Policy processes underpinning universal health insurance in Vietnam.
In almost 30 years since economic reforms or 'renovation' (Doimoi) were launched, Vietnam has achieved remarkably good health results, in many cases matching those in much higher income countries. This study explores the contribution made by Universal Health Insurance (UHI) policies, focusing on the past 15 years. We conducted a mixed method study to describe and assess the policy process relating to health insurance, from agenda setting through implementation and evaluation. The qualitative research methods implemented in this study were 30 in-depth interviews, 4 focus group discussions, expert consultancy, and 420 secondary data review. The data were analyzed by NVivo 7.0. Health insurance in Vietnam was introduced in 1992 and has been elaborated over a 20-year time frame. These processes relate to moving from a contingent to a gradually expanded target population, expanding the scope of the benefit package, and reducing the financial contribution from the insured. The target groups expanded to include 66.8% of the population by 2012. We characterized the policy process relating to UHI as incremental with a learning-by-doing approach, with an emphasis on increasing coverage rather than ensuring a basic service package and financial protection. There was limited involvement of civil society organizations and users in all policy processes. Intertwined political economy factors influenced the policy processes. Incremental policy processes, characterized by a learning-by-doing approach, is appropriate for countries attempting to introduce new health institutions, such as health insurance in Vietnam. Vietnam should continue to mobilize resources in sustainable and viable ways to support the target groups. The country should also adopt a multi-pronged approach to achieving universal access to health services, beyond health insurance.


Adjunctive liraglutide treatment in patients with persistent or recurrent type 2 diabetes after metabolic surgery (GRAVITAS): a randomised, double-blind, placebo-controlled trial.
Many patients with type 2 diabetes do not achieve sustained diabetes remission after metabolic (bariatric) surgery for the treatment of obesity. Liraglutide, a glucagon-like peptide-1 analogue, improves glycaemic control and reduces bodyweight in patients with type 2 diabetes. Our aim was to assess the safety and efficacy of liraglutide 1·8 mg in patients with persistent or recurrent type 2 diabetes after metabolic surgery. In the GRAVITAS randomised double-blind, placebo-controlled trial, we enrolled adults who had undergone Roux-en-Y gastric bypass or vertical sleeve gastrectomy and had persistent or recurrent type 2 diabetes with HbA1c levels higher than 48 mmol/mol (6·5%) at least 1 year after surgery from five hospitals in London, UK. Participants were randomly assigned (2:1) via a computer-generated sequence to either subcutaneous liraglutide 1·8 mg once daily or placebo, both given together with a reduced-calorie diet, aiming for a 500 kcal per day deficit from baseline energy intake, and increased physical activity. The primary outcome was the change in HbA1c from baseline to the end of the study period at 26 weeks, assessed in patients who completed the trial. Safety was assessed in the safety analysis population, consisting of all participants who received either liraglutide or placebo. This trial is registered with EudraCT, number 2014-003923-23, and the ISRCTN registry, number ISRCTN13643081. Between Jan 29, 2016, and May 2, 2018, we assigned 80 patients to receive either liraglutide (n=53) or placebo (n=27). 71 (89%) participants completed the study and were included in the principal complete-cases analysis. In a multivariable linear regression analysis, with baseline HbA1c levels and surgery type as covariates, liraglutide treatment was associated with a difference of -13·3 mmol/mol (-1·22%, 95% CI -19·7 to -7·0; p=0·0001) in HbA1c change from baseline to 26 weeks, compared with placebo. Type of surgery had no significant effect on the outcome. 24 (45%) of 53 patients assigned to liraglutide and 11 (41%) of 27 assigned to placebo reported adverse effects: these were mainly gastrointestinal and in line with previous experience with liraglutide. There was one death during the study in a patient assigned to the placebo group, which was considered unrelated to study treatment. These findings support the use of adjunctive liraglutide treatment in patients with persistent or recurrent type 2 diabetes after metabolic surgery. JP Moulton Foundation.


A method for studying knife tool marks on bone.
The characteristics of knife tool marks retained on hard tissues can be used to outline the shape and angle of a knife. The purpose of this study was to describe such marks on bone tissues that had been chopped with knives. A chopping stage with a gravity accelerator and a fixed bone platform was designed to reconstruct the chopping action. A digital microscope was also used to measure the knife angle (θ) and retained V-shape tool mark angle (ψ) in a pig skull. The κ value (elasticity coefficient; θ/ψ) was derived and recorded after the knife angle (θ) and the accompanied velocity were compared with the proportional impulsive force of the knife and ψ on the bone. The constant impulsive force revealed a correlation between the V-shape tool mark angle (ψ) and the elasticity coefficient (κ). These results describe the tool marks--crucial in the medicolegal investigation--of a knife on hard tissues.


Nitroxyl (HNO) as a vasoprotective signaling molecule.
Nitroxyl (HNO), the one electron reduced and protonated form of nitric oxide (NO(•)), is rapidly emerging as a novel nitrogen oxide with distinct pharmacology and therapeutic advantages over its redox sibling. Whilst the cardioprotective effects of HNO in heart failure have been established, it is apparent that HNO may also confer a number of vasoprotective properties. Like NO(•), HNO induces vasodilatation, inhibits platelet aggregation, and limits vascular smooth muscle cell proliferation. In addition, HNO can be putatively generated within the vasculature, and recent evidence suggests it also serves as an endothelium-derived relaxing factor (EDRF). Significantly, HNO targets signaling pathways distinct from NO(•) with an ability to activate K(V) and K(ATP) channels in resistance arteries, cause coronary vasodilatation in part via release of calcitonin-gene related peptide (CGRP), and exhibits resistance to scavenging by superoxide and vascular tolerance development. As such, HNO synthesis and bioavailability may be preserved and/or enhanced during disease states, in particular those associated with oxidative stress. Moreover, it may compensate, in part, for a loss of NO(•) signaling. Here we explore the vasoprotective actions of HNO and discuss the therapeutic potential of HNO donors in the treatment of vascular dysfunction.


An improved method of intralamellar keratoplasty in rats.
Intralamellar keratoplasty in inbred rats is a valuable method for the investigation of corneal allograft rejection. Present methods of intralamellar corneal grafting are tedious and time-consuming. In addition, induction of neovascularization, a prerequisite to inducing graft rejection, is variable. In an effort to overcome these problems, the authors have developed an improved method of intralamellar keratoplasty. Intralamellar pockets are formed by introducing a 30-g needle into the corneal stroma near the limbus. Approximately 10-25 microliters of lipopolysaccharide (LPS) (100 micrograms/ml) is injected into the stroma, forming a stromal bleb. This bleb is incised to form a pocket, resulting in the leakage of the injected liquid. The intralamellar pocket formation is completed by dissecting any remaining stromal fibers in the area of former bleb. Once the pocket has been formed, grafts of corneal tissue are inserted, and the incision is closed. This method of keratoplasty has the following advantages over previously reported methods: (1) It is more rapid and less tedious because the formation of the corneal bleb protects the anterior chamber from being invaded during the corneal incision; (2) It leads to a reproducible induction of neovascularization in every cornea so treated; (3) It results in a higher frequency of allograft rejection.


The impact of ammonia levels and dialysis on outcome in 202 patients with neonatal onset urea cycle disorders.
Neonatal onset hyperammonemia in patients with urea cycle disorders (UCDs) is still associated with high morbidity and mortality. Current protocols consistently recommend emergency medical and dietary management. In case of increasing or persistent hyperammonemia, with continuous or progressive neurological signs, dialysis is performed, mostly as ultima ratio. It is presently unknown whether the currently defined ammonia threshold (e.g., at 500 μmol/L) to start dialysis is useful to improve clinical outcome. A systematic review of clinical and biochemical data from published neonatal onset UCD patients was performed to identify factors determining clinical outcome and to investigate in which clinical and biochemical setting dialysis was most effective. A total of 202 patients (118 proximal and 84 distal UCDs) described in 90 case reports or case series were included according to predefined inclusion/exclusion criteria. Median age at onset was three days and mean ammonia that triggered start of dialysis was 1199 μmol/L. Seventy-one percent of all patients received any form of dialysis. Total mortality was 25% and only 20% of all patients had a "normal" outcome. In general, patients with higher ammonia levels were more likely to receive dialysis, but this had for most patients no influence on outcome. In conclusion, in severe neonatal onset hyperammonemia, the current practice of dialysis, which effectively clears ammonia, had no impact on outcome. It may be essential for improving outcome to initiate all available treatment options, including dialysis, as early as possible.


Why we should continue to study race...but do a better job: an essay on race, racism and health.
For at least a century, American medical and public health researchers have used race as a marker for biology, and have documented race-associated differences in health and illness behavior. The research has often been inappropriate and has led to abuses and erroneous conclusions about the role played by race in the production of disparities in health status. Consequently, some researchers have begun to advocate the abolition of medical and public health research using race. In this essay, I argue that, although the arguments against continuing to study race have some merit, more rather than fewer studies of race are needed. Researchers should be more careful in the use of race and more conscious of the limitations of the race concept.


Quantifying the sensitivity of scent detection dogs to identify fecal contamination on raw produce.
Consumption of raw produce commodities has been associated with foodborne outbreaks in the United States. In a recent Centers for Disease Control and Prevention report outlining the incidence of food-related outbreaks from 1998 to 2008, produce of all kinds were implicated in 46% of illnesses and 23% of deaths. Methods that quickly identify fecal contamination of foods, including produce, will allow prioritization of samples for testing during investigations and perhaps decrease the time required to identify specific brands or lots. We conducted a series of trials to characterize the sensitivity and specificity of scent detection dogs to accurately identify fecal contamination on raw agricultural commodities (romaine lettuce, spinach, cilantro, and roma tomatoes). Both indirect and direct methods of detection were evaluated. For the indirect detection method, two dogs were trained to detect contamination on gauze pads previously exposed to produce contaminated with feces. For the direct detection method, two dogs were trained to identify fecal contamination on fresh produce. The indirect method did not result in acceptable levels of sensitivity except for the highest levels of fecal contamination (25 g of feces). Each dog had more difficulty detecting fecal contamination on cilantro and spinach than on roma tomatoes. For the direct detection method, the dogs exhibited >75% sensitivity for detecting ≥0.25 g of feces on leafy greens (cilantro, romaine lettuce, and spinach) and roma tomatoes, with sensitivity declining as the amount of feces dropped below 0.025 g. We determined that use of a scent detection dog to screen samples for testing can increase the probability of detecting ≥0.025 g of fecal contamination by 500 to 3,000% when samples with fecal contamination are rare (≤1%).


