DNA barcoding and surveillance sampling strategies for Culicoides biting midges (Diptera: Ceratopogonidae) in southern India

Background: Culicoides spp. biting midges transmit bluetongue virus (BTV), the aetiological agent of bluetongue (BT), an economically important disease of ruminants. In southern India, hyperendemic outbreaks of BT exert high cost to subsistence farmers in the region, impacting on sheep production. Effective Culicoides spp. monitoring methods coupled with accurate species identification can accelerate responses for minimising BT outbreaks. Here, we assessed the utility of sampling methods and DNA barcoding for detection and identification of Culicoides spp. in southern India, in order to provide an informed basis for future monitoring of their populations in the region. Methods: Culicoides spp. collected from Tamil Nadu and Karnataka were used to construct a framework for future morphological identification in surveillance, based on sequence comparison of the DNA barcode region of the mitochondrial cytochrome c oxidase I (COI) gene and achieving quality standards defined by the Barcode of Life initiative. Pairwise catches of Culicoides spp. were compared in diversity and abundance between green (570 nm) and ultraviolet (UV) (390 nm) light emitting diode (LED) suction traps at a single site in Chennai, Tamil Nadu over 20 nights of sampling in November 2013. Results: DNA barcode sequences of Culicoides spp. were mostly congruent both with existing DNA barcode data from other countries and with morphological identification of major vector species. However, sequence differences symptomatic of cryptic species diversity were present in some groups which require further investigation. While the diversity of species collected by the UV LED Center for Disease Control (CDC) trap did not significantly vary from that collected by the green LED CDC trap, the UV CDC significantly outperformed the green LED CDC trap with regard to the number of Culicoides individuals collected. Conclusions: Morphological identification of the majority of potential vector species of Culicoides spp. samples within southern India appears relatively robust; however, potential cryptic species diversity was present in some groups requiring further investigation. The UV LED CDC trap is recommended for surveillance of Culicoides in southern India

Tubular epithelial cell and podocyte apoptosis with de novo sirolimus based immunosuppression in renal allograft recipients with DGF

Sirolimus is associated with prolongeddelayed graft function (DGF) following renaltransplantation and exacerbation of proteinuria. Weassessed renal allograft biopsies from DGF patientstreated with de novosirolimus (n = 10) for renal tubularcell and podocyte apoptosis and expression of activatedcaspase-3, Bcl-2, and mTOR and compared them tobiopsies from DGF patients not receiving sirolimus (n =15). Both groups received mycophenolate mofetil,prednisone and antibody induction. Apoptosis wasassessed using terminal deoxynucleodidyl transferasemediated dUTP nick end labeling (TUNEL) staining.Caspase-3, Bcl-2, and mTOR expression were assessedby immunohistochemistry. Sirolimus treated patients had334±69 TUNEL positive cells per 5 high power fieldscompared to 5.5±2.9 TUNEL positive cells in controlpatients (p<0.001). The number of TUNEL positive cellscorrelated with tubular architectural disruption.Expression of activated caspase-3, Bcl-2, or activatedmTOR did not differ between groups. 60% of biopsiesfrom sirolimus treated patients compared to 7% ofbiopsies from controls showed diffuse podocyteapoptosis (p = 0.007). There was no podocyte expressionof activated mTOR, activated caspase-3, or Bcl-2 ineither group. These data suggest that DGF patientstreated with sirolimus have increased renal tubular cellapoptosis and podocyte apoptosis

Investigating whole‐brain metabolite abnormalities in the chronic stages of moderate or severe traumatic brain injury

Evidence suggests that neurometabolic abnormalities can persist after traumatic brain injury (TBI) and drive clinical symptoms such as fatigue and cognitive disruption. Magnetic resonance spectroscopy has been used to investigate metabolite abnormalities following TBI, but few studies have obtained data beyond the subacute stage or over large brain regions. The purpose of this study was to measure whole-brain metabolites in chronic stages of TBI. Observational study. University. Eleven men with a moderate or severe TBI more than 12 months prior, and 10 age-matched healthy controls completed whole-brain spectroscopic imaging. Ratios of N-acetylaspartate (NAA), Choline (CHO), and myo-inositol (MI) to creatine (CR) were measured in whole brain gray and white matter, as well as 64 brain regions of interest. Arterial spin labeling (ASL) data were also collected to investigate whether metabolite abnormalities were accompanied by differences in cerebral perfusion. There were no differences in metabolite ratios within whole brain gray and white matter ROIs. Linear regression showed lower NAA/CR in the white matter of the left occipital lobe but higher NAA/CR in the grey matter of the left parietal lobe. Metabolite abnormalities were observed in several brain regions in the TBI group including the corpus callosum, putamen, and posterior cingulate. However, none of findings survived correction for multiple comparison There were no differences in cerebral blood flow between patients and controls. Higher MI/CR may indicate ongoing gliosis, and it has been suggested that low CHO/CR at chronic time points may indicate cell death or lack of healthy turnover and repair. However, with the small sample size of this study, we caution against the over-interpretation of our results. None of the findings within ROIs survived correction for multiple comparison. Thus, they may be considered possible avenues for future research in this area. This article is protected by copyright. All rights reserved

