Anastomosing Hemangioma Incidentally Found in Kidney or Adrenal Gland: Study of 10 Cases and Review of Literature.
To describe and report a series of renal and adrenal anastomosing hemangioma (AH) and to investigate its distinctive clinicopathologic features and review its clinical data available in the literature. Clinical data of 10 AHs were retrospectively studied. Imaging and histologic features were re-evaluated and summarized. Immunostaining markers were performed include CD31, CD34, ERG, Fil-1, D2-40, AE1/AE3, SMA, CD10, HHV8, S100, Ki-67. A follow-up of all cases was performed. Other AHs published in PubMed and Web of Science were reviewed. All of 10 AHs were found incidentally, containing 5 female and 5 male patients (median, 48.5 years; mean, 51.7 years) and involved unilateral kidney (n=7) and adrenal glands (n=3) respectively. All lesions were well-defined in Imaging and histologic examination. AHs were morphologically characterized by prominent anastomosing vascular channels without evidence of infiltration to surrounding normal tissues and significant cellular atypia. CD31, CD34, ERG were positive and Ki-67 showed typically low (?3%). All Patients underwent a mass resection and none of them had evidence of recurrence. Together with other cases published, the AHs showed distinctive clinicopathologic features with an excellent prognosis. Renal or adrenal AH is a very rare vascular tumor. They have distinctive histologic features and a favorable prognosis. Its frequently mimicking well-differentiated angiosarcomas easily result in unnecessary overtreatment in clinical practice. To be acquainted with this entity serves to avoid misdiagnosis.


Treatment of pulmonary embolism with rivaroxaban: outcomes by simplified Pulmonary Embolism Severity Index score from a post hoc analysis of the EINSTEIN PE study.
The objective was to assess adverse outcomes in relation to the simplified Pulmonary Embolism Severity Index (PESI) score in patients treated with rivaroxaban or standard therapy in the phase III EINSTEIN PE study and to evaluate the utility of the simplified PESI score to identify low-risk pulmonary embolism (PE) patients. A post hoc analysis of EINSTEIN PE data was performed to assess the efficacy and safety of rivaroxaban in patients with a range of simplified PESI scores. Recurrent venous thromboembolism, fatal PE, all-cause mortality, and major bleeding were stratified by simplified PESI scores of 0, 1, or ≥2 and according to treatment period at 7, 14, 30, and 90 days and at the end of the full intended treatment period. Simplified PESI scores could be calculated in 4,831 of the 4,832 randomized patients; of those, 53.6, 36.7, and 9.7% had PESI scores of 0, 1, and ≥2, respectively. Among patients with simplified PESI scores of 0 or 1, fatal PE, all-cause mortality, and other adverse outcomes were uncommon within the first 7, 14, and 30 days. Patients with simplified PESI scores of ≥2 had more frequent adverse outcomes. Major bleeding was lower in the rivaroxaban group, particularly in those with simplified PESI scores of 1 or ≥2. The findings support using risk stratification with the simplified PESI score to identify low-risk patients with PE.


STXBP2 mutations in children with familial haemophagocytic lymphohistiocytosis type 5.
Familial haemophagocytic lymphohistiocytosis (FHL) is a rare immune deficiency with uncontrolled inflammation; the clinical course usually starts within the first years of life, and is usually fatal unless promptly treated and then cured with haematopoietic stem cell transplant. FHL is caused by genetic mutations resulting in defective cell cytotoxicity; three disease related genes have been identified to date: perforin, Munc13-4 and syntaxin-11. A fourth gene, STXBP2, has been identified very recently as responsible for a defect in Munc18-2 in FHL-5. To describe the result of the screening of families with HLH and previously unassigned genetic defects. Patients with HLH diagnosed according to current diagnostic criteria, and who lacked mutations in the PRF1, Munc13-4, and STX11 genes were sequenced for mutations in STXBP2. Functional study was performed when material was available. Among the 28 families investigated, 4 (14%) with biallelic STXBP2 mutations were identified. They originated from Italy, England, Kuwait and Pakistan. The p.Pro477Leu resulting from c.1430C>T, and p.Arg405Gln resulting from the single c.1214G>A nucleotide change are known, while we contribute two novel mutations: p.Glu132Ala resulting from c.395A>C, and p.Gly541Ser, resulting from c.1621G>A. The detrimental effect of the p.Gly541Ser mutation was documented biochemically and functionally in NK and CD8 cells. Additional polymorphisms are also described. These data expand current knowledge on the genetic heterogeneity of FHL and suggest that patients with FHL5 may have different results in degranulation assays under different conditions.


Identification of COUP-TFII orphan nuclear receptor as a retinoic acid-activated receptor.
The chicken ovalbumin upstream promoter-transcription factors (COUP-TFI and II) make up the most conserved subfamily of nuclear receptors that play key roles in angiogenesis, neuronal development, organogenesis, cell fate determination, and metabolic homeostasis. Although the biological functions of COUP-TFs have been studied extensively, little is known of their structural features or aspects of ligand regulation. Here we report the ligand-free 1.48 A crystal structure of the human COUP-TFII ligand-binding domain. The structure reveals an autorepressed conformation of the receptor, where helix alpha10 is bent into the ligand-binding pocket and the activation function-2 helix is folded into the cofactor binding site, thus preventing the recruitment of coactivators. In contrast, in multiple cell lines, COUP-TFII exhibits constitutive transcriptional activity, which can be further potentiated by nuclear receptor coactivators. Mutations designed to disrupt cofactor binding, dimerization, and ligand binding, substantially reduce the COUP-TFII transcriptional activity. Importantly, retinoid acids are able to promote COUP-TFII to recruit coactivators and activate a COUP-TF reporter construct. Although the concentration needed is higher than the physiological levels of retinoic acids, these findings demonstrate that COUP-TFII is a ligand-regulated nuclear receptor, in which ligands activate the receptor by releasing it from the autorepressed conformation.


Stabilizing the code-methods to preserve RNA prove their worth.
Commercially available platforms to stabilize messenger RNA (mRNA) and microRNA are critically designed to optimize and ensure the quality and integrity of those nucleic acids. This is not only essential for gene expression analyses, but would provide technical utility in providing concordant standard operating procedures in preserving the structural integrity of RNA species in multicenter clinical research programs and biobanking of cells or tissues for subsequent isolation of intact RNA. The major challenge is that the presence of degraded samples may adversely influence the interpretation of expression levels on isolated mRNA or microRNA samples and that in the absence of a concordant operating procedure between multiple collaborating research centers would confound data analysis and interpretation. However, in this issue of Biomarker Insights, Weber et al provide a detailed and critical analysis of two common RNA preservation systems, PAXgene and RNAlater. Such studies are lacking in the literature. However, the authors provide compelling evidence that not all conservation platforms are created equal and only one system proves its worth.


Weightbearing Radiographs Facilitate Functional Treatment of Ankle Fractures of Uncertain Stability.
The most important determinant in the treatment of malleolar fractures is stability. Stable fractures have an intact deep deltoid ligament and do not displace with functional treatment. If the deep deltoid/medial malleolar complex is disrupted, the talus is at risk of displacement. Weber (2010) showed that weightbearing radiographs predicted stability in patients with undisplaced ankle fractures. We developed clinical criteria for potential instability and applied them to a prospective series of patients. The criteria included a medial clear space of <4 mm; medial tenderness, bruising or swelling; a fibular fracture above the syndesmosis; a bimalleolar or trimalleolar fracture; an open fracture; and a high-energy fracture mechanism. A prospectively documented series of 43 patients chose functional brace treatment of the potentially unstable fractures. Weightbearing radiographs were performed with the patient wearing the brace before treatment and free of the brace at clinical union (6 to 9 weeks for all patients). The patients were encouraged to bear full weight and actively exercise their ankles in the brace. All fractures healed without displacement. The risk of displacement was 0% (95% confidence interval 0% to 9.5%). The results of the present preliminary series give support for the use of weightbearing radiographs to guide treatment of undisplaced ankle fractures.


Characterization of the novel amplified in breast cancer-1 (NABC1) gene product.
Positional cloning of the cancer-associated 20q13.2 amplicon identified two genes that display high mRNA levels in breast tumors and here we report the initial characterization of one of these gene products, designated Novel Amplified in Breast Cancer-1 (NABC1). Analysis of the primary structure of the NABC1 protein uncovered two regions of this protein with a high likelihood of forming coiled-coils and assembly of a mouse NABC1 cDNA showed that this protein is conserved between mouse and man. NABC1 antisera showed that, like its transcript, breast tumor lines that harbor amplification of 20q13.2 display high levels of the NABC1 protein compared to normal human fibroblasts or a breast cancer line that does not overexpress the NABC1 mRNA. Further, we conclude from studies using in vivo and in vitro approaches that the NABC1 protein forms detergent stable homodimers, and it is this homodimeric form that accumulates in cells that overexpress this protein. NABC1 mRNA exhibits a limited expression pattern in human tissue with high relative transcript levels observed only in brain and prostate. Immunofluorescence microscopy indicates NABC1 displays a punctate localization pattern in the cytoplasm of cultured cells, but biochemical fractionation indicates that this protein is not an integral component of membranous cytoplasmic organelles. Finally, overexpression of human NABC1 in mouse NIH/3T3 cells did not affect either the growth rate or anchorage-dependent growth properties, suggesting that NABC1 is not a prototypical oncogene.


A Combination of ß-Hydroxybutyrate and Citrate Ameliorates Disease Progression in a Rat Model of Polycystic Kidney Disease.
Our research has shown that interventions producing a state of ketosis are highly effective in rat, mouse, and cat models of polycystic kidney disease (PKD), preventing and partially reversing cyst growth and disease progression. The ketone ß-hydroxybutyrate (BHB) appears to underlie this effect. Additionally, we have demonstrated that naturally formed microcrystals within kidney tubules trigger a renoprotective response that facilitates tubular obstruction clearance in healthy animals but, alternatively, leads to cyst formation in PKD. The administration of citrate prevents microcrystal formation and slows PKD progression. Juvenile Cy/+ rats, a non-orthologous PKD model, were supplemented from 3 to 8 weeks of age with water containing titrated BHB, citrate, or in combination to find minimal effective and optimal dosages, respectively. Adult rats were given a reduced BHB/Citrate combination or equimolar control K/NaCl salts from 8 to 12 weeks of age. Additionally, adult rats were placed in metabolic cages following BHB, citrate, and BHB/Citrate administration to determine the impact on mineral, creatinine, and citrate excretion. BHB or citrate alone effectively ameliorates disease progression in juvenile rats, decreasing markers of cystic disease and, in combination, producing a synergistic effect. BHB/Citrate leads to partial disease regression in adult rats with established cystic disease, inhibiting cyst formation and kidney injury. BHB/Citrate confers benefits via multiple mechanisms, increases creatinine and citrate excretion, and normalizes mineral excretion. BHB and citrate are widely available and generally recognized as safe compounds and, in combination, exhibit high promise for supporting kidney health in polycystic kidney disease.