Malignant carotid body tumors: report of two cases and review of the literature.
Paragangliomas may arise in any area of the body where nonchromaffin paraganglia are situated. These include the glomus jugulare, the carotid body, and the retroperitoneum. Malignant behavior of these paragangliomas has been a controversial subject with most authors, who consider them to be benign tumors. Some authors have diagnosed locally recurrent and infiltrative tumors as malignant, whereas most require the demonstration of metastatic potential as the only criterion for malignancy. Two cases of metastasizing carotid body tumors are reported with a review of the literature on this unusual neoplasm.


Foodproof Salmonella Detection Kit. Performance Tested Method 120301.
The foodproof Salmonella Detection Kit was previously validated in the Performance Tested Methods program for the detection of Salmonella species in a variety of foods, including milk powder, egg powder, coconut, cocoa powder, chicken breast, minced meat, sliced sausage, sausage, smoked fish, pasta, white pepper, cumin, dough, wet pet food, dry pet food, ice cream, watermelon, sliced cabbage, food dye, and milk chocolate. The method was shown to be equivalent to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) and the U.S. Department of Agriculture, Food Safety and Inspection Service's Microbiology Laboratory Guidebook reference culture procedures. In the first Emergency Response Validation (ERV) extension study, peanut butter was inoculated with S. enterica. ser Typhimurium. For the low inoculation level (1.08 CFU/25 g), a Chi-square value of 2.25 indicated that there was no significant performance difference between the foodproof Salmonella Detection Kit and the FDA-BAM reference method. For high-level inoculation (11.5 CFU/25 g) and uninoculated control, there was 100% agreement between the methods. In the second ERV extension study, peanut butter was inoculated with S. enterica. ser Typhimurium. For both inoculation levels (0.1 and 0.5 CFU/25 g by most probable number), Chi-square values of 0 indicated that there was no significant performance difference between foodproof Salmonella Detection Kit and the FDA-BAM reference method.


Cost-effectiveness of pharmacologic and invasive therapies for stroke prophylaxis in atrial fibrillation.
Atrial fibrillation is an abnormal heart rhythm characterized by rapid, disorganized activation (fibrillation) of the left and right atria of the heart, and is responsible for 15% of 700,000 strokes in the United States each year. There are multiple pharmacologic and nonpharmacologic therapies used for stroke prevention in atrial fibrillation, including vitamin K antagonists such as warfarin, antiplatelet agents such as aspirin and clopidogrel, and newer agents such as dabigatran, rivaroxaban and apixaban. Nonpharmacologic therapies involve excluding the left atrial appendage from the systemic circulation by surgical ligation or excision, percutaneous ligation, or endovascular implantation of a left atrial occlusion device. Because atrial fibrillation-related stroke is preventable, a comparison of the value of these interventions by cost-effectiveness analysis (CEA) could inform clinical and health policy recommendations. In this article, we review the principles of CEA and identify 11 articles that examine CEA of stroke prophylaxis strategies in atrial fibrillation. Although most studies evaluate aspirin and warfarin across a variety of atrial fibrillation stroke risk profiles, we also review new literature on new pharmacologic therapies such as direct thrombin inhibitors and discuss the potential value of device-based therapies.


[Intensity of virus multiplication in abortive rabies].
Experiments in random-bred and BALB/c white mice inoculated intraperitoneally with street virus, in Syrian hamsters inoculated intramuscularly with fixed viruses as well as experiments reproducing abortive rabies in 99-100% in random-bred white mice inoculated intramuscularly with fixed virus (ERA strain) showed that in the abortive infection the intensity of virus multiplication in the brain is lower than that in fatal infection, the differences being statistically significant. It was found out that from onset of signs of the disease interferon appeared to have no influence on the outcome of infection.


Improvements in cognition in patients converting from thrice weekly hemodialysis to nocturnal hemodialysis: a longitudinal pilot study.
Cognitive impairment has been documented in uremia with partial improvement after dialysis. Nocturnal daily hemodialysis (NHD) is a novel dialysis modality with multiple benefits. Previous reports have shown marked improvements in quality of life, cardiac function, resolution of peripheral vascular disease, and reversal of central sleep apnea. We hypothesized that patients maintained on NHD would have better cognitive functioning than those receiving conventional therapy. Using a longitudinal study design, patients were tested at baseline and again after >or=6 months NHD. At each of the two time points, a battery of 10 neuropsychological tests were used to evaluate three domains of cognitive functioning--attention and working memory skills, psychomotor efficiency and processing speed, and learning efficiency. Clinical subjective symptoms for cognitive functioning and depression were measured using the Patients Assessment of Own Functioning inventory and the Beck Depression Index. Twelve patients (six males, six females) were recruited. Patients were aged 39.6+/-3.3 years at the time of first testing. Thirty-three percent were diabetic, with a mean Charlson comorbidity score of 3.5+/-2.0. Depression (defined as >16 on the Beck Depression Index score) was not seen in any patient. Over the 6-month period, a 22% reduction in cognitive symptoms (P=0.01), 7% improvement in psychomotor efficiency and processing speed (P=0.02), and 32% improvement in attention and working memory (P=0.04) was seen. Learning efficiency scores were unchanged. NHD may be associated with improved general cognitive efficiency as measured by psychomotor efficiency and attention and working memory.


Nosology and classification of genetic skeletal disorders: 2019 revision.
The application of massively parallel sequencing technology to the field of skeletal disorders has boosted the discovery of the underlying genetic defect for many of these diseases. It has also resulted in the delineation of new clinical entities and the identification of genes and pathways that had not previously been associated with skeletal disorders. These rapid advances have prompted the Nosology Committee of the International Skeletal Dysplasia Society to revise and update the last (2015) version of the Nosology and Classification of Genetic Skeletal Disorders. This newest and tenth version of the Nosology comprises 461 different diseases that are classified into 42 groups based on their clinical, radiographic, and/or molecular phenotypes. Remarkably, pathogenic variants affecting 437 different genes have been found in 425/461 (92%) of these disorders. By providing a reference list of recognized entities and their causal genes, the Nosology should help clinicians achieve accurate diagnoses for their patients and help scientists advance research in skeletal biology.


Effects of soyabean meal- or whey-based diets on lipid metabolism in weaned piglets.
The present study aimed to test the hypothesis that dietary protein source influences lipid metabolism-related parameters weaned piglets. The effects of soyabean meal (SB) and whey proteins (WP) on gene expression of several genes involved in the lipogenic process in liver, visceral (VAT) and subcutaneous (SAT) adipose tissues, plasma insulin concentration and fatty acid (FA) profile were investigated in 18 weaned piglets. Weaned piglets were fed one of two diets containing either SB or WP as the main protein source. Following a 10-h fasting period, plasma insulin concentration and FA profile were assessed at 56 and 72 days of age, whereas gene expression in liver, VAT and SAT was assessed at 72 days of age. Plasma insulin concentration was not affected by diet, although it was 40% lower in SB fed pigs. The SB pigs had lower 14:0 (p < 0.01) and higher 18:3n-3 (p < 0.001) levels in plasma in comparison with WP pigs. However, these changes were attributed to background differences in the dietary FA profile and not to a direct protein source effect. Gene expression of sterol regulatory element-binding protein 1 (SREBP-1) in liver and VAT were lower (p < 0.01 and p < 0.05, respectively) in SB compared to WP fed piglets, but no differences occurred in SAT. No changes were observed in sterol regulatory element-binding protein 2, liver X receptor, peroxisome proliferator-activated receptors α and γ and plasminogen activator inhibitor 1 mRNA levels, either in liver or in adipose tissues. In conclusion, dietary protein source, accompanied likely by side alterations in the dietary composition, affects lipid metabolism in pigs through the downregulation of SREBP-1, which is a crucial determinant of lipogenic process.


Troponin is related to left ventricular mass and predicts all-cause and cardiovascular mortality in hemodialysis patients.
Cardiac troponin T (cTnT) predicts death and cardiovascular outcomes in clinically stable patients with end-stage renal disease. Because this protein is synthesized exclusively in myocardial cells, its predictive power for these outcomes may be because it reflects, besides cardiac ischemia, left ventricular (LV) mass, which is a strong predictor of cardiovascular death in this population per se. We tested the relationship between cTnT level and LV mass and the predictive power of this cardiac protein for all-cause and cardiovascular mortality in a cohort of hemodialysis patients (n = 199) without acute coronary syndrome and heart failure followed up for an average of 35 months (range, 0.8 to 52 months). cTnT was measured by means of a third-generation electrochemiluminescence immunoassay. cTnT level was related directly to interventricular septum (r = 0.36; P < 0.001) and posterior wall thickness (r = 0.40; P < 0.001), as well as LV mass (r = 0.45; P < 0.001). On multivariate analysis, after age, LV mass was the strongest independent predictor of cTnT level (beta = 0.28; P < 0.001). Serum cTnT level was significantly related to all-cause and cardiovascular mortality on univariate analysis (P < 0.001). On multivariate Cox regression analysis, the adjusted risk for all-cause death was 2.39 times (95% confidence interval [CI], 1.13 to 5.06; P = 0.02) greater in patients in the third cTnT tertile than the first tertile, and a similar pattern emerged for cardiovascular mortality (hazard ratio, 2.35; 95% CI, 1.01 to 5.49; P = 0.048). In hemodialysis patients, plasma cTnT level is independently related to LV mass and predicts all-cause and cardiovascular mortality. These data support the hypothesis that this marker can be usefully applied for risk stratification in clinically stable dialysis patients.


[Radiofrequency ablation of lung tumors].
Radiofrequency (RF) ablation is an effective therapy option in the treatment of primary and secondary liver tumors. Percutaneous RF ablation of primary and secondary pulmonary tumors is a possible new indication. Due to the minimally invasive character of percutaneous RF ablation, application in patients who are not candidates for curative radical resection is possible. Moreover, pulmonary RF ablation is a promising palliative therapy option. In this context, an increasing number of results regarding RF ablation of lung tumors were recently published. The present review summarizes and discusses the possible indications, techniques, results, and complications of pulmonary RF ablation.


