Slowly resolving global myocardial inflammation/oedema in Tako-Tsubo cardiomyopathy: evidence from T2-weighted cardiac MRI.
Tako-Tsubo cardiomyopathy (TTC) is associated with regional left ventricular dysfunction, independent of the presence of fixed coronary artery disease. Previous studies have used T2-weighted cardiac MRI to demonstrate the presence of periapical oedema. The authors sought to determine the distribution, resolution and correlates of oedema in TTC. 32 patients with TTC were evaluated at a median of 2 days after presentation, along with 10 age-matched female controls. Extent of oedema was quantified both regionally and globally; scanning was repeated in patients with TTC after 3 months. Correlations were sought between oedema and the extent of hypokinesis, catecholamine release, release of N-terminal prohormone of B-type natriuretic peptide (NT-proBNP), and markers of systemic inflammatory activation (high-sensitivity C-reactive protein and platelet response to nitric oxide). In the acute phase of TTC, T2-weighted signal intensity was greater at the apex than at the base (p<0.0001) but was nevertheless significantly elevated at the base (p<0.0001), relative to control values. Over 3 months, T2-weighted signal decreased substantially, but remained abnormally elevated (p<0.02). The regional extent of oedema correlated inversely with radial myocardial strain (except at the apex). There were also direct correlations between global T2-weighted signal and (1) plasma normetanephrine (r=0.39, p=0.04) and (2) peak NT-proBNP (r=0.39, p=0.03), but not with systemic inflammatory markers. TTC is associated with slowly resolving global myocardial oedema, the acute extent of which correlates with regional contractile disturbance and acute release of both catecholamines and NT-proBNP.


Meta-analysis uncovers genome-wide significant variants for rapid kidney function decline.
Rapid decline of glomerular filtration rate estimated from creatinine (eGFRcrea) is associated with severe clinical endpoints. In contrast to cross-sectionally assessed eGFRcrea, the genetic basis for rapid eGFRcrea decline is largely unknown. To help define this, we meta-analyzed 42 genome-wide association studies from the Chronic Kidney Diseases Genetics Consortium and United Kingdom Biobank to identify genetic loci for rapid eGFRcrea decline. Two definitions of eGFRcrea decline were used: 3 mL/min/1.73m2/year or more ("Rapid3"; encompassing 34,874 cases, 107,090 controls) and eGFRcrea decline 25% or more and eGFRcrea under 60 mL/min/1.73m2 at follow-up among those with eGFRcrea 60 mL/min/1.73m2 or more at baseline ("CKDi25"; encompassing 19,901 cases, 175,244 controls). Seven independent variants were identified across six loci for Rapid3 and/or CKDi25: consisting of five variants at four loci with genome-wide significance (near UMOD-PDILT (2), PRKAG2, WDR72, OR2S2) and two variants among 265 known eGFRcrea variants (near GATM, LARP4B). All these loci were novel for Rapid3 and/or CKDi25 and our bioinformatic follow-up prioritized variants and genes underneath these loci. The OR2S2 locus is novel for any eGFRcrea trait including interesting candidates. For the five genome-wide significant lead variants, we found supporting effects for annual change in blood urea nitrogen or cystatin-based eGFR, but not for GATM or LARP4B. Individuals at high compared to those at low genetic risk (8-14 vs. 0-5 adverse alleles) had a 1.20-fold increased risk of acute kidney injury (95% confidence interval 1.08-1.33). Thus, our identified loci for rapid kidney function decline may help prioritize therapeutic targets and identify mechanisms and individuals at risk for sustained deterioration of kidney function.


[Functional cerebral lateralization: neurobiology and clinical aspects in childhood].
Anatomical and functional evidences of cerebral hemispheric asymmetries are reviewed in reference to fetal life, infancy and childhood. In general, left hemisphere deals with linguistic-sequenced-analytic-logic-deductive cognitive processing, whereas the right hemisphere is involved in spatial-simultaneous-inductive-intuitive tasks; attention and interactional functions predominate on the right hemisphere, too. The clinical and technological means to diagnose the functional lateralization in individual patients are discussed. A critical review is made of abnormal lateralizations and interhemispheric supplencies in various neuropsychiatric conditions of childhood: learning disabilities, epilepsy, attention deficit disorder, mental deficiency with chromosomopathies and infantile autism.


Response of different organs of immune system of mice upon administration bacterial CpG DNA.
CpG DNA are potent immunostimulator and currently being tested as adjuvant in immunotherapy of various diseases. THE AIM of this study was to investigate the effects of a single dose of bacterial CpG DNA challenge on response of murine immune system organs and on a zeta-potential of different lymphoid organ cells and peritoneal macrophages in normal and tumor-bearing mice. Indexes and cellularity of immune organs of mice were evaluated. Z-potential was measured by cellular electrophoresis. Subcutaneous administration of CpG DNA induced local transitory hyperplasia of lymph nodes, moderate increase of splenic index and total spleen cellularity, whereas intraperitoneal injection of CpG DNA caused full-blown spleen enlargement and increase of splenocyte content, and also mice showed transitory aseptic peritonitis. Response of thymus on challenge of CpG DNA was bi-phase: initial phase - response on antigen as stimulus, and late proliferative phase. It was noted that tumor growth does not affect zeta-potential in peritoneal macrophages and mononuclear lymphocytes, but causes increase of zeta-potential on thymocytes and decreases it in lymphocytes from lymph nodes. Furthermore, single administration of CpG DNA normalizes of thymocytes and lymph nodes lymphocytes zeta-potential and increases it in peritoneal macrophages and mononuclear lymphocytes. The findings demonstrate a close correlation between the hyperplasia of lymphoid follicles induced by challenge of CpG DNA and increase of their cellularity. Observed Z-potential alterations of immune system cells after CpG DNA immunization evidence on more significant polyanion accumulation on the surface of splenic macrophages and mononuclear cells.


The Rise Of Cross-Market Hospital Systems And Their Market Power In The US.
Although hospital consolidation within markets has been well documented, consolidation across markets has not, even though economic theory predicts-and evidence is emerging-that cross-market hospital systems raise prices by exerting market power across markets when negotiating with common customers (primarily insurers). This study analyzes hospital systems using the American Hospital Association Annual Survey Database and defines hospital geographic markets as commuting zones that link workers to places of employment. The share of community hospitals in the US that were part of hospital systems increased from 10 percent in 1970 to 67 percent in 2019, resulting in 3,436 hospitals within 368 systems in 2019. Of these systems, 216 (59 percent) owned hospitals in multiple commuting zones, in part because 55 percent of the 1,500 hospitals targeted for a merger or acquisition between 2010 and 2019 were located in a different commuting zone than the acquirer. Based on market-power differences among hospitals in systems, the number of systems in urban commuting zones that could potentially exert enhanced cross-market power increased from thirty-seven systems in 2009 to fifty-seven systems in 2019, an increase of 54 percent. The increase in cross-market hospital systems warrants concern and scrutiny because of the potential anticompetitive impact of hospital systems exerting market power across markets in negotiations with common customers.


Evaluation of Integrated Community Case Management in Eight Districts of Central Uganda.
Evidence is limited on whether Integrated Community Case Management (iCCM) improves treatment coverage of the top causes of childhood mortality (acute respiratory illnesses (ARI), diarrhoea and malaria). The coverage impact of iCCM in Central Uganda was evaluated. Between July 2010 and December 2012 a pre-post quasi-experimental study in eight districts with iCCM was conducted; 3 districts without iCCM served as controls. A two-stage household cluster survey at baseline (n = 1036 and 1042) and end line (n = 3890 and 3844) was done in the intervention and comparison groups respectively. Changes in treatment coverage and timeliness were assessed using difference in differences analysis (DID). Mortality impact was modelled using the Lives Saved Tool. 5,586 Village Health Team members delivered 1,907,746 treatments to children under age five. Use of oral rehydration solution (ORS) and zinc treatment of diarrhoea increased in the intervention area, while there was a decrease in the comparison area (DID = 22.9, p = 0.001). Due to national stock-outs of amoxicillin, there was a decrease in antibiotic treatment for ARI in both areas; however, the decrease was significantly greater in the comparison area (DID = 5.18; p<0.001). There was a greater increase in Artemisinin Combination Therapy treatment for fever in the intervention areas than in the comparison area but this was not significant (DID = 1.57, p = 0.105). In the intervention area, timeliness of treatments for fever and ARI increased significantly higher in the intervention area than in the comparison area (DID = 2.12, p = 0.029 and 7.95, p<0.001, respectively). An estimated 106 lives were saved in the intervention area while 611 lives were lost in the comparison area. iCCM significantly increased treatment coverage for diarrhoea and fever, mitigated the effect of national stock outs of amoxicillin on ARI treatment, improved timeliness of treatments for fever and ARI and saved lives.


Warmth is analgesic in healthy newborns.
This study identifies a behavioral and nonpharmacologic means of preventing and reducing newborn pain. Our objective was to determine whether warmth is analgesic in newborn infants undergoing vaccination-a routine painful hospital procedure. We used a prospective randomized controlled trial of 47 healthy full-term newborn infants. Infants were randomized into 1 of 3 conditions prior to vaccination: warmth exposure, pacifier suckling, or sucrose taste. Crying, grimacing, and heart rate differences were analyzed between groups before, during, and after vaccination as outcome measures. Warmer infants cried significantly less than sucrose taste or pacifier suckling after vaccination. Heart rate patterns reflected this analgesia. Core temperature did not differ between study groups. Providing natural warmth to newborn infants during a painful procedure decreases the crying and grimacing on par with the "gold" standard treatments of sucrose or pacifier.


