Mice humanised for the EGF receptor display hypomorphic phenotypes in skin, bone and heart.
Mice lacking the epidermal growth factor receptor (EGFR) develop epithelial defects and a neurodegenerative disease and die within the first month of birth. By employing a conditional knock-in approach using the human EGFR cDNA mice humanised for EGFR (hEGFRKI/KI) were generated. Homozygous hEGFRKI/KI mice are viable and live up to six months. However, these mice are growth retarded and show skin and hair defects similar to Egfr-/- mutants. Interestingly, the neurodegeneration is fully rescued in hEGFRKI/KI mice, however, they develop a severe heart hypertrophy with semilunar valve abnormalities. Moreover, hEGFRKI/KI mice display accelerated chondrocyte and osteoblast differentiation, a phenotype that is also present in Egfr-/- mice and has not been previously described. The severity of the phenotypes correlates with the expression levels of the hEGFRKI allele, which is not efficiently expressed in epithelial and bone cells, but is expressed at similar and even higher levels as the endogenous Egfr in brain and heart. These results demonstrate that mice humanised for EGFR display tissue-specific hypomorphic phenotypes and describe a novel function for EGFR in bone development.


Reliable evidence for implicit attentional responses to aversive conditioned stimuli.
Evidence for implicit aversive learning effects has been criticized for its lack of experimental rigor and statistical reliability. Here we examine whether attentional emotional responses to aversive conditioned stimuli can occur in the absence of stimulus-outcome contingency awareness, and use a novel Bayesian tool to reliably perform a post hoc categorization of awareness. Across two experiments (n = 40 and 69) participants completed an aversive conditioning task. A novel Bayesian awareness categorization tool was applied to sensitively measure contingency awareness. Finally, attentional and subjective responses toward conditioned stimuli were measured. For participants unaware of contingencies, conditioned stimuli generated attentional avoidance, but only aware participants showed subjective learning effects. For both experiments, awareness scores for unaware participants did not regress above chance level on a subsequent awareness check, revealing a reliable determination of unawareness states. These findings provide evidence for the existence of aversive learning in the absence of contingency awareness, as demonstrated via conditioned attentional responses, and build an analytical framework that can be extrapolated to other implicit paradigms. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Management of adults with paediatric-onset chronic liver disease: strategic issues for transition care.
Advances in the management of children with chronic liver disease have enabled many to survive into adulthood with or without their native livers, so that the most common of these conditions are becoming increasingly common in adult hepatology practice. Because the aetiologies of chronic liver disease in children may vary significantly from those in adulthood, adults with paediatric-onset chronic liver disease may often present with clinical manifestations unfamiliar to their adulthood physician. Transition of medical care to adult practice requires that the adulthood medical staff (primary physicians and subspecialists) have a comprehensive knowledge of childhood liver disease and their implications, and of the differences in caring for these patients. Pending still unavailable Scientific Society guidelines, this article examines causes, presentation modes, evaluation, management, and complications of the main paediatric-onset chronic liver diseases, and discusses key issues to aid in planning a program of transition from paediatric to adult patients.


Cost-effectiveness of a novel molecular test for cytologically indeterminate thyroid nodules.
Determining which patients with thyroid nodules require surgery is limited by cytologically indeterminate findings. A new approach for preoperative molecular classification of cytologically indeterminate thyroid nodules has a reported sensitivity of 91% and specificity of 75%; however, its cost-effectiveness has yet to be assessed. Our objective was to evaluate the 5-yr cost-effectiveness of routine use of a molecular test in adult patients with indeterminate fine-needle aspiration biopsy results from a societal perspective. A 16-state Markov decision model was developed. Probabilities, costs, and quality-adjusted life years (QALY) were estimated from literature review, U.S. Department of Health and Human Services data, Medicare reimbursement schedules, and expert opinion. Decision analysis of a hypothetical group of adult patients with cytologically indeterminate thyroid nodules was conducted. Incremental cost-effectiveness ratio was calculated as incremental cost (measured in U.S. dollars) divided by incremental effectiveness (measured in QALY). Modifying current practice with use of the molecular test resulted in 74% fewer surgeries for benign nodules with no greater number of untreated cancers. Over 5 yr, mean discounted cost estimates were $12,172 for current practice and $10,719 with the molecular test. Current practice and molecular test use produced 4.50 and 4.57 QALY, respectively. Use of this novel molecular test for differential diagnosis of cytologically indeterminate thyroid nodules can potentially avoid almost three fourths of currently performed surgeries in patients with benign nodules. Compared with current practice based on cytological findings alone, use of this test may result in lower overall costs and modestly improved quality of life for patients with indeterminate thyroid nodules.


Patellofemoral joint geometry and osteoarthritis features 3-10 years after knee injury compared with uninjured knees.
In this cross-sectional study, we compared patellofemoral geometry in individuals with a youth-sport-related intra-articular knee injury to uninjured individuals, and the association between patellofemoral geometry and magnetic resonance imaging (MRI)-defined osteoarthritis (OA) features. In the Youth Prevention of Early OA (PrE-OA) cohort, we assessed 10 patellofemoral geometry measures in individuals 3-10 years following injury compared with uninjured individuals of similar age, sex, and sport, using mixed effects linear regression. We also dichotomized geometry to identify extreme (>1.96 standard deviations) features and assessed likelihood of having extreme values using Poisson regression. Finally, we evaluated the associations between patellofemoral geometry with MRI-defined OA features using restricted cubic spline regression. Mean patellofemoral geometry did not differ substantially between groups. However, compared with uninjured individuals, injured individuals were more likely to have extremely large sulcus angle (prevalence ratio [PR] 3.9 [95% confidence interval, CI: 2.3, 6.6]), and shallow lateral trochlear inclination (PR 4.3 (1.1, 17.9)) and trochlear depth (PR 5.3 (1.6, 17.4)). In both groups, high bisect offset (PR 1.7 [1.3, 2.1]) and sulcus angle (PR 4.0 [2.3, 7.0]) were associated with cartilage lesion, and most geometry measures were associated with at least one structural feature, especially cartilage lesions and osteophytes. We observed no interaction between geometry and injury. Certain patellofemoral geometry features are correlated with higher prevalence of structural lesions compared with injury alone, 3-10 years following knee injury. Hypotheses generated in this study, once further evaluated, could contribute to identifying higher-risk individuals who may benefit from targeted treatment aimed at preventing posttraumatic OA.


Biology and treatment of myeloma.
New strategies for the treatment of multiple myeloma, particularly high-dose therapy with peripheral blood stem cell transplantation, can achieve complete response in up to 50% of cases. The patient's condition eventually relapses because transplantation does not completely eliminate myeloma. Posttreatment interferon may prolong responses achieved by transplantation or chemotherapy, but it does not necessarily prolong survival. New approaches are being developed to further eliminate myeloma cells after an incomplete response achieved by transplantation or other chemotherapy. These approaches aim at myeloma cell surface antigens, T-cell immunity, and biological mechanisms of myeloma growth and dissemination. Biological targets include interleukin-6 (the central myeloma growth factor), interleukin-1 beta (an amplifier of stromal and bone cell production of interleukin-6), and serum soluble interleukin-6 receptor (an enhancer of myeloma cell response to interleukin-6). Efforts to eliminate circulating myeloma cells from peripheral blood stem cell harvests use positive selection or purging to provide a product that will engraft only normal stem cells, not myeloma cells. Multidrug resistance, a major factor in treatment failure, can be reversed by agents that inhibit the multidrug resistance protein on the myeloma cell surface. Finally, expanded knowledge of biologic mechanisms has led to the development of new prognostic factors. These aim to identify patients in clinical trials who can benefit from treatment regimens designed to overcome their impact on survival. Translating new biological advances into treatment programs is essential to improving therapy for patients with myeloma.


Cytogenetics in the management of bone marrow failure syndromes: Guidelines from the Groupe Francophone de Cytogénétique Hématologique (GFCH).
Bone marrow failure syndromes are rare disorders characterized by bone marrow hypocellularity and resultant peripheral cytopenias. The most frequent form is acquired, so-called aplastic anemia or idiopathic aplastic anemia, an auto-immune disorder frequently associated with paroxysmal nocturnal hemoglobinuria, whereas inherited bone marrow failure syndromes are related to pathogenic germline variants. Among newly identified germline variants, GATA2 deficiency and SAMD9/9L syndromes have a special significance. Other germline variants impacting biological processes, such as DNA repair, telomere biology, and ribosome biogenesis, may cause major syndromes including Fanconi anemia, dyskeratosis congenita, Diamond-Blackfan anemia, and Shwachman-Diamond syndrome. Bone marrow failure syndromes are at risk of secondary progression towards myeloid neoplasms in the form of myelodysplastic neoplasms or acute myeloid leukemia. Acquired clonal cytogenetic abnormalities may be present before or at the onset of progression; some have prognostic value and/or represent somatic rescue mechanisms in inherited syndromes. On the other hand, the differential diagnosis between aplastic anemia and hypoplastic myelodysplastic neoplasm remains challenging. Here we discuss the value of cytogenetic abnormalities in bone marrow failure syndromes and propose recommendations for cytogenetic diagnosis and follow-up.


A phase-1, open-label, single-dose study of the pharmacokinetics of buparlisib in subjects with mild to severe hepatic impairment.
The pharmacokinetics (PK) and safety of single-dose buparlisib (30 mg) were assessed in subjects with mild to severe hepatic impairment (n = 6 each) relative to healthy controls (n = 13). Blood samples were collected until 336 hours postdose and evaluated by liquid chromatography tandem mass spectrometry. PK parameters (including area under the curve [AUC∞ ] and Cmax ) were derived using noncompartmental analysis. Buparlisib was rapidly absorbed in all groups (median Tmax 1.0-1.3 h). Buparlisib exposure (AUC∞ ) was moderately increased in subjects with mild (geometric mean ratio [GMR] 1.16; 90%CI 0.81, 1.65), moderate (GMR 1.14; 90%CI 0.80, 1.63), or severe (GMR 1.20; 90%CI 0.84, 1.72) hepatic impairment, relative to healthy controls. Apparent oral clearance was similar across groups. Due to a higher unbound fraction in the severe group (0.21) than all other groups (0.17), subjects with severe hepatic impairment had greater exposure to unbound buparlisib (GMR relative to healthy controls: AUC∞ 1.52; 90%CI 1.09, 2.13; Cmax 1.83; 90%CI 1.42, 2.36). The results indicate that a buparlisib dose adjustment may not be necessary for patients with mild to moderate hepatic impairment. The safety and therapeutic indices should be considered before determining if a dose adjustment is appropriate for patients with severe hepatic impairment.


