Thoracoscopic repair of congenital diaphragmatic hernia in neonates: Tips and tricks learned from an institutional experience.

Background: Congenital diaphragmatic hernia (CDH) is an anomaly with significant morbidity in neonates. It has been traditionally managed by an open approach with a recent trend toward a minimally invasive approach. Aim: This is a retrospective study of our institutional experience with neonatal thoracoscopic management of CDH, with the impact of few technical nuances. Patients and Methods: The data was collected on neonatal thoracoscopic CDH repair between January 2015 and December 2018, in terms of the demographics, intra-operative parameters, post-operative status, recurrence, and mortality. While analyzing data, we found few technical modifications adopted by the surgeon such as trimming the margin of the defect, use of prosthetic mesh overlay reinforcement for repairs under tension, and to prefer extra-corporeal knotting along with higher placement of trocar, temporary increase in CO2, maximal use of muscle relaxant, extra-corporeal corner hitch stitch at some point, and continuation for further cases. An internal comparison was made to analyze the technical modifications influencing the outcomes, by dividing them into two groups, those with (group A) and without modifications (group B). The data was analyzed using SPSS software (IBM, Version 23). A P value of <0.05 was considered statistically significant. Results: Out of 45 newborns 64.4% were males with an average birth weight of 2.6 kg. Baseline variables were comparable between the groups. The operating time significantly reduced after a higher-level camera port was used (P-value: 0.0001). The mean follow-up was 30.8 months. There were totally seven recurrences (6 in group A and 1 in group B), all within 12 months. Seven parents gave the overall post-treatment feedback as "unsatisfied". The operating time, recurrence rate, and parental satisfaction feedback were significantly less in group B (P-value: 0.001). Conclusion: We recommend trimming the margin of the defect, use of prosthetic mesh overlay reinforcement for repairs under tension, and to prefer extra-corporeal knotting along with higher placement of trocar, temporary increase in CO2, maximal use of muscle relaxant, extra-corporeal corner hitch stitch to reduce the operating time, and recurrence after thoracoscopic CDH repair.

Pharmacological concentrations of recombinant factor VIIa restore hemostasis independent of tissue factor in antibody-induced hemophilia mice.

UNLABELLED: ESSENTIALS: The role of tissue factor (TF) in recombinant factor VIIa (rFVIIa) therapy in hemophilia is unclear. An acquired mouse hemophilia model with very low or normal levels of human TF was used in the study. rFVIIa is equally effective in correcting the bleeding in mice expressing low or normal levels of TF. Pharmacological doses of rFVIIa restore hemostasis in hemophilia independent of TF. BACKGROUND: Recombinant factor VIIa (rFVIIa) has been used widely for treating hemophilia patients with inhibitory autoantibodies against factor VIII or IX. Its mechanism of action is not entirely known. A majority of in vitro studies suggested that pharmacological concentrations of rFVIIa restore hemostasis in hemophilia in a phospholipid-dependent manner, independent of tissue factor (TF). However, a few studies suggested that a TF-dependent mechanism has a primary role in correction of bleeding by rFVIIa in hemophilia patients. Here, we investigated the potential contribution of TF in rFVIIa-induced hemostasis in hemophilia employing a model system of FVIII antibody-induced hemophilia in TF transgenic mice. METHODS: Mice expressing low levels of human TF (LTF mice), mice expressing relatively high levels of human TF (HTF mice) and wild-type mice (WT mice) had neutralizing anti-FVIII antibodies administered in order to induce hemophilia in these mice. The mice were then treated with varying concentrations of rFVIIa. rFVIIa-induced hemostasis was evaluated with the saphenous vein bleeding model. RESULTS: Administration of FVIII inhibitory antibodies induced the hemophilic bleeding phenotype in all three genotypes. rFVIIa administration rescued the bleeding phenotype in all three genotypes. No significant differences were observed in rFVIIa-induced correction of bleeding between LTF and HTF mice that had FVIII antibodies administered. CONCLUSIONS: Our results provide strong evidence supporting the suggestion that the hemostatic effect of pharmacological doses of rFVIIa stems from a TF-independent mechanism.

Factor VIIa binding to endothelial cell protein C receptor protects vascular barrier integrity in vivo.

BACKGROUND: Recent studies have shown that factor VIIa binds to endothelial cell protein C receptor(EPCR), a cellular receptor for protein C and activated protein C. At present, the physiologic significance of FVIIa interaction with EPCR in vivo remains unclear. OBJECTIVE: To investigate whether exogenously administered FVIIa, by binding to EPCR, induces a barrier protective effect in vivo. METHODS: Lipopolysaccharide(LPS)-induced vascular leakage in the lung and kidney,and vascular endothelial growth factor (VEGF)-induced vascular leakage in the skin, were used to evaluate the FVIIa-induced barrier protective effect. Wild-type, EPCR-deficient, EPCR-overexpressing and hemophilia A mice were used in the studies. RESULTS: Administration ofFVIIa reduced LPS-induced vascular leakage in the lung and kidney; the FVIIa-induced barrier protective effect was attenuated in EPCR-deficient mice. The extent of VEGF-induced vascular leakage in the skin was highly dependent on EPCR expression levels. Therapeutic concentrations of FVIIa attenuated VEGF-induced vascular leakage in control mice but not in EPCR-deficient mice.Blockade of FVIIa binding to EPCR with a blocking mAb completely attenuated the FVIIa-induced barrier protective effect. Similarly, administration of protease activated receptor 1 antagonist blocked the FVIIa induced barrier protective effect. Hemophilic mice showed increased vascular permeability, and administration of therapeutic concentrations of FVIIa improved barrier integrity in these mice. CONCLUSIONS: This is the first study to demonstrate that FVIIa binding to EPCR leads to a barrier protective effect in vivo. This finding may have clinical relevance, as it indicates additional advantages of using FVIIa in treating hemophilic patients.