Christmas, national holidays, sport events, and time factors as triggers of acute myocardial infarction: SWEDEHEART observational study 1998-2013.
To study circadian rhythm aspects, national holidays, and major sports events as triggers of myocardial infarction. Retrospective observational study using the nationwide coronary care unit registry, SWEDEHEART. Sweden. 283 014 cases of myocardial infarction reported to SWEDEHEART between 1998 and 2013. Symptom onset date was documented for all cases, and time to the nearest minute for 88%. Myocardial infarctions with symptom onset on Christmas/New Year, Easter, and Midsummer holiday were identified. Similarly, myocardial infarctions that occurred during a FIFA World Cup, UEFA European Championship, and winter and summer Olympic Games were identified. The two weeks before and after a holiday were set as a control period, and for sports events the control period was set to the same time one year before and after the tournament. Circadian and circaseptan analyses were performed with Sunday and 24:00 as the reference day and hour with which all other days and hours were compared. Incidence rate ratios were calculated using a count regression model. Daily count of myocardial infarction. Christmas and Midsummer holidays were associated with a higher risk of myocardial infarction (incidence rate ratio 1.15, 95% confidence interval 1.12 to 1.19, P<0.001, and 1.12, 1.07 to 1.18, P<0.001, respectively). The highest associated risk was observed for Christmas Eve (1.37, 1.29 to 1.46, P<0.001). No increased risk was observed during Easter holiday or sports events. A circaseptan and circadian variation in the risk of myocardial infarction was observed, with higher risk during early mornings and on Mondays. Results were more pronounced in patients aged over 75 and those with diabetes and a history of coronary artery disease. In this nationwide real world study covering 16 years of hospital admissions for myocardial infarction with symptom onset documented to the nearest minute, Christmas, and Midsummer holidays were associated with higher risk of myocardial infarction, particularly in older and sicker patients, suggesting a role of external triggers in vulnerable individuals.


Susceptibility of laboratory rats to isolates of Borrelia burgdorferi from different geographic areas.
One week old LEW/N rats were inoculated with 15 different isolates of Borrelia burgdorferi from 3 major North American (Northeast, Midwest, and Pacific) and 1 European endemic areas. At 30 days after inoculation, several tissues were cultured for B. burgdorferi and examined for histopathological changes. Sera were tested for IgM and IgG antibody to B. burgdorferi by an enzyme-linked immunosorbent assay. One or more isolates from each geographic region was recovered by culture and caused arthritis in the rats. No differences in pattern or severity of disease were apparent among pathogenic isolates. Several recovered isolates failed to infect rats, or did not cause disease. Rats that were actively infected seroconverted but uninfected rats did not. Infectivity and pathogenicity of B. burgdorferi isolates could not be correlated with molecular weights or expression of outer surface proteins based on agar gel electrophoresis.


Community Resource Connection for Pediatric Caregivers With Unmet Social Needs: A Qualitative Study.
Pediatric health systems are increasingly screening caregivers for unmet social needs. However, it remains unclear how best to connect families with unmet needs to available and appropriate community resources. We aimed to explore caregivers' perceived barriers to and facilitators of community resource connection. We conducted semistructured interviews with caregivers of pediatric patients admitted to one inpatient unit of an academic quaternary care children's hospital. All caregivers who screened positive for one or more unmet social needs on a tablet-based screener were invited to participate in an interview. Interviews were recorded, transcribed, and coded by 2 independent coders using content analysis, resolving discrepancies by consensus. Interviews continued until thematic saturation was achieved. We interviewed 28 of 31 eligible caregivers. Four primary themes emerged. First, caregivers of children with complex chronic conditions felt that competing priorities related to their children's medical care often made it more challenging to establish connection with resources. Second, caregivers cited burdensome application and enrollment processes as a barrier to resource connection. Third, caregivers expressed a preference for geographically tailored, web-based resources, rather than paper resources. Last, caregivers expressed a desire for ongoing longitudinal support in establishing and maintaining connections with community resources after their child's hospital discharge. Pediatric caregivers with unmet social needs reported competing priorities and burdensome application processes as barriers to resource connection. Electronic resources can help caregivers identify locally available services, but longitudinal supports may also be needed to ensure caregivers can establish and maintain linkages with these services.


Epidemiology of bicycle injuries and risk factors for serious injury.
To determine the risk factors for serious injury to bicyclists, aside from helmet use. Prospective case-control study. Seven Seattle area hospital emergency departments and two county medical examiner's offices. Individuals treated in the emergency department or dying from bicycle related injuries. Information collected from injured bicyclists or their parents by questionnaire on circumstances of the crash; abstract of medical records for injury data. Serious injury defined as an injury severity score > 8. Odd ratios computed using the maximum likelihood method, and adjusted using unconditional logistic regression. There were 3854 injured cyclists in the three year period; 3390 (88%) completed questionnaires were returned 51% wore helmets at the time of crash. Only 22.3% of patients had head injuries and 34% had facial injuries. Risk of serious injury was increased by collision with a motor vehicle (odds ratio (OR) = 4.6), self reported speed > 15 mph (OR = 1.2), young age (< 6 years), and age > 39 years (OR = 2.1 and 2.2 respectively, compared with adults 20-39 years). Risk for serious injury was not affected by helmet use (OR = 0.9). Risk of neck injury was increased in those struck by motor vehicles (OR = 4.0), hospitalized for any injury (OR = 2.0), and those who died (OR = 15.1), but neck injury was not affected by helmet use. Prevention of serious bicycle injuries cannot be accomplished through helmet use alone, and may require separation of cyclists from motor vehicles, and delaying cycling until children are developmentally ready.


Right-Angled Traction Bronchiectasis in Differentiating Idiopathic Pulmonary Fibrosis Without Honeycombing From Idiopathic Nonspecific Interstitial Pneumonia.
The aim of this study was to conduct a radiopathologic evaluation of right-angled traction bronchiectasis to differentiate idiopathic pulmonary fibrosis (IPF) without honeycombing from idiopathic nonspecific interstitial pneumonia (NSIP). The derivation cohort included 78 consecutive patients with idiopathic NSIP (n = 39) or IPF (n = 39) without honeycombing who underwent preoperative thin-section computed tomography scans at a single tertiary hospital. The validation cohort comprised 22 patients (14 IPF and 8 NSIP) from another institution. We assessed conventional computed tomography findings, right-angled traction bronchiectasis on minimum intensity projection (MinIP) images, and pathologic features associated with right-angled bronchiectasis. Right-angled traction bronchiectasis was defined as abrupt kinking of a single bronchus by over 90 degrees or an abrupt angle close to 180 degrees of branching bronchi in the background of fibrosis. In the validation cohort, we evaluated the proportion of correct IPF diagnoses and interobserver agreement of 4 radiologists before and after reviewing MinIP images. A probable usual interstitial pneumonia (UIP) pattern (odds ratio [OR], 6.948; 95% confidence interval [CI], 1.525-31.654; P = 0.012) and right-angled traction bronchiectasis (OR, 6.004; 95% CI, 1.980-18.209; P = 0.002) were independently associated with IPF. Patients with right-angled traction bronchiectasis were more likely to have extensive reticular opacity (OR, 1.149; 95% CI, 1.077-1.225; P < 0.001) and pathologically were more likely to have a broad extent of subpleural fibrosis (OR, 4.000; 95% CI, 1.457-10.987; P = 0.007) and relatively thick fibrosis (OR, 7.750; 95% CI, 2.504-23.991; P < 0.001). After reviewing MinIP images, the proportion of correct diagnoses increased from 40.9% to 54.5% to 50.0% to 77.3%. The mean kappa value for right-angled traction bronchiectasis was 0.489 ± 0.192. Right-angled traction bronchiectasis pathologically reflected a subpleural predominance of fibrosis and partly supported the radiologic differentiation of IPF without honeycombing from idiopathic NSIP.


Stimulated photorefractive backscatter leading to six-wave mixing and phase conjugation in iron-doped lithium niobate.
The generation of multiple waves during near-oblique incidence of a 532 nm weakly focused beam on photorefractive iron-doped lithium niobate in a typical reflection geometry configuration is studied. It is shown that these waves are produced through two-wave coupling (self-diffraction) and four-wave mixing (parametric diffraction). One of these waves, the stimulated photorefractive backscatter produced from parametric diffraction, contains the self-phase conjugate. The dynamics of six-wave mixing and its dependence on crystal parameters, angle of incidence, and pump power are analyzed. What we believe to be a novel order analysis of the interaction equations provides further insight into experimental observations in the steady state. The quality of the backscatter is evaluated through image restoration, interference experiments, and visibility measurement. Reduction of two-wave coupling may significantly improve the quality of the self-phase conjugate.


Relationship between betel quid additives and established periodontitis among Bangladeshi subjects.
To determine the relationship between betel quid chewing additives and established periodontitis in Bangladeshi subjects. A total of 864 subjects participated in this study. Among them, 140 pairs of sex- and age-matched case subjects and control subjects were selected. A case was defined as a person who had at least two sites with a clinical attachment level (CAL)> or =6 mm and at least one site with probing depth (PD)> or =5 mm. Subjects who did not fulfill these criteria were considered as controls. Information on sociodemographic variables, psychological stress, dental health behaviour, smoking and betel quid chewing habits was obtained. Multiple logistic regression analysis showed that current betel quid chewers had greater probabilities of having established periodontal disease than did non-chewers (odds ratio=3.97, p<0.05). Mean PD, mean CAL, mean percentage of bleeding on probing and number of missing teeth were significantly higher in chewers of betel quid with tobacco and masala than in chewers of betel quid without such additives adjusting for age, sex, smoking habit, body mass index, dental visit pattern, stress and plaque index. Higher frequency and longer duration of betel quid chewing showed a significant relation to an increase in periodontal parameters. The results indicate that betel quid additives might significantly enhance periodontitis in the population studied.


Stathmin as a marker for malignancy in pheochromocytomas.
Pheochromocytomas of the adrenal medulla may be life-threatening catecholamine-producing tumors which are malignant in about 10% of cases. Differential diagnosis between malignant and benign tumors is dependent on the development of metastasis or extensive local invasion. A number of genetic aberrations have been described in pheochromocytomas, but no marker associated to malignancy has been reported. We applied an expression microarray containing 7770 cDNA clones and analysed the expression profiles in eleven tumors compared to normal adrenal medulla. Stathmin (STMN1, Op18) was most conspiciously overexpressed among the differentially expressed genes. RT-PCR analysis further confirmed mRNA overexpression, 6 to 8-fold for benign and malignant tumors, and 16-fold for metastases. Stathmin protein overexpression was observed by immunohistochemistry, and distinct differential protein expression between benign and malignant/metastasis specimens was confirmed by Western blot analysis. The results introduce stathmin as a possible diagnostic marker for malignant pheochromocytomas, and further evaluations are warranted.