PET/CT and renal pathology: a blind spot for radiologists? Part 2--lymphoma, leukemia, and metastatic disease.
PET/CT with (18)F-FDG is a powerful tool to evaluate patients with hematologic malignancy or to assess the burden of metastatic disease from solid tumors. Metabolically active renal pathology associated with lymphoma, leukemia, or metastatic disease can be missed without close attention to both the PET and CT portions of the study because of physiologic FDG excretion in the kidneys. This article illustrates the appearance of tracer uptake and the key anatomic features of lymphoma, leukemia, and metastatic disease involving the kidney on FDG PET/CT. Close attention to both the FDG PET and CT portions of an FDG PET/CT study is essential to evaluate the kidneys in oncology patients.


The speeding attitude scale and the role of sensation seeking in profiling young drivers at risk.
Seven driving attitude scales representing driving behaviors and beliefs about driving were created and initially validated using 257 undergraduate students (168 females, 89 males) in Study 1. However, the Speeding Attitude Scale (SAS) accounted for most of the strength of the intercorrelations among these scales and discriminant classification analyses showed that SAS dominated the other scales as a sufficient explanation for having speeding tickets. Study 2, using 180 students (75 males, 105 females), replicated findings regarding the significant but low correlation between SAS and speeding tickets, and was significantly correlated with Zuckerman's Sensation Seeking Scale (SSS). Replication also showed that males had higher SAS scores and more speeding tickets. Accidents were typically a function of sex, increasing age, and variables related to recent accident history. Objective sources of speeding attitude confirmation may enhance the future validity of the SAS. Potential interventions for being a safe passenger and attitudinal control in the training of young drivers were discussed.


Nano Silver Fluoride for preventing caries.
Randomised controlled trial, double blind, in a community setting. School children with active caries in primary teeth and no pulpal exposure, fistula or decay in permanent teeth were chosen. Caries and unsupported enamel were left as found and cotton wool rolls were used for isolation. Two drops of NSF or one drop of water were applied to the tooth with a microbrush for two minutes, once in a 12-month period. At one week, five months and 12 months the presence of active caries, as classified using International Caries Detection and Assessment System (ICDAS II) criteria, was measured. If a blunt probe easily penetrated dentine with light force, active caries was recorded and taken to be a failure. One hundred and thirty primary teeth in 60 children with a mean age of 6.31 (± 0.60) were randomised. Sixty-three teeth were in the NSF group and 67 in the control group. After one week there were no losses, at the five-month follow up eight teeth were lost from the NSF group due to exfoliation or extraction and five from the control group. At twelve months a further five teeth were lost from the NSF group and 13 from the control group.At the one week follow up there was a 19% failure rate in the NSF group compared to 100% in the control group. At the five month recall this was 27.3% NSF compared to 72.6% water and at the final recall there was a 33.3% NSF failure rate and 65.3% control failure rate. The preventative fraction at this point was 50% and the NNT 3.12. All were statistically significant results (p= <0.05). The annual application of NSF solution was more effective in hardening and arresting dentine caries in primary teeth than the placebo. The effectiveness of NSF was found to be similar to silver diamine fluoride when applied once a year, but did not stain the dental tissue black and had no metallic taste. The application is simple, does not require a clinical setting and is inexpensive. NSF was demonstrated to be effective in arresting caries in children in poor communities, but further studies are required to investigate alternative protocols and for use in tooth sensitivity and root caries.


Scurvy and rickets masked by chronic neurologic illness: revisiting "psychologic malnutrition".
The North American epidemic of overeating, combined with a sedentary lifestyle, has led to a growing prevalence of obesity, diabetes, and the "metabolic syndrome" in children. Excessive caloric intake does not imply adequate nutrition, and vitamin-deficiency syndromes still occur in some American children. Here we describe cases of scurvy and vitamin D deficiency in 2 children with cognitive disorders. Thorough dietary histories suggested the diagnosis in each patient and, had they been obtained at presentation, would likely have obviated invasive diagnostic workup, unnecessary stress to the patients and their families, and significant functional disability. Overnutrition and malnutrition may coexist, particularly among those with abnormal cognition or autistic spectrum disorders. Classic nutritional deficiencies must not be omitted from the differential diagnosis. A comprehensive dietary history and screening for vitamin deficiencies in at-risk children are important aspects of preventive health care and are essential for prompt diagnosis and treatment.


Nuclear localization of RelB is associated with effective antigen-presenting cell function.
Dendritic cells (DC) are potent APCs that enter resting tissues as precursors and, after Ag exposure, differentiate and migrate to draining lymph nodes. The phenotype of RelB knockout mice implicates this member of the NF kappa B/Rel family in DC differentiation. To further elucidate the role of RelB in DC differentiation, mRNA, intracellular protein expression, and DNA binding activity of RelB were examined in immature and differentiated human DC, as well as other PB mononuclear cell populations. RelB protein and mRNA were detected constitutively in lymphocytes and in activated monocytes, differentiated DC, and monocyte-derived DC. Immunohistochemical staining demonstrated RelB within the differentiated lymph node interdigitating DC and follicular DC, but not undifferentiated DC in normal skin. Active nuclear RelB was detected by supershift assay only in differentiated DC derived from either PB precursors or monocytes and in activated B cells. These RelB+ APC were potent stimulators of the MLR. The data indicate that RelB expression is regulated both transcriptionally and post-translationally in myeloid cells. Within the nucleus, RelB may specifically transactivate genes that are critical for APC function.


Educational Attainment Polygenic Scores: Examining Evidence for Gene-Environment Interplay with Adolescent Alcohol, Tobacco and Cannabis Use.
Genes associated with educational attainment may be related to or interact with adolescent alcohol, tobacco and cannabis use. Potential gene-environment interplay between educational attainment polygenic scores (EA-PGS) and adolescent alcohol, tobacco, and cannabis use was evaluated with a series of regression models fitted to data from a sample of 1871 adult Australian twins. All models controlled for age, age2, cohort, sex and genetic ancestry as fixed effects, and a genetic relatedness matrix was included as a random effect. Although there was no evidence that adolescent alcohol, tobacco or cannabis use interacted with EA-PGS to influence educational attainment, there was a significant, positive gene-environment correlation with adolescent alcohol use at all PGS thresholds (ps <.02). Higher EA-PGS were associated with an increased likelihood of using alcohol as an adolescent (ΔR2 ranged from 0.5% to 1.1%). The positive gene-environment correlation suggests a complex relationship between educational attainment and alcohol use that is due to common genetic factors.


Successful treatment of renal amyloidosis due to familial cold autoinflammatory syndrome using an interleukin 1 receptor antagonist.
Familial cold autoinflammatory syndrome (FCAS) is an autosomal dominant disorder characterized by episodic fever, arthralgias, conjunctivitis, and rash triggered by cold exposure. FCAS is rarely associated with progressive renal insufficiency caused by renal amyloidosis. The genetic defect in patients with this disorder is caused by a mutation in the gene encoding the protein cryopyrin, leading to uninhibited activation of systemic inflammation through specific cellular signaling with increased production of a number of key cytokines, including interleukin 1. We describe the successful treatment of a patient with renal amyloidosis caused by FCAS by using a novel interleukin 1-receptor antagonist. Use of specific anticytokine therapy may be a new paradigm in the treatment of patients with renal amyloidosis caused by systemic inflammatory diseases.


Multi- and univariate analyses of the weekend effect for elective lower-limb joint replacements.
Introduction The weekend effect is a perceived difference in outcome between medical care provided at the weekend when compared to that of a weekday. Clearly multifactorial, this effect remains incompletely understood and variable in different clinical contexts. In this study we analyse factors relevant to the weekend effect in elective lower-limb joint replacement at a large NHS multispecialty academic healthcare centre. Materials and Methods We reviewed the electronic medical records of 352 consecutive patients who received an elective primary hip or knee arthroplasty. Patient, clinical and time-related variables were extracted from the records. The data were anonymised, then processed using a combination of uni- and multivariate statistics. Results There is a significant association between the selected weekend effect outcome measure (postoperative length of stay) and patient age, American Society of Anesthesiologists classification, time to first postoperative physiotherapy and time to postoperative radiography but not day of the week of operation. Discussion We were not able to demonstrate a weekend effect in elective lower-limb joint replacement at our institution nor identify a factor that would require additional weekend clinical medical staffing. Rather, resource priorities would seem to include measures to optimise at-risk patients preoperatively and measures to reduce time to physiotherapy and radiography postoperatively. Conclusions Our findings imply that postoperative length of stay could be minimised by strategies relating to patient selection and access to postoperative services. We have also identified a powerful statistical methodology that could be applied to other service evaluations in different clinical contexts.


The endoplasmic reticulum exerts control over organelle streaming during cell expansion.
Cytoplasmic streaming is crucial for cell homeostasis and expansion but the precise driving forces are largely unknown. In plants, partial loss of cytoplasmic streaming due to chemical and genetic ablation of myosins supports the existence of yet-unknown motors for organelle movement. Here we tested a role of the endoplasmic reticulum (ER) as propelling force for cytoplasmic streaming during cell expansion. Through quantitative live-cell analyses in wild-type Arabidopsis thaliana cells and mutants with compromised ER structure and streaming, we demonstrate that cytoplasmic streaming undergoes profound changes during cell expansion and that it depends on motor forces co-exerted by the ER and the cytoskeleton.


