Aging phenotype(s) in kidneys of diabetic mice are p66ShcA dependent.

The p66ShcA protein controls cellular responses to oxidative stress, senescence, and apoptosis. Here, we test the hypothesis that aging phenotype(s) commonly associated with the broad category of chronic kidney disease are accelerated in diabetic kidneys and linked to the p66ShcA locus. At the organ level, tissue stem cells antagonize senescent phenotypes by replacing old dysfunctional cells. Using established methods, we isolated a highly purified population of stem cell antigen-1-positive mesenchymal stem cells (Sca-1+ MSCs) from kidneys of wild-type (WT) and p66 knockout (p66 KO) mice. Cells were plated in culture medium containing normal glucose (NG) or high glucose (HG). Reactive oxygen species (ROS) metabolism was substantially increased in WT MSCs in HG medium in association with increased cell death by apoptosis and acquisition of the senescent phenotype. DNA microarray analysis detected striking differences in the expression profiles of WT and p66 KO-MSCs in HG medium. Unexpectedly, the analysis for p66 KO-MSCs revealed upregulation of Wnt genes implicated in self-renewal and differentiation. To test the in vivo consequences of constitutive p66 expression in diabetic kidneys, we crossed the Akita diabetic mouse with the p66KO mouse. Homozygous mutation at the p66 locus delays or prevents aging phenotype(s) in the kidney that may be precursors to diabetic nephropathy.

Expression of the gap junction protein connexin36 in small intensely fluorescent (SIF) cells in cardiac parasympathetic ganglia of rodents.

In mammals, several endocrine cell types are electrically coupled by connexin36 (Cx36)-containing gap junctions, which mediate intercellular communication and allow regulated and synchronized cellular activity through exchange of ions and small metabolites via formation of intercellular channels that link plasma membranes of apposing cells. One cell type thought to be endocrine-like in nature are small intensely fluorescent (SIF) cells that store catecholamines in their dense-core vesicles and reside in autonomic ganglia. Here, using immunofluorescence approaches, we examined whether SIF cells located specifically in cardiac parasympathetic ganglia of adult and neonatal mice and adult rats follow patterns of Cx36 expression seen in other endocrine cells. In these ganglia, SIF cells were identified by their distinct small soma size, autofluorescence at 475 nm, and immunolabelling for their markers tyrosine hydroxylase and vesicular monoamine transporter-1. SIF cells were often found in pairs or clusters among principal cholinergic neurons. Immunofluorescence labelling of Cx36 occurred exclusively as fine puncta that appeared at contacts between SIF cell processes and somata or at somato-somatic appositions of SIF cells. These puncta were absent in cardiac parasympathetic ganglia of Cx36 null mice. Transgenic mice expressing enhanced green fluorescent protein reporter for Cx36 expression displayed labelling for the reporter in SIF cells. The results suggest that Cx36-containing gap junctions electrically couple SIF cells, which is consistent with previous suggestions that these may be classified as endocrine-type cells that secrete catecholamines into the bloodstream in a regulated manner.

Opportunities and challenges for biomarker discovery using electronic health record data.

Electronic health records (EHRs) have become increasingly relied upon as a source for biomedical research. One important research application of EHRs is the identification of biomarkers associated with specific patient states, especially within complex conditions. However, using EHRs for biomarker identification can be challenging because the EHR was not designed with research as the primary focus. Despite this challenge, the EHR offers huge potential for biomarker discovery research to transform our understanding of disease etiology and treatment and generate biological insights informing precision medicine initiatives. This review paper provides an in-depth analysis of how EHR data is currently used for phenotyping and identifying molecular biomarkers, current challenges and limitations, and strategies we can take to mitigate challenges going forward.

Characteristics of Electrical Synapses, C-terminals and Small-conductance Ca2+ activated Potassium Channels in the Sexually Dimorphic Cremaster Motor Nucleus in Spinal Cord of Mouse and Rat.