Lung cancer after Hodgkin's disease: a nested case-control study of the relation to treatment.
To investigate the causes of the raised risk of lung cancer in patients who have had Hodgkin's disease, and in particular the relationship to treatment. A nested case-control study was conducted within a cohort of 5,519 patients with Hodgkin's disease treated in Britain during 1963 through 1993. For 88 cases of lung cancer and 176 matched control subjects, information on treatment and other risk factors was extracted from hospital case-notes, and odds ratios for lung cancer in relation to these factors were calculated. Risk of lung cancer was borderline significantly greater in patients treated with mechlorethamine, vincristine, procarbazine, and prednisone (MOPP) chemotherapy than those who did not receive this treatment (relative risk [RR] = 1.66; 95% confidence interval [CI], 0.99 to 2.82), and increased with number of cycles of MOPP (P =.07). Exclusion of lung cancers for which histologic confirmation was not available strengthened these associations (RR = 2.41; 95% CI, 1.33 to 4.51; P =.004 for any MOPP and P =.007 for trend with number of cycles of MOPP). Risks were not raised, however, after chlorambucil, vinblastine, procarbazine, and prednisone treatment. There was evidence that the raised risk of lung cancer occurring in relation to radiotherapy was restricted to histologies other than adenocarcinoma. The results suggest that MOPP chemotherapy may lead to elevated risk of lung cancer, at least in certain subgroups of patients. The role of chemotherapy in the etiology of lung cancer after Hodgkin's disease deserves further investigation.


Comparing the Rat Grimace Scale and a composite behaviour score in rats.
There is a growing interest in the use of voluntarily displayed ongoing behaviours in laboratory animals to assess the pain experience. In rats, two behavioural pain scales, the Rat Grimace Scale (RGS, a facial expression scale) and a composite behaviour score (CBS, a behavioural ethogram reliant on postural changes), are both promising pain assessment methods. Both scales have been used to assess pain in a laparotomy model, however, they have never been compared directly and the knowledge of how different analgesics may affect these two scales is limited. This study aimed to provide a comparison to discriminate the temporal and analgesic response in a laparotomy model. Female Wistar (n = 26) and Sprague Dawley rats (n = 26) were block randomized to receive saline, meloxicam (2 mg/kg) or buprenorphine (0.05 mg/kg) 30 minutes before laparotomy. Rats were video-recorded before surgery (BL) and at 30, 150, 270, and 390 minutes post-operatively. Videos were assessed according to both scales by a trained, blinded observer. Both CBS and RGS scores increased significantly at all post surgical timepoints in the saline group. Both buprenorphine and meloxicam reduced CBS scores to baseline levels following laparotomy; however, RGS scores were only reduced following buprenorphine. RGS scores in the meloxicam group remained similar to scores of the saline group. These findings suggest that the CBS and RGS differ in their sensitivity to discriminating analgesic effects.


Cutting edge: lupus susceptibility interval Sle3/5 confers responsiveness to prolactin in C57BL/6 mice.
Prolactin is of interest in the pathogenesis of systemic lupus erythematosus (SLE) because almost 25% of SLE patients display hyperprolactinemia, and serum prolactin correlates with disease activity in some patients. Furthermore, hyperprolactinemia causes early mortality in lupus-prone mice and induces a lupus-like phenotype in nonspontaneously autoimmune mice. We show here that the immunomodulatory effects of prolactin are genetically determined; hyperprolactinemia breaks B cell tolerance and causes a lupus-like serology in BALB/c mice expressing a transgene encoding the H chain of an anti-DNA Ab but not in C57BL/6 transgenic mice. In C57BL/6 mice that express both the H chain transgene and the lupus susceptibility interval Sle3/5, prolactin induces increased serum titers of anti-DNA Ab and glomerular Ig depositions. The increase in costimulation due to prolactin-mediated up-regulation of both CD40 on B cells and CD40L on T cells would appear to play a central role in lupus induction in this model.


Bosentan in pulmonary hypertension associated with fibrotic idiopathic interstitial pneumonia.
Pulmonary hypertension (PH) associated with fibrotic idiopathic interstitial pneumonia (IIP; idiopathic pulmonary fibrosis and nonspecific interstitial pneumonia) confers important additional morbidity and mortality. To evaluate the safety and clinical efficacy of the dual endothelin-1 receptor antagonist bosentan in this patient group. In a randomized, double-blind, placebo-controlled study, 60 patients with fibrotic IIP and right heart catheter confirmed PH were randomized 2:1 to bosentan (n = 40) or placebo (n = 20). The primary study endpoint was a fall from baseline pulmonary vascular resistance index (PVRi) of 20% or more over 16 weeks. Sixty patients (42 men; mean age, 66.6 ± 9.2 yr), with a mean pulmonary artery pressure of 36.0 (± 8.9) mm Hg, PVRi 13.0 (± 6.7) Wood Units/m(2) and reduced cardiac index of 2.21 (± 0.5) L/min/m(2) were recruited to the study. Accounting for deaths and withdrawals, paired right heart catheter data were available for analysis in 39 patients (bosentan = 25, placebo = 14). No difference in the primary outcome was detected, with seven (28.0%) patients receiving bosentan, and four (28.6%) receiving placebo achieving a reduction in PVRi of greater than or equal to 20% (P = 0.97) at 16 weeks. There was no change in functional capacity or symptoms between the two groups at 16 weeks, nor any difference in rates of serious adverse events or deaths (three deaths in each group). This study shows no difference in invasive pulmonary hemodynamics, functional capacity, or symptoms between the bosentan and placebo groups over 16 weeks. Our data do not support the use of the dual endothelin-1 receptor antagonist, bosentan, in patients with PH and fibrotic IIP. Clinical trial registered with www.clinicaltrials.gov (NCT 00637065).


Increased biting rate and reduced fertility in sporozoite-infected mosquitoes.
Because salivary function and blood location are impaired in sporozoite-infected mosquitoes, we determined whether such pathology also could lead to an increased biting rate. For 5 days, we compared relative daily biting rates of Plasmodium gallinaceum sporozoite-infected mosquitoes (Aedes aegypti) and noninfected mosquitoes with an olfactometer. Mosquitoes then were exposed for 5 min to an anesthetized guinea pig. Infected mosquitoes exhibited a significant increase in olfactometer response which was also reflected in a decreased egg output. We conclude that if duration of contact with a host is limited, then infected mosquitoes may make more attempts at probing before being successful, and thus enhance transmission.


Seasonality of Phosphorus and Nitrate Retention in Riparian Buffers.
Measurement of the retention of dissolved nutrients in riparian areas with snowmelt runoff are much less common than for rainfall runoff, but low rates of uptake or the release of nutrients with snowmelt have been attributed to frozen soils, lower biotic uptake, and release of nutrients from senesced vegetation. In the research presented here, we evaluate whether the potential for uptake of dissolved reactive phosphorus (DRP) and NO differ significantly between snowmelt and summer seasons with flow through 13 riparian buffers downstream of cropland in Manitoba, Canada. Flow-through buffers in small channels are typical in this landscape, and pulsed releases of a conservative tracer and dissolved nutrients were used to measure uptake rates. Although mean uptake rates of NO were higher in summer than for snowmelt, responses varied widely. Aerial uptake rate of DRP showed a significant negative relationships with soil Olsen-P ( = 0.54, < 0.001) and a P saturation index ( = 0.48, < 0.001) across both seasons. Biological processes may be of greater importance for NO retention, but DRP retention appears to be driven by adsorption-desorption regardless of season. Olsen-P is identified as a good indicator of potential for release or retention of DRP in riparian buffers with fine-textured calcareous soils, for both snowmelt and summer seasons. Soil testing may be a good tool to aid in the siting of new buffers and to track the effectiveness of management interventions to remove P from riparian areas, such as harvest of vegetation.


Identification of a common variant with potential pleiotropic effect on risk of inflammatory bowel disease and colorectal cancer.
Although genome-wide association studies (GWAS) have separately identified many genetic susceptibility loci for ulcerative colitis (UC), Crohn's disease (CD) and colorectal cancer (CRC), there has been no large-scale examination for pleiotropy, or shared genetic susceptibility, for these conditions. We used logistic regression modeling to examine the associations of 181 UC and CD susceptibility variants previously identified by GWAS with risk of CRC using data from the Genetics and Epidemiology of Colorectal Cancer Consortium and the Colon Cancer Family Registry. We also examined associations of significant variants with clinical and molecular characteristics in a subset of the studies. Among 11794 CRC cases and 14190 controls, rs11676348, the susceptibility single nucleotide polymorphism (SNP) for UC, was significantly associated with reduced risk of CRC (P = 7E-05). The multivariate-adjusted odds ratio of CRC with each copy of the T allele was 0.93 (95% CI 0.89-0.96). The association of the SNP with risk of CRC differed according to mucinous histological features (P heterogeneity = 0.008). In addition, the (T) allele was associated with lower risk of tumors with Crohn's-like reaction but not tumors without such immune infiltrate (P heterogeneity = 0.02) and microsatellite instability-high (MSI-high) but not microsatellite stable or MSI-low tumors (P heterogeneity = 0.03). The minor allele (T) in SNP rs11676348, located downstream from CXCR2 that has been implicated in CRC progression, is associated with a lower risk of CRC, particularly tumors with a mucinous component, Crohn's-like reaction and MSI-high. Our findings offer the promise of risk stratification of inflammatory bowel disease patients for complications such as CRC.