Drotrecogin alfa (activated) treatment of older patients with severe sepsis.
The incidence of severe sepsis increases dramatically with advanced age, with a mortality rate that approaches 50%. The main purpose of this investigation was to determine both short- and long-term survival outcomes among 386 patients aged >or=75 years who were enrolled in the Protein C Worldwide Evaluation of Severe Sepsis (PROWESS) trial. Subjects who were treated with drotrecogin alfa (activated; DAA) had absolute risk reductions in 28-day and in-hospital mortality of 15.5% and 15.6%, respectively (P=.002 for both), compared with placebo recipients. The relative risk (RR) for 28-day mortality was 0.68 (95% confidence interval [CI], 0.54-0.87), and the in-hospital RR was 0.70 (95% CI, 0.56-0.88). Resource use and patient disposition for DAA-treated patients compared favorably with those for placebo recipients. In addition, long-term follow-up data were available for 375 subjects (97.2%), and survival rates for DAA recipients were significantly higher over a 2-year period (P=.02). The incidences of serious adverse bleeding during the 28-day study period in the DAA and placebo groups were 3.9% and 2.2%, respectively (P=.34). There was no interaction between age and bleeding rates (P=.97). In conclusion, older patients with severe sepsis have higher short- and long-term survival rates when treated with DAA than when treated with placebo but an increased risk of serious bleeding that is not aged related.


Electroencephalography Can Distinguish between Pain and Anaesthetic Intervention in Conscious Lambs Undergoing Castration.
Australian sheep routinely undergo painful surgical husbandry procedures without anaesthesia or analgesia. Electroencephalography (EEG) has been shown to be a successful measure of pain in livestock under a general anaesthetic. The aim of this study was to compare this EEG model to that of conscious lambs undergoing castration with and without local anaesthesia. Sixteen merino crossbred ram lambs 6 to 8 weeks of age (13.81kg ± 1.97) were used in the study. Lambs were randomly allocated to 1 of 4 treatment groups: (1) Conscious EEG and surgical castration with no anaesthetic intervention (CON; n = 4); (2) Conscious EEG and surgical castration with pre-operative applied intra-testicular lignocaine injection (CON + LIG; n = 4); (3) surgical castration under minimal anaesthesia (MAM; n = 4); (4) and surgical castration with pre-operative lignocaine injection (2 mL lignocaine hydrochloride 20 mg/mL, under minimal anaesthesia (MAM + LIG; n = 4). Distinct differences in the EEG parameters Ptot, F50 and F95 between pre-and post-castration in conscious lambs were demonstrated in this study (p < 0.01). Further, CON and CON + LIG treatments were distinguishable using F50 and F95 measures (p = 0.02, p = 0.04, respectively). Significant changes in the EEG output of MAM animals were identified pre- to post-castration (p < 0.01). The EEG output of MAM and MAM + LIG were similar. EEG was successful in differentiating lambs treated with pain relief in a conscious state after castration by examining F50 and F95, which may suggest the suitability of conscious EEG pain measurement.


Possibilities for clinical use of prostacyclin in vascular disease.
Currently available drug therapies for patients suffering severe ischaemia with rest pain and trophic lesions of the limbs remain unsatisfactory. Also vascular reopening procedures are suitable in only about half of the patients. In atherosclerotic disease when the vascular endothelium is damaged prostacyclin synthesis is decreased and thromboxane A2 production increases. Prompted by this knowledge of the importance of prostacyclin in pathogenesis of atherosclerotic disease an attempt was made to employ PGI2 clinically--for treatment of advanced forms of peripheral arterial atherosclerotic disease. Favourable effects of the stable analogue of prostacyclin (Iloprost), were reported in various studies, which included patients with peripheral atherosclerotic arterial disease, thromboangiitis obliterans and Raynaud's phenomenon. The use of Iloprost resulted in a significantly superior response than other drugs and placebo in terms of alleviation of rest pain, ulcer healing and decrease of amputation rate of ischaemic limbs. Therefore prostacyclin provides a therapeutic option in patients with advanced forms of arterial disease--including critical ischaemia.


[Phylogeny of salmonids (Salmoniformes, Salmonidae) and its molecular dating: analysis of nuclear RAG1 gene].
The phylogenetic relationships among 26 species of salmonid fishes (family Salmonidae) were studied using the RAG1 gene as phylogenetic marker. No unambiguous relationships between thymallins, coregonins, and salmonins it was possible to establish. It seems likely, that divergence of these lineages took place during rather short time interval (about 3 to 4 million years). The thymallins are thought to be the first radiated lineage. The genera of the subfamily Salmoninae form two distinct monophyletic groups, represented by (1) Brachymystax and Hucho and (2) Salmo, Parahucho, Salvelinus, Parasalmo and Oncorhynchus. Ancestral forms of these two evolutionary lineages could diverge at the Oligocene-Miocene boundary (about 24 million years ago). It is suggested that diversification of the main lineages within the second group was rather rapid, and took place in middle Miocene (about 19-16 million years ago). Moreover, the lineages of Salvelinus, Parasalmo and Oncorhynchus were the latest to diverge. It seems likely that divergence of the Prosopium and Corergouns ancestral lineages occurred during the same time interval.


Drug checking in the fentanyl era: Utilization and interest among people who inject drugs in San Diego, California.
In North America, overdose rates have steeply risen over the past five years, largely due to the ubiquity of illicitly manufactured fentanyls in the drug supply. Drug checking services (DCS) represent a promising harm reduction strategy and characterizing experiences of use and interest among people who inject drugs (PWID) is a priority. Between February-October 2022, PWID participating in a cohort study in San Diego, CA and Tijuana, Mexico completed structured surveys including questions about DCS, socio-demographics and substance use behaviors. We used Poisson regression to assess factors associated with lifetime DCS use and characterized experiences with DCS and interest in free access to DCS. Of 426 PWID, 72% were male, 59% Latinx, 79% were experiencing homelessness and 56% ever experienced a nonfatal overdose. One third had heard of DCS, of whom 57% had ever used them. Among the latter, most (98%) reported using fentanyl test strips (FTS) the last time they used DCS; 66% did so less than once per month. In the last six months, respondents used FTS to check methamphetamine (48%), heroin (30%) or fentanyl (29%). Relative to White/non-Latinx PWID, those who were non-White/Latinx were significantly less likely to have used DCS [adjusted risk ratio (aRR): 0.22; 95% CI: 0.10, 0.47), as were PWID experiencing homelessness (aRR:0.45; 95% CI: 0.28, 0.72). However, a significant interaction indicated that non-White/Latinx syringe service program (SSP) clients were more likely to have used DCS than non-SSP clients (aRR: 2.79; CI: 1.09, 7.2). Among all PWID, 44% expressed interest in free access to FTS, while 84% (of 196 PWID) expressed interest in advanced spectrometry DCS to identify and quantify multiple substances. Our findings highlight low rates of DCS awareness and utilization, inequities by race/ethnicity and housing situation, high interest in advanced spectrometry DCS versus FTS, and the potential role of SSPs in improving access to DCS, especially among racial/ethnic minorities.


Cerebral Blood Flow after Mild Traumatic Brain Injury: Associations between Symptoms and Post-Injury Perfusion.
Arterial spin labeling (ASL) has emerged as a technique for assessing mild traumatic brain injury (mTBI), as it can noninvasively evaluate cerebrovascular physiology. To date, there is substantial variability in methodology and findings of ASL studies of mTBI. While both increased and decreased perfusion are reported after mTBI, more consistency is emerging when perfusion is examined with regard to symptomology. We evaluated 15 teenage athletes two and six weeks after sports-related concussion (SRC group) using pseudo-continuous ASL. We acquired comparison data from 15 matched controls from a single time point. At each time point, we completed whole-brain contrasts to evaluate differences between the SRC group and controls in relative cerebral blood flow (rCBF). Cluster-level findings directed region of interest (ROI) analyses to test for group differences in rCBF across the left dorsal anterior cingulate cortex (ACC) and left insula. Finally, we evaluated ROI rCBF and symptomology in the SRC group. At two weeks post-injury, the SRC group had significantly higher rCBF in the left dorsal ACC and left insula than controls; at six weeks post-injury, elevated rCBF persisted in the SRC group in the left dorsal ACC. Perfusion in the left dorsal ACC was higher in athletes reporting physical symptoms six weeks post-injury compared with asymptomatic athletes and controls. Overall, these findings are inconsistent with reports of reduced rCBF after mTBI but coherent with studies that report increased perfusion in persons with greater or persistent mTBI-related symptomology. Future work should continue to assess how CBF perfusion relates to symptomology and recovery after mTBI.


Development of the GPSE: a tool to improve feedback on procedural skills in residency.
Learning procedural skills requires supervised practice with feedback. Observation of performance provides an opportunity for feedback, but the quality of feedback is often poor. We developed an instrument to improve feedback during procedural skills training in residency. The Global Procedural Skills Evaluation (GPSE) was drafted based on qualitative date from field notes and literature on procedural skills training. Interview and focus group data described the current state of feedback and guided revisions to the instrument. An iterative process incorporated evidence from education research and input from teachers and learners to optimize the credibility and dependability of the instrument. The final GPSE includes a self-assessment prompt, five criteria (items) assessing performance, a rating scale quantifying the degree of intervention by the teacher, a global assessment, a rating of case difficulty, and suggestions for improvement. The GPSE is designed to improve feedback during procedural skills training and encourage learner self-assessment. Its development process may be of interest to those who use or produce performance assessment tools.


Attitudes and decision-making about early-infant versus early-adolescent male circumcision: Demand-side insights for sustainable HIV prevention strategies in Zambia and Zimbabwe.
As countries approach their scale-up targets for the voluntary medical male circumcision program for HIV prevention, they are strategizing and planning for the sustainability phase to follow. Global guidance recommends circumcising adolescent (below 14 years) and/or early infant boys (aged 0-60 days), and countries need to consider several factors before prioritizing a cohort for their sustainability phase. We provide community and healthcare provider-side insights on attitudes and decision-making process as a key input for this strategic decision in Zambia and Zimbabwe. We studied expectant parents, parents of infant boys (aged 0-60 days), family members and neo-natal and ante-natal healthcare providers in Zambia and Zimbabwe. Our integrated methodology consisted of in-depth qualitative and quantitative one-on-one interviews, and a simulated-decision-making game, to uncover attitudes towards, and the decision-making process for, early adolescent or early infant medical circumcision (EAMC or EIMC). In both countries, parents viewed early infancy and early adolescence as equally ideal ages for circumcision (38% EIMC vs. 37% EAMC in Zambia; 24% vs. 27% in Zimbabwe). If offered for free, about half of Zambian parents and almost 2 in 5 Zimbabwean parents indicated they would likely circumcise their infant boy; however, half of parents in each country perceived that the community would not accept EIMC. Nurses believed their facilities currently could not absorb EIMC services and that they would have limited ability to influence fathers, who were seen as having the primary decision-making authority. Our analysis suggests that EAMC is more accepted by the community than EIMC and is the path of least resistance for the sustainability phase of VMMC. However, parents or community members do not reject EIMC. Should countries choose to prioritize this cohort for their sustainability phase, a number of barriers around information, decision-making by parents, and supply side will need to be addressed.