Protein kinase A RI-alpha predicts for prostate cancer outcome: analysis of radiation therapy oncology group trial 86-10.
The RI-alpha regulatory subunit of protein kinase A type 1 (PKA) is constitutively overexpressed in human cancer cell lines and is associated with active cell growth and neoplastic transformation. This report examined the association between PKA expression and the endpoints of biochemical failure (BF), local failure (LF), distant metastasis (DM), cause-specific mortality (CSM), and overall mortality in men treated with radiotherapy, with or without short-term androgen deprivation in Radiation Therapy Oncology Group trial 86-10. Pretreatment archival diagnostic tissue samples from 80 patients were stained for PKA by immunohistochemical methods from a parent cohort of 456 cases. PKA intensity was scored manually and by image analysis. The Cox proportional hazards model for overall mortality and Fine and Gray's regression models for CSM, DM, LF and BF were then applied to determine the relationship of PKA expression to the endpoints. The pretreatment characteristics of the missing and determined PKA groups were not significantly different. On univariate analyses, a high PKA staining intensity was associated with BF (image analysis, continuous variable, p = 0.022), LF (image analysis, dichotomized variable, p = 0.011), CSM (manual analysis, p = 0.037; image analysis, continuous, p = 0.014), and DM (manual analysis, p = 0.029). On multivariate analyses, the relationships to BF (image analysis, continuous, p = 0.03), LF (image analysis, dichotomized, p = 0.002), and DM remained significant (manual analysis, p = 0.018). In terms of CSM, a trend toward an association was seen (manual analysis, p = 0.08; image analysis, continuous, p = 0.09). PKA overexpression was significantly related to patient outcome and is a potentially useful biomarker for identifying high-risk prostate cancer patients who might benefit from a PKA knockdown strategy.


[Cerebellar disfacilitation after pharmacological destruction of the inferior olive].
The total selective destruction of the inferior olive by 3-acetylpyridine produces a marked increase in the metabolic activity at the intracerebellar nuclei. This increase is locally suppressed by partial ablation or local inactivation of the corresponding regions in the cerebellar cortex, suggesting its localization in the intranuclear presynaptic terminals of the Purkinje cells. In a second series of experiments, it is demonstrated that Purkinje cells of the cerebellar cortex deprived of their climbing fibers and therefore of their complex spikes by destruction of the inferior olive, increase their firing frequency of simple spikes. As a consequence, there is a decreased frequency of discharge of their target neurones in the intracerebellar nuclei; the net result of suppression of the inferior olive is thus an increased cortico-nuclear inhibitory effect.


Autonomous beta-endorphin secretion from the pituitary neurointermediate lobe: in vivo studies.
The existence of independent control mechanisms of beta-endorphin (beta-EP) secretion from the anterior (AP) and intermediate (NIL) pituitary lobes is now ascertained. The aim of this study was to evaluate the effect of surgical separation from the hypothalamus of the two pituitary lobes on beta-EP secretion. Two experimental models of surgical hypothalamo-pituitary disconnection were used: 1) rats with ablation of the medial basal hypothalamus (MBH); 2) rats bearing two entire ectopic pituitaries or two anterior pituitaries (APs) only, transplanted under the kidney capsule. In rats with MBH-ablation plasma beta-EP levels were significantly higher than in sham-operated controls. Plasma beta-EP levels increased in rats transplanted with entire pituitaries 3 days after surgery and were still elevated after 1 week. In rats transplanted with APs only, no significant beta-EP changes in plasma were evident. In both experimental conditions no significant difference was present in beta-LPH plasma levels. Concentrations of beta-EP in the ectopic NILs decreased gradually after transplantation. In all these results indicate that that NIL but not the AP is capable, when is disconnected from the hypothalamus, or secreting autonomously beta-EP.


Porous Poly(ε-caprolactone)-Poly(l-lactic acid) Semi-Interpenetrating Networks as Superior, Defect-Specific Scaffolds with Potential for Cranial Bone Defect Repair.
The treatment of irregular cranial bone defects is currently limited due to the graft resorption that can occur when an ill-fitting interface exists between an autograft and the surrounding tissue. A tissue engineering scaffold able to achieve defect-specific geometries could improve healing. This work reports a macroporous, shape memory polymer (SMP) scaffold composed of a semi-interpenetrating network (semi-IPN) of thermoplastic poly(l-lactic acid) (PLLA) within cross-linked poly(ε-caprolactone) diacrylate (PCL-DA) that is capable of conformal fit within a defect. The macroporous scaffolds were fabricated using a fused salt template and were also found to have superior, highly controlled properties needed for regeneration. Specifically, the scaffolds displayed interconnected pores, improved rigidity, and controlled, accelerated degradation. Although slow degradation rates of scaffolds can limit healing, the unique degradation behavior observed could prove promising. Thus, the described SMP semi-IPN scaffolds overcome two of the largest limitations in bone tissue engineering: defect "fit" and tailored degradation.


Apoptotic signaling clears engineered Salmonella in an organ-specific manner.
Pyroptosis and apoptosis are two forms of regulated cell death that can defend against intracellular infection. When a cell fails to complete pyroptosis, backup pathways will initiate apoptosis. Here, we investigated the utility of apoptosis compared to pyroptosis in defense against an intracellular bacterial infection. We previously engineered Salmonella enterica serovar Typhimurium to persistently express flagellin, and thereby activate NLRC4 during systemic infection in mice. The resulting pyroptosis clears this flagellin-engineered strain. We now show that infection of caspase-1 or gasdermin D deficient macrophages by this flagellin-engineered S. Typhimurium induces apoptosis in vitro. Additionally, we engineered S. Typhimurium to translocate the pro-apoptotic BH3 domain of BID, which also triggers apoptosis in macrophages in vitro. During mouse infection, the apoptotic pathway successfully cleared these engineered S. Typhimurium from the intestinal niche but failed to clear the bacteria from the myeloid niche in the spleen or lymph nodes. In contrast, the pyroptotic pathway was beneficial in defense of both niches. To clear an infection, cells may have specific tasks that they must complete before they die; different modes of cell death could initiate these 'bucket lists' in either convergent or divergent ways.


Dose response for ultraviolet radiation A-induced focal melanocytic hyperplasia and nonmelanoma skin tumors in Monodelphis domestica.
Four groups of 30 dorsally shaved opossums (Monodelphis domestica) were exposed to graded doses of ultraviolet radiation A (UVA) (320-400 nm) three times per week for 90 weeks. Animals were monitored for the appearance of focal melanocytic hyperplasia (FMH) and nonmelanoma skin tumors (NMST) during the course of the exposures and for an additional 20 weeks following termination of exposures. FMH is the putative precursor for melanoma in the opossum. The lowest dose of UVA (2.5 x 10(3) J/m2) used in this study was selected based on the action spectrum for the induction of melanoma in a fish model. The prediction was that 2.5 x 10(3) J/m2 would induce FMH in the opossum if the action spectra for the induction of FMH in the opossum and melanoma in the fish were the same. The highest UVA dose was 2.5 x 10(5) J/m2. Only the highest dose of UVA gave a statistically significant induction of FMH and NMST in the opossum. As in previous studies, the FMH appeared earlier than the NMST during the course of exposures and the final prevalence of FMH was lower than the final prevalence of NMST. Overall, the results of this study indicate that the efficacy of UVA to induce FMH in the opossum is not as great as would be predicted from the action spectrum for melanoma induction in a fish model.


Use of biologics for treatment of autoimmune inner ear disease.
Biologic medications are novel therapeutics in the treatment of Autoimmune Inner Ear Disease (AIED), an etiology of Sensorineural Hearing Loss (SNHL). The goal of this study is to review the currently available literature on the efficacy of biologic medications on autoimmune-mediated hearing loss and associated symptomology among patients with AIED. A systematic review of Pubmed, Scopus, Cochrane, and Web of Science databases was conducted to identify studies investigating the impact of biologic medications on hearing outcomes. Bias assessment was independently conducted by three authors and studies were stratified based on risk of bias. Of 174 unique abstracts screened, 12 articles met inclusion criteria for formal review. One randomized control trial, seven prospective cohort studies, and four retrospective cohort studies were included. Seven biologic medications, Etanercept, Infliximab, Adalimumab, Golimumab, Rituximab, Anakinra, and Canakinumab, were identified targeting three unique molecular targets, TNF-α, CD20, and IL-1. The effects of biologic medications in treating SNHL was highly variable without clear efficacy of a drug or drug category, likely due to rarity of disease, multifactorial etiologies of AIED, and cohort heterogeneity. However, several medications alleviate symptoms associated with AIED, such as vertigo and tinnitus. While biologic medications may be promising therapeutics in AIED patients, the evidence is currently inconclusive. Large-scale randomized control trials and prospective cohort reviews are required to establish the efficacy of biologic medications in treating hearing loss.


Glyoxalase III enhances salinity tolerance through reactive oxygen species scavenging and reduced glycation.
Methylglyoxal (MG) is a metabolically generated highly cytotoxic compound that accumulates in all living organisms, from Escherichia coli to humans, under stress conditions. To detoxify MG, nature has evolved reduced glutathione (GSH)-dependent glyoxalase and NADPH-dependent aldo-keto reductase systems. But both GSH and NADPH have been reported to be limiting in plants under stress conditions, and thus detoxification might not be performed efficiently. Recently, glyoxalase III (GLY III)-like enzyme activity has been reported from various species, which can detoxify MG without any cofactor. In the present study, we have tested whether an E. coli gene, hchA, encoding a functional GLY III, could provide abiotic stress tolerance to living systems. Overexpression of this gene showed improved tolerance in E. coli and Saccharomyces cerevisiae cells against salinity, dicarbonyl, and oxidative stresses. Ectopic expression of the E. coli GLY III gene (EcGLY-III) in transgenic tobacco plants confers tolerance against salinity at both seedling and reproductive stages as indicated by their height, weight, membrane stability index, and total yield potential. Transgenic plants showed significantly increased glyoxalase and antioxidant enzyme activity that resisted the accumulation of excess MG and reactive oxygen species (ROS) during stress. Moreover, transgenic plants showed more anti-glycation activity to inhibit the formation of advanced glycation end product (AGE) that might prevent transgenic plants from stress-induced senescence. Taken together, all these observations indicate that overexpression of EcGLYIII confers salinity stress tolerance in plants and should be explored further for the generation of stress-tolerant plants.


