly all died (median survival of 5 days) following carbon tetrachloride injection while WT littermates experienced only 35% mortality. Elevated levels of TUNEL-positive hepatocyte death on post-injury day 1, increased apoptosis on day 2, and increased areas of centrilobular necrosis on days 2 and 3, were evident in livers from GC-C null mice compared to WT. Collectively these data suggest increased hepatocyte death in the GC-C null mice in the early time period after injury. This corresponds temporally with increased expression of GC-C and its ligands guanylin and uroguanylin in post-injury WT mouse liver. The hepatocyte proliferative response to injury was the same in both genotypes. In contrast, there was no difference in survival between GC-C null and WT mice on the inbred C57BL/6 J background in response to acute liver injury.</p> <p>Conclusions</p> <p>Signalling via GC-C promotes hepatocyte survival <it>in vivo </it>and is required for effective recovery from acute toxic injury to the liver in a strain-specific manner.</p

Lampe1: An ENU-Germline Mutation Causing Spontaneous Hepatosteatosis Identified through Targeted Exon-Enrichment and Next-Generation Sequencing

Using a small scale ENU mutagenesis approach we identified a recessive germline mutant, designated Lampe1 that exhibited growth retardation and spontaneous hepatosteatosis. Low resolution mapping based on 20 intercrossed Lampe1 mice revealed linkage to a ∼14 Mb interval on the distal site of chromosome 11 containing a total of 285 genes. Exons and 50 bp flanking sequences within the critical region were enriched with sequence capture microarrays and subsequently analyzed by next-generation sequencing. Using this approach 98.1 percent of the targeted DNA was covered with a depth of 10 or more reads per nucleotide and 3 homozygote mutations were identified. Two mutations represented intronic nucleotide changes whereas one mutation affected a splice donor site in intron 11–12 of Palmitoyl Acetyl-coenzyme A oxygenase-1 (Acox1), causing skipping of exon 12. Phenotyping of Acox1Lampe1 mutants revealed a progression from hepatosteatosis to steatohepatitis, and ultimately hepatocellular carcinoma. The current approach provides a highly efficient and affordable method to identify causative mutations induced by ENU mutagenesis and animal models relevant to human pathology

Long-term correction of diabetes in rats after lentiviral hepatic insulin gene therapy

Aims/hypothesis: Type 1 diabetes results from the autoimmune destruction of pancreatic beta cells. Exogenous insulin therapy cannot achieve precise physiological control of blood glucose concentrations, and debilitating complications develop. Lentiviral vectors are promising tools for liver-directed gene therapy. However, to date, transduction rates in vivo remain low in hepatocytes, without the induction of cell cycling. We investigated long-term transgene expression in quiescent hepatocytes in vitro and determined whether the lentiviral delivery of furin-cleavable insulin to the liver could reverse diabetes in rats. Materials and methods: To improve transduction efficiency in vitro, we optimised hepatocyte isolation and maintenance protocols and, using an improved surgical delivery method, delivered furin-cleavable insulin alone or empty vector to the livers of streptozotocin-induced diabetic rats by means of a lentiviral vector. Rats were monitored for changes in body weight and blood glucose, and intravenous glucose tolerance tests were performed. Expression of insulin was determined by RT-PCR, immunohistochemistry and electron microscopy. Results: We achieved long-term transgene expression in quiescent hepatocytes in vitro (87 ± 1.2% transduction efficiency), with up to 60 ± 3.2% transduction in vivo. We normalised blood glucose for 500 days-a significantly longer period than previously reported-making this the first successful study using a lentiviral vector. This procedure resulted in the expression of genes encoding several beta cell transcription factors, some pancreatic endocrine transdifferentiation, hepatic insulin storage in granules, and restoration of glucose tolerance. Liver function tests remained normal. Importantly, pancreatic exocrine transdifferentiation did not occur. Conclusions/interpretation: Our data suggest that this regimen may ultimately be employed for the treatment of type 1 diabetes

Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function

Porcine Reproductive and Respiratory Syndrome (PRRS) is a panzootic infectious disease of pigs, causing major economic losses to the world-wide pig industry. PRRS manifests differently in pigs of all ages but primarily causes late-term abortions and stillbirths in sows and respiratory disease in piglets. The causative agent of the disease is the positive-strand RNA PRRS virus (PRRSV). PRRSV has a narrow host cell tropism, limited to cells of the monocyte/macrophage lineage. CD163 has been described as a fusion receptor for PRRSV, whereby the scavenger receptor cysteine-rich domain 5 (SRCR5) region was shown to be an interaction site for the virus in vitro. CD163 is expressed at high levels on the surface of macrophages, particularly in the respiratory system. Here we describe the application of CRISPR/Cas9 to pig zygotes, resulting in the generation of pigs with a deletion of Exon 7 of the CD163 gene, encoding SRCR5. Deletion of SRCR5 showed no adverse effects in pigs maintained under standard husbandry conditions with normal growth rates and complete blood counts observed. Pulmonary alveolar macrophages (PAMs) and peripheral blood monocytes (PBMCs) were isolated from the animals and assessed in vitro. Both PAMs and macrophages obtained from PBMCs by CSF1 stimulation (PMMs) show the characteristic differentiation and cell surface marker expression of macrophages of the respective origin. Expression and correct folding of the SRCR5 deletion CD163 on the surface of macrophages and biological activity of the protein as hemoglobin-haptoglobin scavenger was confirmed. Challenge of both PAMs and PMMs with PRRSV genotype 1, subtypes 1, 2, and 3 and PMMs with PRRSV genotype 2 showed complete resistance to viral infections assessed by replication. Confocal microscopy revealed the absence of replication structures in the SRCR5 CD163 deletion macrophages, indicating an inhibition of infection prior to gene expression, i.e. at entry/fusion or unpacking stages

Synthesis, spectral, thermal studies and antimicrobial activity of cobalt(II) complexes of propantheline bromide

The synthesis, spectral characterization, magnetic, susceptibility and molar conductance studies of new series of cobalt(II)complexes of propantheline bromide (PPB) with general stoichiometry Co(PPB)(2)(H2O)(2)X(2)] are reported, where X = Cl-, Br-, ClO4- and CH3COO-. In all the complexes PPB behaves as a unidentate ligand. Thermal studies (TG/DTA) have been used to evaluate kinetic parameters such as order of the thermal reaction (n) and activation energy (E(a)). These new complexes were found to be more potent as antimicrobial agents than the free ligand