Supplementing lactating dairy cows with a vitamin B12 precursor, 5,6-dimethylbenzimidazole, increases the apparent ruminal synthesis of vitamin B12.
Cobalamin (CBL), the biologically active form of vitamin B12, and its analogs, are produced by bacteria only if cobalt supply is adequate. The analogs differ generally by the nucleotide moiety of the molecule. In CBL, 5,6-dimethylbenzimidazole (5,6-DMB) is the base in the nucleotide moiety. The present study aimed to determine if a supplement of 5,6-DMB could increase utilization of dietary cobalt for synthesis of CBL and change ruminal fermentation, nutrient digestibility, omasal flow of nutrients and ruminal protozoa counts. Eight ruminally cannulated multiparous Holstein cows (mean±standard deviation=238±21 days in milk and 736±47 kg of BW) were used in a crossover design. Cows were randomly assigned to a daily supplement of a gelatin capsule containing 1.5 g of 5,6-DMB via the rumen cannula or no supplement. Each period lasted 29 days and consisted of 21 days for treatment adaptation and 8 days for data and samples collection. Five corrinoids, CBL and four cobamides were detected in the total mixed ration and the omasal digesta from both treatments. The dietary supplement of 5,6-DMB increased (P=0.02) apparent ruminal synthesis of CBL from 14.6 to 19.6 (s.e.m. 0.8) mg/day but had no effect (P>0.1) on apparent ruminal synthesis of the four analogs. The supplement of 5,6-DMB had no effect (P>0.1) on milk production and composition, or on protozoal count, ruminal pH and concentrations of volatile fatty acids and ammonia nitrogen in rumen content. The supplement had also no effect (P>0.1) on intake, omasal flow and apparent ruminal digestibility of dry matter, organic matter, NDF, ADF and nitrogenous fractions. Plasma concentration of CBL was not affected by treatments (P=0.98). Providing a preformed part of the CBL molecule, that is, 5,6-DMB, increased by 34% the apparent ruminal synthesis of CBL by ruminal bacteria but had no effect on ruminal fermentation or protozoa count and it was not sufficient to increase plasma concentrations of the vitamin. Even though the efficiency of cobalt utilization for apparent synthesis of CBL was increased from 2.0% to 2.7% by the 5,6-DMB supplement, this improved efficiency was still very low. Further research is needed to identify the factors affecting efficiency of utilization of cobalt for synthesis of CBL by the bacterial populations in rumen.


Age at onset in bipolar affective disorders: a review.
Bipolar affective disorder (BPAD) is a multifactorial disorder with various clinical presentations. Etiologic heterogeneity may partly underlie the phenotypic heterogeneity. Efforts to dissect BPAD have been based on the course of the disorders (BP I versus BP II or rapid cycling), comorbidity pattern (panic attacks, suicide attempts, addiction or hyperactivity), differences between the sexes, and clinical pattern (cycloid and puerperal psychosis). The present article provides a comprehensive review of the existing data, showing that age at onset (AAO) identifies homogeneous sub-groups of patients with BPAD. Recent work has demonstrated the existence of three--early, intermediate and late--onset bipolar sub-groups based on AAO, following Kendell's criteria for validity (The American Journal of Psychiatry 2003; 160: 999). We will also show how these distinctions may be of use in the search for genetic vulnerability factors and other pathogenic influences. Following Kendell's criteria, we show that AAO of bipolar disorders has been tested with most of the available strategies for establishing the validity of clinical syndromes. We also present data from genetic epidemiologic studies in bipolar disorder, showing that AAO sub-groups may reduce the underlying genetic heterogeneity. No accurate AAO thresholds to define valid sub-groups have been identified precisely. Until recently, studies defined early- and late-onset as corresponding to early or mid-adulthood, not taking into account juvenile-onset bipolar disorder. A recently proposed theoretical model with three AAO sub-groups (onset age 17, 27 and 46) is discussed.


Improved SARS-CoV-2 sequencing surveillance allows the identification of new variants and signatures in infected patients.
Genomic surveillance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the only approach to rapidly monitor and tackle emerging variants of concern (VOC) of the COVID-19 pandemic. Such scrutiny is crucial to limit the spread of VOC that might escape the immune protection conferred by vaccination strategies or previous virus exposure. It is also becoming clear now that efficient genomic surveillance would require monitoring of the host gene expression to identify prognostic biomarkers of treatment efficacy and disease progression. Here we propose an integrative workflow to both generate thousands of SARS-CoV-2 genome sequences per week and analyze host gene expression upon infection. In this study we applied an integrated workflow for RNA extracted from nasal swabs to obtain in parallel the full genome of SARS-CoV-2 and transcriptome of host respiratory epithelium. The RNA extracted from each sample was reverse transcribed and the viral genome was specifically enriched through an amplicon-based approach. The very same RNA was then used for patient transcriptome analysis. Samples were collected in the Campania region, Italy, for viral genome sequencing. Patient transcriptome analysis was performed on about 700 samples divided into two cohorts of patients, depending on the viral variant detected (B.1 or delta). We sequenced over 20,000 viral genomes since the beginning of the pandemic, producing the highest number of sequences in Italy. We thus reconstructed the pandemic dynamics in the regional territory from March 2020 to December 2021. In addition, we have matured and applied novel proof-of-principle approaches to prioritize possible gain-of-function mutations by leveraging patients' metadata and isolated patient-specific signatures of SARS-CoV-2 infection. This allowed us to (i) identify three new viral variants that specifically originated in the Campania region, (ii) map SARS-CoV-2 intrahost variability during long-term infections and in one case identify an increase in the number of mutations in the viral genome, and (iii) identify host gene expression signatures correlated with viral load in upper respiratory ways. In conclusion, we have successfully generated an optimized and cost-effective strategy to monitor SARS-CoV-2 genetic variability, without the need of automation. Thus, our approach is suitable for any lab with a benchtop sequencer and a limited budget, allowing an integrated genomic surveillance on premises. Finally, we have also identified a gene expression signature defining SARS-CoV-2 infection in real-world patients' upper respiratory ways.


Detection of K-ras gene mutation at codon 12 by pancreatic duct brushing for pancreatic cancer.
To assess the diagnostic value of endoscopic pancreatic duct brushing in detecting mutation of the K-ras gene at codon 12 in cytologic specimens from patients with pancreatic cancer. Thirty-five patients treated at Changhai Hospital, Shanghai between 1999 and 2001 were enrolled. Their cells obtained by pancreatic duct brushing during endoscopic retrograde cholangiopancreatography (ERCP) were suspended with phosphate buffer solution (PBS). DNA of the cells was extracted and mutation of the K-ras gene at codon 12 detected by means of PCR-SSCP. The K-ras gene mutation rate of pancreatic cancer was 70%, which was higher than that of chronic pancreatitis (14%, P<0.05). K-ras gene mutation was not found in patients with pancreatic cystocarcinoma and duodenum carcinoma. As to the location of pancreatic cancer, no significant difference was observed between the head, the body and tail. The sensitivity, specificity, accuracy of pancreatic duct brushing in detecting pancreatic cancer was 70%, 94%, and 83%, respectively. K-ras analysis of pancreatic brushing samples is helpful in the diagnosis of patients with early pancreatic cancer.


[Inflammatory myofibroblastic tumor of the lung: problems in differential diagnosis].
Inflammatory myofibroblastic tumor is a neoplasm of intermediate biological potential, a marked inflammatory component, and characteristic genetic changes. Once it was identified as an independent nosological entity from a rather heterogeneous group of inflammatory pseudotumors. This paper describes a case of inflammatory myofibroblastic tumor of the chest in a child, by discussing the criteria for differential diagnosis in the use of up-to-date radiology techniques. When thoracic neoplasms are detected, it is necessary to determine their site and the most likely origin organ as exactly as possible and to assess the X-ray pattern of the pathological process. By applying the state-of-the-art radiodiagnostic techniques, the clarification of the pattern of the disease makes it possible to choose an optimal treatment policy and a surgical procedure and scope, to avoid anesthesia overload, and to decrease the probability of resurgery. Our observation is to demonstrate possible problems in the diagnosis of inflammatory myofibroblastic tumor of the lung in childhood and to recall again the algorithm of radiological approaches to detecting mediastinal and lung masses.


Intestinal ileus as a possible cause of hypobicarbonatemia.
The possible occurrence of metabolic acidosis in patients with intestinal ileus is not well recognized. We describe a patient with acute alcohol-induced pancreatitis and a large transverse colon ileus in which plasma bicarbonate dropped rapidly in the absence of an increase in the plasma anion gap. The urinary anion gap and ammonium excretion were consistent with an appropriate renal response to metabolic acidosis and against the possibility of respiratory alkalosis. The cause of the falling plasma bicarbonate was ascribed to intestinal bicarbonate sequestration owing to the enhancement of chloride-bicarbonate exchange in a dilated paralyzed colon.


Staff-family relationships in the care of older people: a report on a systematic review.
This article reports on a systematic review of the strategies, practices, and organizational characteristics that promote constructive staff-family relationships in the care of older adults in the health care setting. Research evidence points to the need to address power and control issues, communication, and collaborative approaches to care. Interventions designed to promote family-staff relationships are more likely to achieve maximum benefit when implemented with accompanying information sharing, education, and managerial support.


HSV-1 helper virus 5dl1.2 suppresses sodium currents in amplicon-transduced neurons.
The Herpes Simplex Virus-1 (HSV-1) amplicon system is one of several viral-based strategies currently being developed for gene delivery into mammalian neurons for experimental or therapeutic purposes. Amplicon-containing viruses contain no HSV-1 genes and are amplified in titer relative to the helper viruses used to package them. In this way, they are designed to have a minimal impact on the physiology of transduced neurons. We show here, however, that amplicon preparations made using the 5dl1.2 helper virus selectively suppress sodium currents in cultured neurons by approximately 80% within 2 days of transduction and reduce average spike frequency in response to depolarization from 23 +/- 4 to 0.4 +/- 0.4 Hz. We observe similar suppression of Na(+) currents in cells treated with the 5dl1.2 helper virus alone, indicating that the helper virus retains the ability of wild-type HSV-1 to inhibit these currents potently. Staining amplicon-transduced neurons with anti-HSV antibodies, we find that 80% of the neurons express viral proteins, indicating that helper virus typically co-infects these cells. We conclude that Na(+) current suppression by the amplicon preparation results from the preferential coinfection of transduced neurons by the 5dl1.2 helper virus.


[Statins: therapeutic cascade of their effects].
Concept of sequence and time of appearance of various effects of statins is presented. Apart from hypolipidemic action due to inhibition of HMG CoA reductase activity statins exert multiple pleiotropic effects. Combination of these effects makes statins a unique instrument for solution of global tasks of prevention and treatment of atherosclerosis and its consequences (ischemic heart disease etc.). Manifestations of various pleiotropic effects of statins appear after different time intervals and in most cases are not related to suppression of cholesterol synthesis in the body. First 3-4 months (first level of the statin cascade) are characterized mainly by activity of pleiotropic properties aimed at restoration of endothelial function. These properties are responsible for enhanced eNOS expression, antiischemic, antithrombotic and antiatherogenic effects. During same period of time stabilization of unstable atheromas takes place. Manifestations of second level of the cascade of statin action appear after 2 years of treatment. They are represented by retardation of progression and even partial regression of atheromatosis of coronary and peripheral arteries. Third level is signified by achievement of strategic aims of therapy with statins (in 4-5 years) -- lowering of total and cardiac mortality, reduction of number of cardiac complications. Forth level of the cascade is represented by beneficial influences on nonatherogenic cardiological phenomena and comprise hypotensive, antiarrhythmic and cardiotonic effects. And finally some other important properties of statins constitute the fifth level of the therapeutic cascade. These properties are responsible for effects directed at noncardiac pathology (prevention of diabetes, dementia, including dementia associated with Alzheimer's disease, fractures). Immunodepression, ability to reduce saturation of bile with cholesterol belong to this group of effects.