Biochemical correlates of antidepressant response. Results of a trazodone versus amitriptyline trial.
Changes in the uptake of 3H-serotonin into platelets, serotonin receptor activity on platelets, and plasma free and bound tryptophan concentrations were determined in a group of 54 moderately to severely depressed patients (mean Hamilton Rating Scale 17) before and during treatment with either trazodone or amitriptyline. No difference was found between the free and bound tryptophan concentrations of the depressed patients or their controls nor was any change detected during drug treatment. Platelet serotonin receptor activity and uptake were significantly lower in the depressed patients than in the controls and in those patients which subsequently responded to drug treatment. There was no apparent correlation between the ability of trazodone to inhibit the uptake of serotonin into the platelets and the antidepressant response; all patients responding to drug treatment showed an enhanced serotonin uptake. There is evidence from this study that changes in serotonin receptor function and uptake into platelets may correlate with the clinical status of depressed patients.


i-Motif DNA: structural features and significance to cell biology.
The i-motif represents a paradigmatic example of the wide structural versatility of nucleic acids. In remarkable contrast to duplex DNA, i-motifs are four-stranded DNA structures held together by hemi- protonated and intercalated cytosine base pairs (C:C+). First observed 25 years ago, and considered by many as a mere structural oddity, interest in and discussion on the biological role of i-motifs have grown dramatically in recent years. In this review we focus on structural aspects of i-motif formation, the factors leading to its stabilization and recent studies describing the possible role of i-motifs in fundamental biological processes.


Cyclic Tetra-Adenylate (cA4) Recognition by Csa3; Implications for an Integrated Class 1 CRISPR-Cas Immune Response in Saccharolobus solfataricus.
Csa3 family transcription factors are ancillary CRISPR-associated proteins composed of N-terminal CARF domains and C-terminal winged helix-turn-helix domains. The activity of Csa3 transcription factors is thought to be controlled by cyclic oligoadenyate (cOA) second messengers produced by type III CRISPR-Cas surveillance complexes. Here we show that Saccharolobus solfataricus Csa3a recognizes cyclic tetra-adenylate (cA4) and that Csa3a lacks self-regulating "ring nuclease" activity present in some other CARF domain proteins. The crystal structure of the Csa3a/cA4 complex was also determined and the structural and thermodynamic basis for cA4 recognition are described, as are conformational changes in Csa3a associated with cA4 binding. We also characterized the effect of cA4 on recognition of putative DNA binding sites. Csa3a binds to putative promoter sequences in a nonspecific, cooperative and cA4-independent manner, suggesting a more complex mode of transcriptional regulation. We conclude the Csa3a/cA4 interaction represents a nexus between the type I and type III CRISPR-Cas systems present in S. solfataricus, and discuss the role of the Csa3/cA4 interaction in coordinating different arms of this integrated class 1 immune system to mount a synergistic, highly orchestrated immune response.


[The use of Cardioxane in chemotherapy of malignant neoplasms in children].
Treatment with anthracycline antibiotics may have a toxic effect on the myocardium. The report deals with the results of application of a cardioprotector--cardioxan--in 38 children, aged 2-13 years, with malignant tumors treated, among other drugs, with doxorubicin. These data were compared with those on 33 pediatric patients who had received anthracycline antibiotics until 1994. No cardioxan-related complications were recorded whenever the instructions on the use were followed. The results are inconclusive because of the short duration of the study and examination being still incomplete.


Jigless Knotless Internal Brace Versus Other Minimal Invasive Achilles Tendon Repair Techniques in Biomechanical Testing Simulating the Progressive Rehabilitation Protocol.
The jigless knotless internal brace surgery (JKIB), an alternative method for minimal invasive surgery (MIS) repair of acute Achilles tendon rupture, has advantages of preventing sural-nerve injury in MIS and superficial wound infection in open surgery, as previous clinical research demonstrates. However, no comparative study on the biomechanical performance between JKIB and other MIS techniques has been reported until now. In this study, 50 fresh porcine Achilles tendons were used to compare the JKIB with open surgery (two-stranded Krachow suture) with other MIS techniques, including Percutaneus Achilles Repair System (PARS), Speedbridge (SB), and Achillon Achilles Tendon Suture System (ACH), using a biomechanical testing with cyclic loading at 1 Hz. This test was used to simulate a progressive rehabilitation protocol where 20 to 100 N was applied in the first 250 cycles, followed by 20 to 190 N in the second 250 cycles, and then 20 to 369 N in the third 250 cycles. The cyclic displacement after 10, 100 and 250 cycles were recorded. The survived cycles were defined as a sudden drop in measured load. In survived cycles, the JKIB group (552.3 ± 72.8) had significantly higher cycles than the open, PARS, and ACH groups (204.3 ± 33.3, 395.9 ± 96.0, and 397.1 ± 80.9, respectively, p < .01) as analyzed by post hoc analysis, but no significant difference as compared with the SB group (641.6 ± 48.7). In cyclic displacement after 250 cyclic loadings, the JKIB group (11.29 ± 1.29) showed no significant difference as compared with PARS, SB, and ACH groups (12.21 ± 1.18, 9.80 ± 0.80, and 11.57 ± 1.10 mm, respectively) and significant less displacement than the open group (14.50 ± 1.85, p < .01). These findings suggest that JKIB could be an option for acute Achilles tendon repair in the MIS fashion due to no larger cyclic elongation compared with other MIS techniques.


A biomechanical model of swallowing for understanding the influence of saliva and food bolus viscosity on flavor release.
After swallowing a liquid or a semi-liquid food product, a thin film responsible for the dynamic profile of aroma release coats the pharyngeal mucosa. The objective of the present article was to understand and quantify physical mechanisms explaining pharyngeal mucosa coating. An elastohydrodynamic model of swallowing was developed for Newtonian liquids that focused on the most occluded region of the pharyngeal peristaltic wave. The model took lubrication by saliva film and mucosa deformability into account. Food bolus flow rate and generated load were predicted as functions of three dimensionless variables: the dimensionless saliva flow rate, the viscosity ratio between saliva and the food bolus, and the elasticity number. Considering physiological conditions, the results were applied to predict aroma release kinetics. Two sets of conditions were distinguished. The first one was obtained when the saliva film is thin, in which case food bolus viscosity has a strong impact on mucosa coating and on flavor release. More importantly, we demonstrated the existence of a second set of conditions. It was obtained when the saliva film is thick and the food bolus coating the mucosa is very diluted by saliva during the swallowing process and the impact of its viscosity on flavor release is weak. This last phenomenon explains physically in vivo observations for Newtonian food products found in the literature. Moreover, in this case, the predicted thickness of the mix of food bolus with saliva coating the mucosa is approximately of 20 μm; value in agreement with orders of magnitude found in the literature.


Hydrogel based lipid-oligonucleotides: a new route to self-delivery of therapeutic sequences.
Synthetic OligoNucleotides (ON) provide promising therapeutic tools for controlling specifically genetic expression in a broad range of diseases from cancers to viral infections. Beside their chemical stability and intracellular delivery, the controlled release of therapeutic sequences remains an important challenge for successful clinical applications. In this work, Lipid-OligoNucleotide (LON) conjugates stabilizing hydrogels are reported and characterized by rheology and cryo-scanning electron microscopy (cryo-SEM). These studies revealed that lipid conjugation of antisense oligonucleotides featuring partial self-complementarity resulted in entangled pearl-necklace networks, which were obtained through micelle-micelle interaction driven by duplex formation. Owing to these properties, the Lipid AntiSense Oligonucleotide (LASO) sequences exhibited a prolonged release after subcutaneous administration compared to the non-lipidic antisense (ASO) one. The LASO self-assembly based hydrogels obtained without adjuvant represent an innovative approach for the sustained self-delivery of therapeutic oligonucleotides.


The analgesic power of pleasant touch in individuals with chronic pain: Recent findings and new insights.
This mini-review covers recent works on the study of pleasant touch in patients with chronic pain (CP) and its potential use as a treatment. While experiments have demonstrated that pleasant touch, through the activation of CT-afferents and the brain regions involved in its affective value, might reduce the unpleasantness and intensity of induced pain, the interaction between pleasant touch and CP remains under-examined. Some experiments show that CP might disrupt the positive aspects of receiving pleasant touch, while in other studies the perception of pleasantness is preserved. Moreover, only a few attempts have been made to test whether touch can have a modulatory effect on CP, but these results also remain inconclusive. Indeed, while one recent study demonstrated that CT-touch can diminish CP after a short stimulation, another study suggested that pleasant touch might not be sufficient. Future studies should further investigate the psychological and neural interplay between pleasant touch and CP. In the conclusion of this mini-review, we propose a new tool we have recently developed using immersive virtual reality (IVR).


Novel adenoviruses detected in British mustelids, including a unique Aviadenovirus in the tissues of pine martens (Martes martes).
Several adenoviruses are known to cause severe disease in veterinary species. Recent evidence suggests that canine adenovirus type 1 (CAV-1) persists in the tissues of healthy red foxes (Vulpes vulpes), which may be a source of infection for susceptible species. It was hypothesized that mustelids native to the UK, including pine martens (Martes martes) and Eurasian otters (Lutra lutra), may also be persistently infected with adenoviruses. Based on high-throughput sequencing and additional Sanger sequencing, a novel Aviadenovirus, tentatively named marten adenovirus type 1 (MAdV-1), was detected in pine marten tissues. The detection of an Aviadenovirus in mammalian tissue has not been reported previously. Two mastadenoviruses, tentatively designated marten adenovirus type 2 (MAdV-2) and lutrine adenovirus type 1 (LAdV-1), were also detected in tissues of pine martens and Eurasian otters, respectively. Apparently healthy free-ranging animals may be infected with uncharacterized adenoviruses with possible implications for translocation of wildlife.


Relationship between antimicrobial peptides-induced cell membrane damage and bactericidal activity.
Most antimicrobial peptides (AMPs) act by killing bacterial cells. However, there is little information regarding the required interaction time between AMPs and bacterial cells to exert the bactericidal activity. One of the causes of the bactericidal activity is considered to be cell membrane damage, although little direct evidence is available. Here, we investigated the relationship between AMP-induced cell membrane damage in Escherichia coli and AMP-induced cell death at the single-cell level. Magainin 2, lactoferricin B, and PGLa were selected as the AMPs. First, we examined the interaction time (t) of AMPs with cells required to induce cell death using the single-cell analysis. The fraction of microcolonies containing only a single cell, Psingle (t), which indicates the fraction of dead cells, increased with time to reach ∼1 in a short time (≤5 min). Then, we examined the interaction between AMPs and single cells using confocal laser scanning microscopy in the presence of membrane-impermeable SYTOX green. Within a short time interaction, the fluorescence intensity of the cells due to SYTOX green increased, indicating that AMPs induced cell membrane damage through which the dye entered the cytoplasm. The fraction of cells in which SYTOX green entered the cytoplasm among all examined cells after the interaction time (t), Pentry (t), increased with time, reaching ∼1 in a short time (≤5 min). The values of Psingle (t) and Pentry (t) were similar at t ≥ 3 min for all AMPs. The bindings of AMPs to cells were largely reversible, whereas the AMP-induced cell membrane damages were largely irreversible because SYTOX green entered the cells after dilution of AMP concentration. Based on these results, we conclude that the rapid, substantial membrane permeabilization of cytoplasmic contents after a short interaction time with AMPs and the residual damage after dilution induce cell death.


