A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus

Hepatitis E virus (HEV) is transmitted by the fecal-oral route and causes sporadic and epidemic forms of acute hepatitis. Large waterborne HEV epidemics have been documented exclusively in developing countries. At least four major genotypes of HEV have been reported worldwide: genotype 1 (found primarily in Asian countries), genotype 2 (isolated from a single outbreak in Mexico), genotype 3 (identified in swine and humans in the United States and many other countries), and genotype 4 (identified in humans, swine and other animals in Asia). To better detect and quantitate different HEV strains that may be present in clinical and environmental samples, we developed a rapid and sensitive real-time RT-PCR assay for the detection of HEV RNA. Primers and probes for the real-time RT-PCR were selected based on the multiple sequence alignments of 27 sequences of the ORF3 region. Thirteen HEV isolates representing genotypes 1-4 were used to standardize the real-time RT-PCR assay. The TaqMan assay detected as few as four genome equivalent (GE) copies of HEV plasmid DNA and detected as low as 0.12 50% pig infectious dose (PID50) of swine HEV. Different concentrations of swine HEV (120-1.2PID50) spiked into a surface water concentrate were detected in the real-time RT-PCR assay. This is the first reporting of a broadly reactive TaqMan RT-PCR assay for the detection of HEV in clinical and environmental samples.     

Reduced growth of Drosophila neurofibromatosis 1 mutants reflects a non-cell-autonomous requirement for GTPase-Activating Protein activity in larval neurons

Neurofibromatosis type 1 (NF1) is among the most common genetic disorders of humans and is caused by loss of neurofibromin, a large and highly conserved protein whose only known function is to serve as a GTPase-Activating Protein (GAP) for Ras. However, most Drosophila NF1 mutant phenotypes, including an overall growth deficiency, are not readily modified by manipulating Ras signaling strength, but are rescued by increasing signaling through the cAMP-dependent protein kinase A pathway. This has led to suggestions that NF1 has distinct Ras- and cAMP-related functions. Here we report that the Drosophila NF1 growth defect reflects a non-cell-autonomous requirement for NF1 in larval neurons that express the R-Ras ortholog Ras2, that NF1 is a GAP for Ras1 and Ras2, and that a functional NF1-GAP catalytic domain is both necessary and sufficient for rescue. Moreover, a Drosophila p120RasGAP ortholog, when expressed in the appropriate cells, can substitute for NF1 in growth regulation. Our results show that loss of NF1 can give rise to non-cell-autonomous developmental defects, implicate aberrant Ras-mediated signaling in larval neurons as the primary cause of the NF1 growth deficiency, and argue against the notion that neurofibromin has separable Ras- and cAMP-related functions.     

Association of high carotid bifurcation and thyrolinguofacial trunk: a rare variation

Variations in the origins and the branching pattern of the carotid system of arteries are not uncommon. Here we report a rare case of higher bifurcation of the common carotid artery (CCA) (at the level of the greater cornu of the hyoid bone), thyrolinguofacial trunk (TLFT) originating from the CCA, superior laryngeal artery (SLA) arising from the external carotid artery (ECA) on the left side, and linguofacial trunk arising from the ECA on the right side. In the present case, the CCA and carotid bifurcation may have arisen from the second aortic arch. The ECA bud could have developed from parts of the first aortic arch and ventral aorta. Thus, the altered blood flow through these vessels due to high carotid bifurcation could have caused disproportionate growth and shift in the origins of the branches of the ECA. An understanding of the bifurcation of the CCA and the branching pattern of the ECA should prove useful to surgeons performing selective intra-arterial chemotherapy for head and neck cancer.     

Strategies to improve immunization rates and well-child care in a disadvantaged population: a cluster randomized controlled trial

OBJECTIVE: To measure the effect of a multimodal intervention on well-child care visit (WCV) and immunization rates in an inner-city population. DESIGN: Cluster randomized controlled trial. SETTING AND PARTICIPANTS: One-year cohort of 2843 infants born at a hospital in an integrated inner-city health care system. INTERVENTIONS: Eleven clinics were randomly allocated to 1 of 3 study arms: WCV intervention (n = 3), immunization intervention (n = 4), and controls (n = 4). Interventions to improve immunization and WCV rates included both patient-based and clinic-based activities. MAIN OUTCOME MEASURES: Up-to-date status with childhood immunizations and WCVs by age 12 months (primary) and health care utilization and charges (secondary). RESULTS: Compared with the control arm, the WCV and immunization arms had 5% to 6% higher immunization rates and 7% to 8% higher WCV rates. In multivariate analyses that accounted for the clustered nature of the data, the number of immunizations received was greater in the WCV arm than in controls. However, neither the WCV nor the immunization intervention increased WCV or immunization up-to-date rates. The WCV arm had slightly higher health care charges. Neither intervention affected emergency, urgent care or inpatient utilization. CONCLUSIONS: This multimodal intervention produced a small increase in the number of childhood immunizations delivered. However, patient- and clinic-based methods did not lead to significant increases in WCV or immunization up-to-date rates after controlling for other factors. Methods found in some settings to increase immunization up-to-date rates may not be as effective in a population of inner-city socioeconomically disadvantaged children.     

