Purified lymphocyte function-associated antigen-3 and T11 target structure are active in CD2-mediated T cell stimulation.
In this study we have used cells expressing LFA-3 or T11TS, the human and sheep forms of the ligand of CD2, as well as the purified LFA-3 and T11TS molecules themselves to study their effects on T cell activation via the CD2-mediated "alternative pathway". Sheep red blood cells, which bind to CD2 via T11TS in E-rosette formation, and human autologous monocytes, which express the LFA-3 molecule, both induce proliferation of resting T cells in the presence of per se submitogenic concentrations of anti-T11(2) plus anti-T11(3) monoclonal antibodies (mAb). This effect is blocked by mAb to LFA-3, T11TS and CD2 known to inhibit CD2-ligand interaction. In addition, purified LFA-3 and T11TS, when added at ng amounts to cultures containing submitogenic concentrations of anti-T11(2 + 3) mAb, are also strongly mitogenic for resting human T cells. Thus, both LFA-3 and T11TS are potent co-stimulators of the alternative pathway of T cell activation but by themselves do not provide a mitogenic signal. This finding is discussed with regard to a physiological role of CD2-LFA-3 interaction in T cell activation.


Urethral Catheterization Is Not Necessary During Nononcological Laparoscopic Pelvic Surgery.
Expert opinion remains divided regarding whether routine urethral catheterization is required before nononcological laparoscopic pelvic surgery. Catheterization is thought to reduce the incidence of bladder injury when inserting a suprapubic laparoscopic port and prevent obstruction of the view of the pelvis because of bladder filling. However, catheterization comes with a risk of nosocomial infection and harbors financial cost. Moreover, indwelling catheters inhibit early mobilization and increase postoperative discomfort. A systematic review was undertaken using the Meta-Analysis of Observational Studies guidelines to identify eligible publications. End points included bladder injury, positive postoperative urinary microbiology, and postoperative urinary symptoms. The reported incidence rates of laparoscopic bladder injury in included publications ranges from 0% to 1.3%. Importantly, bladder injury has occurred during both catheterized and noncatheterized operations. Our meta-analysis also shows that patients who are catheterized have a 2.33 times relative risk of developing postoperative positive microbiology in their urine (P = 0.01) and a 2.41 times relative risk of postoperative urinary symptoms (P = 0.005), when compared with noncatheterized patients. This meta-analysis indicates that omitting a catheter in emergency and elective nononcological laparoscopic pelvic surgery may be a safe option. Catheterization does not remove the risk of bladder injury but results in more urinary tract infections and symptoms. It may be reasonable to ask a patient to void immediately before anesthesia, after which an on-table bladder scan should be performed. If there is minimal residual volume, a urinary catheter may not be necessary, unless operative time is estimated to be greater than 90 minutes.


Early Autism Screening: A Comprehensive Review.
Autistic spectrum disorder (ASD) refers to a neurodevelopmental condition associated with verbal and nonverbal communication, social interactions, and behavioural complications that is becoming increasingly common in many parts of the globe. Identifying individuals on the spectrum has remained a lengthy process for the past few decades due to the fact that some individuals diagnosed with ASD exhibit exceptional skills in areas such as mathematics, arts, and music among others. To improve the accuracy and reliability of autism diagnoses, many scholars have developed pre-diagnosis screening methods to help identify autistic behaviours at an early stage, speed up the clinical diagnosis referral process, and improve the understanding of ASD for the different stakeholders involved, such as parents, caregivers, teachers, and family members. However, the functionality and reliability of those screening tools vary according to different research studies and some have remained questionable. This study evaluates and critically analyses 37 different ASD screening tools in order to identify possible areas that need to be addressed through further development and innovation. More importantly, different criteria associated with existing screening tools, such as accessibility, the fulfilment of Diagnostic and Statistical Manual of Mental Disorders (DSM-5) specifications, comprehensibility among the target audience, performance (specifically sensitivity, specificity, and accuracy), web and mobile availability, and popularity have been investigated.


Role of the calcium toolkit in cancer stem cells.
Cancer stem cells are a subpopulation of tumor cells that proliferate, self-renew and produce more differentiated tumoral cells building-up the tumor. Responsible for the sustained growth of malignant tumors, cancer stem cells are proposed to play significant roles in cancer resistance to standard treatment and in tumor recurrence. Among the mechanisms dysregulated in neoplasms, those related to Ca2+ play significant roles in various aspects of cancers. Ca2+ is a ubiquitous second messenger whose fluctuations of its intracellular concentrations are tightly controlled by channels, pumps, exchangers and Ca2+ binding proteins. These components support the genesis of Ca2+ signals with specific spatio-temporal characteristics that define the cell response. Being involved in the coupling of extracellular events with intracellular responses, the Ca2+ toolkit is often hijacked by cancer cells to promote notably their proliferation and invasion. Growing evidence obtained during the last decade pointed to a role of Ca2+ handling and mishandling in cancer stem cells. In this review, after a general overview of the concept of cancer stem cells we analyse and discuss the studies and current knowledge regarding the complex roles of Ca2+ toolkit and signaling in these cells. We highlight that numbers of Ca2+ signaling actors promote cancer stem cell state and are associated with cell resistance to current cancer treatments and thus may represent promising targets for potential clinical applications.


Promoting Mental Health and Psychological Thriving in University Students: A Randomized Controlled Trial of Three Well-Being Interventions.
This study aimed to address the decline in mental health on U.S. university campuses by examining the effects of three interventions. University students suffer from high levels of anxiety, depression, and suicide. Counseling centers on university campuses are struggling to meet increased demand. The cost to students and universities could be buffered by offering preventative, psychoeducational, and skill-building training programs that promote mental health and psychological thriving. To date, the research literature has not yielded systematically evaluated and recommendable preventative mental health and well-being programs for university students. In a registered, randomized controlled trial, 131 university students were either placed in a non-intervention control group (N = 47) or received training in one of three 30-hour, eight-week semester-long well-being programs: SKY Campus Happiness ("SKY"; N = 29), Foundations of Emotional Intelligence ("EI"; N = 21) or Mindfulness-Based Stress Reduction ("MBSR"; N = 34). Compared to the control group and controlling for variance of baseline measurements and multiple comparisons, SKY Campus Happiness showed the greatest impact, benefiting six outcomes: depression, stress, mental health, mindfulness, positive affect and social connectedness. EI benefited one outcome: mindfulness. The MBSR group showed no change. Delivering SKY or EI to university students may be a cost-effective and efficient way to proactively and preventatively address mental health for university students and reduce the financial strain on universities.


A rapid and automatic image registration algorithm with subpixel accuracy.
A number of digital imaging techniques in medicine require the combination of multiple images. Using these techniques, it is essential that the images be adequately aligned and registered prior to addition, subtraction, or any other combination of the images. This paper describes an alignment routine developed to register an image of a fixed object containing a global offset error, rotation error, and magnification error relative to a second image. The described routine uses sparsely sampled regional correlation in a novel way to reduce computation time and avoid the use of markers and human interaction. The result is a fast, robust, and automatic alignment algorithm, with accuracy better than about 0.2 pixel in a test with clinical computed radiography images.