Disruption of primary imprinting during oocyte growth leads to the modified expression of imprinted genes during embryogenesis.
Parthenogenetic embryos, which contained one genome from a neonate-derived non-growing oocyte and the other from a fully grown oocyte, developed to day 13.5 of gestation in mice, 3 days longer than previously recorded for parthenogenetic development. To investigate the hypothesis that disruption of primary imprinting during oocyte growth leads to the modified expression of imprinted genes and this parthenogenetic phenotype, we have examined Peg1/Mest, Igf2, Peg3, Snrpn, H19, Igf2r and excess p57KIP2. We show that paternally expressed genes, Peg1/Mest, Peg3 and Snrpn, are expressed in the parthenotes, presumably due to a lack of maternal epigenetic modifications during oocyte growth. In contrast, the expression of Igf2, which is repressed in a competitive manner by transcription of the H19 gene, was very low. Furthermore, we show that the maternally expressed Igf2r and p57KIP2 genes were repressed in the alleles of the non-growing oocyte indicating maternal modifications during oocyte growth are necessary for its expression. Thus, our results show that primary imprinting during oocyte growth exhibits a crucial effect on both the expression and repression of maternal alleles during embryogenesis.


Lifelong Learning of Spatiotemporal Representations With Dual-Memory Recurrent Self-Organization.
Artificial autonomous agents and robots interacting in complex environments are required to continually acquire and fine-tune knowledge over sustained periods of time. The ability to learn from continuous streams of information is referred to as lifelong learning and represents a long-standing challenge for neural network models due to catastrophic forgetting in which novel sensory experience interferes with existing representations and leads to abrupt decreases in the performance on previously acquired knowledge. Computational models of lifelong learning typically alleviate catastrophic forgetting in experimental scenarios with given datasets of static images and limited complexity, thereby differing significantly from the conditions artificial agents are exposed to. In more natural settings, sequential information may become progressively available over time and access to previous experience may be restricted. Therefore, specialized neural network mechanisms are required that adapt to novel sequential experience while preventing disruptive interference with existing representations. In this paper, we propose a dual-memory self-organizing architecture for lifelong learning scenarios. The architecture comprises two growing recurrent networks with the complementary tasks of learning object instances (episodic memory) and categories (semantic memory). Both growing networks can expand in response to novel sensory experience: the episodic memory learns fine-grained spatiotemporal representations of object instances in an unsupervised fashion while the semantic memory uses task-relevant signals to regulate structural plasticity levels and develop more compact representations from episodic experience. For the consolidation of knowledge in the absence of external sensory input, the episodic memory periodically replays trajectories of neural reactivations. We evaluate the proposed model on the CORe50 benchmark dataset for continuous object recognition, showing that we significantly outperform current methods of lifelong learning in three different incremental learning scenarios.


The Modified Arch Landing Areas Nomenclature identifies hostile zones for endograft deployment: a confirmatory biomechanical study in patients treated by thoracic endovascular aortic repair†.
Our goal was to confirm whether the Modified Arch Landing Areas Nomenclature (MALAN) for thoracic endovascular aortic repair, in which each landing area is described by indicating both the proximal landing zone (PLZ) and the type of arch (e.g. 0/I), identifies unfavourable landing zones for endograft deployment in diseased aortas. Preoperative computed tomography angiography scans of 10 patients scheduled for thoracic endovascular aortic repair for aneurysm or penetrating ulcer of the arch and with a potential hostile PLZ were reviewed. Five had proximal deployment planned in MALAN area 3/III and 5, in MALAN area 2/III. The angulation of each PLZ was calculated. Computational fluid dynamics modelling was used to compute magnitude and orientation of pulsatile displacement forces in each PLZ. Normalized values based on PLZ areas (i.e. equivalent surface traction) were calculated. Results were compared to those obtained in healthy controls stratified by the MALAN. Angulation was severe (>60°) in MALAN areas 3/III and 2/III, which was consistent with the findings obtained in healthy controls. Increased magnitude (P = 0.021) and unfavourable orientation (i.e. orthogonal to the longitudinal aortic axis) of equivalent surface traction (P = 0.011) was also found in these areas compared to the adjacent ones, following the same pattern seen in the controls. Adverse events related to proximal endograft performance were reported in 3/10 cases. This study confirms in diseased aortas initial proof-of-concept findings on the predictive value of the MALAN to identify landing areas with a geometric and haemodynamic environment hostile for thoracic endovascular aortic repair. These adverse biomechanical features may entail an increased risk of dismal endograft performance.


Hoxb-5 is expressed in gill arch 5 during pharyngeal arch development of flounder Paralichthys olivaceus embryos.
Hox genes are expressed in domains with clear anterior borders exhibiting 3'-->5' hierarchy in hindbrain and in the pharyngeal area commonly in vertebrate embryos. Teleost embryos form seven pharyngeal arches, the mandibular arch, hyoid arch and the gill arches 1-5. We previously reported that, in Japanese flounder (Paralichthys olivaceus) embryos, Hoxd-4 is expressed from rhombomere 7 to the spinal cord in the central nervous system and at gill arches 2-5. At present, the hierarchy of Hox genes at gill arches 3-5 of teleost fish is unclear. Here, we investigated the expression domains of Hoxb-5 in the flounder embryo by whole-mount in situ hybridization to gain insight into the Hox code at gill arches. The initial signal indicating Hoxb-5 expression was identified in the spinal cord at hatching, corresponding with the prim-5 stage of zebrafish. Then, intense signals were detected from the anterior part of the spinal cord and from the posterior part of the pharyngeal area at 36 h after hatching. By serially sectioning the hybridized embryos, it was found that signal in the pharyngeal area came from the most posterior gill arch 5. Therefore, it is speculated that Hoxb-5 functions in regional identification of gill arch 5 in this teleost.


How the switch to hummingbird pollination has greatly contributed to our understanding of evolutionary processes.
The evolutionary switch to hummingbird pollination exemplifies complex adaptation, requiring evolutionary change in multiple component traits. Despite this complexity, diverse lineages have converged on hummingbird-adapted flowers on a relatively short evolutionary timescale. Here, I review how features of the genetic basis of adaptation contribute to this remarkable evolutionary lability. Large-effect substitutions, large mutational targets for adaptation, adaptive introgression, and concentrated architecture all contribute to the origin and maintenance of hummingbird-adapted flowers. The genetic features of adaptation are likely shaped by the ecological and geographic context of the switch to hummingbird pollination, with implications for future evolutionary trajectories.


Is this patient pregnant? Can you reliably rule in or rule out early pregnancy by clinical examination? .
This review addresses a common problem facing the clinician: "When treating or evaluating a woman of childbearing years, what is the value of historical or physical examination features in determining the probability of early pregnancy?" We focus on the clinical examination findings that may help the clinician rule in or rule out early pregnancy. Generally accepted indicators of pregnancy include amenorrhea, morning sickness, tender or tingling breasts, and, after 8 weeks' gestational age, an enlarged uterus with a soft cervix. We reviewed the value (ie, sensitivity and specificity) of these indicators, as well as home pregnancy test results, as predictors of the diagnosis of early pregnancy. The available evidence suggests that some historical features, when absent, are fair but not reliable for ruling out pregnancy. When diagnosing early pregnancy, the clinician should not rely on the clinical examination or a home pregnancy test-a laboratory test should be requested.


A2B adenosine receptor gene deletion attenuates murine colitis.
The A(2B) adenosine receptor (A(2B)AR) is the predominant adenosine receptor expressed in the colonic epithelia. We have previously shown that A(2B)AR mRNA and protein levels are up-regulated during colitis. In this study, we addressed the role of the A(2B)AR in the development of murine colitis and the potential mechanism underlying its effects. Dextran sodium sulfate (DSS), 2,4,6-trinitrobenzene sulfonic acid (TNBS), and Salmonella typhimurium were used to induce colitis in A(2B)AR-null mice (A(2B)AR(-/-)). Colitis was determined using established clinical and histologic scoring. Keratinocyte-derived chemokine (KC) measurements were performed using an enzyme-linked immunosorbent assay. Colonic inflammation induced by DSS, TNBS, or S typhimurium was attenuated in A(2B)AR(-/-) compared with their wild-type counterparts. Clinical features, histologic score, and myeloperoxidase activity were significantly decreased in A(2B)AR(-/-) mice. However, A(2B)AR(-/-) showed increased susceptibility to systemic Salmonella infection. Tissue levels of the neutrophil chemokine, KC was decreased in colitic A(2B)AR(-/-) mice. In addition, flagellin-induced KC levels were attenuated in A(2B)AR(-/-) mice. Neutrophil chemotaxis in response to exogenous interleukin-8 was preserved in A(2B)AR(-/-) mice, suggesting intact neutrophil migration in response to appropriate stimuli. These data demonstrate, for the first time, that the A(2B)AR plays a proinflammatory role in colitis. A(2B) receptor antagonism may be an effective treatment for acute inflammatory intestinal diseases such as acute flare of inflammatory bowel disease.


Distinct Effects of Heparin and Interleukin-4 Functionalization on Macrophage Polarization and In Situ Arterial Tissue Regeneration Using Resorbable Supramolecular Vascular Grafts in Rats.
Two of the greatest challenges for successful application of small-diameter in situ tissue-engineered vascular grafts are 1) preventing thrombus formation and 2) harnessing the inflammatory response to the graft to guide functional tissue regeneration. This study evaluates the in vivo performance of electrospun resorbable elastomeric vascular grafts, dual-functionalized with anti-thrombogenic heparin (hep) and anti-inflammatory interleukin 4 (IL-4) using a supramolecular approach. The regenerative capacity of IL-4/hep, hep-only, and bare grafts is investigated as interposition graft in the rat abdominal aorta, with follow-up at key timepoints in the healing cascade (1, 3, 7 days, and 3 months). Routine analyses are augmented with Raman microspectroscopy, in order to acquire the local molecular fingerprints of the resorbing scaffold and developing tissue. Thrombosis is found not to be a confounding factor in any of the groups. Hep-only-functionalized grafts resulted in adverse tissue remodeling, with cases of local intimal hyperplasia. This is negated with the addition of IL-4, which promoted M2 macrophage polarization and more mature neotissue formation. This study shows that with bioactive functionalization, the early inflammatory response can be modulated and affect the composition of neotissue. Nevertheless, variability between graft outcomes is observed within each group, warranting further evaluation in light of clinical translation.