Can regenerating axons recapitulate developmental guidance during recovery from spinal cord injury?
The precise wiring of the adult mammalian CNS originates during a period of stunning growth, guidance and plasticity that occurs during and shortly after development. When injured in adults, this intricate system fails to regenerate. Even when the obstacles to regeneration are cleared, growing adult CNS fibres usually remain misdirected and fail to reform functional connections. Here, we attempt to fill an important niche related to the topics of nervous system development and regeneration. We specifically contrast the difficulties faced by growing fibres within the adult context to the precise circuit-forming capabilities of developing fibres. In addition to focusing on methods to stimulate growth in the adult, we also expand on approaches to recapitulate development itself.


mHealth Interventions to Reduce Alcohol Use in Young People: A Systematic Review of the Literature.
Harmful use of alcohol has serious effects on public health and is considered a significant risk factor for poor health. mHealth technology promotes health behavior change and enhances health through increased social opportunities for encouragement and support. It remains unknown whether these types of applications directly influence the health status of young people in reducing harmful levels of alcohol consumption. The purpose of this systematic review is to examine current evidence on the effectiveness of mHealth technology use in positively influencing alcohol-related behaviors of young people without known alcohol addiction. Relevant articles published from 2005 to January 2017 were identified through electronic searches of eight databases. Studies with interventions delivered by mHealth (social networking sites, SMS and mobile phone applications) to young people aged 12-26 years were included. Outcome measures were alcohol use, reduction in alcohol consumption or behavior change. Eighteen studies met the inclusion criteria. Interventions varied in design, participant characteristics, settings, length and outcome measures. Ten studies reported some effectiveness related to interventions with nine reporting a reduction in alcohol consumption. Use of mHealth, particularly text messaging (documented as SMS), was found to be an acceptable, affordable and effective way to deliver messages about reducing alcohol consumption to young people. Further research using adequately powered sample sizes in varied settings, with adequate periods of intervention and follow-up, underpinned by theoretical perspectives incorporating behavior change in young people's use of alcohol, is needed.


FTIR study of thermally induced magnetostructural transitions in breathing crystals.
"Breathing crystals" based on copper(II) hexafluoroacetylacetonates and pyrazolyl-substituted nitronyl nitroxides comprise the exchange-coupled clusters within the polymeric chains. Owing to an interplay of exchange interaction between copper(II) and nitroxide spins and Jahn-Teller nature of copper(II) complex, the breathing crystals demonstrate thermally and light-induced magnetostructural transitions in many aspects similar to the classical spin crossover. Herewith, we report the first application of variable temperature (VT) far/mid Fourier transform infrared (FTIR) spectroscopy and mid FTIR microscopy to breathing crystals. This VT-FTIR study was aimed toward clarification of the transitions mechanism previously debated on the basis of superconducting quantum interference device, X-ray diffraction, and electron paramagnetic resonance data. VT-FTIR showed the onset of new vibrational bands during phase transitions occurring at the expense of several existing ones, whose intensity was significantly reduced. The most pronounced spectral changes were assigned to corresponding vibrational modes using quantum chemical calculations. A clear-cut correlation was found between temperature-dependent effective magnetic moment of studied compounds and the observed VT-FTIR spectra. Importantly, VT-FTIR confirmed coexistence of two types of copper(II)-nitroxide clusters during gradual magnetostructural transition. Such clusters correspond to weakly coupled and strongly coupled spin states, whose relative contribution depends on temperature. The pronounced difference in the VT-FTIR spectra of two states in breathing crystals is a fingerprint of magnetostructural transition, and understanding of these characteristics achieved by us will be useful for future studies of breathing crystals as well as their diamagnetic analogues.


Progress Toward the Validation of Alternative Tests.
ECVAM's role in the practical validation of replacement alternative methods for use in regulatory testing is reviewed, including an outline of the criteria which have been used in determining ECVAM's priorities. Some of the difficulties which have arisen in validation studies are discussed, and solutions to these are proposed, with particular emphasis on ensuring that methods are sufficiently well-developed to enter the validation process, and on the ECVAM prevalidation scheme for encouraging protocol optimisation and the prior assessment of interlaboratory transferability. Comments are made on problems encountered in selecting test materials backed by adequate in vivo data and in undertaking appropriate in vivo/in vitro comparisons.


Microbiological quality of ice and ice machines used in food establishments.
The ice used in the food industry has to be safe and the water used in ice production should have the quality of drinking water. The consumption of contaminated ice directly or indirectly may be a vehicle for transmission of pathogenic bacteria to humans producing outbreaks of gastrointestinal diseases. The objective of this study was to monitor the microbiological quality of ice, the water used in producing ice and the hygienic conditions of ice making machines in various food enterprises. Escherichia coli was detected in seven (6.7%) ice and 23 (21.9%) ice chest samples whereas E. coli was negative in all examined water samples. Psychrophilic bacteria were detected in 83 (79.0%) of 105 ice chest and in 68 (64.7%) of 105 ice samples, whereas Enterococci were detected only in 13 (12.4%) ice samples. Coliforms were detected in 13 (12.4%) water, 71 (67.6%) ice chest and 54 (51.4%) ice samples. In order to improve the microbiological quality of ice, the maintenance, cleaning and disinfecting of ice machines should be carried out effectively and periodically. Also, high quality water should be used for ice production.


Yeast ribosomal protein L7 and its homologue Rlp7 are simultaneously present at distinct sites on pre-60S ribosomal particles.
Ribosome biogenesis requires >300 assembly factors in Saccharomyces cerevisiae. Ribosome assembly factors Imp3, Mrt4, Rlp7 and Rlp24 have sequence similarity to ribosomal proteins S9, P0, L7 and L24, suggesting that these pre-ribosomal factors could be placeholders that prevent premature assembly of the corresponding ribosomal proteins to nascent ribosomes. However, we found L7 to be a highly specific component of Rlp7-associated complexes, revealing that the two proteins can bind simultaneously to pre-ribosomal particles. Cross-linking and cDNA analysis experiments showed that Rlp7 binds to the ITS2 region of 27S pre-rRNAs, at two sites, in helix III and in a region adjacent to the pre-rRNA processing sites C1 and E. However, L7 binds to mature 25S and 5S rRNAs and cross-linked predominantly to helix ES7(L)b within 25S rRNA. Thus, despite their predicted structural similarity, our data show that Rlp7 and L7 clearly bind at different positions on the same pre-60S particles. Our results also suggest that Rlp7 facilitates the formation of the hairpin structure of ITS2 during 60S ribosomal subunit maturation.


Overweight and obesity in the context of heart failure: implications for practice and future research.
Heart failure (HF) is a major cardiovascular disorder, public health concern, and growing epidemic that affects approximately 5,000,000 people in the United States with 550,000 new cases reported annually. The clinical and economic impact of HF is associated with high hospitalization and early readmission rates. Recognizing factors that contribute to increasing the risk for HF, particularly the persistent rise in prevalence of overweight and obesity, may be imperative to reducing the burdens of this poorly prognostic disease process. Overweight and obesity have been associated with increased morbidity and mortality, and have incited extensive interest in therapeutic interventions. However, within the last decade, studies have illustrated the positive effects of overweight and obesity on survival after the onset of HF, which has prompted a variance of opinion within the healthcare community. This article reviews data supporting both the negative and positive effects of overweight and obesity in relationship to HF with implications for future research, and describes recommendations for practice as it relates to lifestyle modification through diet, exercise, and cognitive-behavioral therapy.


Inputs and outcomes: what do staff in services for people with intellectual disabilities perceive they bring to and receive from their work-based relationships?
A number of studies involving staff working in services for people with intellectual disabilities have utilised equity theory as a theoretical framework. According to this theory, people evaluate social relationships through the comparison of inputs and outcomes, respectively, with what a person brings to and receives from a relationship. Little is known about what constitute inputs and outcomes for staff working in services for people with intellectual disabilities. Semi-structured interviews were conducted with 15 staff to find out what constitute inputs and outcomes for staff who work with people with intellectual disabilities. The interviews were conducted in the first half of 2008 in the United Kingdom. Data were analysed using template analysis. A wide range of inputs and outcomes was identified by staff, which were grouped under high-level themes relating to relationships with their employers, their co-workers and the service users. The utility of the findings, in terms of informing future research, is discussed.


Antiretroviral use in Italian children with perinatal HIV infection over a 14-year period.
Information on the use of new antiretroviral drugs in children in the real setting of clinical fields is largely unknown. Data from 2554 combined antiretroviral therapy (cART) regimens administered to 911 children enrolled in the Italian Register for HIV infection in children, between 1996 and 2009, were analysed. Factors potentially associated with undetectable viral load and immunological response to cART were explored by Cox regression analysis. Proportion of protease inhibitor (PI)-based regimens significantly decreased from 88.0% to 51.2% and 54.9%, while proportion on non-nucleoside reverse transcriptase inhibitor (NNRTI)-based regimens increased from 4.5% to 38.8% and 40.2% in 1996-1999, 2000-2004 and 2005-2009, respectively (p < 0.0001). Significant change in the use of each antiretroviral drug occurred over the time periods (p < 0.0001). Factors independently associated with virological and immunological success were as follows: later calendar periods, younger age at regimen (only for virological success) and higher CD4(+) T-lymphocyte percentage at baseline. Use of unboosted PI was associated with lower adjusted hazard ratio (aHR) of virological or immunological success with respect to NNRTI- and boosted PI-based regimens, with no difference among these two latter types. Use of new generation antiretroviral drugs in Italian HIV-infected children is increasing. No different viro-immunological outcomes between NNRTI- and boosted PI-based cART were observed.