Improvements in access to malaria treatment in Tanzania after switch to artemisinin combination therapy and the introduction of accredited drug dispensing outlets - a provider perspective.
To improve access to treatment in the private retail sector a new class of outlets known as accredited drug dispensing outlets (ADDO) was created in Tanzania. Tanzania changed its first-line treatment for malaria from sulphadoxine-pyrimethamine (SP) to artemether-lumefantrine (ALu) in 2007. Subsidized ALu was made available in both health facilities and ADDOs. The effect of these interventions on access to malaria treatment was studied in rural Tanzania. The study was carried out in the villages of Kilombero and Ulanga Demographic Surveillance System (DSS) and in Ifakara town. Data collection consisted of: 1) yearly censuses of shops selling drugs; 2) collection of monthly data on availability of anti-malarials in public health facilities; and 3) retail audits to measure anti-malarial sales volumes in all public, mission and private outlets. The data were complemented with DSS population data. Between 2004 and 2008 access to malaria treatment greatly improved and the number of anti-malarial treatment doses dispensed increased by 78%. Particular improvements were observed in the availability (from 0.24 shops per 1,000 people in 2004 to 0.39 in 2008) and accessibility (from 71% of households within 5 km of a shop in 2004 to 87% in 2008) of drug shops. Despite no improvements in affordability this resulted in an increase of the market share from 49% of anti-malarial sales 2005 to 59% in 2008. The change of treatment policy from SP to ALu led to severe stock-outs of SP in health facilities in the months leading up to the introduction of ALu (only 40% months in stock), but these were compensated by the wide availability of SP in shops. After the introduction of ALu stock levels of the drug were relatively high in public health facilities (over 80% months in stock), but the drug could only be found in 30% of drug shops and in no general shops. This resulted in a low overall utilization of the drug (19% of all anti-malarial sales) The public health and private retail sector are important complementary sources of treatment in rural Tanzania. Ensuring the availability of ALu in the private retail sector is important for its successful uptake.


Recessive mutations in MSTO1 cause mitochondrial dynamics impairment, leading to myopathy and ataxia.
We report here the first families carrying recessive variants in the MSTO1 gene: compound heterozygous mutations were identified in two sisters and in an unrelated singleton case, who presented a multisystem complex phenotype mainly characterized by myopathy and cerebellar ataxia. Human MSTO1 is a poorly studied protein, suggested to have mitochondrial localization and to regulate morphology and distribution of mitochondria. As for other mutations affecting genes involved in mitochondrial dynamics, no biochemical defects typical of mitochondrial disorders were reported. Studies in patients' fibroblasts revealed that MSTO1 protein levels were strongly reduced, the mitochondrial network was fragmented, and the fusion events among mitochondria were decreased, confirming the deleterious effect of the identified variants and the role of MSTO1 in modulating mitochondrial dynamics. We also found that MSTO1 is mainly a cytosolic protein. These findings indicate recessive mutations in MSTO1 as a new cause for inherited neuromuscular disorders with multisystem features.


Circadian Disruptions in the Myshkin Mouse Model of Mania Are Independent of Deficits in Suprachiasmatic Molecular Clock Function.
Alterations in environmental light and intrinsic circadian function have strong associations with mood disorders. The neural origins underpinning these changes remain unclear, although genetic deficits in the molecular clock regularly render mice with altered mood-associated phenotypes. A detailed circadian and light-associated behavioral characterization of the Na+/K+-ATPase α3 Myshkin (Myk/+) mouse model of mania was performed. Na+/K+-ATPase α3 does not reside within the core circadian molecular clockwork, but Myk/+ mice exhibit concomitant disruption in circadian rhythms and mood. The neural basis of this phenotype was investigated through molecular and electrophysiological dissection of the master circadian pacemaker, the suprachiasmatic nuclei (SCN). Light input and glutamatergic signaling to the SCN were concomitantly assessed through behavioral assays and calcium imaging. In vivo assays revealed several circadian abnormalities including lengthened period and instability of behavioral rhythms, and elevated metabolic rate. Grossly aberrant responses to light included accentuated resetting, accelerated re-entrainment, and an absence of locomotor suppression. Bioluminescent recording of circadian clock protein (PERIOD2) output from ex vivo SCN revealed no deficits in Myk/+ molecular clock function. Optic nerve crush rescued the circadian period of Myk/+ behavior, highlighting that afferent inputs are critical upstream mediators. Electrophysiological and calcium imaging SCN recordings demonstrated changes in the response to glutamatergic stimulation as well as the electrical output indicative of altered retinal input processing. The Myshkin model demonstrates profound circadian and light-responsive behavioral alterations independent of molecular clock disruption. Afferent light signaling drives behavioral changes and raises new mechanistic implications for circadian disruption in affective disorders.


Transcriptomic responses to seawater acidification among sea urchin populations inhabiting a natural pH mosaic.
Increasing awareness of spatial and temporal variation in ocean pH suggests some marine populations may be adapted to local pH regimes and will therefore respond differently to present-day pH variation and to long-term ocean acidification. In the Northeast Pacific Ocean, differences in the strength of coastal upwelling cause latitudinal variation in prevailing pH regimes that are hypothesized to promote local adaptation and unequal pH tolerance among resident populations. In this study, responses to experimental seawater acidification were compared among embryos and larvae from six populations of purple sea urchins (Strongylocentrotus purpuratus) inhabiting areas that differ in their frequency of low pH exposure and that prior research suggests are locally adapted to seawater pH. Transcriptomic analyses demonstrate urchin populations most frequently exposed to low pH seawater responded to experimental acidification by expressing genes within major ATP-producing pathways at greater levels than populations encountering low pH less often. Multiple genes within the tricarboxylic acid cycle, electron transport chain and fatty acid beta oxidation pathways were upregulated in urchin populations experiencing low pH conditions most frequently. These same metabolic pathways were significantly over-represented among genes both expressed in a population-specific manner and putatively under selection to enhance low pH tolerance. Collectively, these data suggest natural selection is acting on metabolic gene networks to redirect ATP toward maintaining acid-base homeostasis and enhance tolerance of seawater acidification. As a trade-off, marine populations more tolerant of low pH may have less energy to put towards other aspects of fitness and to respond to additional ocean change.


Docosahexaenoic acid-rich fish oil improves heart rate variability and heart rate responses to exercise in overweight adults.
Dietary fish oil supplementation and regular physical activity can improve outcomes in patients with established CVD. Exercise has been shown to improve heart rate variability (HRV), a predictor of cardiac death, but whether fish oil benefits HRV is controversial. Obese adults at risk of future coronary disease have impaired HRV and may benefit from these interventions. We evaluated the effect of DHA-rich tuna fish oil supplementation with and without regular exercise on HRV in sedentary, overweight adults with risk factors for coronary disease. In a randomised, double-blind, parallel comparison, sixty-five volunteers consumed 6 g fish oil/d (DHA 1.56 g/d, EPA 0.36 g/d) or sunflower-seed oil (placebo) for 12 weeks. Half of each oil group also undertook regular moderate physical activity (3 d/week for 45 min, at 75 % of age-predicted maximal heart rate (HR)). Resting HR and the HR response to submaximal exercise were measured at weeks 0, 6 and 12. In forty-six subjects, HRV was also assessed by power spectrum analysis of 20 min electrocardiogram recordings taken supine at baseline and 12 weeks. Fish oil supplementation improved HRV by increasing high-frequency power, representing parasympathetic activity, compared with placebo (P = 0.01; oil x time interaction). It also reduced HR at rest and during submaximal exercise (P = 0.008; oil x time interaction). There were no significant fish oil x exercise interactions. Dietary supplementation with DHA-rich fish oil reduced HR and modulated HRV in keeping with an improved parasympathetic-sympathetic balance in overweight adults with risk factors for future coronary disease.