The effect of hypocaloric diet enriched in legumes with or without L-arginine and selenium on anthropometric measures in central obese women.
Identifying new ways to decrease adiposity will be very valuable for health. The aim of this study was to find out whether L-Arginine (Arg) and selenium alone or together can increase the effect of hypocaloric diet enriched in legumes (HDEL) on anthropometric measures in healthy obese women. This randomized, double-blind, placebo-controlled trial was undertaken in 84 healthy premenopausal women with central obesity. After 2 weeks of run-in on an isocaloric diet, participants were randomly considered to eat HDEL, Arg (5 g/d) and HDEL, selenium (200 µg/d) and HDEL or Arg, selenium and HDEL for 6 weeks. The following variables were assessed before intervention and 3 and 6 weeks after it: weight, waist circumference, hip circumference, waist to hip ratio (WHR), body mass index (BMI), and fasting nitrite/nitrate (NO(x)) concentrations. Other variables (arm, thigh, calf and breast circumferences, subscapular, triceps, biceps and suprailiac skinfold thicknesses, sum of skinfold thicknesses (SSF), body density (D) and estimated percent of body fat (EPF)) were assessed before and after intervention. HDEL showed a significant effect in reduction of waist, hip, arm, thigh, calf and breast circumferences, triceps, biceps, subscapular and suprailiac skinfold thicknesses, WHR, SSF, D and EPF. HDEL + Arg + selenium significantly reduced suprailiac skinfold thicknesses; and there was no significant effect of HDEL, Arg, selenium and Arg plus selenium on weight, BMI and fasting NO(x). The study indicates that HDEL + Arg + selenium reduce suprailiac skinfold thicknesses which represents the abdominal obesity reduction.


Zero-inflated count distributions for capture-mark-reencounter data.
The estimation of demographic parameters is a key component of evolutionary demography and conservation biology. Capture-mark-recapture methods have served as a fundamental tool for estimating demographic parameters. The accurate estimation of demographic parameters in capture-mark-recapture studies depends on accurate modeling of the observation process. Classic capture-mark-recapture models typically model the observation process as a Bernoulli or categorical trial with detection probability conditional on a marked individual's availability for detection (e.g., alive, or alive and present in a study area). Alternatives to this approach are underused, but may have great utility in capture-recapture studies. In this paper, we explore a simple concept: in the same way that counts contain more information about abundance than simple detection/non-detection data, the number of encounters of individuals during observation occasions contains more information about the observation process than detection/non-detection data for individuals during the same occasion. Rather than using Bernoulli or categorical distributions to estimate detection probability, we demonstrate the application of zero-inflated Poisson and gamma-Poisson distributions. The use of count distributions allows for inference on availability for encounter, as well as a wide variety of parameterizations for heterogeneity in the observation process. We demonstrate that this approach can accurately recover demographic and observation parameters in the presence of individual heterogeneity in detection probability and discuss some potential future extensions of this method.


Monocyte-derived dendritic cells: a potential target for therapy in multiple sclerosis (MS).
Monocytes can differentiate into dendritic cells (DC), cells with a pivotal role in both protective immunity and tolerance. Defects in the maturation or function of DC may be important in the development of autoimmune disease. We sought to establish if there were differences in the cytokine (granulocyte-macrophage colony-stimulating factor and IL-4)-driven maturation of monocytes to DC in patients with MS and whether drugs used to treat MS affected this process in vitro. We have demonstrated that there is no defect in the ability of magnetic activated cell sorting (MACS)-purified monocytes from patients with MS to differentiate to DC, but equally they show no tendency to acquire a DC phenotype without exogenous cytokines. Interferon-beta1a prevents the acquisition of a full DC phenotype as determined by light and electron microscopy and by flow cytometry. Methylprednisolone not only prevents the development of monocyte-derived DC but totally redirects monocyte differentiation towards a macrophage phenotype. Evidence is evolving for a role for DC in central nervous system immunity, either within the brain or in cervical lymph nodes. The demonstrated effect of both drugs on monocyte differentiation may represent an important site for immune therapy in MS.


π-Conjugated Polymer Nanoparticles from Design, Synthesis to Biomedical Applications: Sensing, Imaging, and Therapy.
In the past decade, π-conjugated polymer nanoparticles (CPNs) have been considered as promising nanomaterials for biomedical applications, and are widely used as probe materials for bioimaging and drug delivery. Due to their distinctive photophysical and physicochemical characteristics, good compatibility, and ease of functionalization, CPNs are gaining popularity and being used in more and more cutting-edge biomedical sectors. Common synthetic techniques can be used to synthesize CPNs with adjustable particle size and dispersion. More importantly, the recent development of CPNs for sensing and imaging applications has rendered them as a promising device for use in healthcare. This review provides a synopsis of the preparation and functionalization of CPNs and summarizes the recent advancements of CPNs for biomedical applications. In particular, we discuss their major role in bioimaging, therapeutics, fluorescence, and electrochemical sensing. As a conclusion, we highlight the challenges and future perspectives of biomedical applications of CPNs.


Phlorizin administration increases hepatic gluconeogenic enzyme mRNA abundance but not feed intake in late-lactation dairy cows.
Gluconeogenic capacity may be an important factor regulating dry matter intake (DMI) in lactating dairy cows. To determine whether increased glucose demand affects feed intake and hepatic gene expression, lactating Holstein cows were treated with phlorizin or vehicle (propylene glycol) for 7 d. Multiparous cows (n = 12; 269 +/- 65 d in milk, mean +/- SD) were randomly assigned to treatment sequence in a crossover design and were adapted to a common diet for 7 d before the beginning of the experiment. Phlorizin injected s.c. at 4 g/d caused glucose excretion in urine at the rate of 474 g/d. Although phlorizin decreased lactose synthesis and milk production (both P < 0.01), DMI and 3.5% fat-corrected milk production were not altered by treatment. A net deficit of 383 g glucose/d in milk and urine for phlorizin (relative to control) was likely replaced partially through increased gluconeogenesis. The molar insulin:glucagon ratio was decreased 17% by phlorizin (P < 0.001) and hepatic phosphoenolpyruvate carboxykinase, glucose-6-phosphatase, and pyruvate carboxylase mRNA abundance increased (all P < 0.05). Late-lactation dairy cows adapted quickly to an increase in peripheral glucose demand; adaptation mechanisms likely included enhanced gluconeogenic capacity, whereas DMI was not altered.


Identification of a 16,600-dalton outer membrane protein on nontypeable Haemophilus influenzae as a target for human serum bactericidal antibody.
A 16,600-D outer membrane protein is present in all strains of Haemophilus influenzae and antibodies to this protein are present in human serum. This study was designed to assess the role of this outer membrane protein (P6) in nontypeable H. influenzae as a target for human serum bactericidal antibody. P6 was isolated and coupled to an affinity column. Depleting normal human serum of antibodies to P6 by affinity chromatography resulted in reduced bactericidal activity of that serum for nontypeable H. influenzae. Immunopurified antibodies to P6 from human serum were bactericidal. Finally, preincubation of bacteria with a monoclonal antibody that recognizes a surface epitope on P6, inhibited human serum bactericidal killing. Taken together, these experiments establish that P6 is a target for human bactericidal antibodies. This observation provides evidence that P6 plays a potentially important role in human immunity to infection by nontypeable H. influenzae.


P53, Bcl-2 expression and cell proliferation in benign breast lesions.
The role of proliferation-related markers on breast cancer pathogenesis has been only occasionally investigated. The immunocytochemical expression of P53 and Bcl-2 (using PAb1801 and anti-bcl-2 monoclonal antibodies) and cell proliferation (evaluated as the H-3-thymidine labeling index [H-3-dT LI]) were determined on 62 benign breast lesions at different risk. Accumulation of the P53 protein was never observed; Bcl-2 was detected in 50% of cases and it was unrelated to biologic and clinicopathologic features. Median H-3-dT LI was about three times lower than that observed on large series of invasive breast cancer. It was only slightly higher in lesions from patients younger than 35 years or with a positive family history than in those under 35 or with a negative family history and appeared unrelated to histology or risk classification. Such findings indicate that the investigated biomarkers fail to identify women at increased risk for breast cancer.


Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells.
Classical genetic screens can be limited by the selectivity of mutational targeting, the complexities of anatomically based phenotypic analysis, or difficulties in subsequent gene identification. Focusing on signaling response to the secreted morphogen Hedgehog (Hh), we used RNA interference (RNAi) and a quantitative cultured cell assay to systematically screen functional roles of all kinases and phosphatases, and subsequently 43% of predicted Drosophila genes. Two gene products reported to function in Wingless (Wg) signaling were identified as Hh pathway components: a cell surface protein (Dally-like protein) required for Hh signal reception, and casein kinase 1alpha, a candidate tumor suppressor that regulates basal activities of both Hh and Wg pathways. This type of cultured cell-based functional genomics approach may be useful in the systematic analysis of other biological processes.


Oral carnosine supplementation prevents vascular damage in experimental diabetic retinopathy.
Pericyte loss, vasoregression and neuroglial activation are characteristic changes in incipient diabetic retinopathy. In this study, the effect of the antioxidant and antiglycating dipeptide carnosine was studied on the development of experimental diabetic retinopathy. STZ-induced diabetic Wistar rats were orally treated with carnosine (1g/kg body weight/day). Retinal vascular damage was assessed by quantitative morphometry. Retinal protein extracts were analyzed for markers of oxidative stress, AGE-formation, activation of the hexosamine pathway and changes in the expression of Ang-2, VEGF and heat shock proteins Hsp27 and HO-1. Glial cell activation was analyzed using Western blot analysis and immunofluorescence of GFAP expression and retinal neuronal damage was histologically examined. Oral carnosine treatment prevented retinal vascular damage after 6 months of experimental hyperglycemia. The protection was not caused by ROS- or AGE-inhibition, but associated with a significant induction of Hsp27 in activated glial cells and normalization of increased Ang-2 levels in diabetic retinas. A significant reduction of photoreceptors in retinas of carnosine treated animals was noted. Oral carnosine treatment protects retinal capillary cells in experimental diabetic retinopathy, independent of its biochemical function. The vasoprotective effect of carnosine might be mediated by the induction of protective Hsp27 in activated glial cells and normalization of hyperglycemia-induced Ang-2.