Sexually dimorphic motoneurons (MNs) located in lower lumbar spinal cord are involved in mating and reproductive behaviours and are known to be coupled by electrical synapses. The cremaster motor nucleus in upper lumbar spinal cord has also been suggested to support physiological processes associated with sexual behaviours in addition to its thermoregulatory and protective role in maintaining testes integrity. Using immunofluorescence approaches, we investigated whether cremaster MNs also exhibit features reflecting their potential for electrical synaptic communication and examined some of their other synaptic characteristics. Both mice and rats displayed punctate immunolabelling of Cx36 associated with cremaster MNs, indicative of gap junction formation. Transgenic mice with enhanced green fluorescent protein (eGFP) reporter for connexin36 expression showed that subpopulations of cremaster MNs in both male and female mice express eGFP, with greater proportions of those in male mice. The eGFP+ MNs within the cremaster nucleus vs. eGFP- MNs inside and outside this nucleus displayed a 5-fold greater density of serotonergic innervation and exhibited a paucity of innervation by C-terminals arising from cholinergic V0c interneurons. All MNs within the cremaster motor nucleus displayed prominent patches of immunolabelling for SK3 (K+) channels around their periphery, suggestive of their identity as slow MNs, many though not all of which were in apposition to C-terminals. The results provide evidence for electrical coupling of a large proportion of cremaster MNs and suggest the existence of two populations of these MNs with possibly differential innervation of their peripheral target muscles serving different functions.

Population dynamics of a parasite Pallisentis in two species of fish Channa punctatus and Channa striatus.

In an aquatic environment, there is a profound and inverse relationship between environment quality and disease status of fish. Parasites are one of the most serious limiting factors in aquaculture. Therefore, the present investigation has been undertaken to sort out the population dynamics of Pallisentis (Acanthocephala), in relation to host sex from freshwater fishes, Channa of Rohilkhand region. A total of 517 fishes (Channa punctatus, n = 198 and C. striatus, n = 319) were examined regularly from August 2006 to February 2010. Overall prevalence of Pallisentis in C. striatus was higher in females (67.78%) as compared to males (63.52%). In case of C. punctatus, overall prevalence of Pallisentis was higher in males (53.77%) as compared to females (52.17%) whereas, relative density was higher in females (61.41%) than in males (52.72%). Intensity (2-3 parasite/host, in both sexes), density (1.36 in males and 1.69 in females) and infection index (0.73 in males and 0.88 in females) were recorded.

Total annulus excision tympanoplasty - a solution to recurrent myringitis: double blind randomised trial.

OBJECTIVE: In spite of continuous refinements in tympanoplasty techniques, results are variable, and it is not uncommon to see a discharging eardrum even after a good graft uptake. This study aimed to evaluate the efficacy of total annulus excision tympanoplasty in comparison with conventional underlay tympanoplasty. METHOD: This was a double blinded, randomised, controlled trial performed at a tertiary care centre. After inclusion and exclusion criteria were met, 56 patients were enrolled and randomised, and 28 patients were allocated to each group (group A (conventional tympanoplasty) and group B (total annulus excision)). Patients and evaluators were blind to the procedure performed. RESULTS: Patients in group B (total annulus excision) showed better graft uptake and no discharge with better gains in air conduction thresholds (p < 0.05) when compared with group A (conventional tympanoplasty). CONCLUSION: In view of the advantages it offers, total annulus excision tympanoplasty may be preferred over the conventional techniques in patients with central perforations.

DC-specific ICAM-3-grabbing nonintegrin mediates internalization of HIV-1 into human podocytes.

Human immunodeficiency virus (HIV)-1 has been demonstrated to contribute to the pathogenesis of HIV-associated nephropathy. In renal biopsy studies, podocytes have been reported to be infected by HIV-1. However, the mechanism involved in HIV-1 internalization into podocytes is not clear. In the present study, we evaluated the occurrence of HIV-1 internalization into conditionally immortalized human podocytes and the mechanism involved. Human podocytes rapidly internalized R5 and X4 HIV-1 primary strains via an endocytosis-dependent pathway, without establishing a productive infection. The HIV-1 internalization was dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) receptor mediated. The role of DC-SIGN was confirmed by using specific blocking antibodies and transfection with small interfering (si) RNA/DC-SIGN. Since podocyte HIV-1 trafficking was not altered by pH-modulating agents, it appeared that HIV-1 routing occurred through nonacid vesicular compartments. Interestingly, transfection of podocytes with neither siRNA/caveolin-1 nor siRNA/clathrin heavy chain inhibited podocyte viral accumulation. Thus it appears that clathrin-coated vesicles and caveosomes may not be contributing to HIV-1-associated membrane traffic.