Proton pump inhibitors and risk of fracture: a systematic review and meta-analysis of observational studies.
Proton pump inhibitors (PPIs) are widely used in several acid-related gastrointestinal disorders. In vivo studies have suggested that gastric suppression by PPIs could result in decreased intestinal calcium absorption. Subsequently, there have been concerns that the chronic use of a PPI is associated with an increased risk of bone fracture. However, the results of clinical studies are conflicting. We performed a systematic review and meta-analysis of controlled observational studies to evaluate the risks of PPI use on fracture outcome. All controlled observational studies that compared fracture outcome in patients with PPI therapy with a control group were included. We calculated pooled odds ratios (ORs) using a random-effects model. Of 1,668 identified studies, 10 (4 cohort and 6 case-control) with 223,210 fracture cases were included in our analysis. In PPI users, compared with non/past users, the OR for hip fracture (n=9) was 1.25 (95% confidence interval (CI)=1.14-1.37). The OR for vertebral fracture (n=4) was 1.50 (95% CI=1.32-1.72) and for wrist/forearm fracture (n=3) was 1.09 (95% CI=0.95-1.24). In subgroup analysis of hip fracture, this association was observed in both high-dose and low-dose PPI exposure. When stratified by duration of exposure, the short duration of PPI use was associated with increased risk of developing hip fracture (OR=1.24; 95% CI=1.19-1.28), whereas there was no significant increase in risk of hip fracture in long-term PPI users (OR=1.30; 95% CI=0.98-1.70). There was significant statistical and clinical heterogeneity among studies for the main analysis and most of the subgroup analyses. Our results should be interpreted with caution. We found a modest association between PPI use and increased risk of hip and vertebral fractures, but no evidence of duration effect in subgroup analysis. However, observational studies cannot clarify whether the observed epidemiologic association is a causal effect or a result of unmeasured/residual confounding. Thus, randomized controlled studies are required to confirm or refute these results.


Bisphosphonate therapy for spinal osteoporosis in Hajdu-Cheney syndrome - new data and literature review.
Hajdu-Cheney syndrome (HCS) (#OMIM 102500) is a rare, autosomal dominant condition that presents in early childhood. It is caused by mutations in the terminal exon of NOTCH2, which encodes the transmembrane NOTCH2 receptor. This pathway is involved in the coupled processes of bone formation and resorption. The skeletal features of HCS include acro-osteolysis of the digits and osteoporosis commonly affecting vertebrae and long bones. Fractures are a prominent feature and are associated with significant morbidity. There is no specific treatment, but with both acro-osteolysis and generalized osteoporosis, it is possible that anti-resorptive treatment might be of benefit. However, to date only a few case reports have evaluated the effectiveness of bisphosphonate treatment. We describe the clinical features, treatment regimens and response to bisphosphonate treatment in 7 newly described patients aged 6-39 with HCS, and pooled the data with that from 8 previously published cases (a total of 17 courses of treatment in 15 individuals). The mean lumbar spine bone mineral density (BMD) z-score before treatment was - 2.9 (SD 1.2). In 14 courses of treatment (82%), there was an increase in BMD with bisphosphonate treatment, but the impact (in terms of change in spinal BMD z-score) appeared to be less with advancing age (p = 0.01). There was no evidence that acro-osteolysis was prevented. Although individual response is variable and age-related, the data support a role for bisphosphonates in preventing or treating spinal osteoporosis in HCS, but bone loss from the lumbar spine may be rapid after cessation.


Mutation to overproduction of bacteriophage T4 gene products.
R9 was isolated as one of several mutations that enhanced the growth of a leaky amber (am) mutant of bacteriophage T4 gene 62 (product required for phage DNA synthesis) under conditions of partial suppression by ribosomal ambiguity. R9 also enhanced the growth of leaky am mutants of some, but not all, other T4 "early" gene functions. R9 mapped between mutations in genes 43 and 62. By using assays involving polyacrylamide slab gel electrophoresis in the presence of sodium dodecyl sulfate, we observed the following. (i) R9 resulted in an overproduction of many T4 "early" proteins in infected cells. The most pronounced effects of R9 were observed when phage DNA synthesis and/or the functions of maturation genes 55 and 33 were not expressed. (ii) In rifampintreated infected cells, the capacity to synthesize T4 "early" proteins decayed more slowly in the presence of the R9 mutation than in the presence of the wild-type counterpart of R9. R9 appeared to have no effect on the rates of RNA synthesis either during early or late times after infection. The results suggest that the R9 mutation leads to increased functional stability of T4 "early" messengers.


Reducing Pb poisoning in birds and Pb exposure in game meat consumers: the dual benefit of effective Pb shot regulation.
The use of lead (Pb) ammunition in the form of shot pellets has been identified as a Pb exposure risk in wildlife and their human consumers. We explore the hypothesis that Pb shot ban enforcement reduces the risk of avian Pb poisoning as well as Pb exposure in game meat consumers. We assessed compliance with a partial ban on Pb shot commencing in 2003 by examination of 937 waterbirds harvested by hunters between 2007 and 2012 in the Ebro delta (Spain). Prevalence of Pb shot ingestion was determined, as were Pb concentrations in liver and muscle tissue to evaluate the potential for Pb exposure in game meat consumers. Hunted birds with only embedded Pb shot (no steel) declined from 26.9% in 2007-08 to <2% over the following three hunting seasons after ban reinforcement. Pb shot ingestion in mallards decreased from a pre-ban value of 30.2% to 15.5% in the post-ban period. Liver Pb levels were predominantly defined by the presence of ingested shot, whereas muscle levels were defined by the presence of both ingested and embedded shot. Only 2.5% of mallard muscle tissue had Pb levels above European Union regulations for meat (0.1μg/g wet weight) in the 2008-09 season, when Pb shot ingestion prevalence was also at a minimum (5.1%). Effective restrictions in Pb ammunition use have a dual benefit since this reduces Pb exposure for game meat consumers due to embedded ammunition as well as reducing Pb poisoning in waterbirds.


Relationships between Meteorological Parameters and Particulate Matter in Mae Hong Son Province, Thailand.
Meteorological parameters play an important role in determining the prevalence of ambient particulate matter (PM) in the upper north of Thailand. Mae Hong Son is a province located in this region and which borders Myanmar. This study aimed to determine the relationships between meteorological parameters and ambient concentrations of particulate matter less than 10 µm in diameter (PM10) in Mae Hong Son. Parameters were measured at an air quality monitoring station, and consisted of PM10, carbon monoxide (CO), ozone (O₃), and meteorological factors, including temperature, rainfall, pressure, wind speed, wind direction, and relative humidity (RH). Nine years (2009⁻2017) of pollution and climate data obtained from the Thai Pollution Control Department (PCD) were used for analysis. The results of this study indicate that PM10 is influenced by meteorological parameters; high concentration occurred during the dry season and northeastern monsoon seasons. Maximum concentrations were always observed in March. The PM10 concentrations were significantly related to CO and O₃ concentrations and to RH, giving correlation coefficients of 0.73, 0.39, and -0.37, respectively (p-value < 0.001). Additionally, the hourly PM10 concentration fluctuated within each day. In general, it was found that the reporting of daily concentrations might be best suited to public announcements and presentations. Hourly concentrations are recommended for public declarations that might be useful for warning citizens and organizations about air pollution. Our findings could be used to improve the understanding of PM10 concentration patterns in Mae Hong Son and provide information to better air pollution measures and establish a warning system for the province.


Mitogenesis of vascular smooth muscle cell stimulated by platelet-derived growth factor-bb is inhibited by blocking of intracellular signaling by epigallocatechin-3-O-gallate.
Epigallocatechin gallate (EGCG) is known to exhibit antioxidant, antiproliferative, and antithrombogenic effects and reduce the risk of cardiovascular diseases. Key events in the development of cardiovascular disease are hypertrophy and hyperplasia according to vascular smooth muscle cell proliferation. In this study, we investigated whether EGCG can interfere with PDGF-bb stimulated proliferation, cell cycle distribution, and the gelatinolytic activity of MMP and signal transduction pathways on RAOSMC when it was treated in two different ways-cotreatment with PDGF-bb and pretreatment of EGCG before addition of PDGF-bb. Both cotreated and pretreated EGCG significantly inhibited PDGF-bb induced proliferation, cell cycle progression of the G0/G1 phase, and the gelatinolytic activity of MMP-2/9 on RAOSMC. Also, EGCG blocked PDGF receptor-β (PDGFR-β) phosphorylation on PDGF-bb stimulated RAOSMC under pretreatment with cells as well as cotreatment with PDGF-bb. The downstream signal transduction pathways of PDGFR-β, including p42/44 MAPK, p38 MAPK, and Akt phosphorylation, were also inhibited by EGCG in a pattern similar to PDGFR-β phosphorylation. These findings suggest that EGCG can inhibit PDGF-bb stimulated mitogenesis by indirectly and directly interrupting PDGF-bb signals and blocking the signaling pathway via PDGFR-β phosphorylation. Furthermore, EGCG may be used for treatment and prevention of cardiovascular disease through blocking of PDGF-bb signaling.


