Membrane-bound ß-catenin degradation is enhanced by ETS2-mediated Siah1 induction in Helicobacter pylori-infected gastric cancer cells.

ß-catenin has two different cellular functions: intercellular adhesion and transcriptional activity. The E3 ubiquitin ligase Siah1 causes ubiquitin-mediated degradation of the cytosolic ß-catenin and therefore, impairs nuclear translocation and oncogenic function of ß-catenin. However, the effect of Siah1 on the cell membrane bound ß-catenin has not been studied. In this study, we identified that the carcinogenic bacterium H. pylori increased ETS2 transcription factor-mediated Siah1 protein expression in gastric cancer cells (GCCs) MKN45, AGS and Kato III. Siah1 protein level was also noticeably higher in gastric adenocarcinoma biopsy samples as compared to non-cancerous gastric epithelia. Siah1 knockdown significantly decreased invasiveness and migration of H. pylori-infected GCCs. Although, Siah1 could not increase degradation of the cytosolic ß-catenin and its nuclear translocation, it enhanced degradation of the membrane-bound ß-catenin in the infected GCCs. This loss of membrane-bound pool of ß-catenin was not associated with the proteasomal degradation of E-cadherin. Thus, this work delineated the role of Siah1 in increasing invasiveness of H. pylori-infected GCCs.

Gender-specific brain regional variation of neurons, endogenous estrogen, neuroinflammation and glial cells during rotenone-induced mouse model of Parkinson's disease.

Rotenone (RT) produces reactive oxygen species (ROS) by inhibiting the mitochondrial electron transport chain; causing dopaminergic (DA) cell death in the substantia nigra (SN) and simulates other models of induced Parkinson's disease (PD). There is a sincere dearth of knowledge regarding the status of glial cells, neuroprotective estrogen and the status of neuroinflammatory TNF-α in the different brain regions in either sex during healthy, as well as during PD conditions. In the present study of RT-induced mouse model of PD, we have selected the frontal cortex (FC), hippocampus (HC) and SN from either sex of Swiss albino mice as these are the major regions involved during PD pathogenesis. During non pathogenic conditions, the ROS-scavenging enzyme activity varied among the brain regions and also in between genders. The number of DOPA decarboxylase-positive cells, astrocytes and microglia was similar in the respective regions of the brain in both the sexes. The level of proinflammatory cytokine TNF-α was same in the respective FC and HC in either sex except that of SN. The expression level of estrogen and its receptors varied among the three brain regions. During RT treatment, ROS-scavenging enzyme activities increased, DOPA decarboxylase-positive neurons and fibers in DA as well as in norepinephrinergic (NE) systems become degenerated, number of astrocytes decreased and microglial cells increased in those specific brain regions in either of the sexes except in the SN region of males where astrocyte number remained unaltered and microglial cell percentage decreased. TNF-α increased in the FC and SN but remained unaltered in the HC of both sexes. Estradiol level decreased in the HC and SN but the level unevenly varied in the FC. Similarly, the estrogen bound and nuclear-cytosolic receptor α and ß also varied differentially among the brain regions of the two sexes. Therefore our present study depicts that there exists a clear variation of neuronal and astroglial cell population, estrogen and its receptor levels in different brain regions of both the sexes during control and RT-treated pathogenic condition and these variations have major implication in PD pathogenesis and progression.

Natural resolution or intervention for fluid collections in acute severe pancreatitis.