Angiotensin receptors in myometrium and myometrial vessels from uteri of women during the follicular and luteal phases of the menstrual cycle and in late pregnancy.
1. Receptors for angiotensin II were identified and characterized in membrane fractions from myometrial samples obtained at non-pregnant hysterectomy in women of reproductive age. Specific binding was also measured in paired samples of vascular and "vessel-free' myometrium. 2. A single class of high-affinity receptor sites was identified in the total myometrial preparations (n = 10), with a median equilibrium dissociation constant (Kd) of 0.122 nmol/l (range 0.065-0.465 nmol/l) and concentration of receptor sites (Bmax) of 149 fmol/mg protein (range 105-435 fmol/mg). Receptor characterization studies using losartan (an angiotensin type 1 receptor antagonist) and PD123177 (an angiotensin type 2 receptor antagonist) showed the myometrium to contain almost entirely type 2 receptors. 3. The median Kd and Bmax for specific angiotensin II binding in isolated myometrial vessel homogenates were 0.086 nmol/l (range 0.060-0.433) and 260 fmol/mg protein (range 197-737 fmol/mg) respectively. Vascular specific binding density was always higher in dissected out myometrial vessels than in paired vessel-free myometrium (median 372 compared with 120 fmol/mg protein; n = 20; P < 0.001). The specific binding in the paired samples was strongly correlated (r = 0.8904, P < 0.0001). 4. The specific binding of 125I-labelled angiotensin II in "vessel-free' myometrium was higher in samples from the follicular phase than in samples from the luteal phase (median 144 compared with 37 fmol/mg; P < 0.02). A similar trend was found in the vessels themselves, but this failed to reach statistical significance (459 compared with 225 fmol/mg; P = 0.051). 5. It is suggested that the down-regulation of the angiotensin type 2 receptors in the luteal (secretory) phase is a preparation for pregnancy, removing an inhibitory factor to uterine growth and vascularization and allowing greater prostacyclin synthesis.


AR negative triple negative or "quadruple negative" breast cancers in African American women have an enriched basal and immune signature.
There is increasing evidence that Androgen Receptor (AR) expression has prognostic usefulness in Triple negative breast cancer (TNBC), where tumors that lack AR expression are considered "Quadruple negative" Breast Cancers ("QNBC"). However, a comprehensive analysis of AR expression within all breast cancer subtypes or stratified by race has not been reported. We assessed AR mRNA expression in 925 tumors from The Cancer Genome Atlas (TCGA), and 136 tumors in 2 confirmation sets. AR protein expression was determined by immunohistochemistry in 197 tumors from a multi-institutional cohort, for a total of 1258 patients analyzed. Cox hazard ratios were used to determine correlations to PAM50 breast cancer subtypes, and TNBC subtypes. Overall, AR-negative patients are diagnosed at a younger age compared to AR-positive patients, with the average age of AA AR-negative patients being, 49. AA breast tumors express AR at lower rates compared to Whites, independent of ER and PR expression (p<0.0001). AR-negative patients have a (66.60; 95% CI, 32-146) odds ratio of being basal-like compared to other PAM50 subtypes, and this is associated with an increased time to progression and decreased overall survival. AA "QNBC" patients predominately demonstrated BL1, BL2 and IM subtypes, with differential expression of E2F1, NFKBIL2, CCL2, TGFB3, CEBPB, PDK1, IL12RB2, IL2RA, and SOS1 genes compared to white patients. Immune checkpoint inhibitors PD-1, PD-L1, and CTLA-4 were significantly upregulated in both overall "QNBC" and AA "QNBC" patients as well. Thus, AR could be used as a prognostic marker for breast cancer, particularly in AA "QNBC" patients.


Stimulatory effects of malathion on the key enzymes activities of insulin secretion in langerhans islets, glutamate dehydrogenase and glucokinase.
Previous studies showed that malathion induces hyperglycemia mainly through influence on glucose metabolism in liver and skeletal muscles. The main objective of the present study was to determine what will happen on pancreatic key enzymes of insulin secretion, including glucokinase (GK) and glutamate dehydrogenase (GDH), if animals would be in acute or subchronic exposure to various doses of malathion, an organophosphorous insecticide in rats. In the subchronic study, malathion was administered orally at doses of 100 to 400 ppm for 4 weeks. In the acute experiment, animals received various doses of 3 to 75 mg/kg of malathion intraperitoneally. In each experiment, islets were isolated from the pancreas of rats by a standard collagenase digestion, separation by centrifugation, and hand-picking technique. The activities of the mitochondrial GDH and the nonmitochondrial GK enzymes were determined in islets homogenates spectrophotometrically. Blood sample was taken by cardiac puncture for glucose and insulin assays. In the acute experiment, malathion (3, 15, 75 mg/kg) increased blood glucose and insulin (15 and 75 mg/kg). In the subchronic experiment, malathion (100, 200, 400 ppm) increased blood glucose and insulin (200 and 400 ppm). All doses in both acute and subchronic experiments increased the mitochondrial GDH activity. Acute (15 and 75 ppm) and subchronic (200 and 400 ppm) increased the islets GK activity. It was concluded that pancreatic islet key enzymes are stimulated following acute and subchronic exposure to malathion though not enough to overcome to hyperglycemia. Activation of islets muscarinic receptors by malathion in favor of hyperinsulinemia, overproduction of glutamate/glutathione by GDH, and overproduction of glucose via increased glycogenolysis in counteracting with malathion-induced oxidative stress are possible mechanisms for observed effects. A new NOAEL acceptable daily intake must be established for malathion.


High-intensity focused ultrasound ablation of liver tumours: can radiological assessment predict the histological response?
Cancer therapies usually depend on cross-sectional imaging for the assessment of treatment response. This study was designed to evaluate the ability of MRI to predict zones of necrosis following the use of high-intensity focused ultrasound (HIFU) to treat liver metastases. Patients with liver metastases, who had been scheduled for elective surgical resection of their tumours, were recruited to this non-randomized Phase II study. In each case, a proportion of an index liver tumour target was ablated. The response to HIFU was assessed after 12 days using contrast-enhanced MRI and compared directly with histological analysis at the time of surgery. Eight patients were treated, of whom six were subsequently assessed with both MRI and histology. There were no major complications. MRI predicted complete ablation in three cases. In each case, histological analysis confirmed complete ablation. In one case, the region of ablation observed on MRI appeared smaller than predicted at the time of HIFU, but histology revealed complete ablation of the target region. The predominant characteristic of HIFU-ablated tissue was coagulative necrosis but heat fixation was evident in some areas. Heat-fixed cells appeared normal under haematoxylin and eosin staining, indicating that this is unreliable as an indicator of HIFU-induced cell death. This study demonstrates that HIFU is capable of achieving selective ablation of pre-defined regions of liver tumour targets, and that MRI evidence of complete ablation of the target region can be taken to infer histological success.


Cathepsin S Alterations Induce a Tumor-Promoting Immune Microenvironment in Follicular Lymphoma.
Tumor cells orchestrate their microenvironment. Here, we provide biochemical, structural, functional, and clinical evidence that Cathepsin S (CTSS) alterations induce a tumor-promoting immune microenvironment in follicular lymphoma (FL). We found CTSS mutations at Y132 in 6% of FL (19/305). Another 13% (37/286) had CTSS amplification, which was associated with higher CTSS expression. CTSS Y132 mutations lead to accelerated autocatalytic conversion from an enzymatically inactive profrom to active CTSS and increased substrate cleavage, including CD74, which regulates major histocompatibility complex class II (MHC class II)-restricted antigen presentation. Lymphoma cells with hyperactive CTSS more efficiently activated antigen-specific CD4+ T cells in vitro. Tumors with hyperactive CTSS showed increased CD4+ T cell infiltration and proinflammatory cytokine perturbation in a mouse model and in human FLs. In mice, this CTSS-induced immune microenvironment promoted tumor growth. Clinically, patients with CTSS-hyperactive FL had better treatment outcomes with standard immunochemotherapies, indicating that these immunosuppressive regimens target both the lymphoma cells and the tumor-promoting immune microenvironment.


Are the differences in adulthood ill-health across the north-south divide and between Scotland and England also evident in early childhood health indicators?
Regional differences in adult morbidity and mortality within England (i.e., north-south divide or gradient) and between England and Scotland (i.e., Scottish effect) are only partly explained by adult levels of socioeconomic status or risk factors. This suggests variation in early life, and is supported by the foetal origins and life-course literature which posits that birth outcomes and subsequent, cumulative exposures influence adult health. However, no studies have examined the north-south gradient or Scottish effect in health in the earliest years of life. The aims of the study were: i) to examine health indicators in English and Scottish children at birth and age three to establish whether regional differences exist; and ii) to establish whether observed changes in child health at age three were attributable to birth and/or early life environmental exposures. Respondents included 10,639 biological Caucasian mothers of singleton children recruited to the Millennium Cohort Study (MCS) in the year 2000. Outcome variables were: gestational age and birth weight, and height, body mass index (BMI), and externalising behavioural problems at age three. Region/country was categorised as: South (reference), Midlands, North (England), and Scotland. Respondents provided information on child, maternal, household, and socioeconomic characteristics. Results indicated no significant regional variations for gestational age or birth weight. At age three there was a north-south gradient for externalising behaviour and a north-south divide in BMI which attenuated on adjustment. However, a north-south divide in height was not fully explained by adjustment. There was also evidence of a 'Midlands effect', with increased likelihood of shorter stature and behaviour problems. Results showed a Scottish effect for height and BMI in the unadjusted models, and height in the adjusted model, but a decreased likelihood of behaviour problems. Findings indicated no regional differences in health at birth, but some regional variation at age three supports the cumulative life-course model.


Comparison of three cell-based drug screening platforms for HSV-1 infection.
Acyclovir (ACV) and its derivatives have been highly effective for treating recurrent, lytic infections with Herpes Simplex Virus, type 1 (HSV-1), but searches for additional antiviral drugs are motivated by recent reports of resistance to ACV, particularly among immunocompromised patients. In addition, the relative neurotoxicity of ACV and its inability to prevent neurological sequelae among HSV-1 encephalitis survivors compel searches for new drugs to treat HSV-1 infections of the central nervous system (CNS). Primary drug screens for neurotropic viruses like HSV-1 typically utilize non-neuronal cell lines, but they may miss drugs that have neuron specific antiviral effects. Therefore, we compared the effects of a panel of conventional and novel anti-herpetic compounds in monkey epithelial (Vero) cells, human induced pluripotent stem cells (hiPSCs)-derived neural progenitor cells (NPCs) and hiPSC-derived neurons (N = 73 drugs). While the profiles of activity for the majority of the drugs were similar in all three tissues, Vero cells were less likely than NPCs to identify drugs with substantial inhibitory activity in hiPSC-derived neurons. We discuss the relative merits of each cell type for antiviral drug screens against neuronal infections with HSV-1.