Pharmacovigilance in hospice/palliative care: net effect of gabapentin for neuropathic pain.
Hospice/palliative care patients may differ from better studied populations, and data from other populations cannot necessarily be extrapolated into hospice/palliative care clinical practice. Pharmacovigilance studies provide opportunities to understand the harms and benefits of medications in routine practice. Gabapentin, a γ-amino butyric acid analogue antiepileptic drug, is commonly prescribed for neuropathic pain in hospice/palliative care. Most of the evidence however relates to non-malignant, chronic pain syndromes (diabetic neuropathy, postherpetic neuralgia, central pain syndromes, fibromyalgia). The aim of this study was to quantify the immediate and short-term clinical benefits and harms of gabapentin in routine hospice/palliative care practice. Multisite, prospective, consecutive cohort. 127 patients, 114 of whom had cancer, who started gabapentin for neuropathic pain as part of routine clinical care. 42 centres from seven countries. Data were collected at three time points-at baseline, at day 7 (and at any time; immediate and short-term harms) and at day 21 (clinical benefits). At day 21, the average dose of gabapentin for those still using it (n=68) was 653 mg/24 h (range 0-1800 mg) and 54 (42%) reported benefits, of whom 7 (6%) experienced complete pain resolution. Harms were reported in 39/127 (30%) patients at day 7, the most frequent of which were cognitive disturbance, somnolence, nausea and dizziness. Ten patients had their medication ceased due to harms. The presence of significant comorbidities, higher dose and increasing age increased the likelihood of harm. Overall, 42% of people experienced benefit at a level that resulted in continued use at 21 days.


Effects of emotional disclosure on psychological and physiological outcomes in patients with rheumatoid arthritis: an exploratory home-based study.
The effects of an exploratory, home-based emotional disclosure intervention on psychological and physiological outcomes were assessed in patients with rheumatoid arthritis. Patients were randomly assigned to a disclosure group (n = 19) in which they wrote/talked about traumatic personal experiences, or to a control group (n = 15) in which they wrote/talked about the events of a particular day. Participants undertook these tasks for periods of 20 minutes on 4 consecutive days. The disclosure group demonstrated increases in negative mood and objective markers of disease activity at 1 week post-intervention. However, there were significant trends for the disclosure group to demonstrate minor improvements in mood and stability in disease activity, compared with the control group. These group differences appeared to be due to deteriorations in the control group more than improvements in the disclosure group.


Cancer, COVID-19, and the need for critique.
In this open letter we examine the implications of the coronavirus disease 2019 (COVID-19) pandemic for cancer research and care from the point of view of the social studies of science, technology, and medicine. We discuss how the pandemic has disrupted several aspects of cancer care, underscoring the fragmentation of institutional arrangements, the malleable priorities in cancer research, and the changing promises of therapeutic innovation. We argue for the critical relevance of qualitative social sciences in cancer research during the pandemic despite the difficulties of immersive kinds of fieldwork. Social science research can help understand the ongoing, situated and lived impact of the pandemic, as well as fully underline its socially stratified consequences. We outline the risk that limiting and prioritising research activities according to their immediate clinical outcomes might have in the relational and longitudinal understanding of cancer practices in the UK. Finally, we alert against potential distortions that a "covidization" of cancer research might entail, arguing for the need to maintain a critical point of view on the pandemic.


Occurrence and Relative Incidence of Viruses Infecting Soybeans in Iran.
A survey was conducted to determine the incidence of Alfalfa mosaic virus (AlMV), Bean common mosaic virus (BCMV), Bean yellow mosaic virus (BYMV), Blackeye cowpea mosaic virus (BlCMV), Cucumber mosaic virus (CMV), Pea enation mosaic virus (PEMV), Peanut mottle virus (PeMoV), Soybean mosaic virus (SMV), Tobacco mosaic virus (TMV), Tobacco ringspot virus (TRSV), Tobacco streak virus (TSV), Tomato ringspot virus (ToRSV), and Tomato spotted wilt virus (TSWV) on soybean (Glycine max) in Iran. Totals of 3,110 random and 1,225 symptomatic leaf samples were collected during the summers of 1999 and 2000 in five provinces of Iran, where commercial soybean is grown, and tested by enzyme-linked immunosorbent assay (ELISA) using specific polyclonal antibodies. Serological diagnoses were confirmed by electron microscopy and host range studies. The highest virus incidence among the surveyed provinces was recorded in Mazandaran (18.6%), followed by Golestan (15.7%), Khuzestan (14.2%), Ardabil (13.9%), and Lorestan (13.5%). Incidence of viruses in decreasing order was SMV (13.3%), TSWV (5.4%), TRSV (4.2%), TSV (4.1%), PEMV (2.9%), BYMV (2.2%), ToRSV (2.1%), AlMV (1.3%), BCMV (0.8%), and CMV (0.6%). Additionally, 1.5% of collected leaf samples had positive reactions in ELISA with antiserum to TMV, indicating the possible infection of soybeans in Iran with a Tobamovirus that is related serologically to TMV. Of 195 leaves from plants showing soybean pod set failure syndrome (PSF) in Mazandaran and Lorestan, only 14 (7.2%) samples had viral infection. No correlation was observed between PSF and presence of the 13 viruses tested, suggesting the involvement of other viruses or factors in this syndrome. To investigate the presence of seed-borne viruses, including SMV, TRSV, ToRSV, and TSV, 7,830 soybean seeds were collected randomly at harvesting time from the major sites of soybean seed production located in Mazandaran and Golestan provinces. According to ELISA analyses of germinated seedlings, 7.1 and 8.9% of the seed samples from Golestan and Mazandaran provinces, respectively, transmitted either SMV, TRSV, ToRSV, or TSV through seed. We also showed that SMV and other seed transmissible viruses, as well as TSWV, usually are the most prevalent viruses in soybean fields in Iran. In this survey, natural occurrence of AlMV, BCMV, BlCMV, BYMV, CMV, PEMV, PeMoV, and TSWV was reported for the first time on soybeans in Iran.


The marine natural product manzamine A targets vacuolar ATPases and inhibits autophagy in pancreatic cancer cells.
Manzamine A, a member of the manzamine alkaloids, was originally isolated from marine sponges of the genus Haliclona. It was recently shown to have activity against pancreatic cancer cells, but the precise mechanism of action remained unclear. To further our understanding of the mechanism of action of manzamine A, chemogenomic profiling in the yeast S. cerevisiae was performed, suggesting that manzamine A is an uncoupler of vacuolar ATPases. Fluorescence microscopy confirmed this effect on yeast vacuoles, where manzamine A produced a phenotype very similar to that of the established v-ATPase inhibitor bafilomycin A1. In pancreatic cancer cells, 10 µM manzamine A affected vacuolar ATPase activity and significantly increased the level of autophagosome marker LC3-II and p62/SQSTM1 as observed by western blot analysis. Treatment with manzamine A in combination with bafilomycin A1 (inhibitor of autophagosome-lysosome fusion) did not change the levels of LC3-II when compared to cells treated with bafilomycin A1 alone, suggesting that manzamine A is a potential inhibitor of autophagy by preventing autophagosome turnover. As autophagy is essential for pancreatic tumor growth, blocking this pathway with manzamine A suggests a promising strategy for the treatment of pancreatic cancer.


