Ultra-structural and histopathological features of liver biopsy taken during laparotomy to confirm the diagnosis of biliary atresia

Background: Neonatal cholestasis is caused by a group of diseases that cause jaundice, which can be encountered in the neonatal period. Biliary atresia (BA) and idiopathic neonatal hepatitis (INH) are among neonatal cholestasis diseases. Aims: The aim of this study was to perform histopathological and ultra-structural examinations of liver biopsy tissue samples from BA and INH patients with liver biopsies taken during laparotomy to confirm the diagnosis of biliary atresia. Settings and Design: A total of patients undergoing Kasai surgery before the age of 60 days were included in an “early” group (n = 7), whereas patients undergoing surgery after the age of 60 days were included in a “late” group (n = 11). The control group (n = 11) included INH patients. Materials and Methods: For histopathological examinations, liver tissue samples obtained intra-operatively were subjected to routine histopathological procedures after being stained with caspase-3 and cytokeratin-7 antibodies. Ultra-structural evaluations were also performed. Statistical analysis used: For comparisons between the groups, a one-way analysis of variance (ANOVA) test and the Mann–Whitney U test were used for continuous variables. Results: Histopathological findings reflected the specific liver pathologic findings seen in biliary atresia. Although there was no significant difference between the BA groups, these parameters were not detected in the control group. The histopathological evaluations revealed no significant differences in the findings of liver parenchyma damage between the early, late, and control groups. Electron microscopic examinations showed that the patients in the late group had more severe signs of intra-cellular damage to the liver. Conclusions: Although the histopathological examination revealed no significant differences in liver damage between the three groups, in ultra-structural evaluation, intra-cellular damage was found to be less in groups with better prognosis. Electron microscopy evaluations of intra-cellular damage may be more useful in this respect.

Is Laparoscopy for Non-palpable Testes a Well-known Entity?

ABSTRACT Objective: Laparoscopy has been accepted as the best diagnostic tool and suggested as the treatment of choice for non-palpable testes cases. However, its use in unilateral non-palpable testis cases has been previously debated. Methods: The clinical records of the non-palpable testis cases that were managed with laparoscopy between January 2011 and December 2013 were retrospectively reviewed. Results: Laparoscopy was performed in 29 non-palpable testis cases. The cases were divided into three groups according to the laparoscopic findings. Orchiopexy was performed in cases with viable testes, and the internal inguinal ring was left open in these cases. Conclusion: Laparoscopy provided definitive diagnosis and was helpful in the treatment of unilateral, non-palpable testis cases. Leaving the internal inguinal ring open did not result in subsequent indirect inguinal hernia in our cases.

A Unique Case: Arteriovenous Malformation of the Urinary Bladder Wall

ABSTRACT Arteriovenous malformation (AVM) of urinary bladder is a very rare condition in which a section of blood vessels lacks capillary vessels resulting in blood from an artery being delivered directly to a vein. We report a rare case of AVM of the bladder wall mimicking a bladder tumour presenting with acute abdomen.

A recombinant plasmid of composite cysteine proteinase inhibitor/ glyceraldehyde‑3‑phosphate dehydrogenase gene of periodic Brugia malayi functions on DNA immunity in the host.

Objectives: Both cysteine proteinase inhibitors (CPIs) and glyceraldehyde‑3‑phosphate dehydrogenase (GAPDH) play important roles in the pathogenesis of parasites and their relationship with the hosts. We constructed a new eukaryotic recombinant expression plasmid pcDNA3.1(+)‑BmCPI/BmGAPDH of periodic Brugia malayi for investigation of the DNA vaccine‑elicited immune responses. Materials and Methods: We cloned a gene encoding the CPIs and GAPDH from periodic B. malayi into vector pcDNA3.1. The composited plasmid or the control was injected into the tibialis anterior muscle of the hind leg in BALB/c mice, respectively. The target genes were detected by reverse transcription‑polymerase chain reaction in muscle tissues. The stimulation index (SI) of T‑lymphocyte proliferation and the levels of interferon‑gamma (INF‑γ) and interleukin‑4 (IL‑4) in serum were detected by thiazolyl blue tetrazolium blue and enzyme‑linked immunosorbent assays. Results: The pcDNA3.1(+)‑BmCPI/ BmGAPDH was amplified from muscle tissues of the mice after immunisation. The SI of the immunised group was significantly higher than that of the two control groups (P < 0.05). The levels of INF‑γ and IL‑4 of pcDNA3.1(+)‑BmCPI/BmGAPDH group were both higher than those of the two control groups (P < 0.05). The level of INF‑γ of pcDNA3.1(+)‑BmCPI/BmGAPDH group was significantly higher than that of pcDNA3.1(+)‑BmCPI/CpG group (P < 0.05). Conclusions: We conclude that the recombinant plasmid pcDNA3.1(+)‑BmCPI/BmGAPDH could elicit specific humoural and cellular immune responses in mice.

Construction of a recombinant plasmid harbouring the glyceraldenyde-3-phosphate dehydrogenase gene of periodic Brugia malayi and observation on DNA immunity.