Accuracy of intraocular pressure measurements in dogs using two different tonometers and plano therapeutic soft contact lenses.
To compare and evaluate the accuracy of intraocular pressure (IOP) measured through a therapeutic contact lens, using applanation (TonoPen XL(®)) and rebound (TonoVet(®)) tonometers in enucleated dog eyes. A total of 30 enucleated eyes from 15 beagle dogs. To measure accurate IOP, the anterior chamber of each enucleated eye was cannulated with two 26-gauge needles and two polyethylene tubes were connected vertically to an adjustable reservoir bag of normal saline and a pressure transducer. IOP was measured by the TonoPen XL(®) followed by the TonoVet(®) without a contact lens. After a contact lens was applied to the cornea, IOP was re-measured in the same order. Three consecutive IOP measurements were performed using both tonometers. Without the contact lens, the IOP values obtained by both tonometers correlated well according to the regression analysis (TonoVet(®): γ(2) = 0.98, TonoPen XL(®): γ(2) = 0.97, P < 0.001). The TonoPen XL(®) consistently underestimated values as transducer IOP increased; however, IOP values measured with the TonoPen XL(®) were in close agreement and were less variable than those determined with the TonoVet(®) when a contact lens was applied to the cornea. Bland-Altman analysis was used to determine the lower and upper limits of agreement (TonoVet(®): -29.7 and +21.1 mmHg, TonoPen XL(®): -3.9 and +3.6 mmHg) between the two devices. This study suggests that the TonoPen XL(®) is a useful tonometer for dogs wearing therapeutic contact lenses, and importantly, contact lenses would not need to be removed prior to IOP measurement.


[Protein metabolism in the newborn lamb. II. Influence of diet on protein synthesis and turnover].
Leucine turnover and protein fractional synthesis rates were measured in fed and unfed newborn lambs. The fed group (4 animals) was fed hourly with cow colostrum throughout the experiment which started at birth; the unfed group (5 animals) was given saline (NaCl 9 0/00). Two hours after birth, L-(1-14C) leucine was infused into the jugular vein at a constant rate for 8 h. Eight blood samples were taken during the infusion period; at the end of the experiment, the liver, skin and three muscles (supraspinatus, longissimus dorsi, tensor fasciae latae) were sampled. The specific activity of free and protein-bound leucine was measured in these samples and in the whole body (table 2). The specific activity of free leucine reached a quasi-steady state in the plasma within 2 h after the beginning of infusion (fig. 1). The irreversible loss rate of plasma leucine and the leucine catabolic rate were higher in fed than in unfed lambs. The highest fractional rates of protein synthesis occurred in the liver, skin and whole body (table 3). They were similar in both groups. In contrast, fractional rates of protein synthesis in muscles were higher in fed than in unfed animals. Thus, the proportion of whole body protein synthesis, accounted for by muscle protein synthesis, was higher in fed than in fasting newborn lambs (fig. 2). The leucine flux from whole body protein breakdown was unaffected by nutritional status (table 1). Feeding colostrum stimulated muscle protein synthesis, which was in keeping with body weight gain and protein accretion.


Effect of carbon source availability and growth phase on expression of Corynebacterium glutamicum genes involved in the tricarboxylic acid cycle and glyoxylate bypass.
The effect of different carbon sources on the expression of tricarboxylic acid (TCA) cycle genes, along with glyoxylate bypass genes, in Corynebacterium glutamicum was determined. All TCA cycle genes were coordinately expressed in medium containing acetate. Growth in the presence of acetate gave rise to abundant expression of most TCA cycle genes, with the level of gltA transcript being the highest. However, when the cells entered the stationary phase triggered by acetate exhaustion, all genes were repressed, except sucCD and mdhB, which were slightly induced. Acetate withdrawal from the growth medium during the exponential phase also led to rapid repression of most TCA cycle genes and a corresponding twofold increase in the expression of sucCD, which were strongly induced by citrate and succinate. In addition, glucose depletion during the stationary phase led to a corresponding 8-20-fold induction of the sucCD, aceA and aceB genes. Addition of glucose to acetate medium resulted in about 10-fold induction of sucCD. The strong dependence of TCA cycle sucCD and glyoxylate bypass aceA and aceB expression on carbon source availability was confirmed and the regulatory system will be studied precisely.


Mitochondrial dysfunction and oxidative stress contribute to the pathogenesis of spinocerebellar ataxia type 12 (SCA12).
Spinal cerebellar ataxia type 12 (SCA12) has been attributed to the elevated expression of ppp2r2b. To better elucidate the pathomechanism of the neuronal disorder and to search for a pharmacological treatment, Drosophila models of SCA12 were generated by overexpression of a human ppp2r2b and its Drosophila homolog tws. Ectopic expression of ppp2r2b or tws caused various pathological features, including neurodegeneration, apoptosis, and shortened life span. More detailed analysis revealed that elevated ppp2r2b and tws induced fission of mitochondria accompanied by increases in cytosolic reactive oxygen species (ROS), cytochrome c, and caspase 3 activity. Transmission electron microscopy revealed that fragmented mitochondria with disrupted cristae were engulfed by autophagosomes in photoreceptor neurons of flies overexpressing tws. Additionally, transgenic flies were more susceptible to oxidative injury induced by paraquat. By contrast, ectopic Drosophila Sod2 expression and antioxidant treatment reduced ROS and caspase 3 activity and extended the life span of the SCA12 fly model. In summary, our study demonstrates that oxidative stress induced by mitochondrial dysfunction plays a causal role in SCA12, and reduction of ROS is a potential therapeutic intervention for this neuropathy.


[Effect of change of plasma K+ and pH value induced by exercise on muscle fatigue and surface EMG].
This study was undertaken to explore the relationship between plasma K+, H+ and muscle fatigue as well as electromyogram changes. Fourteen male students enrolled as the subjects of this study performed two exercise tests of different intensity (200 W; 120 W) on a cycle ergometer till being fatigued. The plasma K+, pH value and electromyography (EMG) of active and inactive muscles during 10 s isometric contraction fixed load were measured before exercise and at 0, 5, 10, 20 minutes after exercise. Both plasma K+ and mean power frequency (MPF) of the active muscles changed significantly (P<0.01) after exercise, but recovered in 20 minutes and were highly negatively correlated. Plasma pH at 20 minutes after 200 W exercise was significantly lower than that at rest (P<0.01), but did not change after 120 W exercise, and no correlation was found between pH and EMG. Integral electromyography (IEMG) of inactive muscles at 20 minutes after 120 W exercise was significantly lower than that at rest (P<0.05). The results showed that high extra cellular K+ concentration for exercise is one of the main factors inducing fatigue and MPF decrease of the active muscles as well as IEMG decrease of the inactive muscle, but H+ is not a main factor for fatigue and it has no direct effect on EMG.


Action observation circuits in the macaque monkey cortex.
In both monkeys and humans, the observation of actions performed by others activates cortical motor areas. An unresolved question concerns the pathways through which motor areas receive visual information describing motor acts. Using functional magnetic resonance imaging (fMRI), we mapped the macaque brain regions activated during the observation of grasping actions, focusing on the superior temporal sulcus region (STS) and the posterior parietal lobe. Monkeys viewed either videos with only the grasping hand visible or videos with the whole actor visible. Observation of both types of grasping videos activated elongated regions in the depths of both lower and upper banks of STS, as well as parietal areas PFG and anterior intraparietal (AIP). The correlation of fMRI data with connectional data showed that visual action information, encoded in the STS, is forwarded to ventral premotor cortex (F5) along two distinct functional routes. One route connects the upper bank of the STS with area PFG, which projects, in turn, to the premotor area F5c. The other connects the anterior part of the lower bank of the STS with premotor areas F5a/p via AIP. Whereas the first functional route emphasizes the agent and may relay visual information to the parieto-frontal mirror circuit involved in understanding the agent's intentions, the second route emphasizes the object of the action and may aid in understanding motor acts with respect to their immediate goal.


Severe stress and adjustment disorder diagnoses in the population of Denmark.
We created a registry of Danish-born citizens of Denmark with incident International Classification of Diseases (10th ed.; ICD-10) severe stress and adjustment disorder diagnoses between 1995 and 2011. A unique personal identifier was used to retrieve and merge data on demographic characteristics and diagnoses (ICD-10 codes F43.x). Here we report on the incidence of these disorders and the demographic characteristics of the subset of the Danish population who have received 1 of these diagnoses: 111,844 adults and children received a first diagnosis between 1995 and 2011. More women than men (60.1% vs. 39.9%) received a diagnosis. Diagnoses increased during the late teens through early 30s. Adjustment disorder was the most common diagnosis (65.7% of adults and 64% of children). Reaction to severe stress unspecified was the second most common (19.8% of adults and 23.8% of children), and there was a large increase in both, as well as acute stress reaction diagnoses, in 2007 (3,717-5,141, 1,248-2,520, and 348-1,024 in 2006 to 2007, respectively). Findings regarding gender and age of onset are similar to other westernized countries. This registry can be used for future research programs, contributing to the study of stress and trauma.


Conservative treatment of upper urinary tract carcinoma: Long-term results.
We sought to assess the long-term results of conservative treatment for upper urinary tract carcinoma (UUTC) with regard to tumour recurrence and preservation of renal unit. From October 1987 to January 2014, 65 patients (median age 68 years) were diagnosed with UUTC and underwent endoscopic and open surgical techniques. Thirteen patients had bilateral disease and one had a single kidney. The primary approach was endoscopic in 37 reno-ureteral units (20 percutaneous resections, 17 ureteroscopies ). Open surgery was performed in 19 cases. A total of 20 patients received mitomycin C. Superficial stage pTa or T1 was noted in 37 patients, infiltrating stage pT2 and pT3 in seven and inverted papilloma in one. The stage of the tumour was impossible to classify in 20. With a median followup of 75.12 months (interquartile range [IQR] 144.71-17.41), the kidney preservation, recurrence, specific survival, and global survival rates were 78.5% (51/65), 40.0% (26/65), 92.3% (60/65), and 69.2% (45/65), respectively. From the patients who had recurrence, 15 were salvaged with radical nephroureterectomy (RNU). The bladder tumour recurrence rate after the surgery was 30.76% (20/65). At the end of the followup, five patients had died of UUTC progression and 16 from other causes. Postoperative complications included one case of fistula, one case of stricture, and one case of nephrectomy due to bleeding. In selected cases, conservative management is a safe and feasible alternative to RNU, with the advantage of renal unit preservation.