BACKGROUND: Revisions in terminology of fluid collections in acute pancreatitis have necessitated reanalysis of their evolution and outcome. The course of fluid collections in patients with acute pancreatitis was evaluated prospectively. METHODS: Consecutive adults with acute pancreatitis, who had contrast-enhanced CT (CECT) within 5-7 days of symptom onset, were enrolled in a prospective cohort study in a tertiary-care centre. Patients were treated according to standard guidelines. Follow-up transabdominal ultrasonography was done at 4-week intervals for at least 6 months. CECT was repeated at 6-10 weeks, or at any time if there were new or persistent symptoms. Asymptomatic collections were followed until spontaneous resolution. Risk factors for pancreatic pseudocysts or walled-off necrosis (WON) were assessed in multivariable analyses. RESULTS: Of 122 patients with acute pancreatitis, 109 were analysed. Some 91 patients (83·5 per cent) had fluid collections at baseline. Eleven of 29 with interstitial oedematous pancreatitis had acute peripancreatic fluid collections, none of which evolved into pseudocysts. All 80 patients with acute necrotizing pancreatitis had at least one acute necrotizing collection (ANC); of these, five patients died (2 after drainage), three underwent successful drainage within 5 weeks, and collections resolved spontaneously in 33 and evolved into WON in 39. By 6 months' follow-up, WON had required drainage in eight patients, resolved spontaneously in 23 and was persistent but asymptomatic in seven. Factors associated with increased risk of WON were blood urea nitrogen 20 mg/dl or more (odds ratio (OR) 10·96, 95 per cent c.i. 2·57 to 46·73; P = 0·001) and baseline ANC diameter greater than 6 cm (OR 14·57, 1·60 to 132·35; P = 0·017). Baseline ANC diameter over 6 cm was the only independent predictor of either the need for drainage or persistence of such collections beyond 6 months (hazard ratio 6·61, 1·77 to 24·59; P = 0·005). CONCLUSION: Pancreatic pseudocysts develop infrequently in oedematous acute pancreatitis. Only one-quarter of ANCs either require intervention or persist beyond 6 months, whereas more than one-half of WONs resolve without any intervention within 6 months of onset. Baseline diameter of ANC(s) is an important predictor of outcome.

Mal-united fracture of calcaneum treated with lateral decompression.

Fracture calcaneum is commonly complicated with mal-union. Mal-union is usually presented with broadening of the lateral wall of the calcaneum, irregularity of the sub-talar joint and loss of the vertical height of the body of calcaneum with varus rotation. In this study twenty three (n=23) patients underwent a lateral decompression for pain on lateral aspect of foot on walking and tip toeing, secondary to a mal-united calcaneal fracture with or without sub-talar intra-articular involvement. Almost every patient had ballooned out body of calcaneum with impingement of peroneal tendon sheath with its content and lateral heel pain as a result of deformity. So in current study the operative procedure selected was excision of a lateral calcaneal bony mass and decompression of the peroneal tendon to relieve pain. Peroneal tenogram was a routine procedure before surgery to detect presence and site of blockade within tendon sheath. The average follow-up was for a period of 31.6 months. Result was assessed according to American Orthopaedic Foot and Ankle Society (AOFAS) Ankle-Hind foot Scale. Two patients (n=2, 8.69%) had excellent (Category 1) result, seventeen (n=17, 73.9%) were good (Category 2), two (n=2, 8.69%) were fair (Category 3) and rest two (n=2, 8.69%) were with poor (Category 4) outcome. Every four out of five patients got back painless Heel and Ankle after surgery. The results suggest that the lateral decompression procedure is a technically simpler yet viable alternative to late sub-talar fusion for lateral pain following a mal-united fracture of calcaneum.

Splenocyte apoptosis in Plasmodium berghei ANKA infection: possible role of TNF-α and TGF-ß.

Cerebral malaria is associated with the circulating levels of tumour necrosis factor alpha (TNF-α) and transforming growth factor ß (TGF-ß), but association between these two cytokines and implications in splenocyte apoptosis remain largely obscured. We have evaluated the outcome of TGF-ß and TNF-α production in the context of splenocyte apoptosis during Plasmodium berghei ANKA (PbA) infection. Blood-stage PbA infection confirmed blood-brain barrier disruption, disarray of white pulp, increase in percentage of sub-G0/G1 and splenocyte apoptosis. Flow cytometric analysis reveals up-regulation of Fas-L followed by caspase-8 and caspase-3 activation and signifies possible involvement of Fas-L-mediated splenocyte apoptosis. We have observed down-regulation of TGF-ß and up-regulation of TNF-α in tissue and serum level, respectively, during PbA infection. Association between the production of TGF-ß and the severity of malaria infection in splenocytes was verified with TGF-ß inhibitor that exacerbated the apoptotic process. In contrary, TNF-α inhibitor causes significant delay in apoptotic process, but could not alter the lethality of parasite. Thus, results from this study suggest that the critical balance between TGF-ß and TNF-α might have a key role on Fas-L-mediated splenocyte apoptosis during experimental cerebral malaria.