Purpose: Controlling and eliminating lymphatic filariasis will require further research of preventative measures and implementation. Parasite is dependent on glycolysis for ATP production. The glycolytic enzyme glyceraldenyde-3-phosphate dehydrogenase (GAPDH) plays an important role in glycolysis and therefore is either a potential target for anti-parasite drug development or a vaccine candidate. Therefore, we tried to investigate the DNA vaccine-elicited immune responses in BALB/c mice. Materials and Methods: We cloned a gene encoding the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from periodic Brugia malayi into vector pcDNA3.1. Mice were injected at a dosage of 100 μg recombinant plasmid DNA with CpG intramuscular injection and immunized three times at 2-week intervals. pcDNA3.1 and normal saline were used as control. The tissue of muscles at the 4 weeks after the third injection was collected and target genes were detected using RT-PCR. The humoral responses elicited in mice by inoculation with the recombinant plasmid pcDNA3.1-BmGAPDH were detected using a standard ELISA. Two weeks after the third immunization, stimulation index (SI) was measured using the MTT method and the level of secreted IL-4 and INF-g were detected using ELISA. Results: Specific gene fragment coding GAPDH was amplified and the recombinant plasmid pcDNA3.1-BmGAPDH was constructed. Post-challenge sera from the mice immunized with the DNA vaccine had specific antibody titres of 1:1600 to 1:6400, and the highest titre was observed in the mice that were inoculated by pcDNA3.1-BmGAPDH/CpG at 6 weeks. At 4 weeks after immunization, the spleens of the mice were obviously enlarged. The proliferation of spleen T lymphocytes seen on the MTT assay was higher in the pcDNA3.1-BmGAPDH group than in the control group (P value <0.05). The levels of IL-4 and INF-g in serums from the immunized mice were significantly higher than that of the control (P value <0.05). Conclusions: We conclude that the recombinant eukaryotic plasmid pcDNA3.1-BmGAPDH could elicit humoral and cellular immune responses in mice.

NutriSim[copyright sign] protects-against apoptosis in hippocampus induced by transient ischemia in mongolian gerbils [meriones unguiculatus]

Mongolian gerbil model has been extensively used for the study of neuroprotective drugs since transient bilateral common carotid artery occlusion induces neuronal cell death to selectively vulnerable regions, including the CA1 sector of the hippocampus. Previously we have shown that NutriSim[copyright sign], a nutritive supplement used empirically in the treatment of several degenerative disorders protects against brain damage induced by ischemia-reperfusion in Mongolian gerbils. These effects are partly attributed to its antioxidant action. The purpose of this study was to further investigate the potential neuroprotective effects of NutriSim[copyright sign] with histological measures of global ischemia in gerbils. We found that a single dose of NutriSim[copyright sign] was able to prevent significantly the ischemia-induced pyramidal cell loss as well as the number of hyperchromatic cells and glial cells after a week of treatment. In consonance with these data, increased TUNEL positive cells after ischemia is reduced in NutriSim[copyright sign] treated animal after a week

Microsatellites in Portuguese autochthonous horse breeds: usefulness for parentage testing

The use of DNA technology for parentage testing is increasing every day. Most laboratories have improved their exclusion probabilities (PE) by the addition of DNA microsatellite loci to standard blood-typing results. The efficacy of each locus depends on the number of alleles detected and their frequencies in the breeds tested. Here we analyzed the usefulness of six microsatellites for routine parentage testing in three Portuguese autochthonous horse breeds: Lusitano, Sorraia and Garrano. The DNA loci analyzed - ASB2, HMS3, HMS7, HTG4, HTG10, and VHL20 - were chosen based upon the polymorphism detected in other breeds with Iberian horse influence. The estimated probability of exclusion of wrongly named parents (PE) was high, with values ranging from 88.5 per cent to 99.6 per cent

Hybridization between Brugia malayi and Brugia pahangi from south Kalimantan, Indonesia.

Hybridization experiments were carried out between sub-periodic Brugia malayi and B. pahangi recovered from cats originating from the same area in South Kalimantan (Borneo), Indonesia. Five to six week-old immature males and females of both species from Mongolian gerbil (Meriones unguiculatus) intraperitoneal infections were inoculated into the peritoneal cavity of other gerbils. Mating occurred between the species and microfilariae developed into infective third-stage larvae in mosquitoes. Larvae from the mosquitoes developed into adults in the peritoneal cavity of gerbils but microfilariae were not produced since the males were sterile. Back-crossing between hybrids and males and females of the parental species were also done and female hybrids produced microfilariae when crossed with parental males but females from the parental species did not produce microfilariae when mated with the hybrid males. This confirmed that the males were sterile but the females were not. If these species are hybridized in nature and humans are exposed to hybrid third-stage larvae, could this lead to occult filarial infection? These results confirmed previous hybridization experiments of these species orginating from other geographic areas.

Susceptibility of Oncomelania hupensis subspecies to infection with geographic strains of Schistosoma japonicum.

Subspecies of Oncomelania hupensis were exposed to infection with zoophilic and anthropophilic strains of Schistosoma japonicum. The snails were Philippine O. h. quadrasi from the islands of Bohol, Leyte and Mindanao, O. h. formosana from Changhua and Ilan, and O. h. chiui from Taiwan, O. h. hupensis from China, O. h. nosophora from Japan and O. h. lindoensis from Indonesia. Anthropophilic strains of S. japonicum were from Bohol and Leyte in the Philippines, China, Japan and Indonesia. The zoophilic strains were from Changhua and Ilan, Taiwan. All geographic strains of the parasite developed exceptionally well and produced cercariae in 32-50% of O. h. chiui and 5-43% of the natural snail hosts. Development in other subspecies of snails with geographic strains of the parasite were variable. Only a few O. h. formosana (Changhua) could be infected with the parasites from other geographic areas and only one O. h. quadrasi each from Mindanao became infected with the Bohol and Changhua strains of S. japonicum.