What DNA methylation modifications and/or genetic variations interact with childhood maltreatment in the development of depression: A systematic review.
Child maltreatment predicts a significant risk factor for depression. The relationship between child maltreatment and depression has been shown to vary as a function of genetic factors. There have been very few systematic reviews conducted to date to synthesize what DNA methylations and/ or genetic variations interact with childhood maltreatment in the course of depression. This systematic review aimed to provide an overview of DNA methylation modifications with/without genetic variations associated with childhood maltreatment in depression. Computerized and manual search on six databases (EMBASE, HealthStar, PsychoInfo, Medline, PubMed and Cochrane Library) and grey literature up to June 30th 2018 were conducted. Studies were critically evaluated for their eligibility and study quality. The initial search resulted in 196 articles. Five articles met the eligibility criteria being included in this review. All the selected studies were from the United States and published within the last five years. Changes in ID3, TPPP, GRIN1, and OXTR DNA methylation sites were found to be involved in the childhood maltreatment-depression relationship. The number of eligible articles included in this review was small. Selected articles had small sample sizes. A high degree of heterogeneity was found. It is difficult to conclude what the roles of DNA methylation modifications are in the relationship between maltreatment and depression. Population stratification has not been extensively studied so far and should be considered in the further research. This review synthesizes an overview of the interaction between childhood maltreatment, DNA methylation modifications and genetic variations in depression. Findings of this review highlight an urgent need for genetic and epigenetic research in the area of childhood maltreatment and depression. Future etiological explorations should target on the above identified sites.


Methylation Linear Discriminant Analysis (MLDA) for identifying differentially methylated CpG islands.
Hypermethylation of promoter CpG islands is strongly correlated to transcriptional gene silencing and epigenetic maintenance of the silenced state. As well as its role in tumor development, CpG island methylation contributes to the acquisition of resistance to chemotherapy. Differential Methylation Hybridisation (DMH) is one technique used for genome-wide DNA methylation analysis. The study of such microarray data sets should ideally account for the specific biological features of DNA methylation and the non-symmetrical distribution of the ratios of unmethylated and methylated sequences hybridised on the array. We have therefore developed a novel algorithm tailored to this type of data, Methylation Linear Discriminant Analysis (MLDA). MLDA was programmed in R (version 2.7.0) and the package is available at CRAN 1. This approach utilizes linear regression models of non-normalised hybridisation data to define methylation status. Log-transformed signal intensities of unmethylated controls on the microarray are used as a reference. The signal intensities of DNA samples digested with methylation sensitive restriction enzymes and mock digested are then transformed to the likelihood of a locus being methylated using this reference. We tested the ability of MLDA to identify loci differentially methylated as analysed by DMH between cisplatin sensitive and resistant ovarian cancer cell lines. MLDA identified 115 differentially methylated loci and 23 out of 26 of these loci have been independently validated by Methylation Specific PCR and/or bisulphite pyrosequencing. MLDA has advantages for analyzing methylation data from CpG island microarrays, since there is a clear rational for the definition of methylation status, it uses DMH data without between-group normalisation and is less influenced by cross-hybridisation of loci. The MLDA algorithm successfully identified differentially methylated loci between two classes of samples analysed by DMH using CpG island microarrays.


The efficacy and safety of a new enteric-coated formulation of fluoxetine given once weekly during the continuation treatment of major depressive disorder.
A simple, once-weekly dosing regimen could be a convenient alternative for many patients during long-term treatment of depression. Such a strategy might also be effective for improving medication compliance and the outcome of continuation treatment. The safety and effectiveness of a new formulation of enteric-coated fluoxetine (90 mg) given once weekly was tested during the continuation treatment of major depressive disorder. Patients meeting DSM-IV criteria for major depressive disorder with modified 17-item Hamilton Rating Scale for Depression (HAM-D-17) scores > or = 18 and Clinical Global Impressions-Severity of Illness scale (CGI-S) scores > or = 4 were treated 13 weeks with open-label 20 mg/day of fluoxetine in a multicenter U.S. study. Responders (N = 501) were randomly assigned to receive 20 mg of fluoxetine daily, placebo, or 90 mg of enteric-coated fluoxetine weekly for 25 weeks of double-blind continuation treatment. The primary efficacy measure was the percentage of patients who relapsed. Time to relapse was tested over the 25-week continuation period using log-rank analyses of the Kaplan-Meier estimates of relapse rates. Additional analyses of efficacy included comparison of change from baseline to endpoint for the HAM-D-17, CGI-S, and HAM-D-28 subscales by last observation carried forward (LOCF). Safety measures included comparison of treatment-emergent adverse events, both spontaneous and solicited (using the Association for Methodology of Documentation in Psychiatry-Module 5), vital signs, and laboratory measures. Relapse rates for patients assigned to fluoxetine, either 20 mg daily or 90 mg weekly, were significantly lower than for placebo by log-rank analysis and LOCF analyses of secondary efficacy measures. Efficacy did not significantly differ between the 2 active drug groups by these measures. Enteric-coated fluoxetine at a once-weekly dose of 90 mg was well tolerated, and its safety profile was similar to that of daily 20 mg of fluoxetine. The formulation of enteric-coated fluoxetine taken once weekly is effective, safe, and well tolerated for continuation treatment of depression in patients who responded to acute treatment with 20 mg/day of fluoxetine. Monitoring during long-term treatment for evidence of sustained remission is important regardless of dosing regimen.


Infarcts of undetermined cause: the NINCDS Stroke Data Bank.
In a prospective study of 1,805 hospitalized patients in the Stroke Data Bank of the National Institute of Neurological and Communicative Disorders and Stroke, the 1,273 with infarction were classified into diagnostic subtypes. Diagnosis was based on the clinical history, examination, and laboratory tests including computed tomography, noninvasive vascular imaging, and where safe and relevant, angiography. Five hundred and eight cases (fully 40%) were labeled as infarcts of undetermined cause (IUC), of which 138 (27%) were evaluated with both computed tomography and angiography. The clinical syndrome and computed tomographic and angiographic findings in 91 (65.9%) of these 138 IUC cases were clearly not attributable to large-artery thrombosis and could permit reclassification of the infarct as due to some form of embolism. Failure to define a source of embolus kept them in the category of IUC. Thirty-one cases (22.5%) could be reclassified as due to stenosis or thrombosis of a large artery, and 16 (11.6%) as lacunar infarction. To determine if those selected for angiography among the IUC patients differed from those with other final diagnoses, a stepwise multiple logistic model was used. The most important characteristics were young age, presence of a superficial infarct, prior transient ischemic attack, low weakness score, and presentation with a nonlacunar syndrome. The results of the model suggest that angiography use was determined by clinical characteristics uniformly across centers and not by final diagnosis. Continued use of the category IUC may help clarify risk factors and stroke subtypes, allow new mechanisms of ischemic stroke to be uncovered, and prevent classification categories of stroke used in clinical trials from becoming too broad.


Velocity profiles and shear strain rate variability in the USP Dissolution Testing Apparatus 2 at different impeller agitation speeds.
The fluid velocity profiles at different locations inside a standard USP Dissolution Testing Apparatus 2 were experimentally obtained via Laser Doppler Velocimetry (LDV) at three impeller agitations speeds, namely 50rpm, 75rpm and 100rpm. The experimental results were compared with the predictions obtained with Computational Fluid Dynamics (CFD) where the κ-ω model with low Reynolds number correction was used to account for turbulence effects. In general, good agreement was found between the experimental LDV velocity measurements and the CFD simulation predictions. The non-dimensional tangential, axial and radial velocity profiles (scaled with the impeller tip speed) and the flow pattern were found to be nearly independent of the agitation speed in most regions of the vessel, implying that increasing the agitation speed generally produced a corresponding increase in the local values of the velocity. However, the velocity profiles and flow pattern in the inner core region just below the impeller, where the dissolving tablet is usually located, were found to be much less sensitive to agitation speed. In this region, the axial and radial velocities were especially low and were not significantly affected by agitation increases. This inner core region at the center of the vessel bottom persisted irrespective of agitation intensity. The CFD predictions also indicated that increasing the agitation speed resulted in a higher shear strain rate distribution along the vessel bottom, although the strain rate was always very low at the center of the vessel bottom, even when the agitation speed was increased.


Analytical fractionation of homogenates from cultured rat embryo fibroblasts.
Homogenates of cultured rat embryo fibroblasts have been assayed for acid phosphatase, N-acetyl-beta-glucosaminidase, cathepsin D, acid deoxyribonuclease, cytochrome oxidase, NADH cytochrome c reductase, 5'-nucleotidase, inosine diphosphatase, acid pyrophosphatase, neutral pyrophosphatase, esterase, catalase, cholesterol, and RNA. The validity of the assay conditions was checked. Neutral pyrophosphatase is a readily soluble enzyme. Acid hydrolases, except acid pyrophosphatase, are particle-bound enzymes, which exhibit a high degree of structural latency. They are activated and solubilized in a parallel fashion by mechanical treatments and tensio-active agents. Catalase is also particle-bound and latent; activating conditions stronger than those for hydrolases are required to activate the enzyme. Acid pyrophosphatase, 5'-nucleotidase and inosine diphosphatase are firmly particle-bound, but not latent; they are not easily solubilized. In differential and isopycnic centrifugation, the latent hydrolases, cytochrome oxidase and catalase dissociate largely from each other; this suggests the occurrence of lysosomes and peroxisome-like structures besides mitochondria. The distribution patterns of 5'-nucleotidase and cholesterol are largely similar; digitonin influences their equilibrium density to the same extent; these two constituents are thought to be related to the plasma membrane. Inosine diphosphatase and acid pyrophosphatase are also partially associated with the plasma membrane, although some part of these enzymic activities probably belongs to other structures. NADH cytochrome c reductase is associated partly with the endoplasmic reticulum, partly with mitochondria.