Stevioside induced ROS-mediated apoptosis through mitochondrial pathway in human breast cancer cell line MCF-7.

Stevioside is a diterpene glycoside found in the leaf of Stevia rebaudiana, a traditional oriental medicinal herb, which has been shown to have various biological and ethno-medicinal activities including antitumor activity. In this study, we investigated the effects of stevioside on the cytotoxicity, induction of apoptosis, and the putative pathways of its action in human breast cancer cells (MCF-7). For the analysis of apoptotic pathway, measurement of reactive oxygen species (ROS) and assessment of mitochondrial transmembrane potential (MTP) were achieved. We showed that stevioside was a potent inducer of apoptosis and it conveyed the apoptotic signal via intracellular ROS generation; thereby inducing change in MTP and induction of mitochondrial mediated apoptotic pathway. Taken together, our data indicated that stevioside induces the ROS-mediated mitochondrial permeability transition and results in the increased expression of apoptotic proteins such as Bax, Bcl-2 and Caspase-9. Effect of stevioside on stress-related transcription factors like NF-E2-related factor-2 opens up a new vista for further studies. This is the first report on the mechanism of the antibreast cancer (in vitro) activity of stevioside.

Immunomodulatory effect of leaf extract of Murraya koenigii in diabetic mice.

CONTEXT: Extracts of different parts of Murraya koenigii were traditionally used as medicine in many countries for the treatment of various diseases including diabetes. OBJECTIVE: To delineate whether aqueous (AEM) and 50% methanol (MEM) leaf extracts have immunomodulatory properties to regulate oxidative stress metabolism and fight against the adverse effect of diabetes in diabetic mice. MATERIALS AND METHODS: Alloxan-induced diabetic mice were received oral doses of extract (≥ 200 mg/kg body weight). Fasting blood glucose, total cholesterol (TC), triglycerides (TG), phospholipids (PL), and antioxidant enzymes activity were estimated by biochemical processes. Western blotting, flow cytometry study, cell count, and histology were performed to justify different aspects of diabetes-related pathology. RESULTS: Alloxanized mice showed higher blood glucose that was further reduced after treatment of extracts for 30 days. Extract-treated diabetic mice were found lower TC, TG, and PL than diabetic mice. Rising of glutathione (GSH) and superoxide dismutase (SOD) enzyme activities compared with diabetic mice showed antioxidant property of the extracts. We found anti-inflammatory response as it was evident by interleukin (IL)-2, 4, 10, and tumor necrosis factor alpha (TNF-α) expression. In addition, the reduction of apoptosis in pancreatic cells was found upon extract-treated diabetic mice. DISCUSSION: The leaf extracts of the plant not only have hypoglycemic property but also have certain effects to regulate mice immunology related to oxidative stress metabolism. Moreover, AEM gave better response than MEM. CONCLUSION: Thus, the work reveals about immunomodulatory property of the leaf extracts of M. koenigii on diabetes and diabetes-related pathology in mice.

Thiazolidinedione precipitated thyroid associated ophthalmopathy.

Thyroid associated ophthalmopathy (TAO), a cardinal clinical pointer to diagnose Graves' disease (GD), is seen less frequently in our country than in the West, but can have sight threatening consequences. Smoking, diabetes, male gender, increasing age and radioactive iodine treatment for thyrotoxicosis are known precipitating factors for TAO. We report four cases of thiazolidinediones (TZD) precipitated TAO. All were male, had autoimmune thyroid disease (three had Graves' disease and one had Hashimoto's thyroiditis) and type 2 diabetes mellitus (T2DM). They developed eye symptoms three to four months after taking TZDs for glycaemic control. Two of them responded to medical treatment, the other two underwent surgical decompression.

CASA-based sperm kinematics of environmental risk factor-exposed human semen samples designated as normozoospermic in conventional analysis.