Dichloroacetate Prevents Cisplatin-Induced Nephrotoxicity without Compromising Cisplatin Anticancer Properties.
Cisplatin is an effective anticancer drug; however, cisplatin use often leads to nephrotoxicity, which limits its clinical effectiveness. In this study, we determined the effect of dichloroacetate, a novel anticancer agent, in a mouse model of cisplatin-induced AKI. Pretreatment with dichloroacetate significantly attenuated the cisplatin-induced increase in BUN and serum creatinine levels, renal tubular apoptosis, and oxidative stress. Additionally, pretreatment with dichloroacetate accelerated tubular regeneration after cisplatin-induced renal damage. Whole transcriptome sequencing revealed that dichloroacetate prevented mitochondrial dysfunction and preserved the energy-generating capacity of the kidneys by preventing the cisplatin-induced downregulation of fatty acid and glucose oxidation, and of genes involved in the Krebs cycle and oxidative phosphorylation. Notably, dichloroacetate did not interfere with the anticancer activity of cisplatin in vivo. These data provide strong evidence that dichloroacetate preserves renal function when used in conjunction with cisplatin.


A comparison of nefazodone, the cognitive behavioral-analysis system of psychotherapy, and their combination for the treatment of chronic depression.
Patients with chronic forms of major depression are difficult to treat, and the relative efficacy of medications and psychotherapy is uncertain. We randomly assigned 681 adults with a chronic nonpsychotic major depressive disorder to 12 weeks of outpatient treatment with nefazodone (maximal dose, 600 mg per day), the cognitive behavioral-analysis system of psychotherapy (16 to 20 sessions), or both. At base line, all patients had scores of at least 20 on the 24-item Hamilton Rating Scale for Depression (indicating clinically significant depression). Remission was defined as a score of 8 or less at weeks 10 and 12. For patients who did not have remission, a satisfactory response was defined as a reduction in the score by at least 50 percent from base line and a score of 15 or less. Raters were unaware of the patients' treatment assignments. Of the 681 patients, 662 attended at least one treatment session and were included in the analysis of response. The overall rate of response (both remission and satisfactory response) was 48 percent in both the nefazodone group and in the psychotherapy group, as compared with 73 percent in the combined-treatment group. (P<0.001 for both comparisons). Among the 519 subjects who completed the study, the rates of response were 55 percent in the nefazodone group and 52 percent in the psychotherapy group, as compared with 85 percent in the combined-treatment group (P<0.001 for both comparisons). The rates of withdrawal were similar in the three groups. Adverse events in the nefazodone group were consistent with the known side effects of the drug (e.g., headache, somnolence, dry mouth, nausea, and dizziness). Although about half of patients with chronic forms of major depression have a response to short-term treatment with either nefazodone or a cognitive behavioral-analysis system of psychotherapy, the combination of the two is significantly more efficacious than either treatment alone.


Liquid-phase extraction coupled with metal-organic frameworks-based dispersive solid phase extraction of herbicides in peanuts.
Liquid-phase extraction coupled with metal-organic frameworks-based dispersive solid phase extraction was developed and applied to the extraction of pesticides in high fatty matrices. The herbicides were ultrasonically extracted from peanut using ethyl acetate as extraction solvent. The separation of the analytes from a large amount of co-extractive fat was achieved by dispersive solid-phase extraction using MIL-101(Cr) as sorbent. In this step, the analytes were adsorbed on MIL-101(Cr) and the fat remained in bulk. The herbicides were separated and determined by high-performance liquid chromatography. The experimental parameters, including type and volume of extraction solvent, ultrasonication time, volume of hexane and eluting solvent, amount of MIL-101(Cr) and dispersive solid phase extraction time, were optimized. The limits of detection for herbicides range from 0.98 to 1.9 μg/kg. The recoveries of the herbicides are in the range of 89.5-102.7% and relative standard deviations are equal or lower than 7.0%. The proposed method is simple, effective and suitable for treatment of the samples containing high content of fat.


Dietary tryptophan alleviates dextran sodium sulfate-induced colitis through aryl hydrocarbon receptor in mice.
Ulcerative colitis is the typical progression of chronic inflammatory bowel disease. Amino acids, particularly tryptophan, have been reported to exert a protective effect against colitis induced by dextran sodium sulfate (DSS), but the precise underlying mechanisms remain incompletely clarified. Tryptophan metabolites are recognized to function as endogenous ligands for aryl hydrocarbon receptor (Ahr), which is a critical regulator of inflammation and immunity. Thus, we conducted this study to investigate whether dietary tryptophan supplementation protects against DSS-induced colitis by acting through Ahr. Female wild-type (WT) and Ahr-deficient (knockout; KO) mice (10-12 weeks old) were divided into four groups and fed either a control or 0.5% tryptophan diet. The tryptophan diet ameliorated DSS-induced colitis symptoms and severity in WT mice but not in KO mice, and the diet reduced the mRNA expression of Il-6, Tnfα, Il-1β and the chemokines Ccl2, Cxcl1 and Cxcl2 in the WT groups. Furthermore, Il-22 and Stat3 mRNA expression in the colon was elevated in WT mice fed with the tryptophan diet, which mainly protected epithelial layer integrity, and Ahr also modulated immune homeostasis by regulating Foxp3 and Il-17 mRNA expression. These data suggest that tryptophan-containing diet might ameliorate DSS-induced acute colitis and regulate epithelial homeostasis through Ahr. Thus, tryptophan could serve as a promising preventive agent in the treatment of ulcerative colitis.


Identifying yoga-based knee strengthening exercises using the knee adduction moment.
This study aimed to compare muscle activations, co-contraction indices, and the knee adduction moment between static standing yoga postures to identify appropriate exercises for knee osteoarthritis. Healthy young women (24.4 (5.4) years, 23.1 (3.7) kg/m(2)) participated. Primary outcome variables were electromyographic activations of the vastus lateralis, rectus femoris, vastus medialis, biceps femoris, and semitendinosus; co-contraction between the biceps femoris and rectus femoris, and vastus lateralis and vastus medialis; and knee adduction moments of both legs during six static, standing yoga postures (two squatting postures, two lunging postures, a hamstring stretch, and a single-leg balance posture). A two-factor repeated measures analysis of variance was used to identify differences in muscle amplitudes, co-contractions, and knee adduction moment between postures and legs. Quadriceps activations were highest during squat and lunge postures (p≤0.001). Hamstring activations were highest during the hamstring stretch (p≤0.003). Squat and lunge postures produced higher co-contraction indices than other postures (p≤0.011). The wide legged squat (Goddess) and lunge with trunk upright (Warrior) produced the lowest knee adduction moments (p≤0.006), while the single-leg balance posture elicited a higher knee adduction moment than all other postures (p<0.05). Squatting and lunging postures could improve leg strength while potentially minimizing exposure to high knee adduction moments. Future work should evaluate whether these exercises are useful in people with knee osteoarthritis.


Abortion reporting in the era of medical procedures: why is it important?
Food and Drug Administration approval of mifepristone and the subsequent widespread use of medical abortion will change the patterns and practice of abortion services in this country. Accurate monitoring of new, nonsurgical abortion techniques will be critical as this change takes place. Providers will want to know which women will be the predominant users of medical versus surgical methods. If medical procedures are used widely, the national trend in gestational age at the time of abortion is likely to change because medical procedures are usually performed early (at less than eight weeks' gestation). New and existing abortion providers must be informed about the reporting statutes and requirements in their areas. Information and instructions for reporting abortions are available from the vital statistics offices in each state health department and the health departments of New York City and the District of Columbia. Ongoing comprehensive monitoring of legal induced abortion is needed in all states to determine the number of procedures performed, the characteristics of women who obtain them, and the evolving trends in procedures.


Pattern of paediatric blood pressure in rural, semi-urban and urban communities in Ilorin, Nigeria.
The blood pressures of 2,526 children aged between 1 and 14 years were measured in their home environments in rural, semi-urban and urban communities in Ilorin. Blood pressure increased with age, but there was no significant difference between the communities. Girls had higher blood pressures than boys between the ages of 9 and 13 years, in the semi-urban and rural communities. The prevalence of high blood pressure in the urban community (9.5%) was higher, though not significantly so, than the rural community (6.3%). It is concluded that the prevalence of paediatric high blood pressure in the African is higher than previously reported.


α2β1 integrin regulates Th17 cell activity and its neutralization decreases the severity of collagen-induced arthritis.
Th17 cells play a critical role in the pathogenesis of rheumatoid arthritis (RA), but the mechanisms by which these cells regulate the development of RA are not fully understood. We have recently shown that α2β1 integrin, the receptor of type I collagen, is the major collagen-binding integrin expressed by human Th17 cells. In this study, we examined the role of α2β1 integrin in Th17-mediated destructive arthritis in the murine model of collagen-induced arthritis (CIA). We found that α2β1 integrin is expressed on synovial Th17 cells from CIA mice and its neutralization with a specific mAb significantly reduced inflammation and cartilage degradation, and protected the mice from bone erosion. Blockade of α2β1 integrin led to a decrease in the number of Th17 cells in the joints and to a reduction of IL-17 levels in CIA mice. This was associated with an inhibition of receptor activator of NF-κB ligand levels and osteoclast numbers, and reduction of bone loss. We further show that α2β1 integrin is expressed on synovial Th17 cells from RA patients, and that its ligation with collagen costimulated the production of IL-17 by polarized human Th17 cells by enhancing the expression of retinoic acid receptor-related orphan receptor C through ERK and PI3K/AKT. Our findings provide the first evidence, to our knowledge, that α2β1 integrin is an important pathway in Th17 cell activation in the pathogenesis of CIA, suggesting that its blockade can be beneficial for the treatment of RA and other Th17-associated autoimmune diseases.


Fundamentals of affinity cell separations.
Cell separations using affinity methods continue to be an enabling science for a wide variety of applications. In this review, we discuss the fundamental aspects of affinity separation, including the competing forces for cell capture and elution, cell-surface interactions, and models for cell adhesion. Factors affecting separation performance such as bond affinity, contact area, and temperature are presented. We also discuss and demonstrate the effects of nonspecific binding on separation performance. Metrics for evaluating cell separations are presented, along with methods of comparing separation techniques for cell isolation using affinity capture.