Cognitive impairments in patients with intractable temporal lobe epilepsy.
Cognitive impairment associated with temporal lobe epilepsy (TLE) has been recognized in multiple studies. We designed this study to find a specific cognitive profile in patients with TLE who were candidates for epilepsy surgery. We also sought to find if neuropsychological assessment could differentiate left TLE, right TLE and normal subjects. The sample of this study consisted of 29 patients with right TLE, 31 with left TLE, and 32 subjects without history of seizure as the control group. For all recruited patients and controls, demographic questionnaire, Wechsler Memory Scale-III (WMS-III) and Wechsler Adult Intelligence Scale-R (WAIS-R) were administered. Multivariate analysis of variance was carried out to reveal differences in memory and intelligence performance between the three groups. All of the mean scores of the WMS-III indexes were significantly higher in the control group in comparison with the right or the left TLE groups (p < 0.001). There were not any significant differences between mean scores of WMS-III indexes of the right and the left. The WAIS-R also showed significantly better mean scores of full scale intelligence quotient (FSIQ) and performance intelligence quotient (PIQ) in the control groups than both of the right and left TLE patients (p < 0.001). Although the verbal intelligence quotient (VIQ) mean scores were significantly different between the left TLE and the control group (p = 0.037), there were not any significant differences between the right TLE patients and the control group. These findings indicated that WMS-III and WAIS-R can differentiate patients with refractory temporal lobe epilepsy from normal subjects. However, the obtained cognitive profile could not differentiate between the right and the left TLE.


Peripheral Sympathectomy Alters Neuroinflammatory and Microglial Responses to Sleep Fragmentation in Female Mice.
Sleep loss, either induced by obstructive sleep apnea or other forms of sleep dysfunction, induces an inflammatory response, as commonly measured by increased circulating levels of pro-inflammatory cytokines. Increased catecholamines from sympathetic nervous system (SNS) activation regulates this peripheral inflammation. However, the role that catecholamines play in mediating neuroinflammation from sleep perturbations is undescribed. The aims of this study were to determine (i) the effect of peripheral SNS inhibition upon neuroinflammatory responses to sleep fragmentation (SF) and (ii) whether homeostasis can be restored after 1 week of recovery sleep. We measured gene expression levels of pro- and anti-inflammatory cytokines and microglial activity in brain (prefrontal cortex, hippocampus and hypothalamus) of female mice that were subjected to acute SF for 24 hours, chronic SF for 8 weeks, or 7 days of recovery after chronic SF. In each experiment, SF and control mice were peripherally sympathectomized with 6-hydroxydopamine (6-OHDA) or injected with vehicle. SF elevated cytokine mRNA expression in brain and increased microglial density and cell area in some regions. In addition, chronic SF promoted hyper-ramification in resting microglia upon exposure to chronic, but not acute, SF. Effects of chronic SF were more pronounced than acute SF, and 1 week of recovery was not sufficient to alleviate neuroinflammation. Importantly, 6-OHDA treatment significantly alleviated SF-induced inflammation and microglial responses. This study provides evidence of SNS regulation of neural inflammation from SF, suggesting a potential role for therapeutics that could mitigate neuroinflammatory responses to sleep dysfunction.


Effects of a Snoezelen room on the behavior of three autistic clients.
The effect of a Snoezelen room on the disruptive and prosocial behavior of three male, autistic inpatients was examined. In an ABAB reversal design, specific disruptive and prosocial behaviors were recorded for each client throughout the four 28-day periods of the study. Results indicated that the three clients had different responses to the room, but no client showed a decrease in disruptive behaviors while in the Snoezelen condition compared to baseline, and one client showed a clear pattern of increased disruptive behavior during the Snoezelen periods. There was a slight tendency for clients to engage in more prosocial behaviors while in Snoezelen. These findings do not support the contention that Snoezelen rooms are effective interventions for aggressive behavior in this client population.


The trade-off dilemma in pharmacotherapy of COVID-19: systematic review, meta-analysis, and implications.
The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected millions of people worldwide and has contributed to over 650,000 deaths. This review synthesizes the literature on COVID-19 pharmacotherapy to inform practice and policymaking. The authors systematically review the published literature on COVID-19 therapeutics, grouping candidate treatments into repurposed, adjunct, and experimental agents. They conducted meta-analysis where appropriate and provide recommendations based on compilation from real-time/interim therapeutic guidelines. They then advise on how to navigate and advance the evidence in the current context of uncertainty and urgency and provide expert opinion on suggested framework. Current evidence does not support a clear role for pharmacotherapy in COVID-19. While promising signals have been found through limited number of RCTs, these must be interpreted with caution. Without proper protection from bias and confounding we risk exposing patients to treatments where the potential for benefit is at best unclear, yet the potential for harm from adverse effects is high leading to a trade-off dilemma in decision making. Advancing the evidence requires a coordinated effort to design and conduct robust trials and to systematically synthesize and critically evaluate findings. Therapies should be reserved for use in clinical trials, emergency or compassionate access until we gain more confidence in the balance of benefit and harm.


Pomegranate polyphenols and extract inhibit nuclear factor of activated T-cell activity and microglial activation in vitro and in a transgenic mouse model of Alzheimer disease.
Alzheimer disease (AD) brain is characterized by extracellular plaques of amyloid β (Aβ) peptide with reactive microglia. This study aimed to determine whether a dietary intervention could attenuate microgliosis. Memory was assessed in 12-mo-old male amyloid precursor protein/presenilin 1 (APP/PS1) transgenic mice via Barnes maze testing followed by division into either a control-fed group provided free access to normal chow and water or a treatment group provided free access to normal chow and drinking water supplemented with pomegranate extract (6.25 mL/L) for 3 mo followed by repeat Barnes maze testing for both groups. Three months of pomegranate feeding decreased the path length to escape of mice compared with their initial 12-mo values (P < 0.05) and their control-fed counterparts (P < 0.05). Brains of the 3-mo study pomegranate-fed mice had lower tumor necrosis factor α (TNF-α) concentrations (P < 0.05) and lower nuclear factor of activated T-cell (NFAT) transcriptional activity (P < 0.05) compared with controls. Brains of the 3-mo pomegranate or control mice were also compared with an additional control group of 12-mo-old mice for histologic analysis. Immunocytochemistry showed that pomegranate- but not control-fed mice had attenuated microgliosis (P < 0.05) and Aβ plaque deposition (P < 0.05) compared with 12-mo-old mice. An additional behavioral study again used 12-mo-old male APP/PS1 mice tested by T-maze followed by division into a control group provided with free access to normal chow and sugar supplemented drinking water or a treatment group provided with normal chow and pomegranate extract-supplemented drinking water (6.25 mL/L) for 1 mo followed by repeat T-maze testing in both groups. One month of pomegranate feeding increased spontaneous alternations versus control-fed mice (P < 0.05). Cell culture experiments verified that 2 polyphenol components of pomegranate extract, punicalagin and ellagic acid, attenuated NFAT activity in a reporter cell line (P < 0.05) and decreased Aβ-stimulated TNF-α secretion by murine microglia (P < 0.05). These data indicate that dietary pomegranate produces brain antiinflammatory effects that may attenuate AD progression.


Esophagitis associated with the use of alendronate.
Alendronate, an aminobisphosphonate and a selective inhibitor of osteoclast-mediated bone resorption, is used to treat osteoporosis in postmenopausal women and Paget's disease of bone. Aminobiphosphonates can irritate the upper gastrointestinal mucosa. We describe three patients who had severe esophagitis shortly after starting to take alendronate and also analyze adverse esophageal effects reported to Merck, the manufacturer, through postmarketing surveillance. As of March 5, 1996, alendronate had been prescribed for an estimated 475,000 patients worldwide, and 1213 reports of adverse effects had been received. A total of 199 patients had adverse effects related to the esophagus; in 51 of these patients (26 percent), including the 3 we describe in case reports, adverse effects were categorized as serious or severe. Thirty-two patients (16 percent) were hospitalized, and two were temporarily disabled. Endoscopic findings generally indicated chemical esophagitis, with erosions or ulcerations and exudative inflammation accompanied by thickening of the esophageal wall. Bleeding was rare, and stomach or duodenal involvement unusual. In patients for whom adequate information was available, esophagitis seemed to be associated with swallowing alendronate with little or no water, lying down during or after ingestion of the tablet, lying down during or after ingestion of the tablet, continuing to take alendronate after the onset of symptoms, and having preexisting esophageal disorders. Alendronate can cause chemical esophagitis, including severe ulcerations, in some patients. Recommendations to reduce the risk of esophagitis include swallowing alendronate with 180 to 240 ml (6 to 8 oz) of water on arising in the morning, remaining upright for at least 30 minutes after swallowing the tablet and until the first food of the day has been ingested, and discontinuing the drug promptly if esophageal symptoms develop.


Stray field nuclear magnetic resonance of soil water: development of a new, large probe and preliminary results.
Development, characterization, and preliminary results of a recent technique capable of local measurements of pore-size distribution by a spatially resolved low resolution nuclear magnetic resonance (NMR) technique are described. Potential environmental uses include studying the change in pore-size distribution caused by surface compaction, which influences surface runoff, and obtaining information on the physical state of non-aqueous compounds in porous materials, which should aid the selection of appropriate soil remediation methods. Stray field (STRAFI) imaging is an NMR technique that allows distortion-free imaging of materials with short NMR relaxation times. The sample is placed in the strong axial fringe field gradient of a superconducting NMR magnet. We report on a new, unique, large 5-cm-diameter STRAFI probe, and its use for three preliminary test cases: water in ceramics of known pore size, paraffin wax and oil in sandstone rock, and water in soil at different matric potentials. The imaging is confined to one dimension with a spatial resolution of the order of 100 microm for protons. The optimum position for imaging occurs at 2.62 T and a gradient of 12.1 T/m. Water relaxation decay curves can be measured at any position in the 8-cm-long sample. These curves are decomposed into a series of terms each corresponding to a different pore size. Preliminary results show continuum fits to decay curves for a soil drained to three different matric potentials. Such information will be useful for interpreting water retention curves and will lead to understanding of the behavior of fluids in the vadose zone.


Evaluation of the Role of Human DNAJAs in the Response to Cytotoxic Chemotherapeutic Agents in a Yeast Model System.
Heat-shock proteins (HSPs) play a crucial role in maintaining protein stability for cell survival during stress-induced insults. Overexpression of HSPs in cancer cells results in antiapoptotic activity contributing to cancer cell survival and restricting the efficacy of cytotoxic chemotherapy, which continues to play an important role in the treatment of many cancers, including triple-negative breast cancer (TNBC). First-line therapy for TNBC includes anthracycline antibiotics, which are associated with serious dose-dependent side effects and the development of resistance. We previously identified YDJ1, which encodes a heat-shock protein 40 (HSP40), as an important factor in the cellular response to anthracyclines in yeast, with mutants displaying over 100-fold increased sensitivity to doxorubicin. In humans, the DNAJA HSP40s are homologues of YDJ1. To determine the role of DNAJAs in the cellular response to cytotoxic drugs, we investigated their ability to rescue ydj1Δ mutants from exposure to chemotherapeutic agents. Our results indicate that DNAJA1 and DNAJA2 provide effective protection, while DNAJA3 and DNAJA4 did not. The level of complementation was also dependent on the agent used, with DNAJA1 and DNAJA2 rescuing the ydj1Δ strain from doxorubicin, cisplatin, and heat shock. DNAJA3 and DNAJA4 did not rescue the ydj1Δ strain and interfered with the cellular response to stress when expressed in wild type background. DNAJA1 and DNAJA2 protect the cell from proteotoxic damage caused by reactive oxygen species (ROS) and are not required for repair of DNA double-strand breaks. These data indicate that the DNAJAs play a role in the protection of cells from ROS-induced cytotoxic stress.