Uptake of glutamate into synaptic vesicles by an inorganic phosphate transporter.
Previous work has identified two families of proteins that transport classical neurotransmitters into synaptic vesicles, but the protein responsible for vesicular transport of the principal excitatory transmitter glutamate has remained unknown. We demonstrate that a protein that is unrelated to any known neurotransmitter transporters and that was previously suggested to mediate the Na(+)-dependent uptake of inorganic phosphate across the plasma membrane transports glutamate into synaptic vesicles. In addition, we show that this vesicular glutamate transporter, VGLUT1, exhibits a conductance for chloride that is blocked by glutamate.


Single cell transcriptome analysis of developing arcuate nucleus neurons uncovers their key developmental regulators.
Despite the crucial physiological processes governed by neurons in the hypothalamic arcuate nucleus (ARC), such as growth, reproduction and energy homeostasis, the developmental pathways and regulators for ARC neurons remain understudied. Our single cell RNA-seq analyses of mouse embryonic ARC revealed many cell type-specific markers for developing ARC neurons. These markers include transcription factors whose expression is enriched in specific neuronal types and often depleted in other closely-related neuronal types, raising the possibility that these transcription factors play important roles in the fate commitment or differentiation of specific ARC neuronal types. We validated this idea with the two transcription factors, Foxp2 enriched for Ghrh-neurons and Sox14 enriched for Kisspeptin-neurons, using Foxp2- and Sox14-deficient mouse models. Taken together, our single cell transcriptome analyses for the developing ARC uncovered a panel of transcription factors that are likely to form a gene regulatory network to orchestrate fate specification and differentiation of ARC neurons.


Functional and molecular analysis of Escherichia coli strains lacking multiple DEAD-box helicases.
DEAD-box RNA helicases are enzymes that unwind RNA duplexes and are found in virtually all organisms. Most organisms harbor multiple DEAD-box helicases, suggesting that these factors participate in distinct aspects of RNA metabolism. To define the individual and collective contribution of the five DEAD-box helicases in the bacterium Escherichia coli (E. coli), nonpolar deletion mutants lacking single or multiple DEAD-box genes were constructed. An analysis of the single-deletion strains indicated that the absence of either the DeaD or SrmB RNA helicase causes growth and/or ribosomal defects under typical laboratory growth conditions. The analysis of strains lacking multiple DEAD-box genes showed cumulative growth defects at low temperatures. A strain deleted for all five DEAD-box genes was also constructed for these studies, representing the first time all DEAD-box genes have been removed in any organism. Additional investigations revealed that the growth and ribosomal defects of such a DEAD-box deficient strain can be sharply attenuated under alternative conditions, indicating that the defects caused by a lack of DEAD-box genes are modulated by growth context.


Endogenous ghrelin and 5-HT regulate interdigestive gastrointestinal contractions in conscious rats.
Endogenous ghrelin causes interdigestive contractions of the stomach in rats. In contrast, previous studies showed that 5-HT(3) and 5-HT(4) receptors were involved in regulating intestinal interdigestive contractions. We studied the possible role of endogenous ghrelin and 5-HT regulating interdigestive gastrointestinal (GI) contractions in rats. Four strain gauge transducers were implanted on the antrum, duodenum, and proximal and distal jejunum. After an overnight fast, GI contractions were recorded in freely moving conscious rats and ghrelin receptor antagonists [(d-lys3)GHRP6; 1 micromol/kg], 5-HT(3) antagonists (Ondansetron; 0.5 mg/kg) and 5-HT(4) antagonists (GR 125,487; 1 mg/kg) were administered (bolus iv). To evaluate the relationship between the luminal concentrations of 5-HT and phase III-like contractions of the duodenum, duodenal juice was collected via the intraduodenal catheter. 5-HT content of the duodenal juice was measured by HPLC. (d-lys3)GHRP6 significantly attenuated the occurrence and amplitude of phase III-like contractions of the antrum, but not the duodenum and jejunum. 5-HT(4) antagonists significantly reduced spontaneous phase III-like contractions of the jejunum, without affecting those of the antrum and duodenum. In contrast, 5-HT(3) antagonists did not affect phase III-like contractions in GI tract. Luminal concentration of 5-HT at the phase III-like contraction (36.0 +/- 13.3 ng/ml, n = 9) was significantly higher than that at the phase I-like contractions of the duodenum (4.9 +/- 1.6 ng/ml, n = 9, P < 0.05). It is suggested that released ghrelin from the gastric mucosa mediates gastric phase III-like contractions, whereas 5-HT released from enterochromaffin cells of the duodenal mucosa mediates intestinal phase III-like contractions via 5-HT(4) receptors.


Role of SOX2 in the etiology of embryonal carcinoma, based on analysis of the NCCIT and NT2 cell lines.
The transcription factor SOX2, associated with amongst others OCT3/4, is essential for maintenance of pluripotency and self-renewal of embryonic stem cells. SOX2 is highly expressed in embryonal carcinoma (EC), the stem cell component of malignant nonseminomatous germ cell tumors, referred to as germ cell cancer (GCC). In fact, OCT3/4 together with SOX2 is an informative diagnostic tool for EC in a clinical setting. Several studies support the hypothesis that SOX2 is a relevant oncogenic factor in various cancers and recently, SOX2 has been suggested as a putative therapeutic target for early stage EC. We demonstrate the presence of genomic amplification of SOX2 in an EC cell line, NCCIT, using array comparative genome hybridization and fluorescence in situ hybridization. Down-regulation of SOX2 by targeted siRNA provokes NCCIT cells towards apoptosis, while inhibition of OCT3/4 expression induced differentiation, with retained SOX2 levels. Mice pluripotent xenografts from NCCIT (N-NCCIT and N2-NCCIT) show a consistent SOX2 expression, in spite of loss of the expression of OCT3/4, and differentiation, with retained presence of genomic amplification. No SOX2 amplification has been identified in primary pure and mixed EC in vivo patient samples so far. The data presented in this study are based on a single EC cell line with a SOX2 amplification, with NT2 as control EC cell line, showing no profound induction of apoptosis upon SOX2 downregulation. The findings are of relevance to identify mechanisms involved in the pathogenesis of EC tumors, and support the model of SOX2-oncogene dependency of EC, which however, does not exclude induction of differentiation. This finding is likely related to the presence of wild type p53 in GCC, resulting in expression of downstream target genes, amongst others miR-34a, miR-145 and SOX2, associated to the unique sensitivity of GCC to DNA damaging agents.


Selective loss and preservation of lake water dissolved organic matter fluorescence during long-term dark incubations.
The biogeochemical processing of dissolved organic matter (DOM) in inland waters is inherently related to its molecular structure and ecological function. Controlled bioassays are a valuable tool to analyze these relationships, but are seldom conducted and compared at temporal scales that typically prevail in natural inland waters. Here we incubated water from six boreal lakes in the dark and examined changes to the initial fluorescence and absorbance after 3.5 years. We identified five fluorescence components with parallel factor (PARAFAC) analysis (C(C,) C(M), C(A), C(X) and C(T)) and found a consistent change in the relative intensity of two dominant PARAFAC components (increase in C(A):C(C), corresponding to Peak A:Peak C), commonly found in lake water, that represent terrestrially-derived DOM. Surprisingly, we only found minor changes to specific absorbance (SUVA), and did not find any changes to other spectral indexes including the fluorescence index, humification index and freshness index. By incorporating lakes spanning a wide range of initial total organic carbon concentrations (3.7 to 32.5 mg L(-1)), water residence times, and spectral characteristics (e.g. SUVA 1.13 to 3.77 L·mg C(-1)·m(-1)), we found that the relative intensities of two humic-like peaks were the most revealing of changes to DOM structure during dark incubations. We also verified that inner filter effects were adequately corrected within the concentration range of incubated samples. Thus, the processing of DOM under dark conditions, including microbial decomposition and flocculation, may have a greater influence on the humic-like peaks, particularly C(C) (Peak C), with negligible changes to more commonly used spectral indexes.


Adrenoreceptors and renal function.
The effects of beta- and alpha-adrenoreceptor blockade on renal function are reviewed briefly, and a detailed analysis of the effects of labetalol, an antihypertensive drug that blocks both alpha and beta adrenoreceptors, is presented. In most published studies, no significant change in any parameter of renal function is seen during labetalol therapy. Data from our institution are presented in detail demonstrating the effects of labetalol on renal function in 17 patients with either normal renal function or varying degrees of renal impairment. No consistent change in glomerular filtration rate, renal plasma flow, filtration fraction, maximal concentrating ability, or free-water clearance is observed. Renal vascular resistance tends to fall in those patients with relatively normal renal function and tends to change inconsistently in patients with more impaired renal function. Labetalol appears, in summary, to have little negative impact on renal function.


Concise Review: Induced Pluripotent Stem Cell-Based Drug Discovery for Mitochondrial Disease.
High attrition rates and loss of capital plague the drug discovery process. This is particularly evident for mitochondrial disease that typically involves neurological manifestations and is caused by nuclear or mitochondrial DNA defects. This group of heterogeneous disorders is difficult to target because of the variability of the symptoms among individual patients and the lack of viable modeling systems. The use of induced pluripotent stem cells (iPSCs) might significantly improve the search for effective therapies for mitochondrial disease. iPSCs can be used to generate patient-specific neural cell models in which innovative compounds can be identified or validated. Here we discuss the promises and challenges of iPSC-based drug discovery for mitochondrial disease with a specific focus on neurological conditions. We anticipate that a proper use of the potent iPSC technology will provide critical support for the development of innovative therapies against these untreatable and detrimental disorders. Stem Cells 2017;35:1655-1662.