Radiation intensity (CTDIvol) and visibility of anatomical structures in head CT examinations.
The purpose of this study was to quantify how changing the amount of radiation used to perform routine head CT examinations (CTDIvol) affects visibility of key anatomical structures. Eight routine noncontrast head CT exams were selected from six CT scanners, each of which had a different CTDIvol setting (60 to 75 mGy). All exams were normal and two slices were selected for evaluation, one at the level of basal ganglia and the other at the fourth ventricle. Three experienced neuroradiologists evaluated the visibility of selected structures, including the putamen, caudate nucleus, thalamus, internal capsule, grey/white differentiation, and brainstem. Images were scored on a five-point scoring scheme (1, unacceptable, 3, satisfactory, and 5, excellent). Reader scores, averaged over the cases obtained from each scanner, were plotted as a function of the corresponding CTDIvol. Average scores for the fourth ventricle were 3.06 ± 0.83 and for the basal ganglia were 3.20 ± 0.86. No image received a score of 1. Two readers showed no clear trend of an increasing score with increasing CTDIvol. One reader showed a slight trend of increasing score with increasing CTDIvol, but the increase in score from a 25% increase in CTDIvol was a fraction of the standard deviation associated average scores. Collectively, results indicated that there were no clear improvements in visualizing neuroanatomy when CTDIvol increased from 60 to 75 mGy in routine head CT examinations. Our study showed no apparent benefit of using more than 60 mGy when performing routine noncontrast head CT examinations.


Renal glycosphingolipid metabolism is dysfunctional in lupus nephritis.
Nearly one half of patients with lupus develop glomerulonephritis (GN), which often leads to renal failure. Although nephritis is diagnosed by the presence of proteinuria, the pathology of nephritis can fall into one of five classes defined by different forms of tissue injury, and the mechanisms involved in pathogenesis are not completely understood. Glycosphingolipids are abundant in the kidney, have roles in many cellular functions, and were shown to be involved in other renal diseases. Here, we show dysfunctional glycosphingolipid metabolism in patients with lupus nephritis and MRL/lpr lupus mice. Specifically, we found that glucosylceramide (GlcCer) and lactosylceramide (LacCer) levels are significantly higher in the kidneys of nephritic MRL/lpr lupus mice than the kidneys of non-nephritic lupus mice or healthy controls. This elevation may be, in part, caused by altered transcriptional regulation and/or activity of LacCer synthase (GalT5) and neuraminidase 1, enzymes that mediate glycosphingolipid metabolism. We show increased neuraminidase 1 activity early during the progression of nephritis (before significant elevation of GlcCer and LacCer in the kidney). Elevated levels of urinary LacCer were detected before proteinuria in lupus mice. Notably, LacCer levels were higher in the urine and kidneys of patients with lupus and nephritis than patients with lupus without nephritis or healthy controls. Together, these results show early and significant dysfunction of the glycosphingolipid metabolic pathway in the kidneys of lupus mice and patients with lupus nephritis and suggest that molecules in this pathway may serve as early markers in lupus nephritis.


Novel canine anti-Coccidioides immunoglobulin G enzyme immunoassay aids in diagnosis of coccidioidomycosis in dogs.
The diagnosis of coccidioidomycosis (CM) in dogs is typically based on clinical presentation, serology, and (less frequently) spherule identification. Agar gel immunodiffusion (AGID) is the most commonly employed serological method, but AGID is slow (requiring up to a week for titer). A Coccidioides antigen enzyme immunoassay (EIA) is also available; however, sensitivity is low in CM dogs. An antibody EIA was developed to detect canine immunoglobulin G (IgG) reacting to Coccidioides antigens. Serum was evaluated from dogs with pathology proven CM and/or AGID positive CM, as well as dogs with histoplasmosis, blastomycosis, non-fungal infections, or healthy dogs. A standard curve was used to convert optical density (OD) values into EIA units (EU). Serum and urine samples from CM dogs were also tested in the antigen EIA. Sensitivity and specificity for IgG were 89.2% and 97.2%, respectively, upon evaluation of dogs with proven or probable CM and control dogs. Cross-reactivity was observed in 7.7% and in 6.4% of dogs with histoplasmosis or blastomycosis, respectively. The antigen EIA alone was insensitive (33.8%). Combined IgG and antigen testing increased sensitivity to 93.2%, as three dogs were IgG-negative but had detectable serum or urine antigen. In 22 dogs with proven CM, sensitivity was statistically similar for antibody EIA and AGID (86% and 73%; P = .487). The MiraVista® canine Coccidioides antibody IgG EIA may aid in the diagnosis of CM by improving turnaround time with comparable sensitivity to AGID. Serial or concurrent testing by antibody and antigen EIAs may be beneficial when screening dogs for CM.


[A 62-year-old man with familial parkinsonism with the onset at 24 years of the age].
We report a right-handed 62-year-old man with early onset familial parkinsonism. The patient was well until 24 years of the age when he noted an onset of resting tremor in his right hand. During the next four years, he noted rigidity, bradykinesia, and difficulty in walking. He was seen in another hospital at 28 years of the age, where he received left pallidotomy. Rigidity on the left side showed marked improvement. He received right pallidotomy at age 30 years. He developed right hemiplegia after this surgery. He was admitted to our hospital in March, 1983 when he was 51 years of the age. He was treated with levodopa but improvement was rather of minor degree. He was transferred to another hospital, but his motor disturbance progressed slowly, and was admitted again to our hospital in November 1990. He had 6 siblings 4 of whom including himself suffered from parkinsonism. No consanguinity was noted in parents. On admission, he appeared chronically ill but the general physical examination was unremarkable. Neurologic examination revealed an alert and mentally sound man. Hasegawa dementia scale was 28.5/32.5. Upward gaze was slightly restricted (3/5). Cranial nerve examination revealed oculogyric crisis, apraxia of eyelid opening, masked face, and small voice. He was able to stand with support; his posture showed left-ward leaning. He had right hemiparesis with moderate weakness. He showed marked bradykinesia and moderate rigidity in his left upper extremity. Fine postural tremor was noted in the left hand. Deep tendon reflexes were diminished in the upper extremities. No Babinski sign was noted. Pain sensation was somewhat diminished on the right side. Results of routine laboratory examination were unremarkable. Cranial CT scan revealed atrophy in the frontal lobe, particularly in the prefrontal area. In addition, MRI revealed T1-and-T2-low signal intensity lesions in the right ventral pallidal region and in the left ventrolateral thalamic-hypothalamic areas. He was treated with 600 mg of levodopa with benserazide and 22.5 mg of bromocriptine with mild to moderate improvement in his bradykinesia and rigidity. He was discharged in January 1991. His clinical course was complicated by intestinal obstruction in October, 1994. He was admitted to another hospital where he was operated on the obstruction on November 5, 1994. The sigmoid colon was markedly dilated but no mass was found. Postoperative course was uneventful until November 18, 1994 when he was found dead in his hospital room shortly after 4 am. The patient was discussed in neurological CPC, and the chief discussant arrived at the conclusion that the patient had young-onset familial Lewy body-negative parkinsonism. Opinions were divided between Lewy body-positive familial Parkinson's disease and Lewy body negative young onset parkinsonism. Postmortem examination revealed aspiration pneumonia, which appeared to be the cause of his death, in the right lung. Neuropathologic examination revealed loss of malanized neurons in the substantia nigra and the locus coeruleus. In the substantia nigra, neuronal loss was particularly severe in the ventrolateral area. No Lewy bodies were seen. The dorsal motor nucleus of the vagal nerve was well preserved. Stereotaxic lesions involved bilateral thalamic areas. This patient appears to represent a case of autosomal recessive juvenile parkinsonism (AR-JP). Early onset, superb response to levodopa, sleep effect, and easy development of dyskinesias and motor fluctuations characterize AR-JP. The reason why this patient did not show these clinical features is probably bilateral sterotaxic surgeries. Particularly, the second surgery was complicated by right hemiparesis. His siblings who developed parkinsonism showed typical clinical features of AR-JP.


Characterization of novel CD19-specific VHHs isolated from a camelid immune library by phage display.
Monoclonal antibody (mAb)-based immunotherapies have achieved promising outcomes in the treatment of immunological and oncological indications. CD19 is considered one of the most qualified antigens in the treatment of B-cell neoplasms. VHHs (nanobodies) are known for their physicochemical advantages over conventional mAbs rendering them suitable therapeutics and diagnostic tools. Herein, we aimed to isolate CD19-specific VHHs from a novel immune library using phage display. An immune VHH gene library was constructed. Using phage display and after five biopanning rounds, two monoclonal CD19-specific VHHs were isolated. The selected VHHs were expressed, purified, and characterized in terms of their affinity, specificity, sensitivity, and ability to target CD19-positive cell lines. Moreover, in silico analyses were employed for further characterization. A VHH library was developed, and because the outputs of the 4th biopanning round exhibited the most favorable characteristics, a panel of random VHHs was selected from them. Ultimately, two of the most favorable VHHs were selected and DNA sequenced (designated as GR37 and GR41). Precise experiments indicated that GR37 and GR41 exhibited considerable specificity, sensitivity, and affinity (1.15 × 107 M-1 and 2.08 × 107 M-1, respectively) to CD19. Flow cytometric analyses revealed that GR37 and GR41 could bind CD19 on the surface of cell lines expressing the antigen. Moreover, in silico experiments predicted that both VHHs target epitopes that are distinct from that targeted by the CD19-specific single-chain variable fragment (scFv) FMC63. The selected VHHs can be used as potential targeting tools for the development of CD19-based immunotherapeutics.