Clinical Assessment of the Effects of Low-Level Laser Therapy on Coronally Advanced Flap Procedure in the Management of Isolated Gingival Recession

The aim of this randomized, controlled split-mouth clinical trial was to assess the effect of LLLT on wound healing after modified coronally advanced flap (MCAF) procedure for treatment of isolated recession-type defects. Fifteen patients with isolated bilaterally symmetrical gingival recessions (Miller’s Class I or Class II, or a combination of both) were enrolled in this study. After a modified, coronally advanced flap technique was implemented, a diode laser (810 nm) with a power of 120 mW irradiated the inner surface of the flap and the outer surface of the flap (low-level laser therapy—LLLT) after suturing for 5 min. This was repeated for the following four consecutive days. Descriptive statistics, a Kruskal–Wallis test and a Mann–Whitney test were performed to analyze the data. A p-value of less than 0.05 was considered statistically significant. The mean recession depth decreased from 3.33 ± 0.9 mm (baseline) to 0.2 ± 0.3 mm (3 months) and 0.4 ± 0.2 mm (6 months) in the test group. The mean recession width decreased from 3.8 ± 0.7 mm (baseline) to 0.2 ± 0.3 mm (3 months) and 0.5 ± 0.3 mm (6 months) in the test group. Due to minimal pain and discomfort, patient acceptability was quite high

Role of mTOR in podocyte function and diabetic nephropathy in humans and mice

Chronic glomerular diseases, associated with renal failure and cardiovascular morbidity, represent a major health issue. However, they remain poorly understood. Here we have reported that tightly controlled mTOR activity was crucial to maintaining glomerular podocyte function, while dysregulation of mTOR facilitated glomerular diseases. Genetic deletion of mTOR complex 1 (mTORC1) in mouse podocytes induced proteinuria and progressive glomerulosclerosis. Furthermore, simultaneous deletion of both mTORC1 and mTORC2 from mouse podocytes aggravated the glomerular lesions, revealing the importance of both mTOR complexes for podocyte homeostasis. In contrast, increased mTOR activity accompanied human diabetic nephropathy, characterized by early glomerular hypertrophy and hyperfiltration. Curtailing mTORC1 signaling in mice by genetically reducing mTORC1 copy number in podocytes prevented glomerulosclerosis and significantly ameliorated the progression of glomerular disease in diabetic nephropathy. These results demonstrate the requirement for tightly balanced mTOR activity in podocyte homeostasis and suggest that mTOR inhibition can protect podocytes and prevent progressive diabetic nephropathy

Pancreas Allograft Biopsies with Positive C4d Staining and Anti-Donor Antibodies Related to Worse Outcome for Patients

C4d+ antibody-mediated rejection following pancreas transplantation has not been well characterized. Therefore, we assessed the outcomes of 27 pancreas transplantation patients (28 biopsies), with both C4d staining and donor-specific antibodies (DSA) determined, from a cohort of 257 patients. The median follow-up was 50 (interquartile range [IQR] 8-118) months. Patients were categorized into 3 groups: group 1, patients with minimal or no C4d staining and no DSA (n = 13); group 2, patients with either DSA present but no C4d, diffuse C4d+ and no DSA or focal C4d+ and DSA (n = 6); group 3, patients with diffuse C4d+ staining and DSA (n = 9). Active septal inflammation, acinar inflammation and acinar cell injury/necrosis were significantly more abundant in group 3 than in group 2 (respective p-values: 0.009; 0.033; 0.025) and in group 1 (respective p-values: 0.034; 0.009; 0.002). The overall uncensored pancreas graft survival rate for groups 1, 2 and 3 were 53.3%, 66.7% and 34.6%, respectively (p = 0.044). In conclusion, recipients of pancreas transplants with no C4d or DSA had excellent long-term graft survival in comparison with patients with both C4d+ and DSA present. Hence, C4d should be used as an additional marker in combination with DSA in the evaluation of pancreas transplant biopsies.Nephrolog

Pig-to-Baboon Liver Xenoperfusion Utilizing GalTKO.hCD46 Pigs and Glycoprotein Ib Blockade

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