Genomically Informed Strain-Specific Recovery of Shiga Toxin-Producing Escherichia coli during Foodborne Illness Outbreak Investigations.
Next-generation sequencing plays an important role in the characterization of clinical bacterial isolates for source attribution purposes during investigations of foodborne illness outbreaks. Once an illness cluster and a suspect food vehicle have been identified, food testing is initiated for confirmation and to determine the scope of a contamination event so that the implicated lots may be removed from the marketplace. For biochemically diverse families of pathogens such as Shiga toxin-producing Escherichia coli (STEC), the ability to detect specific strains may be hampered by the lack of a universal selective enrichment approach for their recovery against high levels of background microbiota. The availability of whole genome sequence data for a given outbreak STEC strain prior to commencement of food testing may provide food microbiologists an opportunity to customize selective enrichment techniques favoring the recovery of the outbreak strain. Here we demonstrate the advantages of using the publicly available ResFinder tool in the analysis of STEC model strains belonging to serotypes O111 and O157 to determine antimicrobial resistance traits that can be used in formulating strain-specific enrichment media to enhance recovery of these strains from microbiologically complex food samples. The improved recovery from ground beef of model STEC strains with various antimicrobial resistance profiles was demonstrated using three classes of antibiotics as selective agents, suggesting the universal applicability of this new approach in supporting foodborne illness investigations.


Median to radial nerve transfer after traumatic radial nerve avulsion in a pediatric patient.
This case report describes an isolated radial nerve avulsion in a pediatric patient, treated by combination sensory and motor median to radial nerve transfers. After traumatic avulsion of the proximal radial nerve, a 12-year-old male patient underwent end-to-end transfer of median nerve branches to flexor carpi radialis and flexor digitorum superficialis to the posterior interosseous nerve and extensor carpi radialis nerve, respectively. He underwent end-to-side sensory transfer of the superficial radial sensory to the median sensory nerve. Pronator teres to extensor carpi radialis brevis tendon transfer was simultaneously performed to power short-term wrist extension. Within months after surgery, the patient had regained 9-10/10 sensation in the hand and forearm. In the following months and years, he regained dexterity, independent fine-finger and thumb motions, and 4-5/5 strength in all extensors except the abductor pollicis longus muscle. He grew 25 cm without extremity deformity or need for secondary orthopedic procedures. In appropriate adult and pediatric patients with proximal radial nerve injuries, nerve transfers have advantages over tendon transfers, including restored independent fine finger motions, regained sensation, and reinnervation of multiple muscle groups with minimal donor sacrifice.


Allografting in total knee replacement arthroplasty.
Revision, and complicated primary total knee arthroplasty (TKA), may present with significant structural bony defects. Allograft offers blocks of material that can be shaped to the defect. The opportunity to gain a biological bond may theoretically enhance loading conditions and enhance any subsequent surgery. However, the biomechanical and biological issues of immune response, infection, graft incorporation, resorption, and load transfer render allografting a powerful but problematic technology.


Paper-based platform for detection by hybridization using intrinsically labeled fluorescent oligonucleotide probes on quantum dots.
A paper-based platform was investigated in which the selective detection of oligonucleotide targets by hybridization was accomplished via the enhancement of fluorescence emission from intrinsically labeled DNA probes that were immobilized on the surface of quantum dots (QDs). Multiple copies of a derivative of thiazole orange, an intercalating dye known to form non-emissive dimers, were conjugated to single-stranded oligonucleotide probes. Dimerization resulted in the formation of H-aggregates where excitonic interactions led to the suppression of fluorescence. The hybridization of the oligonucleotide probe with a complementary target resulted in the enhancement of fluorescence emission as the dimers dissociated and the dyes preferentially intercalated with the duplex. The detection of oligonucleotide targets using this configuration eliminated the need for labeling the target strands, and fluorescence intensity was proportional to the extent of hybridization. In addition, the dye molecules were excited using Foerster Resonance Energy Transfer (FRET) from QD donors, which resulted in improved selectivity and allowed for ratiometric detection. A solution-phase hybridization assay based on similar operational principles has been previously reported, and this new work investigated the advantages offered for this transduction scheme using paper-based solid-phase substrates. QD-probe conjugates were immobilized in sufficient density on the paper matrix to provide for multiple-donor-multiple-acceptor interactions that resulted in a 20-fold enhancement of acceptor emission compared to the solution-based assay, providing a limit of detection of 0.1 pmol. The paper-based assay provided for the reduction of the time needed for sample preparation and data acquisition, demonstrated that transduction was possible in a complex matrix (goat serum) without compromising on the performance observed in buffer solution, and that oligonucleotides generated from standard PCR amplification could be detected.


Preoperative hormonal stimulation effect on hypospadias repair complications: Meta-analysis of observational versus randomized controlled studies.
Preoperative hormonal stimulation (PHS) is commonly administered to improve hypospadias outcomes. However, current literature provides no clear evidence to support its use, and controversies exist regarding its effect on postoperative complications. Herein we systematically evaluate the effect of PHS on postoperative complications rates following hypospadias repair. Comprehensive literature search performed on April 2016 including OVID MEDLINE, EMBASE, Web of Science, Cochrane Library, Clinicaltrials.gov, World Health Organization (WHO) International Clinical Trial Registry Platform (ICTRP), and ProQuest. Study quality assessment followed the Cochrane collaboration recommendation using risk of bias assessment, ROBINS-I and NOQAS. Effect estimates were extracted as relative risk (RR) with 95% confidence interval (CI) and pooled using Mantel-Haenzel method. Evidence quality was assessed using GRADE approach. Protocol Registration: PROSPERO CRD42016037881. Twenty-one publications from 16 trials examining the effect of androgenic PHS on postoperative outcome were included. Subgroup analysis based on study design (including nine cohort studies with moderate to high risk of bias for patient selection) showed no significant difference on postoperative outcomes (RR 1.31, 95% CI 0.95-1.81) (table). Pooled effect estimates with moderate quality of evidence from three randomized controlled trials suggest that significant lesser postoperative complications occur among patients exposed to PHS (RR 0.36, 95% CI 0.20-0.65). Uniformly, the reported adverse effects related to PHS were mild and transient, with good overall tolerance. Effect estimates generated from the better quality of evidence showed that PHS may decrease postoperative complications following hypospadias repair. Future studies should aim at identifying indicated subgroup of patients that will benefit from it and standardizing the dose and mode of delivery for best clinical result.


Evaluation and comparison of three different separation techniques for analysis of retroamide enantiomers and their biological evaluation against h-P2X7 receptor.
The P2X receptors are seven-transmembrane domain G protein-coupled receptors and the 7 subtypes of P2X receptors identified in humans, and named P2X1 to P2X7, are channel receptors whose endogenous ligand is ATP. New antagonists of the P2X7 receptor were developed, since this purinergic receptor was highlighted to be involved in many diseases such as different types of pain, cancer, ischemia, neurodegenerative diseases (including Parkinson's and Alzheimer's diseases) characterized by inflammatory processes. With the aim of evaluate the impact of chirality on the pharmacological activity of a new P2X7R antagonist, a semi-preparative method was developed in supercritical fluid chromatography (SFC). Among four polysaccharide based chiral stationary phases: Chiralcel OD-H and OJ-H and Chiralpak AS-H and AD-H, the last one namely amylose tris (3,5-dimethylphenylcarbamate) with a mobile phase consisted of carbon dioxide-ethanol (80:20, v/v), led to the successful separation of the enantiomers in short run time and with good resolution. Limits of detection and quantification were calculated and were found equal for compound 1, to 1.37 μM and 4.57 μM respectively, for peak 1 and were equal to 1.60 μM and 5.30 μM respectively, for peak 2 at λ=210 nm. Before carrying out the pharmacological evaluation of each enantiomer, two complementary methodologies, e.g. liquid chromatography and capillary electrophoresis were performed in parallel to improve the limits of detection and quantification to assess the enantiomeric purity. HPLC using a Chiralpak AD stationary phase led to four times lower limits of detection and quantification with regard to SFC. In the same time, capillary electrophoresis involving dual cyclodextrins system constituted of a SBE-β-CD and a MM-β-CD mixture enhanced the signal-to-noise ratio and led to similar limits of detection and quantification with regard to SFC. No trace of the other enantiomer was found in the isolated one. Biological activities of individual enantiomers were then evaluated and revealed no cytotoxicity against cell lines and a significant difference in terms of their IC50 values with respect to the investigated racemate (6.43 μM): 3.49 μM for the (R)-enantiomer and >10(-4)μM for the (S)-enantiomer, for compound 1, showing that, this antagonist activity is stereospecific.


Enhanced Tumor Targeting and Penetration of Proteolysis-Targeting Chimeras through iRGD Peptide Conjugation: A Strategy for Precise Protein Degradation in Breast Cancer.
Proteolysis-targeting chimeras (PROTACs) have recently emerged as a promising technology for drug development. However, poor water solubility, limited tissue selectivity, and inadequate tumor penetration pose significant challenges for PROTAC-based therapies in cancer treatment. Herein, we developed an iRGD-PROTAC conjugation strategy utilizing tumor-penetrating cyclic peptide iRGD (CRGDK/RGPD/EC) to deliver PROTACs deep into breast cancer tissues. As a conceptual validation study, iRGD peptides were conjugated with a bromodomain-containing protein 4 (BRD4) PROTAC through a GSH-responsive linker. The resulting iRGD-PROTAC conjugate iPR showed enhanced water solubility, tumor-targeting capability, and penetration within tumor tissues, resulting in increased antibreast cancer efficacy in animal models and patient-derived organoids. This study demonstrates the advantages of combining iRGD and PROTACs in improving drug delivery and highlights the importance of tissue selectivity and penetration ability in PROTAC-based therapeutics.


Ligand close packing, molecular compactness, the methyl tilt, molecular conformations, and a new model for the anomeric effect.
All the atoms in a molecule attract each other until they reach their equilibrium positions at which point the repulsive forces between the atoms just balance the attractive forces and there are no resultant forces acting on any of the atoms in the molecule. Thus, we can consider that in the equilibrium geometry the atoms in a molecule are arranged as compactly as possible. This is the basis of the ligand close packing (LCP) model according to which three or four monatomic ligands X, such as F, Cl or O (formally =O or O(-)) pack as closely as possible around a small central atom such as a boron or carbon atom giving a truly close-packed equilateral triangular AX(3) molecule or a tetrahedral AX(4) molecules. Such monatomic ligands can, to a good approximation, be described as having a spherical shape with a single ligand radius r(X). In contrast, ligands with donor atoms with lone pairs such as the oxygen atom in an OX group have a less symmetrical electron density requiring two ligand radii, r(O(lp)) in the lone pair direction, and r(O(b)) in the bonding direction, where r(O(lp)) < r(O(b)) for an approximate description. On this basis we propose an explanation for the "methyl tilt", in methanol and many related molecules, and in conjunction with the concept of compactness, a model for explaining the relative energies of the conformations of molecules containing OH and OMe ligands, including molecules that exhibit the anomeric effect. We compare our model for the anomeric effect with the widely accepted "hyperconjugation" model. We also discuss the relationship between the concept of compactness and the concept of hardness.