Implementation findings from an effectiveness-implementation trial of tablet-based parent training in pediatric primary care.
The use of mobile delivery to deliver parent training can address barriers to access and improve the translation of interventions in existing settings like pediatric primary care. Studying implementation provides critical information to identify and address barriers and facilitators and inform sustainability efforts. This study was a descriptive evaluation using the RE-AIM (Reach, Effectiveness, Adoption, Implementation, and Maintenance) framework as part of a Hybrid Type I Effectiveness-Implementation trial of the ezParent Program within 4 pediatric primary care clinics. We collected data before, during, and after implementation to evaluate provider implementation and their perspectives on program benefits and barriers to implementation. On average, 14% of eligible parents were introduced to the study. Of these parents, 78% expressed interest in participating and 37% enrolled in the study. Seventy-eight percent of staff providers (n = 36) implemented the procedures at least once, and among those who implemented 3 or more times (n = 24), 79% maintained implementation for 6 months. Barriers to implementation include limited time, lack of information, and full practice buy-in and engagement. Implementation fidelity may improve with additional education and training of the interdisciplinary team, clear messaging regarding the purpose and content of the ezParent program, defining roles within the care team, identifying practice champions, and use of the electronic health record. Findings from this evaluation, including data from the randomized controlled trial and literature to support intervention effectiveness and implementation, will be used to develop an implementation toolkit to include specific strategies for implementation and ideas for local adaptations. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Association between single nucleotide polymorphisms in non-coding regions of the insulin (INS) gene and schizophrenia.
Schizophrenia is a psychotic disorder with high heritability. There are also indications that impaired cellular signalling via the insulin receptor-A and the insulin-like growth factor 1 receptor may play a role in its pathogenesis. Insulin, and possibly also C-peptide, are ligands to these receptors. The insulin gene, coding both insulin and C-peptide, has however not been genetically studied in schizophrenia. Therefore, this study was undertaken to investigate the involvement of this gene in schizophrenia susceptibility. For identification of single nucleotide polymorphisms (SNPs) of interest, the whole insulin gene and parts of its promoter region were first DNA sequenced in two subgroups of the study population (37 schizophrenia patients with heredity for schizophrenia or related psychosis, and 25 controls), and mapped to the reference sequence. Then, 7 identified SNPs of potential interest were typed by TaqMan® SNP Genotyping Assays in the whole study population, consisting of 94 patients with schizophrenia and 60 controls. Allele frequencies tended to differ between patients and controls for two of the 7 SNPs, rs5505 and rs3842749 (p=0.077 and p=0.078, respectively), whereas subgroup analyses of diabetes mellitus (type 1 or 2) and/ or heredity for diabetes mellitus (type 1 or 2) in patients and controls showed overall significant differences in genotype/ allele frequencies solely for rs5505 (p=0.021/ 0.023). These findings are of interest, as the two SNPs - rs5505 and rs3842749 - may have regulatory function on the coding of insulin and C-peptide, against which increased antibody reactivity has been previously reported in schizophrenia.


Structural segregation of gut microbiota between colorectal cancer patients and healthy volunteers.
Despite a long-suspected role in the development of human colorectal cancer (CRC), the composition of gut microbiota in CRC patients has not been adequately described. In this study, fecal bacterial diversity in CRC patients (n=46) and healthy volunteers (n=56) were profiled by 454 pyrosequencing of the V3 region of the 16S ribosomal RNA gene. Both principal component analysis and UniFrac analysis showed structural segregation between the two populations. Forty-eight operational taxonomic units (OTUs) were identified by redundancy analysis as key variables significantly associated with the structural difference. One OTU closely related to Bacteroides fragilis was enriched in the gut microbiota of CRC patients, whereas three OTUs related to Bacteroides vulgatus and Bacteroides uniformis were enriched in that of healthy volunteers. A total of 11 OTUs belonging to the genera Enterococcus, Escherichia/Shigella, Klebsiella, Streptococcus and Peptostreptococcus were significantly more abundant in the gut microbiota of CRC patients, and 5 OTUs belonging to the genus Roseburia and other butyrate-producing bacteria of the family Lachnospiraceae were less abundant. Real-time quantitative PCR further validated the significant reduction of butyrate-producing bacteria in the gut microbiota of CRC patients by measuring the copy numbers of butyryl-coenzyme A CoA transferase genes (Mann-Whitney test, P<0.01). Reduction of butyrate producers and increase of opportunistic pathogens may constitute a major structural imbalance of gut microbiota in CRC patients.


Performance of commonly used respiratory questionnaire items in a cohort of infants born preterm.
Items from respiratory questionnaires validated in older children are often used in research studies of preterm infants, although they have not been validated in this population. We aimed to assess both test-retest reliability and convergent validity of a group of commonly used respiratory questionnaire items in a cohort of preterm infants. The health status of 300 preterm infants was assessed by telephone questionnaire as part of a prospective cohort study. The questionnaire items analyzed in this study included six commonly used respiratory questions. The questionnaire responses used in this analysis were from the telephone follow-up in this cohort at six months of age adjusted for prematurity. A repeat interview one to two weeks after this interview was performed in a subset of subjects to assess test-retest reliability. The convergent validity of the respiratory items was also assessed by calculating the associations among the responses to the respiratory questions. A total of 43 infants were singletons that met the criteria for test-retest reliability analysis. All of the respiratory questions demonstrated fair to strong test-retest reliability. Among 206 respondents, respiratory questionnaire items also demonstrated strong convergent validity, in that caretakers reporting wheezing or whistling in the chest were significantly more likely to also report other respiratory events. This selection of standard respiratory questionnaire items performed well for research purposes in this population.


Quality of emergency medical care in Gondar University Referral Hospital, Northwest Ethiopia: a survey of patients' perspectives.
Ethiopia has fairly good coverage but very low utilization of health care services. Emergency medical care services require fast, correct and curious services to clients as they present with acute problems. In Ethiopia and Gondar in particular, the quality of emergency medical care has not been studied. The main aim of this study was to assess the disease profile and patients' satisfaction in Gondar University Referral Hospital (GURH). A facility based cross-sectional study was conducted among patients visiting GURH for emergency care. Ethical clearance was obtained from the Institutional Review Board of University of Gondar. Patients were selected by systematic random sampling, using patient flow list in the day and night emergency services. Data were collected using a standard Press Ganey questionnaire by BSc health science graduates. Data were entered in to Epi Info 3.5.3 software and exported to SPSS version 20.0 for windows for analysis. A total of 963 patients (response rate = 96.8%) were studied. The mean (+ s.d.) age of patients was 28.4 (+17.9) years. The overall satisfaction using the mean score indicates that 498 (51.7%) 95%CI: (48.4% - 54.9%) were satisfied with the service, the providers and the facility suitability whereas 465(48.3%) 95%CI: (45.1%- 51.6%) were not satisfied. Seven hundred and six (73.3%) 95%CI: 70.4%-76.1%, patients reported that they have been discriminated or treated badly during the service provision in the hospital. OPD site visited (p < 0.0001), visiting days of the week (P < 0.049), medical condition on arrival (P < 0.0001), degree of confidence in the hospital (AOR = 1.9, 95%CI: 1.1, 3.1), reported discrimination/bad treatment of patients with service (AOR = 0.4, 95%CI: 0.2, 0.7), were significantly associated determinants of patient satisfaction. Non-communicable disease emergencies like injuries and cardiovascular diseases are common. There is a low level of patient satisfaction related to lack of confidence in the hospital for treatment, discrimination towards patient care, and under and delayed treatment of patients who were not in serious medical conditions. Hospitals shall prepare themselves to address the increasing challenge of non-communicable disease emergencies. It is important to revise the service delivery in the emergency department to improve staff courtesy and politeness, commitment, reduce discrimination and bad treatment and proper triage of emergencies at all points of care to increase patient satisfaction giving emphasis to earlier working days.


Socio-economic status, social support, social network, dental status, and oral health reported outcomes in adolescents.
This study explored the relationships between sex, socio-economic status, social support, social network, dental clinical status, dental pain, oral health-related quality of life (OHRQoL), and self-rated oral health (SROH) in adolescents. A cross-sectional study involving 542 adolescents, aged 12-14 yr, was conducted in Dourados, Brazil, to collect dental clinical measures (dental caries, missing teeth, and dental trauma), as well as measures of social support, social network, dental pain, OHRQoL, and SROH. Information on family income and parental education were collected from participant's parents. Structural equation modeling showed that higher income predicted better dental status and better SROH. Greater social support was linked to better dental status and better OHRQoL. Having more social networks was directly linked to better dental status. Poor dental status was linked to dental pain and poor OHRQoL. Dental pain predicted poor OHRQoL and worse SROH. Poor OHRQoL predicted worse SROH. Family income, social support, and social networks indirectly predicted dental pain via dental status. The latter was indirectly linked to OHRQoL and SROH via dental pain. Social support and social networks indirectly predicted OHRQoL and SROH via dental status and dental pain. Socio-economic factors and social relationships should be considered when planning health promotion and dental care provision to improve an adolescent's oral health.


Absence of miRNA-146a Differentially Alters Microglia Function and Proteome.
Background: MiR-146a is an important regulator of innate inflammatory responses and is also implicated in cell death and survival. Methods: By sorting CNS resident cells, microglia were the main cellular source of miR-146a. Therefore, we investigated microglia function and phenotype in miR-146a knock-out (KO) mice, analyzed the proteome of KO and wild-type (WT) microglia by LC-MS/MS, and examined miR-146a expression in different brain lesions of patients with multiple sclerosis (MS). Results: When stimulated with LPS or myelin in vitro, microglia from KO mice expressed higher levels of IL-1β, TNF, IL-6, IL-10, CCL3, and CCL2 compared to WT. Stimulation increased migration and phagocytosis of WT but not KO microglia. CD11c+ microglia were induced by cuprizone (CPZ) in the WT mice but less in the KO. The proteome of ex vivo microglia was not different in miR-146a KO compared to WT mice, but CPZ treatment induced differential and reduced protein responses in the KO: GOT1, COX5b, CRYL1, and cystatin-C were specifically changed in KO microglia. We explored discriminative features of microglia proteomes: sparse Partial Least Squares-Discriminant Analysis showed the best discrimination when control and CPZ-treated conditions were compared. Cluster of ten proteins separated WT and miR-146a KO microglia after CPZ: among them were sensomes allowing to perceive the environment, Atp1a3 that belongs to the signature of CD11c+ microglia, and proteins related to inflammatory responses (S100A9, Ppm1g). Finally, we examined the expression of miR-146a and its validated target genes in different brain lesions of MS patients. MiR-146 was upregulated in all lesion types, and the highest expression was in active lesions. Nineteen of 88 validated target genes were significantly changed in active lesions, while none were changed in NAWM. Conclusion: Our data indicated that microglia is the major source of miR-146a in the CNS. The absence of miR-146a differentially affected microglia function and proteome, and miR-146a may play an important role in gene regulation of active MS lesions.