This study was conducted after an initial epidemiological survey of patients in and around Calcutta, India, concerning their lifestyle history, degree of risk exposure and semen analysis based on conventional WHO criteria. It was found that a large group of exposed patients were showing normozoospermic semen parameters in conventional semen analysis. Hence, a selected group of subjects, designated as normozoospermic in routine analysis, but under risk factor exposure, were selected for a repeat computer aided semen analysis (CASA) and were compared with a control group. The parameters considered among CASA results were: curvilinear velocity (VCL), straight-line velocity, average path velocity (VAP), straightness index (STR), lateral head displacement (ALH) and beat cross frequency. The results depict a significant decline in the mean values of VCL (P = 0.029) and STR (P = 0.007) in the tobacco-exposed group when compared with the unexposed group. On the other hand, there was a significant decline in the mean values of VCL (P = 0.014) and ALH (P = 0.040) in the heavy metal-exposed group when compared with the unexposed group. The other parameters did not show significant change in either group. Semen samples that had been designated normozoospermic in conventional analysis were seen to be influenced by risk factors at the level of sperm motion kinetics.

Selective separation of radio-cesium from acidic solutions using supported liquid membrane containing chlorinated cobalt dicarbollide (CCD) in phenyltrifluoromethyl sulphone (PTMS).

A supported liquid membrane method was developed using chlorinated cobalt dicarbollide (CCD) in phenyltrifluoromethyl sulphone (PTMS) as the carrier, impregnated in PTFE flat sheet membranes for the selective separation of Cs(I) from nitric acid feed solution. Solvent extraction studies were carried out for optimizing the feed as well as strip conditions. >95% Facilitated transport of Cs(I) was observed in about 3h when 1M HNO(3) and 8M HNO(3) were used as the feed and strip solutions, respectively while 0.025M CCD in PTMS was used as the carrier extractant. Selectivity studies, carried out using a mixture of radiotracers viz. (51)Cr, (59)Fe, (99)Mo, (99m)Tc, (106)Ru, (137)Cs, (152)Eu and (241)Am, indicated selective transport of Cs(I) with DF values >100. Though reproducibility of the transport data was excellent when carried out in two successive transport experiments with freshly loaded carrier solvent, the stability of the membrane was poor which restricts its long term use.

Inhibition of a protein tyrosine phosphatase using mesoporous oxides.

The feasibility of utilizing mesoporous matrices of alumina and silica for the inhibition of enzymatic activity is presented here. These studies were performed on a protein tyrosine phosphatase by the name chick retinal tyrosine phosphotase-2 (CRYP-2), a protein that is identical in sequence to the human glomerular epithelial protein-1 and involved in hepatic carcinoma. The inhibition of CRYP-2 is of tremendous therapeutic importance. Inhibition of catalytic activity was examined using the sustained delivery of p-nitrocatechol sulfate (pNCS) from bare and amine functionalized mesoporous silica (MCM-48) and mesoporous alumina (Al(2)O(3)). Among the various mesoporous matrices employed, amine functionalized MCM-48 exhibited the best release of pNCS and also inhibition of CRYP-2. The maximum speed of reaction v(max) (=160 +/- 10 micromol/mnt/mg) and inhibition constant K(i) (=85.0 +/- 5.0 micromol) estimated using a competitive inhibition model were found to be very similar to inhibition activities of protein tyrosine phosphatases using other methods.

A systems biology approach to Down syndrome: identification of Notch/Wnt dysregulation in a model of stem cells aging.

Stem cells are central to the development and maintenance of many tissues. This is due to their capacity for extensive proliferation and differentiation into effector cells. More recently it has been shown that the proliferative and differentiative ability of stem cells decreases with age, suggesting that this may play a role in tissue aging. Down syndrome (DS), is associated with many of the signs of premature tissue aging including T-cell deficiency, increased incidence of early Alzheimer-type, Myelodysplastic-type disease and leukaemia. Previously we have shown that both hematopoietic (HSC) and neural stem cells (NSC) in patients affected by DS showed signs of accelerated aging. In this study we tested the hypothesis that changes in gene expression in HSC and NSC of patients affected by DS reflect changes occurring in stem cells with age. The profiles of genes expressed in HSC and NSC from DS patients highlight pathways associated with cellular aging including a downregulation of DNA repair genes and increases in proapoptotic genes, s-phase cell cycle genes, inflammation and angiogenesis genes. Interestingly, Notch signaling was identified as a potential hub, which when deregulated may drive stem cell aging. These data suggests that DS is a valuable model to study early events in stem cell aging.