Interactive Effects of CO2 , Temperature, Irradiance, and Nutrient Limitation on the Growth and Physiology of the Marine Diatom Thalassiosira pseudonana (Coscinodiscophyceae).
The marine diatom Thalassiosira pseudonana was grown in continuous culture systems to study the interactive effects of temperature, irradiance, nutrient limitation, and the partial pressure of CO2 (pCO2 ) on its growth and physiological characteristics. The cells were able to grow at all combinations of low and high irradiance (50 and 300 μmol photons · m-2 · s-1 , respectively, of visible light), low and high pCO2 (400 and 1,000 μatm, respectively), nutrient limitation (nitrate-limited and nutrient-replete conditions), and temperatures of 10-32°C. Under nutrient-replete conditions, there was no adverse effect of high pCO2 on growth rates at temperatures of 10-25°C. The response of the cells to high pCO2 was similar at low and high irradiance. At supraoptimal temperatures of 30°C or higher, high pCO2 depressed growth rates at both low and high irradiance. Under nitrate-limited conditions, cells were grown at 38 ± 2.4% of their nutrient-saturated rates at the same temperature, irradiance, and pCO2 . Dark respiration rates consistently removed a higher percentage of production under nitrate-limited versus nutrient-replete conditions. The percentages of production lost to dark respiration were positively correlated with temperature under nitrate-limited conditions, but there was no analogous correlation under nutrient-replete conditions. The results suggest that warmer temperatures and associated more intense thermal stratification of ocean surface waters could lower net photosynthetic rates if the stratification leads to a reduction in the relative growth rates of marine phytoplankton, and at truly supraoptimal temperatures there would likely be a synergistic interaction between the stresses from temperature and high pCO2 (lower pH).


Impact of HLA-driven HIV adaptation on virulence in populations of high HIV seroprevalence.
It is widely believed that epidemics in new hosts diminish in virulence over time, with natural selection favoring pathogens that cause minimal disease. However, a tradeoff frequently exists between high virulence shortening host survival on the one hand but allowing faster transmission on the other. This is the case in HIV infection, where high viral loads increase transmission risk per coital act but reduce host longevity. We here investigate the impact on HIV virulence of HIV adaptation to HLA molecules that protect against disease progression, such as HLA-B*57 and HLA-B*58:01. We analyzed cohorts in Botswana and South Africa, two countries severely affected by the HIV epidemic. In Botswana, where the epidemic started earlier and adult seroprevalence has been higher, HIV adaptation to HLA including HLA-B*57/58:01 is greater compared with South Africa (P = 7 × 10(-82)), the protective effect of HLA-B*57/58:01 is absent (P = 0.0002), and population viral replicative capacity is lower (P = 0.03). These data suggest that viral evolution is occurring relatively rapidly, and that adaptation of HIV to the most protective HLA alleles may contribute to a lowering of viral replication capacity at the population level, and a consequent reduction in HIV virulence over time. The potential role in this process played by increasing antiretroviral therapy (ART) access is also explored. Models developed here suggest distinct benefits of ART, in addition to reducing HIV disease and transmission, in driving declines in HIV virulence over the course of the epidemic, thereby accelerating the effects of HLA-mediated viral adaptation.


An Avian Connection as a Catalyst to the 1918-1919 Influenza Pandemic.
The 1918 Influenza pandemic was one of the most virulent strains of influenza in history. This strain quickly dispatched previously held theories on influenza. World War One introduced new environmental stresses and speed of dissemination logistics never experienced by humans. In light of new phylogenic evidence the cause of this influenza outbreak is now being considered to have linkage to the avian influenza. Animals act as reservoirs for this influenza virus and research indicates the influenza virus often originates in the intestines of aquatic wildfowl. The virus is shed into the environment, which in turns infects domestic poultry, which in turn infects mammalian hosts. These animals, usually pigs, act as a transformer or converters; creating a strain that can more readily infect humans. Therefore swine can be infected with both avian and human influenza A viruses and serve as a source for infection for a number of species as the incidents of direct infection from birds to humans have been rare. Increased human habitation near poultry and swine raising facilities pose greater influenza outbreak risk. It was this combination of environmental factors that may have contributed to the greatest pandemic of recent times, and, moreover, similar conditions exist throughout Southeast Asia today.


Isolation and characterization of α-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors.
A new α-conotoxin LsIA was isolated from the crude venom of Conus limpusi using assay-guided RP-HPLC fractionation. Synthetic LsIA was a potent antagonist of α3β2, α3α5β2 and α7 nAChRs, with half-maximal inhibitory concentrations of 10, 31 and 10 nM, respectively. The structure of LsIA determined by NMR spectroscopy comprised a characteristic disulfide bond-stabilized α-helical structure and disordered N-terminal region. Potency reductions of up to 9-fold were observed for N-terminally truncated analogues of LsIA at α7 and α3β2 nAChRs, whereas C-terminal carboxylation enhanced potency 3-fold at α3β2 nAChRs but reduced potency 3-fold at α7 nAChRs. This study gives further insight into α-conotoxin pharmacology and the molecular basis of nAChR selectivity, highlighting the influence of N-terminal residues and C-terminal amidation on conotoxin pharmacology.


Tumefactive multiple sclerosis lesions in two patients after cessation of fingolimod treatment.
Fingolimod (FTY) is the first oral medication approved for multiple sclerosis therapy. Until now, little has been known about the effects of FTY withdrawal regarding disease activity and development of tumefactive demyelinating lesions (TDLs), as already described in patients who discontinue natalizumab. In this study we present the clinical and radiological findings of two patients who had a severe rebound after FTY withdrawal and compare these with patients identified by a PubMed data bank analysis using the search term 'fingolimod rebound'. In total, 10 patients, of whom three developed TDLs, are presented. Patients suffering from TDLs were free of clinical and radiological signs of disease activity under FTY therapy (100% versus 57%, compared with patients without TDLs) and had rebounds after a mean of 14.6 weeks (standard deviation 11.5) [patients without TDLs 11.7 (standard deviation 3.4)]. We propose that a good therapeutic response to FTY might be predisposing for a severe rebound after withdrawal. Consequently, therapy switches should be planned carefully with a short therapy free interval.


Three-Dimensional Imaging of the Developing Vasculature within Stem Cell-Seeded Scaffolds Cultured in ovo.
Successful tissue engineering requires functional vascularization of the three-dimensional constructs with the aim to serve as implants for tissue replacement and regeneration. The survival of the implant is only possible if the supply of oxygen and nutrients by developing capillaries from the host is established. The chorioallantoic membrane (CAM) assay is a valuable tool to study the ingrowth and distribution of vessels into scaffolds composed by appropriate biomaterials and stem cell populations that are used in cell-based regenerative approaches. The developing vasculature of chicken embryos within cell-seeded scaffolds can be visualized with microcomputed tomography after intravenous injection of MicroFil®, which is a radiopaque contrast agent. Here, we provide a step-by-step protocol for the seeding of stem cells into silk fibroin scaffolds, the CAM culture conditions, the procedure of MicroFil® perfusion, and finally the microcomputed tomography scanning. Three-dimensional imaging of the vascularized tissue engineered constructs provides an important analytical tool for studying the potential of cell seeded scaffolds to attract vessels and form vascular networks, as well as for analyzing the number, density, length, branching, and diameter of vessels. This in ovo method can greatly help to screen implants that will be used for tissue regeneration purposes before their in vivo testing, thereby reducing the amount of animals needed for pre-clinical studies.


Acute otitis externa: an update.
Acute otitis externa is a common condition involving inflammation of the ear canal. The acute form is caused primarily by bacterial infection, with Pseudomonas aeruginosa and Staphylococcus aureus the most common pathogens. Acute otitis externa presents with the rapid onset of ear canal inflammation, resulting in otalgia, itching, canal edema, canal erythema, and otorrhea, and often occurs following swimming or minor trauma from inappropriate cleaning. Tenderness with movement of the tragus or pinna is a classic finding. Topical antimicrobials or antibiotics such as acetic acid, aminoglycosides, polymyxin B, and quinolones are the treatment of choice in uncomplicated cases. These agents come in preparations with or without topical corticosteroids; the addition of corticosteroids may help resolve symptoms more quickly. However, there is no good evidence that any one antimicrobial or antibiotic preparation is clinically superior to another. The choice of treatment is based on a number of factors, including tympanic membrane status, adverse effect profiles, adherence issues, and cost. Neomycin/polymyxin B/hydrocortisone preparations are a reasonable first-line therapy when the tympanic membrane is intact. Oral antibiotics are reserved for cases in which the infection has spread beyond the ear canal or in patients at risk of a rapidly progressing infection. Chronic otitis externa is often caused by allergies or underlying inflammatory dermatologic conditions, and is treated by addressing the underlying causes.


Persistence of transgene expression influences CD8+ T-cell expansion and maintenance following immunization with recombinant adenovirus.
Previous studies determined that the CD8(+) T-cell response elicited by recombinant adenovirus exhibited a protracted contraction phase that was associated with long-term presentation of antigen. To gain further insight into this process, a doxycycline-regulated adenovirus was constructed to enable controlled extinction of transgene expression in vivo. We investigated the impact of premature termination of transgene expression at various time points (day 3 to day 60) following immunization. When transgene expression was terminated before the maximum response had been attained, overall expansion was attenuated, yielding a small memory population. When transgene expression was terminated between day 13 and day 30, the memory population was not sustained, demonstrating that the early memory population was antigen dependent. Extinction of transgene expression at day 60 had no obvious impact on memory maintenance, indicating that maintenance of the memory population may ultimately become independent of transgene expression. Premature termination of antigen expression had significant but modest effects on the phenotype and cytokine profile of the memory population. These results offer new insights into the mechanisms of memory CD8(+) T-cell maintenance following immunization with a recombinant adenovirus.