Visit Content Analysis: Doctor-Patient Communication in Patients with Type 2 Diabetes.
The primary care visit is an important opportunity to discuss and modify diabetes management. To gain insight into doctor-patient communication during primary care visits among English and Spanish speaking patients with type 2 diabetes and suboptimal glycemic control (HbA1c > 7%). We conducted a quantitative content analysis of audiotaped primary care visits in 2 patient cohorts. In Study 1 (31 English-speaking patients), we examined factors associated with management changes, and in Study 2 (20 Spanish-speaking patients and their Spanish-speaking providers), we examined the association of question asking with HbA1c control. This study was conducted between November 2017 and January 2020 across 8 primary care practices within Kaiser Permanente Northern California. In Study 1, the only factor significantly associated with a diabetes management change was patient identification of diabetes as a priority prior to the visit (91.7% had a management change vs 52.6% of patients who did not identify diabetes as a priority; p = 0.02). In Study 2, patients with poorer glycemic control (HbA1c ≥ 10.0) asked significantly fewer questions (3.4 ± 1.8 vs 10.7 ± 6.9 questions per 15 minutes; p = 0.004). Overall, despite receiving primary care from language-concordant providers, Spanish-speaking Study 2 patients asked fewer questions than English-speaking Study 1 patients (4.5 ± 2.9 vs 7.5 ± 3.7 questions per 15 minutes, respectively; p = 0.004). Our results highlight 2 potential strategies (preparing patients for their visits through identifying priorities and learning how to ask more questions during visits) for improving diabetes primary care.


Kinetic and kinematic compensations in amputee vertical jumping.
A unilateral transtibial amputation causes a disruption to the musculoskeletal system, which results in asymmetrical biomechanics. The current study aimed to assess the movement asymmetry and compensations that occur as a consequence of an amputation when performing a countermovement vertical jump. Six unilateral transtibial amputees and 10 able-bodied (AB) participants completed 10 maximal vertical jumps, and the highest jump was analyzed further. Three-dimensional lower limb kinematics and normalized (body mass) kinetic variables were quantified for the intact and prosthetic sides. Symmetry was assessed through the symmetry index (SI) for each individual and statistically using the Mann-Whitney U test between the intact and prosthetic sides for the amputee group. A descriptive analysis between the amputee and AB participants was conducted to explore the mechanisms of amputee jumping. The amputee jump height ranged from 0.09 to 0.24 m. In the countermovement, all ankle variables were asymmetrical (SI > 10%) and statistically different (p < .05) for the amputees. At the knee and hip, there was no statistical difference between the intact and prosthetic sides range of motion, although there was evidence of individual asymmetry. The knees remained more extended compared with the AB participants to prevent collapse. In propulsion, the prosthesis did not contribute to the work done and the ankle variables were asymmetrical (p < .05). The knee and hip variables were not statistically different between the intact and prosthetic sides, although there was evidence of functional asymmetry and the contribution tended to be greater on the intact compared with the prosthetic side. The lack of kinetic involvement of the prosthetic ankle and both knees due to the limitation of the prosthesis and the altered musculoskeletal mechanics of the joints were the reason for the reduced height jumped.


Placental alkaline phosphatase immunoreactivity in testicular germ-cell neoplasms.
We analyzed the sensitivity of a polyclonal antibody to placental alkaline phosphatase (PLAP) in labelling testicular germ-cell neoplasms, by utilizing the peroxidase-antiperoxidase technique. The immunoreactivity of 89 germ-cell tumors for PLAP was as follows: 98% of cases with seminomatous elements were PLAP-positive; 97% of embryonal carcinomas and 85% of endodermal sinus tumors also showed reactivity. Cytotrophoblastic cells were focally immunoreactive in one of two cases with choriocarcinomatous elements. Staining for PLAP was strongest and most diffuse in seminomas. Intratubular germ-cell neoplasia (ITGCN) or carcinoma in situ was present in 53 (84%) of 63 specimens that had adjacent seminiferous tubules available for evaluation; PLAP was demonstrated in 98% of these. In addition, the germ-cell elements in 11 gonadoblastomas were immunoreactive for PLAP. One of 17 cases of undescended testes had ITGCN that was also strongly immunoreactive, but the remaining 16 cases were negative. Five dysgenetic gonads without ITGCN were studied, and one was immunoreactive for PLAP. Three testicular biopsy specimens from infertile men without ITGCN were PLAP-negative. Our findings indicate that PLAP is a highly sensitive marker for seminomas, for the majority of embryonal carcinomas and endodermal sinus tumors of the testis, and for ITGCN.


Optical single-channel recording: imaging Ca2+ flux through individual ion channels with high temporal and spatial resolution.
Developments in imaging technology now enable visualization of the functioning of individual ion channels in living cells: something previously possible only by the electrophysiological patch-clamp technique. We review techniques that track channel gating via changes in intracellular [Ca2+] resulting from openings of Ca(2+)-permeable channels. Spatial and temporal resolution are optimized by monitoring Ca2+ close to the channel mouth, and we describe the use of two imaging modalities: confocal laser scan microscopy (linescan CLSM) and total internal reflection fluorescence microscopy (TIRFM). Both currently achieve a kinetic resolution of <10 ms, provide a simultaneous and independent readout from many channels, and enable their locations to be mapped with submicrometer resolution. TIRFM provides 2-D images from a very thin (approximately 100 nm) optical section, but it is restricted to channels in the plasma membrane of cells adhering close to a cover glass. In contrast, CLSM can image channels in intracellular membranes but, to achieve good temporal resolution, has been utilized only in a linescan mode with limited spatial information. We anticipate that imaging techniques will develop as a useful adjunct to patch-clamping for single-channel studies, with capabilities including simultaneous readout from multiple channels, high-resolution mapping of channel location, and mobility that is inaccessible by electrophysiological means. Optical single-channel recording is applicable to diverse voltage- and ligand-gated Ca(2+)-permeable channels and has potential for high-throughput functional analysis.


Prevalence of disability among US- and foreign-born Arab Americans: results from the 2000 US Census.
Although the prevalence of disability for various racial and ethnic groups has been documented, little attention has been paid to Arab Americans in the United States. We estimated the age- and sex-adjusted prevalence of disability among older Arab Americans and examined the association between nativity status and self-reported physical and self-care disability before and after controlling for covariates. We used data from the 5% Public Use Microdata Samples of the 2000 US Census. Our sample included 4,225 individuals 65 years of age and older who identified with an Arab ancestry. Of these, 2,280 were foreign-born and 1,945 were US-born. The age- and sex-adjusted prevalence of having a physical disability was 31.2% for foreign- and 23.4% for US-born older Arab Americans, and the age- and sex- adjusted prevalence of having a self-care disability was 13.5% for foreign- and 6.8% for US-born Arab Americans. Iraqis reported the highest estimates for both disabilities (physical, 36.2%; self-care, 19.8%) compared to other Arab ethnic groups. In the crude model, foreign-born Arab Americans were more likely (OR=1.32; 95% CI=1.28, 1.36) to report a physical disability compared to US-born Arab Americans. When adjusting for English language ability in the final model, the odds of having a physical disability for foreign-born Arab Americans was protective compared to US-born Arab Americans (OR=0.92; 95% CI=0.88, 0.96). In the crude model, foreign-born Arab Americans were 1.82 times (95% CI=1.74, 1.90) more likely to report a self-care disability compared to US-born Arab Americans. In the fully adjusted model, this association was slightly attenuated (OR=1.32; 95% CI=1.24, 1.41). These findings indicate English language ability is associated with variations in reporting a physical disability. Future studies should include better measures of acculturation. Arab Americans are heterogeneous and should be disaggregated both by subgroups and from the white category in order to reveal a more accurate health and disease status profile for these groups. These efforts will assist in tailoring more effective interventions in reducing or preventing disability among Arab Americans 65 years of age and older.


An uncertainty-based model of the effects of fixation on choice.
When people view a consumable item for a longer amount of time, they choose it more frequently; this also seems to be the direction of causality. The leading model of this effect is a drift-diffusion model with a fixation-based attentional bias. Here, we propose an explicitly Bayesian account for the same data. This account is based on the notion that the brain builds a posterior belief over the value of an item in the same way it would over a sensory variable. As the agent gathers evidence about the item from sensory observations and from retrieved memories, the posterior distribution narrows. We further postulate that the utility of an item is a weighted sum of the posterior mean and the negative posterior standard deviation, with the latter accounting for risk aversion. Fixating for longer can increase or decrease the posterior mean, but will inevitably lower the posterior standard deviation. This model fits the data better than the original attentional drift-diffusion model but worse than a variant with a collapsing bound. We discuss the often overlooked technical challenges in fitting models simultaneously to choice and response time data in the absence of an analytical expression. Our results hopefully contribute to emerging accounts of valuation as an inference process.


[Pleural sarcoidosis - a report of three cases].
Three patients with pleural sarcoidosis are reported. Pleural effusion in two patients and a massive pleural thickening that mimicked a tumour were observed. Histological examination of pleural biopsies revealed sarcoidosis. None of the patients received treatment. No recurrence of the pleural effusion was observed after a year of follow-up and the massive pleural thickening remained stable.


[The number of carotid artery operations in the Twente Medical Spectrum before and after the publication of relevant international research on the benefits of the relief of carotid stenosis].
To determine the effect of adequate scientific research on the treatment of extracranial stenosis of the internal carotid artery. Retrospective and comparative. Twenty Medical Spectrum, Enschede, the Netherlands. A comparison was made of the relevant data from 2 years of carotid artery surgery before (1989-1990; period I) and after the publication of two randomized multicentre studies (1994-1995; period II). The number of patients treated surgically and the number of carotid artery desobstructions had increased during period II by 339% and 319%, respectively. In period I, 25% of the patients had an asymptomatic ipsilateral stenosis of the internal carotid artery; in period II, this had decreased to 11%. In period I, 65% of the patients had a stenosis in excess of 70% of the diameter of the vessel; in period II this was 85%. The combined mortality and permanent disabling morbidity after 30 days was 6% in period I and 3% in period II. After the publication of two high-quality studies in 1991, the number of carotid artery operations increased by over 300%. The indications for the surgical treatment of stenosis were stricter rather than less strict in period II. The increase of the number of carotid desobstructions can be explained by the fact that GPs' and neurologists' referral to the vascular surgeon has changed. This change in the referring pattern may be the consequences of use of 'evidence-based' medicine.