Endovascular management of infective intracranial aneurysms with acrylic glue. A report of two cases.

SUMMARY: Cerebral mycotic aneurysms (MAs) also called infective aneurysms, are uncommon and are usually encountered in patients with infective endocarditis. These aneurysms often present with intracranial hemorrhage. MAs may resolve on treatment with antibiotics alone. However prognosis with medical management alone is unpredictable. Good prognosis with surgery has been reported for single accessible ruptured MAs. However surgery is associated with significant morbidity. Endovascular treatment of MAs along with appropriate antibiotics is emerging as an acceptable option for these patients. We describe two cases of infective endocarditis complicated by ruptured MA treated successfully by liquid embolic glue material.

Dark after cholecystectomy.

We report a patient who complained of becoming darker after an abdominal surgery. The index patient not only had a darker complexion after cholecystectomy, but his glycaemic control was also getting better after operation to the extent that he could stop insulin, which he had been taking for five years. Also, he had lost significant weight after operation. Later, we found that he had developed primary hypocortisolism due to unrecognized bilateral adrenal haemorrhage in the immediate postoperative period.

Evidence-based review of aetiopathogenic theories of congenital and acquired cholesteatoma.

Cholesteatoma is a non-neoplastic, keratinising lesion which has two forms: congenital and acquired. Congenital cholesteatoma develops behind a normal, intact tympanic membrane, whilst acquired cholesteatoma is associated with a defect in the tympanic membrane. The pathological substrate of cholesteatoma is keratinising stratified squamous epithelium, but the origin of this epidermal tissue in the middle ear is controversial. Here, we review the most relevant and recent evidence for the principal aetiopathogenic theories of both forms of cholesteatoma, in the light of recent otopathological findings. Congenital cholesteatoma is most plausibly explained by the persistence of fetal epidermoid formation. Conclusive 'proof' awaits the unambiguous demonstration of the metamorphosis of an epidermoid nidus into a lesion in vivo. Acquired cholesteatoma may develop by various mechanisms: immigration, basal hyperplasia, retraction pocket and/or trauma (iatrogenic or non-iatrogenic). However, squamous metaplasia of the normal cuboidal epithelium of the middle ear is a highly unlikely explanation. Chronic inflammation seems to play a fundamental role in multiple aetiopathogenic mechanisms of acquired cholesteatoma. Therefore early treatment of inflammatory conditions might reduce their sequelae, perhaps by preventing the development of hyperplastic papillary protrusions. Continued otopathological, cellular and molecular research would enhance our limited understanding of cholesteatoma and may lead to new therapeutic strategies for this erosive disease, which often defies surgical treatment.

Water precautions and ear surgery: evidence and practice in the UK.

BACKGROUND: Patients are traditionally advised to refrain from exposing their ears to water after most otological procedures. However, recent evidence suggests that water does not adversely affect the outcome for children with tympanostomy tubes. The evidence behind the potential harmful effects of water on the outcome of other otological procedures is scarce. OBJECTIVE: The study was done to determine the current practice of otolaryngology consultants in the UK on the advice given to patients regarding swimming, diving and bathing in soapy water after myringotomy and tympanostomy tube insertion, mastoidectomy and myringoplasty. METHOD: Questionnaire based survey mailed to 382 members (consultants only) of the British Association of Otolaryngologists--Head & Neck Surgeons in the UK. RESULTS: A total of 195 responses were received (reply rate 51 per cent). In all, 95.6 per cent of the respondents allowed their patients to swim after insertion of tympanostomy tubes, with 32.9 per cent insisting on the use of earplugs until extrusion of the tympanostomy tubes. However, 61.6 per cent of the respondents restricted diving in these patients. In comparison, the respondents were more conservative with water precautions following myringoplasty and mastoidectomy. More than half the respondents recommended earplugs for bathing after all three operations. CONCLUSION: This study reveals current national practice among UK otolaryngologists. There is no general consensus in post-operative advice following otolaryngological procedures, indicating a need for national guidelines.