Safe and rapid resolution of severe hypertriglyceridaemia in two patients with intravenous insulin.

AIM: To rapidly reduce serum triglyceride to a safe serum level. Severe hypertriglyceridaemia is associated with uncontrolled diabetes, obesity and poor physical activity. Even moderate increases in triglyceride levels (> 5mmol/L) confer an increased risk of pancreatitis and coronary artery disease. We present two patients with diabetes and serum triglyceride levels of greater than 85mmol/L despite polypharmacy intervention. METHOD: 72-hour intravenous insulin infusion was administered. RESULTS: Serum triglyceride levels fell to 9.4 and 4.6 mmol/L respectively, without adverse events and sustained effect over several months. CONCLUSION: We suggest the use of intravenous insulin infusion where lifestyle and oral drug therapies have failed can impact on severe hypertriglyceridaemia.

Right atrial pacing for prevention of postoperative atrial fibrillation following coronary artery bypass grafting: a prospective observational trial.

INTRODUCTION: Atrial fibrillation (AF) is the most common complication after coronary artery bypass grafting (CABG). Conventional clinical practice is to treat AF after its onset in the postoperative period. However effective prophylaxis can result in reduced morbidity and hospital stay. This prospective study was designed to assess the efficacy of right atrial pacing in the prevention of postoperative AF following CABG. METHODS: All patients who had isolated first time CABG on cardiopulmonary bypass between January 2007 and June 2007, operated by one surgeon were included in this study (study group). During the same period, isolated CABG carried out by other surgeons in the same unit and not paced served as control (control group). In the study group (n=24) right atrial pacing was continued for 96h in the postoperative period. The end points of the study were occurrence of AF, death in postoperative period or discharge from hospital. RESULTS: In the study group 5 patients could not be paced because of persistent tachycardia or failure of atrial electrodes. There was a statistically significant reduction in the incidence of postoperative atrial fibrillation from 31.1% in control group to 5.2% (p-value 0.03) in patients who were paced. CONCLUSIONS: The preliminary results of this study suggest that prophylactic right atrial pacing significantly reduced the incidence of AF following CABG. However these results need to be confirmed in a larger cohort of patients in a prospective randomised trial.

Biotechnological advancement in isolation of anti-neoplastic compounds from natural origin: a novel source of L-asparaginase.

L-asparaginase catalyzes the hydrolysis of L-asparagine into aspartate and ammonia, which is used as an anti-neoplastic agent. Isolation of asparaginase from microorganisms may be cardinal for producing this anticancer agent at industrial level. A total of three hundred fungal isolates were screened for L-asparaginase production. These fungal isolates were growing on various fruits and vegetables. Among these, the Fusariumn sp. isolate that was growing on green chilly showed highest enzyme production. This study may give an outstanding contribution for finding organisms with high yielding L-asparaginase. Fungal L-asparaginase is superior in terms of its eukaryotic origin that may be responsible for its lesser toxicity.

Ascending aortic tear with severe aortic regurgitation following rugby injury.

Blunt ascending aortic injuries are rare in clinical practice. Aortic valve regurgitation is an uncommon consequence of closed chest injury. It is caused by either ruptured valve cusp or when subadventitial rupture of the ascending aorta causes prolapse of subjacent valve cusp. Most of the blunt aortic injuries are secondary to high speed motor vehicle accident or falls from significant height. We report on a 33-year-old man who sustained an injury during a rugby game and presented with severe aortic regurgitation secondary to subadventitial rupture of ascending aorta on a background of asymptomatic dilated aortic root. He underwent emergency operation for torrential aortic regurgitation and severe left ventricular failure.

Tooth brushing skills for the children aged 3-11 years.