Evaluating process in child and family interventions: aggression prevention as an example.
This article reports on 2 studies designed to develop and validate a set of measures for use in evaluating processes of child and family interventions. In Study 1 responses from 187 families attending an outpatient clinic for child behavior problems were factor analyzed to identify scales, consistent across sources: Alliance (Satisfactory Relationship with Interventionist and Program Satisfaction), Parenting Skill Attainment, Child Cooperation During Session, Child Prosocial Behavior, and Child Aggressive Behavior. Study 2 focused on patterns of scale scores among 78 families taking part in a 22-week preventive intervention designed to affect family relationships, parenting, and child antisocial and prosocial behaviors. The factor structure identified in Study 1 was replicated. Scale construct validity was demonstrated through across-source convergence, sensitivity to intervention change, and ability to discriminate individual differences. Path analysis validated the scales' utility in explaining key aspects of the intervention process. Implications for evaluating processes in family interventions are discussed.


PVB exerts anti-inflammatory effects by inhibiting the activation of MAPK and NF-κB signaling pathways and ROS generation in neutrophils.
Pinaverium bromide (PVB) has been shown to protect mice against sepsis, which is predominantly attributed to PVB-mediated anti-inflammatory effects by inhibiting primed neutrophils to produce proinflammatory cytokines. However, the underlying mechanism(s) by which PVB affects neutrophils remains unknown. In this study, we report that treatment with PVB either before or after LPS stimulation attenuated IL-1β and TNF-α expression at both mRNA and protein levels in LPS-activated murine neutrophils. Further experiments revealed that PVB inhibited the phosphorylation of ERK, JNK, and IκBα in LPS-stimulated murine neutrophils. Moreover, PVB reduced reactive oxygen species (ROS) levels via regulating NADPH oxidase 2 (NOX2) activity, as represented by inhibiting p47phox translocation from the cytoplasm to the cellular membrane. Importantly, PVB significantly attenuated IL-1β, TNF-α, IL-6, CXCL1 production in both LPS-stimulated low density neutrophils (LDNs) and normal density neutrophils (NDNs) isolated from septic patients. Collectively, we demonstrated that PVB exerts anti-inflammatory effect by attenuating ROS generation and suppressing the activation of MAPK and NF-κB signaling pathways, suggesting that PVB may act as a potential therapeutic agent for sepsis by inhibiting neutrophil priming and activation.


A homozygous SCN5A mutation in a severe, recessive type of cardiac conduction disease.
Cardiac sodium channels are key players in the generation and propagation of action potentials in the human heart. Heterozygous mutations in the SCN5A gene have been found to be associated with long QT syndrome, Brugada syndrome, and sinus node dysfunction (SND). Recently, overlapping arrhythmia phenotypes have been reported as well. Here we describe a novel recessive SCN5A mutation in a family originating from the German minority in White Russia. Four affected children with a history of early cardiac arrhythmia encompassing SND, conduction disease, and severe ventricular arrhythmias, are homozygous carriers of a novel SCN5A missense mutation (p.I230T) in the channel protein. Interestingly, the heterozygous mutation carriers had neither significant ECG abnormalities nor a history of cardiac events. Heterologous expression of SCN5A(I230T) channels revealed normal protein transport but altered biophysical sodium channel properties. Voltage range of both activation and inactivation were shifted in a way that resulted in decreased sodium current and loss of channel function. In conclusion, we describe a rare clinical condition with a novel SCN5A mutation causing a new type of complex cardiac arrhythmia. Unlike most previously reported sodium channelopathies, this overlap syndrome displays recessive inheritance characteristics and does not seem to follow simple Mendelian rules.


Arterio-venous fistula causing acute right heart failure: A rare complication of veno-arterial extracorporeal membrane oxygenation.
Introduction: Vascular complications are a major issue in V-A ECMO and can affect long term outcome. Among these, Artero-Venous (AV) fistulas may lead to right heart failure.Case History: A 73-years patient supported with V-A ECMO for post-cardiotomy biventricular failure developed right heart failure after V-A ECMO decannulation, requiring V-A ECMO recannulation. The presence of an AV femoral fistula was incidentally revealed during femoral vein cannulation, from which oxygenated blood was unexpectedly drawn. The angiographic assessment confirmed the presence of a fistula between superficial femoral artery and vein, at the site of the first venous ECMO cannulation. This was caused by the venous cannula that inadvertently passed across the artery and created an AV fistula that was opened by the venous cannula removal. The exclusion of the endovascular fistula allowed the right heart failure resolution.Conclusions: In the presence of right heart failure after recent vascular manipulation, AV fistula should be ruled out.


Intralocus tactical conflict: genetic correlations between fighters and sneakers of the dung beetle Onthophagus taurus.
Males and females differ in their phenotypic optima for many traits, and as the majority of genes are expressed in both sexes, some alleles can be beneficial to one sex but harmful to the other (intralocus sexual conflict; ISC). ISC theory has recently been extended to intrasexual dimorphisms, where certain alleles may have opposite effects on the fitness of males of different morphs that employ alternative reproductive tactics (intralocus tactical conflict; ITC). Here, we use a half-sib breeding design to investigate the genetic basis for ISC and ITC in the dung beetle Onthophagus taurus. We found positive heritabilities and intersexual genetic correlations for almost all traits investigated. Next, we calculated the intrasexual genetic correlation between males of different morphs for horn length, a sexually selected trait, and compared it to intrasexual correlations for naturally selected traits in both sexes. Intrasexual genetic correlations did not differ significantly between the sexes or between naturally and sexually selected traits, failing to support the hypothesis that horns present a reduction of intrasexual genetic correlations due to ITC. We discuss the implications for the idea of developmental reprogramming between male morphs and emphasize the importance of genetic correlations as constraints for the evolution of dimorphisms.


A national survey of the primary and acute care pediatric nurse practitioner educational preparation.
The past decade has been marked by a gradual expansion of the traditional primary care role of the pediatric nurse practitioner (PNP) into practice arenas that call for more acute and critical care of children. The purpose of the study was to explore the educational programming needs of dual (combined) track PNP programs that prepare graduates to provide care to children and adolescents across the continuum of health and illness. A two-phase, exploratory, mixed method design was utilized. An electronic survey was completed by 65% of PNP program directors in the country. Semi-structured telephone interviews were conducted with hospital-based PNPs who were practicing in roles that met a range of health care needs across the primary and acute care continuum. Primary care and acute care programs have more common than unique elements, and the vast majority of clinical competencies are common to both types of program. Only three competencies appear to be unique to acute care programs. The Association of Faculties of Pediatric Nurse Practitioner Programs should utilize existing evidence and develop guidelines for dual PNP programs that focus on the provision of care to children across a wide continuum of health and illness.


Mechanisms of recurrent varicocele after balloon occlusion or surgical ligation of the internal spermatic vein.
Clinical recurrence of varicocele developed in eight of 70 patients (11%) who underwent balloon occlusion of the internal spermatic vein (ISV) at the level of the third or fourth lumbar vertebra. Five patients also underwent venography for postoperative recurrence of varicocele. Recurrence was due to either collateral veins that bypassed the balloon occlusion or surgical ligation. The ISV was reconstituted in the pelvis of five of the eight patients following balloon occlusion, and in all patients following surgical ligation. Two patients who had recurrence after balloon embolization had clinically undetected right sided varicoceles. The technique of balloon occlusion of the spermatic vein should be modified: in most cases the balloon should be placed in the inguinal segment of the vein below the point where the collateral veins enter the ISV. The best site for balloon detachment can be determined by test occlusion and ISV venography to observe for blood flow within the collateral veins beyond the balloon. Venography is performed prior to detachment, and if the anatomy suggests a likely recurrence, the position of the balloon is shifted, which offers an advantage over other methods of treatment.


Fatty acids and immune responses--a new perspective in searching for clues to mechanism.
Dietary essential fatty acids are the precursors for eicosanoids. Among the eicosanoids derived from arachidonic acid, prostaglandin (PG) E2 is known to possess immunosuppressive actions. Thus, it has been a prevailing hypothesis that the immuno-modulatory roles of dietary fatty acids are mediated at least in part through the alteration of PG biosynthesis. PGs exert their biological effects through their cognate receptors. There are four subtypes of PGE receptors (EP1, EP2, EP3, and EP4) so far identified. Although the association of EP receptors with G proteins coupled to adenylate cyclase and the mobilization of intracellular calcium are well documented, downstream signaling pathways for these receptors are virtually unknown. Identification of downstream signaling pathways for each subtype of EP receptors and target genes regulated by the activation of the receptor will help with our understanding of the mechanism by which dietary fatty acids affect immune responses through the modulation of PGE2 biosynthesis. Emerging evidence suggests that fatty acids can additionally act as second messengers, regulators of signal transducing molecules or transcription factors. Acylation with long-chain fatty acids can occur on a variety of signaling molecules and can affect their membrane translocation and functions. Dietary fatty acids can alter functional properties of lipid mediators by changing the composition of acyl moieties of these molecules. Evidence accumulated recently indicates that long-chain unsaturated fatty acids and their metabolites bind and activate peroxisome proliferator-activated receptors (PPARs). PPARs are nuclear hormone receptors and transcription factors that regulate the expression of broad arrays of genes involved not only in lipid and glucose metabolism, but also in immune and inflammatory responses. PPARs may therefore be important cellular targets that mediate modulation of immune responses by dietary fatty acids. Together, it becomes clear now that multiple steps in various receptor-mediated signaling pathways can be modulated by dietary fatty acids. It will be a challenging task to quantitatively determine how different fatty acids alter functional properties of multitude of signaling components and final cellular responses. Elucidating the mechanism of actions of fatty acids on receptor-mediated signaling pathways in immuno-competent cells will provide a new insight for understanding the immuno-modulatory roles of dietary fatty acids.


Ligand- and weak base-induced redistribution of asialoglycoprotein receptors in hepatoma cells.
The receptor for asialoglycoproteins (ASGPR) was localized in human hepatoma Hep G2 cells by means of quantitative immunoelectron microscopy. Without ligand added to the culture medium, we found 34% of the total cellular receptors on the plasma membrane, 37% in compartment of uncoupling receptor and ligand (CURL), and 21% in a trans-Golgi reticulum (TGR) that was defined by the presence of albumin after immuno-double labeling. A small percent of the ASGPR was associated with coated pits, the Golgi stacks, and lysosomes. After incubation of the cells with saturating concentrations of the ligand asialo-orosomucoid (ASOR), the number of cell surface receptors decreased to 20% of total cellular receptors, whereas the receptor content of CURL increased by a corresponding amount to 50%. The ASGPR content of TGR remained constant. In contrast, after treatment of the cells with 300 microM of the weak base primaquine (PMQ), cell surface ASGPR had decreased dramatically to only 4% of total cellular receptors whereas label in the TGR had increased to 42%. ASGPR labeling of CURL increased only to 47%. The labeling of other organelles remained unchanged. This affect of PMQ was independent of the presence of additional ASOR. Implications for the intracellular pathway of the ASGPR are discussed.