Improvement of Psoriasis During Sunitinib Therapy for Renal Cell Carcinoma

Sunitinib is an oral tyrosine kinase inhibitor, which is indicated for the treatment of renal cell carcinoma and gastrointestinal stromal tumors. The authors report the case of a patient who underwent treatment for renal cell carcinoma and noted additional benefit by improvement in his psoriatic skin lesions. This may be attributed to the antiangiogenic activity of sunitinib by inhibition of vascular endothelial growth factor receptors.     

Gastric bypass and On-Q pump: effectiveness of Soaker Catheter system on recovery of bariatric surgery patients

BackgroundThe continuous infusion of ropivacaine is effective in controlling pain for a wide variety of surgical procedures and reducing opioid adverse effects and dependency. The present study assessed the efficacy of ropivacaine infusion using the I-Flow dual Soaker Catheter system at the surgical site for bariatric surgery recovery at the Dallas Veterans Affairs Medical Center Hospital (Dallas, TX). We hypothesized that patients receiving ropivacaine would report lower levels of morphine requirement and pain, would have shorter hospital stays, and would return to ambulating faster than patients in the control group.MethodsA total of 45 patients undergoing Roux-en-Y gastric bypass surgery were randomized to 1 of 2 treatment groups, with a target study population of 50 patients, receiving either .2% ropivacaine (n = 24) or saline solution (n = 21). Before incision closure, the surgeon infiltrated the surrounding tissues with 30 mL of ropivacaine (.5%) or saline solution. The catheter was then placed in both the subfascial space and subcutaneously. Next, the infusion pump was connected to the Soaker Catheters to complete the system design and deliver solution to the surgical site.ResultsNo significant differences were found in the pain scores, morphine requirement, or length of stay between the 2 groups. The ropivacaine group interval to sitting up was one half day shorter than that of patients receiving saline (P = .038).ConclusionsPatients receiving ropivacaine were found to ambulate much more quickly than did the control group patients. This could be very beneficial in reducing the complications from blood clots and improving patient recovery and overall well-being after surgery by assisting with a quicker return to activities of daily living and reducing the dependence on the nursing staff.     

Construction of a rhamnose mutation in Bacillus anthracis affects adherence to macrophages but not virulence in guinea pigs

Carbohydrate analyses of whole-spore extracts have confirmed the presence of rhamnose in the spore of the fully virulent Ames strain of Bacillus anthracis. A gene cluster containing loci with high homology to the rhamnose biosynthetic genes, rmlACBD, was identified within the B. anthracis chromosome. The first gene of this cluster, rmlA, was inactivated by forming a merodiploid cointegrate using an internal fragment of the gene within the Ames strain of B. anthracis to construct the mutant strain Ames-JAB1. Carbohydrate analysis of spores from this mutant demonstrated the loss of rhamnose. When assaying for spore infection of macrophages, we detected a significant decrease in the recovery with the Ames-JAB1 strain compared to the recovery with the Ames wild-type strain. When pre-treating macrophages with cytochalasin-D, spores of the mutant were further hindered in recovery, indicating that the spores were not able to bind as well to the macrophages. However, in guinea pigs challenge experiments, no difference in virulence was observed between the mutant and wild-type strains. These results suggest that the incorporation of rhamnose into the spore coat of B. anthracis is required for optimal interaction with macrophages but is not required for full virulence in this animal model.     

Dipeptidyl peptidase 4 inhibitors in COVID-19: Beyond glycemic control

Coronavirus disease 2019 (COVID-19) is associated with a high risk of mortality and complications in patients with diabetes mellitus. Achieving good glycemic control is very important in diabetic patients to reduce complications and mortality due to COVID-19. Recent studies have shown the mortality benefit and anti-inflammatory effects of Dipeptidyl-peptidase-4 inhibitors (DPP-4i) in diabetic patients with COVID-19. DPP-4i may have a beneficial role in halting the severity of infection primarily by three routes, namely viral entry inhibition, anti-inflammatory and anti-fibrotic effects and glycemic control. This has raised the pro-mising hypothesis that DPP-4i might be an optimal strategy for treating COVID-19 in patients with diabetes. This review aims to summarise the possible therapeutic non-glycemic effects of DPP-4i in diabetic patients diagnosed with COVID-19 in the light of available evidence.