A preliminary sketch of horn cancer transcriptome in Indian zebu cattle.

Horn cancer, a type of squamous cell carcinoma, in zebu cattle is an expensive affair in Indian agriculture sector, which accounts for 83.34% of total tumors found. In general, cancer tissue confirms considerably different expression patterns when compared to a normal stage. This includes not only up/down regulation, but also, the aberrant gene expression, the presence of different non-coding RNAs (ncRNAs), pseudogenes expression and genes involved in unusual pathways. We employed Roche 454 next generation sequencing platform to sequence Bos indicus cancerous and normal horn tissue transcripts. This resulted into a total of 909,345 high-confidence deep sequencing reads and detected a range of unusual transcriptional events including tumor associated genes. We also validated expression of two of the four tested genes in five other similar tissue samples by RT-qPCR. Further, seven cancer specific non-coding transcripts were accessed and a few of them have been suggested as cancer specific markers. This study for the first time provides primary transcriptome sketch of Bos indicus horn cancer tissue, and also demonstrates the suitability of the 454 sequencer for transcriptome analysis, which supports the concept of varied gene expression in cancerous condition.

Decreased calcineurin activity in circulation of Duchenne muscular dystrophy.

OBJECTIVES: In an analysis of enzymes in easily accessible tissues like blood cells, serum can provide a valuable information and a simple tool for disease and carrier detection. In the study presented we have analyzed calcineurin activity in Duchenne muscular dystrophy (DMD) and carrier sera and lymphocytes for its diagnostic value and its status in DMD pathology. DESIGN AND METHODS: We have monitored calcineurin activity in sera and lymphocytes of DMD, in carriers and in controls using colorimetric method by following the p-nitrophenol released in the presence and absence of Trifluoperazine (TFP), an inhibitor of calcineurin. RESULTS: Results showed a significant decrease in serum and lymphocyte calcineurin activity in DMD (p<0.001) without alteration in carriers compared to normal. CONCLUSION: Further studies are required to understand possible alterations mediated by calcineurin with reference to DMD lymphocytes as any alteration in phosphorylation/dephosphorylation pathway can disturb the normal functioning of these cells. The decreased calcineurin activity observed in DMD serum compared with controls could be further examined for its diagnostic utility.

Altered expression, intracellular distribution and activity of lymphocyte calpain II in Duchenne muscular dystrophy.

BACKGROUND: Calpain II is an calcium-dependent cysteine protease involved in essential regulatory or processing functions of the cell, mediated by physiological concentrations of Ca(2+). However, in an environment of abnormal intracellular calcium as in Duchenne muscular dystrophy (DMD), calpain is suggested to cause membrane alterations. METHODS: Twelve individuals with dystrophin gene deletion and an equal number of age and sex matched controls were chosen for the study. The expression pattern of calpain II (both at RNA and protein levels), its cellular location upon activation and its activity in lymphocytes were specifically assessed to know if our earlier report of increased calpain activity in DMD lymphocytes is a result of de novo synthesis or is due to basic defect in calcium handling. RESULTS: We found a significant increase in the expression, alteration in calpain II distribution and increased activity of this enzyme. CONCLUSION: Membrane abnormalities and altered signaling pathways observed in DMD lymphocytes may be due to increased association of calpain II onto membrane and cytosol.

Induction of cancer-specific cytotoxicity towards human prostate and skin cells using quercetin and ultrasound.

Bioflavonoids, such as quercetin, have recently emerged as a new class of chemotherapeutic drugs for the treatment of various cancer types, but are marred by their low potency and poor selectivity. We report that a short application of low-frequency ultrasound selectively sensitises prostate and skin cancer cells against quercetin. Pretreatment of cells with ultrasound (20 kHz, 2 W cm(-2), 60 s) selectively induced cytotoxicity in skin and prostate cancer cells, while having minimal effect on corresponding normal cell lines. About 90% of the viable skin cancer cell population was lost within 48 h after ultrasound-quercetin (50 microM) treatment. Ultrasound reduced the LC50 of quercetin for skin cancer cells by almost 80-fold, while showing no effect on LC50 for nonmalignant skin cells.

Default Among Patients with Gonorrhoea-the Role of Health Education.

Factors relating to default among 100 health educated and 63 uneducated patients with gonorrhoea were studied. Default was not significantly associated with age, marital status or level of education. Repeaters were not particularly more prone to default than non-repeaters. Health education did not significantly influence default. Default was significantly low (p.001) among a small group of patients with concomitant diseases detected at the initial or follow - up visits. A high incidence of trichomoniasis (14.5%) in the present study detected in the follow-up period emphasizes the need for thorough clinical and laboratory examination at each follow up visit.