Lipid balance must be just right to prevent development of severe liver damage.
Nonalcoholic fatty liver disease (NAFLD) is a major health concern that often associates with obesity and diabetes. Fatty liver is usually a benign condition, yet a fraction of individuals progress to severe forms of liver damage, including nonalcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). Elevated sterol regulatory element-binding protein-driven (SREBP-driven) hepatocyte lipid synthesis is associated with NAFLD in humans and mice. In this issue of the JCI, Kawamura, Matsushita, et al. evaluated the role of SREBP-dependent lipid synthesis in the development of NAFLD, NASH, and HCC in the phosphatase and tensin homolog-knockout (PTEN-knockout) NASH model. Deletion of the gene encoding SREBP cleavage-activating protein (SCAP) from the liver resulted in decreased hepatic lipids, as expected. However, SCAP deletion accelerated progression to more severe liver damage, including NASH and HCC. This study provides a note of caution for those pursuing de novo fat biosynthesis as a therapeutic intervention in human NASH.


The eyes of the deep diving hooded seal (Cystophora cristata) enhance sensitivity to ultraviolet light.
The mammalian visual range is approximately 400-700 nm, although recent evidence suggests varying ultraviolet (UV) extensions in diverse terrestrial species. UV sensitivity may have advantages in the dim, blue light shifted environment experienced by submerged marine mammals. It may also be advantageous when seals are on land as UV is reflected by snow and ice but absorbed by fur, enhancing visual contrast. Here we show that the pelagic hooded seal (Cystophora cristata) has a highly UV permissive cornea and lens. Seals like other carnivores have a tapetum lucidum (TL) reflecting light back through the retina increasing sensitivity. The TL in this seal is unusual being white and covering almost the entire retina unlike that in other carnivores. Spectral reflectance from its surface selectively increases the relative UV/blue components >10 times than other wavelengths. Retinal architecture is consistent with a high degree of convergence. Enhanced UV from a large TL surface with a high degree of retinal convergence will increase sensitivity at a cost to acuity. UV electrophysiological retina responses were only obtained to dim, rod mediated stimuli, with no evidence of cone input. As physiological measurements of threshold sensitivity are much higher than those for psychophysical detection, these seals are likely to be more UV sensitive than our results imply. Hence, UV reflections from the TL will afford increased sensitivity in dim oceanic environments.


Microplastics and mesoplastics in fish from coastal and fresh waters of China.
Plastic pollution is a growing global concern. In the present study, we investigated plastic pollution in 21 species of sea fish and 6 species of freshwater fish from China. All of the species were found to ingest micro- or mesoplastics. The average abundance of microplastics varied from 1.1 to 7.2 items by individual and 0.2-17.2 items by gram. The average abundance of mesoplastics varied from 0.2 to 3.0 items by individual and 0.1-3.9 items by gram. Microplastics were abundant in 26 species, accounting for 55.9-92.3% of the total number of plastics items in each species. Thamnaconus septentrionalis contained the highest abundance of microplastics (7.2 items/individual). The average abundance of plastics in sea benthopelagic fishes was significantly higher than in freshwater benthopelagic fishes by items/individual. The plastics were dominanted by fiber in shape, transparent in color and cellophane in composition. The proportion of plastics in the stomach to the intestines showed great variation in different species, ranging from 0.5 to 1.9 by items/individual. The stomach of Harpodon nehereus and intestines of Pampus cinereus contained the highest number of plastics, (3.3) and (2.7), respectively, by items/individual. Our results suggested that plastic pollution was widespread in the investigated fish species and showed higher abundance in comparison with worldwide studies. The ingestion of plastics in fish was closely related to the habitat and gastrointestinal tract structure. We highly recommend that the entire gastrointestinal tract and digestion process be used in future investigations of plastic pollution in fish.


Experiences of families who help young rural men with emergent mental health problems in a rural community in New South Wales, Australia.
A qualitative study was conducted in rural New South Wales, Australia, to understand the barriers to help-seeking among young rural men with emergent mental health problems. Participants who had real life experiences of these problems within their families were interviewed. Themes emerged from the data which explained some barriers to early intervention. Despite these barriers, families had developed skills in helping and in providing early mental health help to their sons. The findings of this study showed that a substantial burden on the emotional and social integrity of the family, combined with diminished psychological well-being, caused some parents to question how long they could cope before they reached 'the end of their strings'. This downward spiralling trajectory of mental health and well-being for both the young men and their families has implications for clinical practice. Current models of mental health service delivery do not adequately capture the early help-seeking dynamics of young rural men and their families. A more flexible approach is needed to identify and help the family and the young men, without the pre-requisite for a formal medical diagnosis. Future research should involve health and well-being solution focused service delivery.


Sonic hedgehog is essential for first pharyngeal arch development.
The secreted protein sonic hedgehog (Shh) is essential for normal development of many organs. Targeted disruption of Shh in mouse leads to near complete absence of craniofacial skeletal elements at birth, and mutation of SHH in human causes holoprosencephaly (HPE), frequently associated with defects of derivatives of pharyngeal arches. To investigate the role of Shh signaling in early pharyngeal arch development, we analyzed Shh mutant embryos using molecular markers and found that the first pharyngeal arch (PA1) was specifically hypoplastic and fused in the midline, and remaining arches were well formed at embryonic day (E) 9.5. Molecular analyses using specific markers suggested that the growth of the maxillary arch and proximal mandibular arch was severely defective in Shh-null PA1, whereas the distal mandibular arch was less affected. TUNEL assay revealed an increase in the number of apoptotic signals in PA1 of Shh mutant embryos. Ectodermal expression of fibroblast growth factor (Fgf)-8, a cell survival factor for pharyngeal arch mesenchyme, was down-regulated in the PA1 of Shh mutants. Consistent with this observation, downstream transcriptional targets of Fgf8 signaling in neural crest-derived mesenchyme, including Barx1, goosecoid, and Dlx2, were also down-regulated in Shh-null PA1. These results demonstrate that epithelial-mesenchymal signaling and transcriptional events coordinated by Shh, partly via Fgf8, is essential for cell survival and tissue outgrowth of the developing PA1.


Identification of 50 class D β-lactamases and 65 Acinetobacter-derived cephalosporinases in Acinetobacter spp.
Whole-genome sequencing of a collection of 103 Acinetobacter strains belonging to 22 validly named species and another 16 putative species allowed detection of genes for 50 new class D β-lactamases and 65 new Acinetobacter-derived cephalosporinases (ADC). All oxacillinases (OXA) contained the three typical motifs of class D β-lactamases, STFK, (F/Y)GN, and K(S/T)G. The phylogenetic tree drawn from the OXA sequences led to an increase in the number of OXA groups from 7 to 18. The topologies of the OXA and RpoB phylogenetic trees were similar, supporting the ancient acquisition of blaOXA genes by Acinetobacter species. The class D β-lactamase genes appeared to be intrinsic to several species, such as Acinetobacter baumannii, Acinetobacter pittii, Acinetobacter calcoaceticus, and Acinetobacter lwoffii. Neither blaOXA-40/143- nor blaOXA-58-like genes were detected, and their origin remains therefore unknown. The phylogenetic tree analysis based on the alignment of the sequences deduced from blaADC revealed five main clusters, one containing ADC belonging to species closely related to A. baumannii and the others composed of cephalosporinases from the remaining species. No indication of blaOXA or blaADC transfer was observed between distantly related species, except for blaOXA-279, possibly transferred from Acinetobacter genomic species 6 to Acinetobacter parvus. Analysis of β-lactam susceptibility of seven strains harboring new oxacillinases and cloning of the corresponding genes in Escherichia coli and in a susceptible A. baumannii strain indicated very weak hydrolysis of carbapenems. Overall, this study reveals a large pool of β-lactamases in different Acinetobacter spp., potentially transferable to pathogenic strains of the genus.


Soluble PD-L1 generated by endogenous retroelement exaptation is a receptor antagonist.
Immune regulation is a finely balanced process of positive and negative signals. PD-L1 and its receptor PD-1 are critical regulators of autoimmune, antiviral and antitumoural T cell responses. Although the function of its predominant membrane-bound form is well established, the source and biological activity of soluble PD-L1 (sPD-L1) remain incompletely understood. Here, we show that sPD-L1 in human healthy tissues and tumours is produced by exaptation of an intronic LINE-2A (L2A) endogenous retroelement in the CD274 gene, encoding PD-L1, which causes omission of the transmembrane domain and the regulatory sequence in the canonical 3' untranslated region. The alternatively spliced CD274-L2A transcript forms the major source of sPD-L1 and is highly conserved in hominids, but lost in mice and a few related species. Importantly, CD274-L2A-encoded sPD-L1 lacks measurable T cell inhibitory activity. Instead, it functions as a receptor antagonist, blocking the inhibitory activity of PD-L1 bound on cellular or exosomal membranes.


The clinical potential of ademetionine (S-adenosylmethionine) in neurological disorders.
This review focuses on the biochemical and clinical aspects of methylation in neuropsychiatric disorders and the clinical potential of their treatment with ademetionine (S-adenosylmethionine; SAMe). SAMe is required in numerous transmethylation reactions involving nucleic acids, proteins, phospholipids, amines and other neurotransmitters. The synthesis of SAMe is intimately linked with folate and vitamin B12 (cyanocobalamin) metabolism, and deficiencies of both these vitamins have been found to reduce CNS SAMe concentrations. Both folate and vitamin B12 deficiency may cause similar neurological and psychiatric disturbances including depression, dementia, myelopathy and peripheral neuropathy. SAMe has a variety of pharmacological effects in the CNS, especially on monoamine neurotransmitter metabolism and receptor systems. SAMe has antidepressant properties, and preliminary studies indicate that it may improve cognitive function in patients with dementia. Treatment with methyl donors (betaine, methionine and SAMe) is associated with remyelination in patients with inborn errors of folate and C-1 (one-carbon) metabolism. These studies support a current theory that impaired methylation may occur by different mechanisms in several neurological and psychiatric disorders.