Integration of 16S rRNA gene sequencing and LC/MS-based metabolomic analysis of early biomarkers of acute ischaemic stroke in Tibetan miniature pigs.
Acute ischaemic stroke (AIS) is a complex, systemic, pathological, and physiological process. Systemic inflammatory responses and disorders of the gut microbiome contribute to increased mortality and disability following AIS. We conducted 16S high-throughput sequencing and ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry-based non-targeted metabolomic analyses of the plasma from a Tibetan miniature pig middle cerebral artery occlusion (MCAO) model. A significant decrease in the abundance of Firmicutes and a significant increase in the abundance of Actinobacteria were observed after the onset of AIS. Among the plasma metabolites, the levels of phospholipids and amino acids were considerably altered. Loading values and differential metabolite-bacterial group association analyses of the metabolome and microbiome indicated a correlation between the microbiome and metabolome of Tibetan miniature pigs after MCAO. Furthermore, significant changes were observed in the ABC transporter pathway and purine metabolism in the gut microbiome-plasma metabolome during the early stage of AIS. Kyoto Encyclopaedia of Genes and Genomes enrichment analysis showed that arginine, proline, and cyanoamino acid metabolism was upregulated while ABC transporter metabolism pathway and carbohydrate digestion and absorption were substantially downregulated. The results of this study suggest that AIS affects the gut microbiota and plasma metabolites in Tibetan miniature pigs and that faecal microbiota transplantation could be a potential therapeutic approach for AIS.


Prioritizing Seafloor Mapping for Washington's Pacific Coast.
Remote sensing systems are critical tools used for characterizing the geological and ecological composition of the seafloor. However, creating comprehensive and detailed maps of ocean and coastal environments has been hindered by the high cost of operating ship- and aircraft-based sensors. While a number of groups (e.g., academic research, government resource management, and private sector) are engaged in or would benefit from the collection of additional seafloor mapping data, disparate priorities, dauntingly large data gaps, and insufficient funding have confounded strategic planning efforts. In this study, we addressed these challenges by implementing a quantitative, spatial process to facilitate prioritizing seafloor mapping needs in Washington State. The Washington State Prioritization Tool (WASP), a custom web-based mapping tool, was developed to solicit and analyze mapping priorities from each participating group. The process resulted in the identification of several discrete, high priority mapping hotspots. As a result, several of the areas have been or will be subsequently mapped. Furthermore, information captured during the process about the intended application of the mapping data was paramount for identifying the optimum remote sensing sensors and acquisition parameters to use during subsequent mapping surveys.


Deep venous thrombosis after renal transplantation.
Four hundred and eighty consecutive renal transplant operations performed upon adults over a ten year period were reviewed to obtain the incidence of deep venous thrombosis (DVT) or pulmonary embolism (PE), or both. Using strict objective diagnostic criteria, 40 (8.3 per cent) thrombotic events were diagnosed, comprising 25 lower limb DVT alone, 11 DVT with PE and four PE alone. Four deaths were directly attributable to PE which was the fourth major cause of death in the review period. DVT was more common on the side of the transplant but the difference was not significant. The low incidence of thrombotic events (1.7 per cent) in the first month of transplantation does not suggest that chemical prophylaxis should be used during this period. However, the peak incidence occurred in the fourth month after transplantation which may be associated with a significant rise in mean hemoglobin concentration in both the thrombotic population and a control group of transplant recipients. Patient age at the time of transplantation and predisposing events, such as prolonged bedrest, further surgical treatment and pelvic pathology, were significant risk factors. All of the patients in the thrombotic population were receiving steroids at the time of diagnosis and no thrombotic events were demonstrated in patients receiving cyclosporin alone. The results of this review suggest that chemical prophylaxis is indicated for patients more than 40 years of age, with stable renal function and receiving steroids, who undergo periods of prolonged bedrest or further surgical treatment at some time after renal transplantation.


Diverse environmental perturbations reveal the evolution and context-dependency of genetic effects on gene expression levels.
There is increasing appreciation that, in addition to being shaped by an individual's genotype and environment, most complex traits are also determined by poorly understood interactions between these two factors. So-called "genotype × environment" (G×E) interactions remain difficult to map at the organismal level but can be uncovered using molecular phenotypes. To do so at large scale, we used TM3'seq to profile transcriptomes across 12 cellular environments in 544 immortalized B cell lines from the 1000 Genomes Project. We mapped the genetic basis of gene expression levels across environments and revealed a context-dependent genetic architecture: The average heritability of gene expression levels increased in treatment relative to control conditions, and on average, each treatment revealed new expression quantitative trait loci (eQTLs) at 11% of genes. Across our experiments, 22% of all identified eQTLs were context-dependent, and this group was enriched for trait- and disease-associated loci. Further, evolutionary analyses suggested that positive selection has shaped G×E loci involved in responding to immune challenges and hormones but not to man-made chemicals. We hypothesize that this reflects a reduced opportunity for selection to act on responses to molecules recently introduced into human environments. Together, our work highlights the importance of considering an exposure's evolutionary history when studying and interpreting G×E interactions, and provides new insight into the evolutionary mechanisms that maintain G×E loci in human populations.


Effects of electroconvulsive therapy and magnetic seizure therapy on acute memory retrieval.
Electroconvulsive therapy (ECT) is currently the most effective treatment for severe depression. However, it is frequently associated with negative cognitive side effects. Magnetic seizure therapy (MST) depicts an alternative, although experimental, convulsive treatment for major depression. Initial results suggest comparable antidepressant effects accompanied by a better side effect profile. However, no studies up to now have addressed acute retrieval disruption after MST in comparison to ECT. Therefore, we intended to broaden insight into the side effect profile of MST compared to ECT by examining the disruption of acute verbal memory processes after treatment. Twenty depressed patients were randomly assigned to either MST (10 patients) or ECT (10 patients) treatment. On 2 treatment days and 2 treatment-free days, the patients memorized words using a controlled learning paradigm derived from the Batchelder and Riefer storage retrieval model. Four hours after memorization, the patients were asked to retrieve words freely (delayed recall) and a second time with the help of an additional cue constructed out of a hypernymic category (cued recall). By comparing memory performance on treatment days to control days, treatment-induced memory disruption was evaluated. After ECT, delayed recall was disturbed, whereas after MST, it was not. However, this difference in performance was no longer apparent upon cue application (cued recall). This study demonstrates that ECT-induced acute memory disruption measured by delayed recall is absent after MST, confirming its superior side effect profile. We hope that confirming advantages of MST over ECT will improve therapy options for patients with severe depression.


Cerebral perfusion pressure between 50 and 60 mm Hg may be beneficial in head-injured patients: a computerized secondary insult monitoring study.
To describe the occurrence of secondary insults using a computerized monitoring data collecting system and to investigate their relationship to outcome when the neurointensive care was dedicated to avoiding secondary insults. Patients 16 to 79 years old admitted to the neurointensive care unit between August 1998 and December 2002 with traumatic brain injury and 54 hours or more of valid monitoring within the first 120 hours after trauma (one value/min) were included. Monitoring of intracranial pressure (ICP), cerebral perfusion pressure (CPP), systolic blood pressure (BPs), and mean blood pressure (BPm) was required, and insult levels were defined (ICP >25/>35, BPs <100/<90/>160/>180, BPm <80/<70/>110/>120, and CPP <60/<50/>70/>80 mm Hg). Insults were quantified as proportion of valid monitoring time at the insult level. Logistic regression analyses were performed with admission and secondary insult variables as explanatory variables and favorable outcome as dependent variable. Eighty-one patients, 63 men and 18 women, with a mean age of 43.0 years, fulfilled the inclusion criteria. Seventy-two patients (89%) had Glasgow Coma Scale scores of 8 or less. Thirty-one patients (38%) had diffuse injury, and 50 (62%) had mass lesions. Mean Injury Severity Score was 26.6. After 6 months, 54% of the patients had achieved a favorable outcome. Most patients spent 5% or less of the monitoring time at the insult level except for CPP greater than 70 mm Hg. Low age, high Glasgow Coma Scale motor score, low Injury Severity Score, and CPP less than 60 mm Hg insults were significant predictors of favorable outcome in the final multiple logistic regression model. Overall, the secondary insults were rare, except for high CPP. The results suggest that patients with traumatic brain injury might benefit from a CPP slightly less than 60 mm Hg.


Resolution of G(s)alpha and G(q)alpha/G(11)alpha proteins in membrane domains by two-dimensional electrophoresis: the effect of long-term agonist stimulation.
Low-density membrane-domain fractions were prepared from S49 lymphoma cells and clone e2m11 of HEK293 cells expressing a large number of thyrotropin-releasing hormone receptor (TRH-R) and G(11)alpha by flotation on sucrose density gradients. The intact cell structure was broken by detergent-extraction, alkaline-treatment or drastic homogenization. Three types of low-density membranes were resolved by two-dimensional electrophoresis and analyzed for G(s)alpha (S49) or G(q)alpha/G11) (e2m11) content. Four individual immunoblot signals of Gsalpha protein were identified in S49 lymphoma cells indicating complete resolution of the long G(s)alpha L+/-ser and short G(s)alpha S+/-ser variants of G(s)alpha. All these were diminished by prolonged agonist (isoprenaline) stimulation. In e2m11-HEK cells, five different immunoblot signals were detected indicating post-translational modification of G proteins of G(q)alpha/G(11)alpha family. The two major spots corresponding to exogenously (over)expressed G(11)alpha and endogenous G(q)alpha were reduced; the minor spots diminished by hormonal stimulation. Parallel analysis by silver staining of the total protein content indicated that no major changes in protein composition occurred under these conditions. Our data thus indicate that agonist-stimulation of target cells results in down-regulation of all different members of G(s) and G(q)/G(11) families. This agonist-specific effect may be demonstrated in crude membrane as well as domain/raft preparations and it is not accompanied by changes in overall protein composition.