OBJECTIVES: The objective of this study was to evaluate tooth brushing management and ability of children in relation to age and gender. MATERIALS AND METHODS: The study population consisted of 45 children, who attended Department of Pedodontics and Preventive Dentistry at V.S Dental College and Hospital Bangalore. Children were divided into three groups according to age: Group I: 3-5 years, Group II: 6-8 years, and Group III: 9-11 years. Each child selected his/her favorite toothbrush and brushed his/her teeth under supervision of one instructor. The grip type during tooth brushing was recorded on a videotape and duration of tooth brushing also was noted. The data were analyzed in relation to the age and gender using the chi-square test. ANOVA was used to find if there is any significant difference between the mean brushing times in the different groups. RESULTS: The results of statistical analysis showed that in all cases P> 0.05 for grip preferences and P. CONCLUSION: The required manual dexterity for tooth brushing was present in younger age groups.

Penetrating atheromatous ulcer of ascending aorta: a case report and review of literature.

Penetrating atherormatous ulcer (PAU) is the condition in which ulceration of an aortic atherosclerotic lesion penetrates the internal elastic lamina into the media. Penetrating atheromatous ulcers usually involve the descending thoracic aorta and are very uncommon in ascending aorta. Differentiation of PAU from other causes of Acute Aortic Syndrome (AAS) such as intramural haematoma (IMH) and aortic dissection is difficult. The diagnosis of these ulcers is made on computerised tomography (CT) scan, magnetic resonance imaging (MRI), or transoesophageal echocardiography (TOE). Once diagnosed, PAU of ascending aorta should be treated surgically on emergency basis. We report on a patient of penetrating atherosclerotic aortic ulcer in ascending aorta with a review of the natural history, imaging diagnosis and management of the condition. She underwent ascending aortic interposition graft using 26mm Dacron graft successfully.

Mitral valve replacement in patient with absent right superior vena cava, pectus excavatum, Marfan syndrome and severe mitral regurgitation.

Combination of persistent left superior vena cava (PLSVC) draining into right atrium via the coronary sinus with a normal right superior vena cava (RSVC) is the most common systemic venous anomaly in visceroatrial situs solitus. On the other hand, a PLSVC in absence of RSVC is an extremely rare anomaly. Cardiovascular abnormalities are frequently encountered in patients with Marfan's syndrome but absence of RSVC with PLSVC is not reported in these patients. We report on a 52-year-old man with Marfan syndrome, an absence of right superior vena cava, severe pectus excavatum and severe mitral regurgitation who underwent successful mitral valve replacement. CT scan chest revealed marked cardiac deviation to the left associated with severe pectus excavatum, absent right superior vena cava and persistent left superior vena cava. During surgery replacement of mitral valve was performed through midsternotomy successfully.

Intrapericardial haematoma: a rare cause of congestive cardiac failure after three years of coronary artery bypass grafting.

We present a rare case of organised intrapericardial haematoma after three years of coronary artery bypass grafting. Delayed cardiac tamponade after open heart surgery usually occurs within first few weeks of surgery. Intrapericardial organised haematoma causing localised compression of cardiac chambers after three years of open heart surgery is very rare. Under cardiopulmonary bypass and cardioplegic cardiac arrest, we removed the haematoma. It is difficult to make a clinical diagnosis. However once diagnosis is made and haematoma removed, it gives good symptomatic relief to the patient.

Optimal surgical management of descending necrotising mediastinitis: our experience and review of literature.