Glomerular Hypertrophy Is a Risk Factor for Relapse in Minimal Change Disease Patients.
Patients with minimal change disease (MCD) have a high relapse rate, which results in many complications. Identifying the risk factors for relapse is crucial, but little is known about these factors. Therefore, we performed the current study to determine the factors related to relapse in this patient population. We retrospectively analyzed 51 adult patients with biopsy-proven primary MCD treated between 2003 and 2013. The demographic, physiologic, laboratory and therapeutic data were gathered from the electronic medical records database. Lesions of the glomerulus, tubulointerstitium and vasculature were analyzed for associations with relapse. During a median 50.9 months, 96.1% (49 of 51) of patients had achieved complete remission, and the rest ultimately achieved at least partial remission. A total of 56.9% (29 of 51) patients experienced at least 1 episode of relapse after the first remission. Patients with relapse had a higher rate of glomerular hypertrophy (GH; 34.5%) than those without relapse (9.1%; p < 0.05). After adjusting for confounders, GH was associated with increased odds of relapse (OR 15.992; 95% CI 1.537-166.362; p = 0.02). In a subgroup analysis according to median age, sex and tubulointerstitial (TI) lesions, the association between GH and relapse was evident only in men and in the group with TI lesions. GH is associated with relapse in adult patients with MCD, particularly in men and in those with TI lesions. Frequent monitoring and early intervention are needed in these groups. Future large prospective cohort studies are needed to confirm the study results.


Newer intranasal migraine medications.
Two new intranasal migraine medications, sumatriptan and dihydroergotamine mesylate, may offer specific advantages for patients who are seeking alternatives to various oral or parenteral migraine abortive therapies. Placebo-controlled clinical studies demonstrate that both intranasal forms are effective in relieving migraine headache pain, but published clinical trial information comparing these two intranasal medications with current abortive therapies is lacking. Both agents are generally well tolerated by patients, with the exception of mild, local adverse reactions of the nose and throat.


Striatal Mutant Huntingtin Protein Levels Decline with Age in Homozygous Huntington's Disease Knock-In Mouse Models.
Huntington's disease (HD) is a progressive neurodegenerative disorder associated with aging, caused by an expanded polyglutamine (polyQ) repeat within the Huntingtin (HTT) protein. In HD, degeneration of the striatum and atrophy of the cortex are observed while cerebellum is less affected. To test the hypothesis that HTT protein levels decline with age, which together with HTT mutation could influence disease progression. Using whole brain cell lysates, a unique method of SDS-PAGE and western analysis was used to quantitate HTT protein, which resolves as a monomer and as a high molecular weight species that is modulated by the presence of transglutaminase 2. HTT levels were measured in striatum, cortex and cerebellum in congenic homozygous Q140 and HdhQ150 knock-in mice and WT littermate controls. Mutant HTT in both homozygous knock-in HD mouse models and WT HTT in control striatal and cortical tissues significantly declined in a progressive manner over time. Levels of mutant HTT in HD cerebellum remained high during aging. A general decline in mutant HTT levels in striatum and cortex is observed that may contribute to disease progression in homozygous knock-in HD mouse models through reduction of HTT function. In cerebellum, sustained levels of mutant HTT with aging may be protective to this tissue which is less overtly affected in HD.


Clinical and Laboratory Profile of Zika and Dengue Infected Patients: Lessons Learned From the Co-circulation of Dengue, Zika and Chikungunya in Brazil.
The current triple epidemic caused by dengue, zika and chikungunya constitutes a serious health problem in Brazil. The aim of this study was to investigate acute samples (up to the 7 days of symptoms) from patients presenting acute fever syndrome suspected as arboviral infection and characterize the clinical and laboratorial profile during the co-circulation of dengue, zika and chikungunya in Campo Grande, Mato Grosso do Sul (MS), midwest region of Brazil. All suspected cases (n=134) were tested by using serological and molecular diagnostic tests including DENV, ZIKV and CHIKV RT-PCR, Dengue nonstructural protein 1 (NS1) antigen capture ELISA, anti- DENV IgM ELISA and anti-CHIKV IgM ELISA. In addition, clinical, hematological and biochemical parameters of infected patients were analyzed. It was observed that 79.1% of the blood samples were confirmed for ZIKV and/or DENV infection Of those, 38.0% patients were DENV monoinfected, 26.8% were ZIKV monoinfected and 13.4% were DENV/ZIKV co-infected. Seven patients presented Chikungunya IgM antibodies indicating a previous CHIKV infection. Common symptoms included fever, rash, arthralgia, myalgia, prostration, headache and conjunctivitis. Statistical analysis showed that pruritus and edema were associated with ZIKV infection while prostration and vomiting were more associated with dengue. Additionally, total protein and ALT levels were significantly different in DENV patients compared to ZIKV ones. Some DENV infected patients as well as co-infected needed hospitalization and venous hydration. Otherwise, most ZIKV infected patients presented mild clinical course. Among the pregnant women studied (n=11), three were ZIKV monoinfected while four were DENV monoinfected and two were DENV-1/ZIKV coinfected. In general, normal birth outcomes were observed except for the death due to respiratory insufficiency of one baby born to a mother coinfected with DENV-1/ZIKV. Herein, we provide evidence of the co-circulation of DENV, ZIKV and CHIKV infections in the Campo Grande, MS, Brazil, with a high frequency of DENV-1/ZIKV coinfection. Laboratorial diagnosis poses a challenge where those arboviruses are endemic and differential diagnosis proved to imperative for cases characterization. The knowledge about disease severity during arbovirus coinfections is still scarce and there are several issues emphasizing the importance of adequate management of patients at risk areas.


[Prognostic factors of hospital mortality from diarrhea or pneumonia in infants younger than 1 year old. A case-control study].
Diarrhea and pneumonia are common diseases in children aged under one year, for which there are simple therapeutic measures. However, infant mortality due to these diseases is still very high, varying markedly according to socio-economic status. The characteristics of children who died (cases) and of those who were hospitalized with diarrhea or pneumonia, but survived (controls), were studied. The following groups of variables were studied: socio-economic, environmental and biological conditions, nutritional status and breast-feeding. Information on cases and controls was collected from hospital records and through home interviews. Important losses occurred in the latter: 40% of cases and 50% of controls were not interviewed. There were no significant differences between cases who were included and those who were not, in terms of age, sex or place of residence. To estimate relative risks of prognostic factors unconditional Logistic Regression was used to calculate the odds ratios and their 95% confidence intervals. Prematurity, low birth weight, weight/age deficit, presence of edema and poor general status at hospital admission were prognostic factors for hospital case-fatality. In relation to the anthropometric variables, it was not possible to conclude for certain whether the increased case-fatality was linearly or non-linearly (threshold) associated with nutritional deficit. The duration of breast-feeding was only associated with case-fatality for pneumonia. Socio-economic factors were not important for the prognosis of children admitted to hospital with diarrhea or pneumonia. Some of the expected risk factor associations were not detected, maybe due to the small sample size (resulting from the high losses) which was insufficient to show small differences. In this study the biological conditions of children with diarrhea or pneumonia appeared to be the important prognostic factors for hospital case-fatality.