Simulated seizures and spreading depression in a neuron model incorporating interstitial space and ion concentrations.
Sustained inward currents in neuronal membranes underlie tonic-clonic seizure discharges and spreading depression (SD). It is not known whether these currents flow through abnormally operating physiological ion channels or through pathological pathways that are not normally present. We have now used the NEURON simulating environment of Hines, Moore, and Carnevale to model seizure discharges and SD. The geometry and electrotonic properties of the model neuron conformed to a hippocampal pyramidal cell. Voltage-controlled transient and persistent sodium currents (I(Na,T) and I(Na,P)), potassium currents (I(K,DR) and I(K,A)), and N-methyl-D-aspartate (NMDA) receptor-controlled currents (I(NMDA)), were inserted in the appropriate regions of the model cell. The neuron was surrounded by an interstitial space where extracellular potassium and sodium concentration ([K(+)](o) and [Na(+)](o)) could rise or fall. Changes in intra- and extracellular ion concentrations and the resulting shifts in the driving force for ionic currents were continuously computed based on the amount of current flowing through the membrane. A Na-K exchange pump operated to restore ion balances. In addition, extracellular potassium concentration, [K(+)](o), was also controlled by a "glial" uptake function. Parameters were chosen to resemble experimental data. As long as [K(+)](o) was kept within limits by the activity of the Na-K pump and the "glial" uptake, a depolarizing current pulse applied to the cell soma evoked repetitive firing that ceased when the stimulating current stopped. If, however, [K(+)](o) was allowed to rise, then a brief pulse provoked firing that outlasted the stimulus. At the termination of such a burst, the cell hyperpolarized and then slowly depolarized and another burst erupted without outside intervention. Such "clonic" bursting could continue indefinitely maintained by an interplay of the rise and fall of potassium and sodium concentrations with membrane currents and threshold levels. SD-like depolarization could be produced in two ways, 1) by a dendritic NMDA-controlled current. Glutamate was assumed to be released in response to rising [K(+)](o). And 2) by the persistent (i.e., slowly inactivating) Na-current, I(Na,P). When both I(NMDA) and I(Na,P) were present, the two acted synergistically. We conclude that epileptiform neuronal behavior and SD-like depolarization can be generated by the feedback of ion currents that change ion concentrations, which, in turn, influence ion currents and membrane potentials. The normal stability of brain function must depend on the efficient control of ion activities, especially that of [K(+)](o).


[Production of monospecific serum against secretory IgA (anti-SC) and a standard for quantitative analysis].
The authors describe a method of obtaining monospecific serum against secretory IgA and the corresponding standard. An immunochemically pure (11.6S) secretory human IgA was extracted from the colostrum by salt fractionation and gel-filtration through Sephadex G-200 and Sepharose 6B; this IgA was used as an antigen for the immunization and the standard for the quantitative determination of SIgA in the secretions. Monospecific anti-SC-serum was obtained by successive exhaustion of the antiserum against the S IgA immunosorbents prepared from normal human serum and the serum of a patient suffering from A myeloma containing polymeric IgA forms.


Molecularly imprinted beads with double thermosensitive gates for selective recognition of proteins.
A new approach is reported on the use of poly(N-isopropylacrylamide) (PNIPAM)-coated molecularly imprinted beads (coated MIP beads) for controlling the release of protein. The coated MIP beads were composed of double layers, an internal thermosensitive lysozyme-imprinted layer, and an external PNIPAM layer. The coated MIP beads were prepared by two-step surface-initiated living-radical polymerization (SIP). In this systemic study, the coated MIP beads had good selectivity to the template protein (lysozyme) and temperature stimulus-responsive behavior, both of which were superior to those of MIP beads having a layer of thermosensitive lysozyme-imprinted polymer only. Using the coated MIP beads, reference proteins and the template lysozyme could be released separately at 38 °C and at 23 °C. The corresponding coated non-imprinted beads (coated NIP beads) did not have such double thermosensitive "gates" with specific selectivity for a particular protein. The proposed smart controlled imprinted system for protein is attractive for chemical carriers, drug-delivery system, and sensors.


Mass distribution of azithromycin for trachoma control is associated with increased risk of azithromycin-resistant Streptococcus pneumoniae carriage in young children 6 months after treatment.
Emerging evidence suggests that the mass distribution of azithromycin for trachoma control (MDA) may increase circulation of macrolide resistance in bacteria associated with severe pediatric infections in treated communities. We examined the effect of MDA on nasopharyngeal carriage of antibiotic-resistant Streptococcus pneumoniae among 1015 young children living in rural Tanzania. MDA with a single dose of oral azithromycin was provided in 4 of 8 communities where trachoma prevalence was ≥10%. Isolates were tested for susceptibility to azithromycin (AZM) and commonly used antibiotics by disk diffusion and Etest. We calculated the proportion of antibiotic-resistant S. pneumoniae carriage at baseline and again 1, 3, and 6 months after treatment, and at comparable intervals in the untreated villages. The proportion of AZM-resistant isolates was similar between groups at baseline (MDA: 35.8% vs non-MDA: 35.4%), however, this proportion was greater in the MDA group in all subsequent surveys. At 6 months, the percentage of AZM-resistant isolates was significantly higher in the MDA group (81.9% vs 46.9%, P < .001). The odds of AZM-resistant carriage was 5-fold greater in the MDA group (odds ratio, 4.95 [95% confidence interval, 3.23-7.61]). The proportion of isolates clinically resistant to AZM (minimum inhibitory concentration ≥16 µg/mL) was also significantly greater in the MDA group at 6 months (35.3% vs 12.4%, P < .006). Mass distribution of a single dose of oral azithromycin for trachoma was associated with increased circulation of macrolide-resistant S. pneumoniae carriage among young children in the 6 months following treatment. It is crucial that changes in antibiotic resistance patterns and their clinical significance in the treatment of severe pediatric infections be assessed in future MDA trials.


Health and safety in small workplaces: refocusing upstream.
Small workplaces have particular injury risks and are enduringly difficult for the occupational health and safety (OHS) system to reach. This paper puts forward an "upstream" perspective on OHS in small workplaces that moves beyond the attributes of the workplace and those who work there. The paper draws on and synthesizes ideas and findings from emerging upstream OHS research, our own empirical investigations in Ontario and Quebec, and our collected research experience in small workplace health. Upstream structures and processes (regulations, policies, services, interventions, professional practices) are often misaligned with the conditions of work and social relations of small workplaces. Key upstream factors include regulatory exemption, subcontracting, unionization levels, the changing character of small enterprise, joint management, service and inspection constraints, competing institutional accountabilities, institutional orientation to large business, and inappropriate service and policy. Misalignment of the OHS system with the nature and practical realities of small workplaces can undermine prevention and the management of ill health and injury. To address such misalignments, the paper calls for: 1) restructuring of data collection and consultation processes to increase the visibility, voice and credibility of small workplaces; 2) "audits" of OHS-related legislation, policy and interventions to assess and address implications for small workplaces; 3) reflection on current terms and concepts that render workers invisible and capture poorly the essence and (increasing) diversity of these workplaces; and 4) extension of the upstream gaze to the global level.


[Pneumococcal vaccination in adults].
Pneumococcal pneumonia and bacteraemia are an important cause of morbidity and mortality, especially in a number of risk groups. On the basis of data from available literature, there is no convincing evidence that vaccination of such risk groups, including all people over 65, with a polyvalent vaccine of pneumococcal capsular polysaccharides in addition to influenza vaccination offers any additional protection against the risk of acquiring pneumococcal pneumonia. There is adequate evidence that pneumococcal vaccination does protect against invasive infections and that in this respect vaccination of all elderly persons could be cost-effective. However, there are insufficient data on whether pneumococcal vaccination provides additional protection against invasive infections in people already receiving influenza vaccination. Therefore a well-considered assessment of the cost-effectiveness of applying such a strategy in the Netherlands is not yet possible. Vaccination of (imminent) immuno-compromised persons is only effective and of value if an adequate antibody response can be expected. There is as yet no proven advantage of vaccination with a conjugate vaccine in adults.


A pan-cancer atlas of somatic mutations in miRNA biogenesis genes.
It is a well-known and intensively studied phenomenon that the levels of many miRNAs are differentiated in cancer. miRNA biogenesis and functional expression are complex processes orchestrated by many proteins cumulatively called miRNA biogenesis proteins. To characterize cancer somatic mutations in the miRNA biogenesis genes and investigate their potential impact on the levels of miRNAs, we analyzed whole-exome sequencing datasets of over 10 000 cancer/normal sample pairs deposited within the TCGA repository. We identified and characterized over 3600 somatic mutations in 29 miRNA biogenesis genes and showed that some of the genes are overmutated in specific cancers and/or have recurrent hotspot mutations (e.g. SMAD4 in PAAD, COAD and READ; DICER1 in UCEC; PRKRA in OV and LIN28B in SKCM). We identified a list of miRNAs whose level is affected by particular types of mutations in either SMAD4, SMAD2 or DICER1 and showed that hotspot mutations in the RNase domains in DICER1 not only decrease the level of 5p-miRNAs but also increase the level of 3p-miRNAs, including many well-known cancer-related miRNAs. We also showed an association of the mutations with patient survival. Eventually, we created an atlas/compendium of miRNA biogenesis alterations providing a useful resource for different aspects of biomedical research.