INTRODUCTION: Descending necrotising mediastinitis is a form of mediastinitis caused by odontogenic infection or deep cervical infections, which spreads to the mediastinum via the cervical fascial planes. Despite the increased use of CT scan as a diagnostic aid and the improvement in antibiotics, mortality in patients with descending necrotising mediastinitis remains high, reported between 25 and 40% in the literature. Surgical management and optimal form of mediastinal drainage remain controversial. PATIENTS AND METHODS: We have treated three patients with descending necrotising mediastinitis at our institution. Two patients were male and one was female with mean age 54.3+/-12.5 years. One of the patients was a known diabetic. The primary oropharyngeal infection was Ludwig's angina, odontogenic abscess and parapharyngeal abscess. All patients underwent mediastinal drainage, one through midsternotomy and two through right thoracotomy in addition to cervical drainage. All the three patients had mixed aerobic and anaerobic infection. RESULTS: There was no perioperative mortality. Mean ICU stay was 32 days (12-53 days). All three patients had septicaemic shock requiring prolonged inotropic support. Two patients required tracheostomy because of prolonged ventilation. All the patients had recurrent abscesses and collections either in neck or in chest requiring drainage either surgically or percutaneously under CT scan or ultrasound guidance, thus decreasing the need of repeated surgical procedures. CONCLUSION: Descending necrotising mediastinitis is a potentially fatal condition. Early diagnosis, prompt surgical drainage, monitoring of disease process, appropriate medical management in an intensive care unit and a multi-disciplinary approach can significantly reduces the mortality in this otherwise fatal condition. Percutaneous drainage of recurrent abscesses and collections can decrease the need of repeated surgical procedures in these critically ill patients.

Evidence for immunoglobulin Fc receptor-mediated prostaglandin2 and platelet-activating factor formation by cultured rat mesangial cells.

The possibility of Fc-dependent uptake of IgG immune complexes was examined in subcultured rat mesangial cells free of monocytes. 195Au-labeled colloidal gold particles were coated either with BSA only or with BSA followed by rabbit anti-BSA-IgG or the F(ab')2 fragment of the IgG. Mesangial cells preferentially took up 195Au particles covered with BSA-anti-BSA-IgG over those covered with BSA or the F(ab')2 fragment. This uptake was a time-dependent and saturable process inhibitable by sodium azide or cytochalasin B. Using phase-contrast microscopy in the light reflectance mode, it was established that essentially all mesangial cells took up IgG-coated gold particles. By electron microscopy the process was shown to consist of vesicular uptake with delivery to endosomes. Mesangial binding-uptake of the IgG-covered particles was associated with stimulation of PGE2 synthesis and production of platelet-activating factor, a lipid mediator of inflammation. To characterize the potential Fc receptor for IgG we used the rosetting technique with sheep red blood cells coated with IgG subclass-specific mouse monoclonal antibodies. 50% of mesangial cells exhibited rosetting with red cells coated with mouse IgG2a, whereas negligible rosetting was observed with IgG2b or IgG1. Competition experiments confirmed the specificity of IgG2a binding. We conclude that cultured rat mesangial cells exhibit specific receptors for IgG and that occupancy of Fc receptors results in endocytosis and is associated with generation of PGE2 and platelet-activating factor. These observations may be of significance for immune-mediated glomerular diseases.

Primary fallopian tube carcinoma: a retrospective clinicopathologic study.

INTRODUCTION: Primary fallopian tube carcinoma is a rare tumor. The aim of this study was to evaluate clinical characteristics and management of fallopian tube malignancies at a large tertiary care cancer institute. METHODS: A retrospective review of the Tumor Registry was conducted to identify all primary fallopian tube carcinomas between 1980 and 2001. Medical charts were retrospectively reviewed. Primary endpoints were overall survival and disease recurrence. RESULTS: Thirty-five patients had histology consistent with fallopian tube carcinoma. The median age at diagnosis was 56 years. The most common signs or symptoms were abnormal vaginal bleeding (29%) and abdominal/pelvic mass (26%). The most common histology was adenocarcinoma in 16 (46%) patients. Five patients (14%) were Stage I, seven patients (20%) Stage II, 17 patients (49%) Stage III and six patients (17%) Stage IV. Thirty-two (91%) patients received adjuvant chemotherapy and 77% received platinum-based chemotherapy. Twenty-seven (77%) patients underwent second-look surgery, of which 17 patients (63%) were positive for disease. The 5-year survival rate was 64% for Stage I, 42% for Stage II, 32% for Stage III, and 17% for Stage IV. CONCLUSIONS: Fallopian tube malignancies are rare and carry a poor prognosis. More extensive research needs to be performed to have definitive etiologic, diagnostic and treatment guidelines.