FDX1 regulates cellular protein lipoylation through direct binding to LIAS.
Ferredoxins are a family of iron-sulfur (Fe-S) cluster proteins that serve as essential electron donors in numerous cellular processes that are conserved through evolution. The promiscuous nature of ferredoxins as electron donors enables them to participate in many metabolic processes including steroid, heme, vitamin D, and Fe-S cluster biosynthesis in different organisms. However, the unique natural function(s) of each of the two human ferredoxins (FDX1 and FDX2) are still poorly characterized. We recently reported that FDX1 is both a crucial regulator of copper ionophore-induced cell death and serves as an upstream regulator of cellular protein lipoylation, a mitochondrial lipid-based post-translational modification naturally occurring on four mitochondrial enzymes that are crucial for TCA cycle function. Here we show that FDX1 directly regulates protein lipoylation by binding the lipoyl synthase (LIAS) enzyme promoting its functional binding to the lipoyl carrier protein GCSH and not through indirect regulation of cellular Fe-S cluster biosynthesis. Metabolite profiling revealed that the predominant cellular metabolic outcome of FDX1 loss of function is manifested through the regulation of the four lipoylation-dependent enzymes ultimately resulting in loss of cellular respiration and sensitivity to mild glucose starvation. Transcriptional profiling established that FDX1 loss-of-function results in the induction of both compensatory metabolism-related genes and the integrated stress response, consistent with our findings that FDX1 loss-of-function is conditionally lethal. Together, our findings establish that FDX1 directly engages with LIAS, promoting its role in cellular protein lipoylation, a process essential in maintaining cell viability under low glucose conditions.


Fine structure constant defines visual transparency of graphene.
There are few phenomena in condensed matter physics that are defined only by the fundamental constants and do not depend on material parameters. Examples are the resistivity quantum, h/e2 (h is Planck's constant and e the electron charge), that appears in a variety of transport experiments and the magnetic flux quantum, h/e, playing an important role in the physics of superconductivity. By and large, sophisticated facilities and special measurement conditions are required to observe any of these phenomena. We show that the opacity of suspended graphene is defined solely by the fine structure constant, a = e2/hc feminine 1/137 (where c is the speed of light), the parameter that describes coupling between light and relativistic electrons and that is traditionally associated with quantum electrodynamics rather than materials science. Despite being only one atom thick, graphene is found to absorb a significant (pa = 2.3%) fraction of incident white light, a consequence of graphene's unique electronic structure.


Reversed diurnal blood pressure rhythm in hypertensive pregnancies.
1. Reversal of the normal diurnal blood pressure pattern has been demonstrated in women with severe hypertension and renal impairment in pregnancy (pre-eclampsia). 2. Maximum arterial pressure occurred at night in these women. The phenomenon was not due to hypotensive drug therapy or posture. Patients with uncomplicated essential hypertension in pregnancy retained a normal diurnal blood pressure pattern. 3. Nocturnal hypertension in pre-eclampsia is of theoretical interest and practical importance.


Etiology of Macular Edema Defined by Deep Learning in Optical Coherence Tomography Scans.
To develop an automated method based on deep learning (DL) to classify macular edema (ME) from the evaluation of optical coherence tomography (OCT) scans. A total of 4230 images were obtained from data repositories of patients attended in an ophthalmology clinic in Colombia and two free open-access databases. They were annotated with four biomarkers (BMs) as intraretinal fluid, subretinal fluid, hyperreflective foci/tissue, and drusen. Then the scans were labeled as control or ocular disease among diabetic macular edema (DME), neovascular age-related macular degeneration (nAMD), and retinal vein occlusion (RVO) by two expert ophthalmologists. Our method was developed by following four consecutive phases: segmentation of BMs, the combination of BMs, feature extraction with convolutional neural networks to achieve binary classification for each disease, and, finally, multiclass classification of diseases and control images. The accuracy of our model for nAMD was 97%, and for DME, RVO, and control were 94%, 93%, and 93%, respectively. Area under curve values were 0.99, 0.98, 0.96, and 0.97, respectively. The mean Cohen's kappa coefficient for the multiclass classification task was 0.84. The proposed DL model may identify OCT scans as normal and ME. In addition, it may classify its cause among three major exudative retinal diseases with high accuracy and reliability. Our DL approach can optimize the efficiency and timeliness of appropriate etiological diagnosis of ME, thus improving patient access and clinical decision making. It could be useful in places with a shortage of specialists and for readers that evaluate OCT scans remotely.


The only African wild tobacco, Nicotiana africana: alkaloid content and the effect of herbivory.
Herbivory in some Nicotiana species is known to induce alkaloid production. This study examined herbivore-induced defenses in the nornicotine-rich African tobacco N. africana, the only Nicotiana species indigenous to Africa. We tested the predictions that: 1) N. africana will have high constitutive levels of leaf, flower and nectar alkaloids; 2) leaf herbivory by the African bollworm Helicoverpa armigera will induce increased alkaloid levels in leaves, flowers and nectar; and 3) increased alkaloid concentrations in herbivore-damaged plants will negatively affect larval growth. We grew N. africana in large pots in a greenhouse and exposed flowering plants to densities of one, three and six fourth-instar larvae of H. armigera, for four days. Leaves, flowers and nectar were analyzed for nicotine, nornicotine and anabasine. The principal leaf alkaloid was nornicotine (mean: 28 µg/g dry mass) followed by anabasine (4.9 µg/g) and nicotine (0.6 µg/g). Nornicotine was found in low quantities in the flowers, but no nicotine or anabasine were recorded. The nectar contained none of the alkaloids measured. Larval growth was reduced when leaves of flowering plants were exposed to six larvae. As predicted by the optimal defense theory, herbivory had a localized effect and caused an increase in nornicotine concentrations in both undamaged top leaves of herbivore damaged plants and herbivore damaged leaves exposed to one and three larvae. The nicotine concentration increased in damaged compared to undamaged middle leaves. The nornicotine concentration was lower in damaged leaves of plants exposed to six compared to three larvae, suggesting that N. africana rather invests in new growth as opposed to protecting older leaves under severe attack. The results indicate that the nornicotine-rich N. africana will be unattractive to herbivores and more so when damaged, but that potential pollinators will be unaffected because the nectar remains alkaloid-free even after herbivory.


The tyrosine kinase Lyn is required for B cell development beyond the T1 stage in the spleen: rescue by over-expression of Bcl-2.
We have analyzed the effects of deficiency in the tyrosine kinase Lyn on B cell development using transgenic mice that express a B cell antigen receptor (BCR) of defined specificity (3-83,anti-H-2K(k or b)). In the absence of Lyn, immature B cells are abundant in the bone marrow and spleen up until the T1 stage (IgM(hi) IgD(-) CD21(-)CD23(-)), after which B cell development is severely impaired. The small number of mature B cells that do develop in Lyn-deficient mice express normal levels of the transgenic BCR and lack expression of CD80 and CD86, suggesting they are not activated. In Lyn-deficient animals the presence of a Bcl-2 transgene leads to a dramatic increase in B cell numbers and restores T2 stage (IgM(hi) IgD(hi) CD21(hi) CD23(int)) and mature populations. In 3-83 lyn-/- Bcl-2 Tg mice, a population of lambda-positive cells that also express the 383 idiotype is evident, suggesting that in the absence of lyn isotype exclusion by the transgenic BCR is less efficient. The results indicate that Lyn plays a positive role in the selection and survival of mature B cells in addition to its previously documented negative role in tolerance and B cell activation.


Mechanoelectrical transduction of adult outer hair cells studied in a gerbil hemicochlea.
Sensory receptor cells of the mammalian cochlea are morphologically and functionally dichotomized. Inner hair cells transmit auditory information to the brain, whereas outer hair cells (OHC) amplify the mechanical signal, which is then transduced by inner hair cells. Amplification by OHCs is probably mediated by their somatic motility in a mechanical feedback process. OHC motility in vivo is thought to be driven by the cell's receptor potential. The first steps towards the generation of the receptor potential are the deflection of the stereociliary bundle, and the subsequent flow of transducer current through the mechanosensitive transducer channels located at their tips. Quantitative relations between transducer currents and basilar membrane displacements are lacking, as well as their variation along the cochlear length. To address this, we simultaneously recorded OHC transducer currents (or receptor potentials) and basilar membrane motion in an excised and bisected cochlea, the hemicochlea. This preparation permits recordings from adult OHCs at various cochlear locations while the basilar membrane is mechanically stimulated. Furthermore, the stereocilia are deflected by the same means of stimulation as in vivo. Here we show that asymmetrical transducer currents and receptor potentials are significantly larger than previously thought, they possess a highly restricted dynamic range and strongly depend on cochlear location.