Effects of oxycodone on immune function in patients undergoing radical resection of rectal cancer under general anesthesia.
This study aims to explore the effect of oxycodone hydrochloride injection on the immune function of patients who underwent radical resection of rectal cancer under general anesthesia.Eighty patients were enrolled and randomly divided into group A and B (n = 40, each). All patients underwent general intravenous anesthesia. At the end of surgery, each patient in group A was injected with 5 mg (5 mL) of oxycodone hydrochloride, while 5 mg (5 mL) of morphine hydrochloride in group B. Venous blood was withdrawn in both groups at different time points. Changes in the numbers of T lymphocyte subsets and natural killer (NK) cells were determined by flow cytometry.First the numbers of T lymphocyte subsets and NK cells at T1, T2, T3, and T4 decreased in both groups, compared with those at T0, and the differences were statistically significant. Furthermore, the numbers reduced to a minimum at T2 and began to recover at T3. Second the differences between group A and B at T1, T2, T3, and T4 were statistically significant; and the numbers of T lymphocytes and NK cells were higher in group A than in group B at corresponding time points.Oxycodone hydrochloride and morphine hydrochloride both have inhibitory effects on immune function in patients undergoing radical resection of rectal cancer after surgery. However, oxycodone hydrochloride has a smaller effect compared to morphine hydrochloride.


Ancient Metagenomic Studies: Considerations for the Wider Scientific Community.
Like modern metagenomics, ancient metagenomics is a highly data-rich discipline, with the added challenge that the DNA of interest is degraded and, depending on the sample type, in low abundance. This requires the application of specialized measures during molecular experiments and computational analyses. Furthermore, researchers often work with finite sample sizes, which impedes optimal experimental design and control of confounding factors, and with ethically sensitive samples necessitating the consideration of additional guidelines. In September 2020, early career researchers in the field of ancient metagenomics met (Standards, Precautions & Advances in Ancient Metagenomics 2 [SPAAM2] community meeting) to discuss the state of the field and how to address current challenges. Here, in an effort to bridge the gap between ancient and modern metagenomics, we highlight and reflect upon some common misconceptions, provide a brief overview of the challenges in our field, and point toward useful resources for potential reviewers and newcomers to the field.


A role for caspases in lens fiber differentiation.
There is increasing evidence that programmed cell death (PCD) depends on a novel family of intracellular cysteine proteases, called caspases, that includes the Ced-3 protease in the nematode Caenorhabditis elegans and the interleukin-1beta-converting enzyme (ICE)-like proteases in mammals. Some developing cells, including lens epithelial cells, erythroblasts, and keratinocytes, lose their nucleus and other organelles when they terminally differentiate, but it is not known whether the enzymatic machinery of PCD is involved in any of these normal differentiation events. We show here that at least one CPP32 (caspase-3)-like member of the caspase family becomes activated when rodent lens epithelial cells terminally differentiate into anucleate lens fibers in vivo, and that a peptide inhibitor of these proteases blocks the denucleation process in an in vitro model of lens fiber differentiation. These findings suggest that at least part of the machinery of PCD is involved in lens fiber differentiation.


Athletes' expectations with regard to officiating competence.
This study aimed to identify the cues upon which athletes rely when developing their expectations with regard to the competence of sports officials and to examine the sources of information, which are given priority in different kinds of sport (i.e. team, racquet and fighting sports). A questionnaire - the Athlete Perception of Sports Officials Questionnaire (APSO-Q) - was developed in which athletes (N=472) were asked to indicate the influence of 32 cues on their impressions of sports officials. Exploratory factor analysis (EFA) yielded a four-factor model (i.e. static cues, psychological, communicational and performance cues). Athletes mostly rely on psychological and personal communication attributes when evaluating officiating competence. Moreover, team players perceived that static cues were more influential when forming their expectations of sports officials than racquet players and fighting contestants. Such findings may have implications for athlete-official relationships and training of sports officials.


Dopamine action on hippocampal pyramidal cells.
Dopamine (DA) was applied to CA1 region pyramidal cells in slices of guinea pig hippocampus maintained in vitro in order to examine its electrophysiological effect on CNS neurons. DA induced hyperpolarization of membrane potential and an increased conductance in 75% the 21 CA1 neurons to which it was applied. DA also augmented the afterhyperpolarizations and increased conductance which normally follow spike trains in these neurons. These effects were not altered by intracellular injections of Cl- but were blocked when slices were bathed in Mn2+ solutions. The Mn2+ blockade of DA-induced hyperpolarizations could be overcome when large amounts of agonists were applied. The DA effects were long lasting, were mimicked by the dopamine agonists apomorphine and Epinine, and were blocked by the dopamine antagonists flupenthixol and chlorpromazine. Extracellular or intracellular application of cyclic AMP mimicked the effects of DA. The results suggest that DA-induced hyperpolarization and conductance changes are mediated by a Ca2+-activated K+ conductance. DA may increase the intracellular Ca2+ concentration through effects on one of the Ca2+ buffering mechanisms. The long duration of these effects suggest that DA works though some intracellular intermediary, perhaps cyclic AMP, considering that the actions of cyclic AMP on membrane properties are similar to those of DA. The dopaminergic projection to the hippocampus should have a powerful inhibitory action, which would be most effective in modulating the activities of neurons exhibiting high levels of excitability, particularly cells involved in cyclical burst generation.


Neopterin as a new biochemical marker for diagnosis of allograft rejection. Experience based upon evaluation of 100 consecutive cases.
The use of daily urinary neopterin evaluation to detect immunological complications has been tested in 96 consecutive cadaveric kidney recipients, three liver recipients, and one pancreas recipient. In 29 of these patients an immunologically uncomplicated posttransplant course was associated with stable or low neopterin levels, or both. In only 5% of daily determinations on these patients were increasing or high neopterin levels seen. On the other hand, major immunological complications, such as acute rejection episodes (38 cases), viral infections (17 cases), or both problems (8 cases), were preceded by increasing or high neopterin levels or both--on the average by one day. Withdrawal of cyclosporine was also found to be followed by increase of urinary neopterin levels. Neopterin evaluation enabled reliable and accurate prediction of immunological complications in 95% of patients with acute rejections and in 100% of patients with viral infections. It thus appears that daily assessment of urinary neopterin levels represents a useful tool for biochemical detection of immunological complications in allograft recipients.


Role of the cytoplasmic domain of the beta-subunit of integrin alpha(v)beta6 in infection by foot-and-mouth disease virus.
Field isolates of foot-and-mouth disease virus (FMDV) are believed to use RGD-dependent integrins as cellular receptors in vivo. Using SW480 cell transfectants, we have recently established that one such integrin, alpha(v)beta6, functions as a receptor for FMDV. This integrin was shown to function as a receptor for virus attachment. However, it was not known if the alpha(v)beta6 receptor itself participated in the events that follow virus binding to the host cell. In the present study, we investigated the effects of various deletion mutations in the beta6 cytoplasmic domain on infection. Our results show that although loss of the beta6 cytoplasmic domain has little effect on virus binding, this domain is essential for infection, indicating a critical role in postattachment events. The importance of endosomal acidification in alpha(v)beta6-mediated infection was confirmed by experiments showing that infection could be blocked by concanamycin A, a specific inhibitor of the vacuolar ATPase.


Development of Gut Microbiota in a Mouse Model of Ovalbumin-induced Allergic Diarrhea under Sub-barrier System.
This study aimed to present a mouse model of ovalbumin (OVA) induced allergic diarrhea under a sub-barrier system and investigate the development of gut microbiota in this model. Male BALB/c mice were systemically sensitized with OVA or sham-sensitized with saline, and followed by oral OVA intubation, leading to OVA-specific acute diarrhea. Compared with sham-sensitized mice, sera OVA-specific IgG1 and total IgE in OVA-sensitized mice were dramatically elevated, and the number of mast cells was greatly increased in the jejunum of the OVA-sensitized mice. Principle component analysis of the DGGE profile showed that samples from group of OVA-sensitized mice and group of sham-sensitized mice were scattered into two different regions. Real-time PCR analysis showed that the number of 16S rRNA gene copies of Lactobacillus in the colon of OVA-sensitized mice decreased significantly, while there was no significant difference in the number of Bifidobacterium and total bacteria. In conclusion, OVA-specific allergic diarrhea was successfully induced under a sub-barrier system, and changes of allergic reactions during induction was coupled with changes in gut microbiota, especially the number of colonic Lactobacillus, but the role of gut microbiota in the development of food allergic reactions needs to be further evaluated.


Sex Ratio at Birth in Vietnam: Results From Data in CHILILAB HDSS, 2004 to 2013.
This study aimed to explore the association of demographic and socioeconomic characteristics and imbalanced sex ratio at birth (SRB) in Chi Linh district, Hai Duong. The data were collected from a longitudinal study using a community-based periodic, referred as Chi Linh Health and Demographic Surveillance System (CHILILAB HDSS) during 2004 to 2013. A total of 7568 children were analyzed. Results showed that SRB in Chi Linh dramatically increased to the imbalanced sex ratio (114.6 boys to 100 girls) by 2013. SRB was associated with birth order and sex of preceding siblings. SRB was extremely high among families without any sons (136/100). SRB was highest among families having third or more children (175/100). Imbalanced SRB was more likely to occur among women working in small business/homemakers and others, women who attained high education level, and women in wealthy households. We suggested further efforts to tackle imbalanced SRB in periurban areas in Vietnam.


Decreased hexosamine biosynthesis in GH-deficient dwarf rat muscle. reversal with GH, but not IGF-I, therapy.
Enhanced glucose flux via the hexosamine biosynthesis pathway (HNSP) has been implicated in insulin resistance. We measured L-glutamine:D-fructose-6-phosphate amidotransferase activity (GFAT, a rate-limiting enzyme) and concentrations of UDP-N-acetyl hexosamines (UDP-HexNAc, major products of HNSP) in muscle and liver of growth hormone (GH)-deficient male dwarf (dw) rats. All parameters measured, except body weight, were similar in 5-wk-old control and dw rats. Muscle GFAT activity declined progressively with age in controls and dw rats but was consistently 30-60% lower in 8- to 14-wk-old dw rats vs. age-matched controls; UDP-HexNAc concentrations in muscle were concomitantly 30% lower in dw rats vs. controls (P < 0.01). Concentrations of UDP-hexoses, GDP-mannose, and UDP in muscle were similar in control and dw rats. Muscle HNSP activity was similarly diminished in fed and fasted dw rats. In liver, only a small difference in GFAT activity was evident between controls and dw rats, and no differences in UDP-HexNAc concentrations were observed. Treatment with recombinant human GH (rhGH) for 5 days restored UDP-HexNAc to control levels in dw muscles (P < 0.01) and partially restored GFAT activity. Insulin-like growth factor I treatment was ineffective. We conclude that GH participates in HNSP regulation in muscle.