Acute obstructive jaundice: a possible clinical manifestation of IPMT. Case report and review of the literature.
Pancreatic masses causing acute obstructive jaundice still pose diagnostic difficulties and their characterization can often be complex as there is significant overlap in their imaging features. We describe a case of Intraductal Papillary Mucinous Tumor (IPMT) presenting with acute obstructive jaundice in a patient with history of recurrent mild pancreatitis. Clinical evaluation, abdominal ultrasonography (US) and CT-scan posed suspicion of adenocarcinoma with cystic degeneration of the pancreatic head or mucinous cystadenocarcinoma; magnetic resonance (MR) with magnetic resonance cholangiopancreatography (MRCP) demonstrated the communication of the mass with the main pancreatic duct, posing differential diagnosis between main-duct-IPMT and mucinous cystadenocarcinoma. Endoscopic retrograde cholangiopancreatography (ERCP) demonstrated the presence of a mucus-secreting lesion inside duodenum and duodenal biopsies showed no evidence of neoplastic cells. The patient underwent spleen preserving total pancreatectomy that led to histological diagnosis of intraductal papillary mucinous with carcinoma in situ. The international guidelines for management of IPMT, reported in 2006 and revised in 2012, establish that the resectability and the absence of an invasive carcinoma are the most important prognostic factors in IPMT. Therefore an early diagnosis and a radical resection are crucial to improve the patient survival and reduce the recurrence rate. When an IPMT is suspected, the imaging modalities are essential to pose the diagnosis, maximise the chance to select the right surgical candidate and to perform the best treatment for each patient.


Hormonal role of adenosine in maintaining patency of the ductus arteriosus in fetal lambs.
The hypothesis that endogenously released adenosine plays an important role in maintaining patency of the fetal lamb ductus arteriosus was tested. The design of the study was (1) to determine the effect, if any, of exogenous adenosine on blood flow through the ductus arteriosus and (2) to evaluate the relationship among the partial pressure of oxygen in arterial blood, circulating endogenous plasma adenosine concentration, and the rate of blood flow through the ductus. When exogenous adenosine (5 mumoles) was administered during oxygen-induced ductal constriction, ductal blood flow increased from 101 +/- 6 ml/min to 153 +/- 4 ml/min (p less than 0.01). When fetal blood adenosine concentrations were measured during nonventilation and ventilation with 100% oxygen, endogenous adenosine concentrations fell to less than one-half of the preventilation levels, i.e., from 1.12 +/- 0.17 to 0.49 +/- 0.03 microM (p less than 0.01). Finally, when fetal lambs were ventilated with increasing concentrations of oxygen (0%, 10%, 20%, 60%, and 100%) and measurements obtained simultaneously at each level, there was a significant monoexponential relationship among the rise in PO2, the fall in plasma adenosine concentration, and the decrease in ductal blood flow. These data suggest that: (1) adenosine is a potent vasodilator of the lamb ductus arteriosus during oxygen-induced vasoconstriction; (2) fetal endogenous plasma adenosine levels fall significantly when PO2 is increased; and (3) the fall in adenosine concentrations parallels a decrease in ductal blood flow. The findings suggest that the endogenous vasodilator adenosine plays an important role in maintaining ductal patency in utero.


Chloride channel and chloride conductance regulator domains of CFTR, the cystic fibrosis transmembrane conductance regulator.
CFTR is a cyclic AMP (cAMP)-activated chloride (Cl-) channel and a regulator of outwardly rectifying Cl- channels (ORCCs) in airway epithelia. CFTR regulates ORCCs by facilitating the release of ATP out of cells. Once released from cells, ATP stimulates ORCCs by means of a purinergic receptor. To define the domains of CFTR important for Cl- channel function and/or ORCC regulator function, mutant CFTRs with N- and C-terminal truncations and selected individual amino acid substitutions were created and studied by transfection into a line of human airway epithelial cells from a cystic fibrosis patient (IB3-1) or by injection of in vitro transcribed complementary RNAs (cRNAs) into Xenopus oocytes. Two-electrode voltage clamp recordings, 36Cl- efflux assays, and whole cell patch-clamp recordings were used to assay for the Cl- channel function of CFTR and for its ability to regulate ORCCs. The data showed that the first transmembrane domain (TMD-1) of CFTR, especially predicted alpha-helices 5 and 6, forms an essential part of the Cl- channel pore, whereas the first nucleotide-binding and regulatory domains (NBD1/R domain) are essential for its ability to regulate ORCCs. Finally, the data show that the ability of CFTR to function as a Cl- channel and a conductance regulator are not mutually exclusive; one function could be eliminated while the other was preserved.


Utilization of chopped and long alfalfa by dairy heifers.
Two physical forms of alfalfa hay, chopped and long, were fed at maintenance and for ad libitum consumption to dairy heifers to determine effects upon digestibility and energy utilization. Ad libitum intake was associated with lower digestibility of dry matter (56 versus 58%), neutral detergent fiber (45 versus 58%), and acid detergent fiber (45 versus 52%). Energy balance was 1.77 Mcal/day at ad libitum versus .08 Mcal/day at maintenance. Dry matter intake was similar for chopped (5.4 kg/day) versus long alfalfa (5.5 kg/day). Digestibilities of dry matter, neutral detergent fiber, acid detergent fiber, and cellulose were 7 to 10 percentage units lower when heifers were fed the chopped alfalfa. Metabolizable energy intake generally reflected digestible energy intake, which was about 2 Mcal/day less for heifers consuming chopped alfalfa. Heat production was similar for the two physical forms. Consequently, heifers consuming chopped alfalfa had lower energy balance. Reducing particle size of the alfalfa resulted in lower dry matter and energy digestibility, presumably because of increased rate of passage. Increased intake to compensate for depressed energy utilization of the chopped alfalfa did not occur; effects upon animal gain could not be assessed.


Culture of Borrelia persica and its flagellar antigen in vitro.
Borrelia persica is a strain seen only in the Middle East and responsible for relapsing fever. These spirochetes are notable for multiphasic antigenic variation of polymorphic outer membrane lipoproteins, a phenomenon responsible for immune evasion. Diagnosis of the disease is a problem and requires a fixed antigen like the flagellar antigen. In vitro culture of B. persica was carried out for the first time and flagellar antigen was purified from culture. 10% SDS was added to the mixture to dissolve the cell wall and then the solution was sheared in an Omni mixer. Electron microscopy confirmed the purity of a 42 KDa periplasmic antigen as revealed by SDS-PAGE. Indirect haemagglutination kits were designed using the pure flagella and tested for cross reactivity with another relapsing fever spirochaete Borrelia microtii positive serum. The kit showed 98% sensitivity and 95% specificity.


Gold Nanoparticles Supported on Mesoporous Titania Thin Films with High Loading as a CO Oxidation Catalyst.
In the present work, 2.4 nm gold nanoparticles (Au NPs) are uniformly dispersed on mesoporous titania thin films which are structurally tuned by controlling the calcination temperature. The gold content of the catalyst is as high as 27.8 wt %. To our knowledge, such a high loading of Au NPs with good dispersity has not been reported until now. Furthermore, the reaction rate of the gold particles is enhanced by one order of magnitude when supported on mesoporous titania compared to non-porous titania. This significant improvement can be explained by an increase in the diffusivity of the substrate due to the presence of mesopores, the resistance to agglomeration, and improved oxygen activation.


Nutrition of Patients with Severe Neurologic Impairment.
Commercial enteral formulas are generally recommended for gastrostomy feeding in patients with severe neurologic impairment. However, pureed food diets are still widely used and even gaining popularity among certain groups. We tried to compare the effectiveness of gastrostomy feeding for treatment of severe malnutrition with either enteral formulas or pureed feeds. A 6-month nutritional intervention was made with 37 malnourished children, adolescents and young adults (2-26 years old) with severe neurologic impairment (Gross Motor Function Classification system [GMFCS] grade V). The individual needs were calculated. Participants were fed by gastrostomy with either enteral formulas (n = 17) or pureed food (n = 20). Measurements to assess nutritional status were made at the beginning and at the end of intervention. The Z scores for weight-for-age and for the body-mass index increased more in enteral formula than in pureed food group (2.07 vs. 0.70, p = 0.0012; and 3.75 vs. 0.63, p = 0.0014, respectively). Fat mass index increased more in enteral formula than in pureed food group (1.12 kg/m2vs. 0.38 kg/m2; p = 0.0012). Patients in the enteral formula group showed increase in lean body mass expressed as fat-free mass index (0.70 kg/m2), while those in pureed food group did not (-0.06 kg/m2) (p = 0.0487). The results suggest that even professionally planned pureed food diet is less effective than commercial enteral formula for nutritional rehabilitation of malnourished patients with severe neurologic impairment. However, larger and if possible randomised clinical studies should be made to confirm our findings.


Quantification of polyphenols and pharmacological analysis of water and ethanol-based extracts of cultivated agarwood leaves.
Mangiferin (3) and genkwanin 5-O-β-primeveroside (5) are the two major bioactive polyphenols with laxative property present in the extracts of agarwood (Aquilaria sinensis) leaves (AL). Here we developed an HPLC method to determine these bioactive components and four other major polyphenols in AL extracts and evaluated the pharmacological equivalence of organic and water extracts. Using mobile phase gradient conditions combined with UV detection at 330 nm, all six compounds were separated and we determined the relative extraction ratios of the six compounds present in A. sinensis extracts that were prepared under different conditions and compared the contents of the two laxative polyphenols present in the 60% ethanol extracts of A. sinensis and A. crassna. The polyphenols present in water extracts of 13 commercially cultivated A. crassna plants have also been analyzed. The laxative properties of 60% ethanol and four water extracts of A. crassna were evaluated by the frequency and weight of stools in loperamide-induced constipation model mice. The pharmacological equivalence of 60% ethanol extract and hot water (95°C) extract was identified in mice.


Propagation properties of the circular Airy beam with a Gaussian envelope in Fourier space.
We theoretically and experimentally study the propagation properties of the circular Airy beam (CAB) with a Gaussian envelope in Fourier space. The two parameters of the Gaussian function can be used to control the distribution of the spatial frequency, and thus to tune the propagation properties of this modified CAB. When the two parameters are chosen appropriately, the size of focal spot will be reduced, the maximum focal intensity and especially the abruptly autofocusing property will be greatly enhanced. Meanwhile, the focal position can remain almost the same with the common CAB. The experimental results show that the proposed beam can be generated conveniently by using the same Fourier transform method as used to generate the common CAB.