The technology - activities of daily living questionnaire: a version with a technology-related subscale.
Information and communication technology (ICT) has become an increasingly important part of daily life. The ability to use technology is becoming essential for autonomous functioning in society. Current functional scales for patients with cognitive impairment do not evaluate the use of technology. The objective of this study was to develop and validate a new version of the Activities of Daily Living Questionnaire (ADLQ) that incorporates an ICT subscale. A new technology-based subscale was incorporated into the Spanish version of the ADLQ (SV-ADLQ), entitled the Technology version of the ADLQ (T-ADLQ). The T-ADLQ was administered to 63 caregivers of dementia patients, 21 proxies of mild cognitive impairment patients and 44 proxies of normal elderly subjects (mean age of the sample ± SD: 73.5 ± 8.30 years). We analysed the convergent validity, internal consistency, reliability cut-off point, sensitivity and specificity of the T-ADLQ. The results of the T-ADLQ were compared to the SV-ADLQ. The T-ADLQ showed significant correlations with the Mini-Mental State Examination (MMSE), the Frontal Assessment Battery (FAB) as well as other measures of functional impairment and dementia severity (MMSE: r = -0.70; FAB: r = -0.65; Functional Assessment Questionnaire: r = 0.77; Instrumental Activities of Daily Living Scale: r = -0.75; Clinical Dementia Rating Scale: r = 0.72; p < 0.001). The T-ADLQ showed a good reliability with a relatively high Cronbach's α-coefficient (Cronbach's α = 0.861). When considering a functional impairment cut-off point greater than 29.25%, the sensitivity and specificity of the T-ADLQ were 82 and 90%, respectively. The area under the receiver-operating characteristic curve was 0.937 for the T-ADLQ and 0.932 for the original version of the test. The T-ADLQ revealed adequate indicators of validity and reliability for the functional assessment of activities of daily living in dementia patients. However, the inclusion of technology items in the T-ADLQ did not improve the performance of the scale, which may reflect the lack of widespread use of technology by elderly individuals. Thus, although it appeared reasonable to add technology use questions to the ADLQ, our experience suggested that this has to be done cautiously, since the sensitivity of these additional items could vary in different populations. The T-ADLQ needs to be validated in a different population of dementia subjects.


Structure of signaling-competent neurotensin receptor 1 obtained by directed evolution in Escherichia coli.
Crystallography has advanced our understanding of G protein-coupled receptors, but low expression levels and instability in solution have limited structural insights to very few selected members of this large protein family. Using neurotensin receptor 1 (NTR1) as a proof of principle, we show that two directed evolution technologies that we recently developed have the potential to overcome these problems. We purified three neurotensin-bound NTR1 variants from Escherichia coli and determined their X-ray structures at up to 2.75 Å resolution using vapor diffusion crystallization experiments. A crystallized construct was pharmacologically characterized and exhibited ligand-dependent signaling, internalization, and wild-type-like agonist and antagonist affinities. Our structures are fully consistent with all biochemically defined ligand-contacting residues, and they represent an inactive NTR1 state at the cytosolic side. They exhibit significant differences to a previously determined NTR1 structure (Protein Data Bank ID code 4GRV) in the ligand-binding pocket and by the presence of the amphipathic helix 8. A comparison of helix 8 stability determinants between NTR1 and other crystallized G protein-coupled receptors suggests that the occupancy of the canonical position of the amphipathic helix is reduced to various extents in many receptors, and we have elucidated the sequence determinants for a stable helix 8. Our analysis also provides a structural rationale for the long-known effects of C-terminal palmitoylation reactions on G protein-coupled receptor signaling, receptor maturation, and desensitization.


Life cycle sustainability assessment of a light rail transit system: Integration of environmental, economic, and social impacts.
The transition toward sustainable urban transportation has gained importance in recent decades. However, urban transportation has not been addressed for all dimensions of sustainability. This study presents a life cycle sustainability assessment of a light rail transit system in Kayseri, Turkey, by integrating environmental, economic, and social aspects. The sustainability performance of the light rail transit system is evaluated using a cradle-to-grave approach to assess three aspects of sustainability. For the environmental evaluation, a life cycle assessment was applied using SimaPro 8.4.1 PhD version based on ISO 14040 and 14044. The method, which includes nine environmental impact categories, was employed to assess the environmental performance of the light rail transit system with a functional unit of 1 passenger-km. For the economic assessment, life cycle costing was utilized with the functional unit of USD for 1 passenger-km. A social life cycle assessment was applied to assess the social performance of the light rail transit system based on guidelines published by the United Nations Environment Programme in collaboration with the Society of Environmental Toxicology and Chemistry. For the determination of social impacts, 11 subcategories and 18 social indicators were selected. The results showed that the global warming potential and abiotic depletion potential of the light rail system per passenger-km were 2.4E - 02 kg CO2 eq. and 2.7E - 01 MJ, respectively, with a service life of 50 years. The total life cycle cost of the light rail system was calculated as 0.046 USD for 1 passenger-km. The results also revealed that the main contributor to the total life cycle cost was energy cost, with 92% (2.88E + 08 USD) of the total cost. In the social performance evaluation, it is found that the industry performs well for society, the local community, and workers but has a weaker social performance for the consumer due to a weak feedback mechanism. Integr Environ Assess Manag 2021;17:1070-1082. © 2021 SETAC.


Coronary artery bypass grafting: quality of life of patients in Karachi.
In the Third World, cardiac patients often suffer not only from the nature of their illnesses, but also the insufficiency of the facilities available. Although the effectiveness of coronary artery bypass grafting (CABG) is being evaluated in terms of mortality, complications or recurrence of symptoms, empirical studies assessing the change in patients' quality of life (QOL) after CABG within the Pakistani cultural context are lacking. The aims of the present study are to assess and compare the change in health-related quality of life before and 1 month after CABG and to assess the differences in QOL with respect to age, gender and cardiac rehabilitation programme attendance. A prospective single group pre- and post-study design was used, whereby patients completed a questionnaire before and 1 month after surgery. Questions considered the person's physical capacity, psychological wellbeing, social relationships and satisfaction from their health condition. The results indicated a significant improvement in patients' physical and psychological health, satisfaction with social relationships and overall health status. Younger patients showed significantly lower scores on the social domain (reflecting greater dissatisfaction with their sex life and personal relationships). Female participants showed significantly lower scores in psychological health post surgery. Lastly, participants of the cardiac rehabilitation programme scored significantly higher in psychological health compared with non-participants. QOL improved from prior to and 1 month after CABG while differing (in specific domains) with respect to age, gender and cardiac rehabilitation programme attendance. The findings can be used to develop interventions to improve health and QOL in specific domains with respect to specific groups.


Exposure of the vertebrobasilar artery junction with traction of the dentate ligament for the treatment of large vertebral artery aneurysms: technical note.
A technique for exposing the vertebrobasilar junction with traction of the dentate ligament is described for treatment of large vertebral artery (VA) aneurysms via the far lateral suboccipital approach with partial condylar resection. The most rostral attachment of the dentate ligament is divided above the site where the VA pierces the dura mater. A traction suture is placed into the dentate ligament and gently retracted using mosquito forceps. As a result, the medulla oblongata is lifted dorsally and slightly rotated by the divided and retracted dentate ligament, allowing an approach from a more superior or inferior direction. The present technique is useful for the treatment of large thrombosed VA aneurysms.


The mucosal inflammatory response to non-typhoidal Salmonella in the intestine is blunted by IL-10 during concurrent malaria parasite infection.
Coinfection can markedly alter the response to a pathogen, thereby changing its clinical presentation. For example, non-typhoidal Salmonella (NTS) serotypes are associated with gastroenteritis in immunocompetent individuals. In contrast, individuals with severe pediatric malaria can develop bacteremic infections with NTS, during which symptoms of gastroenteritis are commonly absent. Here we report that, in both a ligated ileal loop model and a mouse colitis model, malaria parasites caused a global suppression of gut inflammatory responses and blunted the neutrophil influx that is characteristic of NTS infection. Further, malaria parasite infection led to increased recovery of Salmonella enterica serotype Typhimurium from the draining mesenteric lymph node (MLN) of mice. In the mouse colitis model, blunted intestinal inflammation during NTS infection was independent of anemia but instead required parasite-induced synthesis of interleukin (IL)-10. Blocking of IL-10 in coinfected mice reduced dissemination of S. Typhimurium to the MLN, suggesting that induction of IL-10 contributes to development of disseminated infection. Thus IL-10 produced during the immune response to malaria in this model contributes to suppression of mucosal inflammatory responses to invasive NTS, which may contribute to differences in the clinical presentation of NTS infection in the setting of malaria.


Nitric oxide, but not vasodilating prostaglandins, contributes to the improvement of exercise hyperemia via ascorbic acid in healthy older adults.
Acute ascorbic acid (AA) administration increases muscle blood flow during dynamic exercise in older adults, and this is associated with improved endothelium-dependent vasodilation. We directly tested the hypothesis that increase in muscle blood flow during AA administration is mediated via endothelium-derived vasodilators nitric oxide (NO) and prostaglandins (PGs). In 14 healthy older adults (64 ± 3 yr), we measured forearm blood flow (FBF; Doppler ultrasound) during rhythmic handgrip exercise at 10% maximum voluntary contraction. After 5-min steady-state exercise with saline, AA was infused via brachial artery catheter for 10 min during continued exercise, and this increased FBF ∼25% from 132 ± 16 to 165 ± 20 ml/min (P < 0.05). AA was infused for the remainder of the study. Next, subjects performed a 15-min exercise bout in which AA + saline was infused for 5 min, followed by 5 min of the nitric oxide synthase (NOS) inhibitor N(G)-monomethyl-l-arginine (l-NMMA) and then 5 min of the cyclooxygenase inhibitor ketorolac (group 1). The order of inhibition was reversed in eight subjects (group 2). In group 1, independent NOS inhibition reduced steady-state FBF by ∼20% (P < 0.05), and subsequent PG inhibition had no impact on FBF (Δ 3 ± 5%). Similarly, in group 2, independent PG inhibition had little effect on FBF (Δ -4 ± 4%), whereas subsequent NO inhibition significantly decreased FBF by ∼20% (P < 0.05). In a subgroup of five subjects, we inhibited NO and PG synthesis before AA administration. In these subjects, there was a minimal nonsignificant improvement in FBF with AA infusion (Δ 7 ± 3%; P = nonsignificant vs. zero). Together, our data indicate that the increase in muscle blood flow during dynamic exercise with acute AA administration in older adults is mediated primarily via an increase in the bioavailability of NO derived from the NOS pathway.


Viral Aggregation: Impact on Virus Behavior in the Environment.
Aggregates of viruses can have a significant impact on quantification and behavior of viruses in the environment. Viral aggregates may be formed in numerous ways. Viruses may form crystal like structures and aggregates in the host cell during replication or may form due to changes in environmental conditions after virus particles are released from the host cells. Aggregates tend to form near the isoelectric point of the virus, under the influence of certain salts and salt concentrations in solution, cationic polymers, and suspended organic matter. The given conditions under which aggregates form in the environment are highly dependent on the type of virus, type of salts in solution (cation, anion. monovalent, divalent) and pH. However, virus type greatly influences the conditions when aggregation/disaggregation will occur, making predictions difficult under any given set of water quality conditions. Most studies have shown that viral aggregates increase the survival of viruses in the environment and resistance to disinfectants, especially with more reactive disinfectants. The presence of viral aggregates may also result in overestimation of removal by filtration processes. Virus aggregation-disaggregation is a complex process and predicting the behavior of any individual virus is difficult under a given set of environmental circumstances without actual experimental data.


Recoding between two types of STM representation revealed by the dynamics of memory search.
Visual STM (VSTM) is thought to be related to visual attention in several ways. Attention controls access to VSTM during memory encoding and plays a role in the maintenance of stored information by strengthening memorized content. We investigated the involvement of visual attention in recall from VSTM. In two experiments, we measured electrophysiological markers of attention in a memory search task with varying intervals between VSTM encoding and recall, and so we were able to track recoding of representations in memory. Results confirmed the involvement of attention in VSTM recall. However, the amplitude of the N2pc and N3rs components, which mark orienting of attention and search within VSTM, decreased as a function of delay. Conversely, the amplitude of the P3 and sustained posterior contralateral negativity components increased as a function of delay, effectively the opposite of the N2pc and N3rs modulations. These effects were only observed when verbal memory was not taxed. Thus, the results suggested that gradual recoding from visuospatial orienting of attention into verbal recall mechanisms takes place from short to long retention intervals. Interestingly, recall at longer delays was faster than at short delays, indicating that verbal representation is coupled with faster responses. These results extend the orienting-of-attention hypothesis by including an account of representational recoding during short-term consolidation and its consequences for recall from VSTM.


Pentapeptide (proctolin) associated with an identified neuron.
Individual neurons can be recognized and identified anatomically, physiologically, and biochemically in the insect central nervous system. Biochemical analyses of extracts prepared from one such identified neuron show it to be associated with a bioactive pentapeptide called proctolin. This peptide may be a neurotransmitter, and a preparation is established in which its physiological action can be studied at the cellular level.


Stimulation with a class A CpG oligonucleotide enhances resistance to infection with feline viruses from five different families.
Domestic cats are commonly affected by viral pathogens that induce lengthy infections with fatal outcomes. Prevention of viral propagation is of primordial importance in shelters and catteries, where cats from different backgrounds have narrow contacts. Oligonucleotides (ODN) containing cytosine-phosphate-guanosine motifs of class A (CpG-A) are highly potent synthetic inducers of innate antiviral mechanisms. The aim of this study was to test their ability to modulate innate immune responses and prevent viral replication as stand-alone agents in the domestic cat. CpG-A stimulation of feline peripheral blood mononuclear cells (PBMCs) enhanced their proliferation, increased the presence of co-stimulatory molecules on their surface and influenced their gene expression profiles in an antiviral orientation. Incubation of the supernatants of CpG-A stimulated PBMCs with feline cell lines of epithelial and fibroblastic origin induced expression of the antiviral myxovirus resistance (Mx) gene in these target cells, which also showed enhanced resistance to feline viruses from five distinct families, namely Coronaviridae, Herpesviridae, Caliciviridae, Parvoviridae, and Retroviridae. Most importantly, subcutaneous administration of CpG-A in domestic cats systemically increased the expression of Mx, reaching maximal levels within 24 h. Plasma from treated cats could furthermore inhibit viral replication in vitro. Altogether, our data highlight the promising potential of CpG-A to induce a preventive antiviral state in the cat and to protect feline populations against a broad range of virus infections.


On-line EM algorithm for the normalized gaussian network.
A normalized gaussian network (NGnet) (Moody & Darken, 1989) is a network of local linear regression units. The model softly partitions the input space by normalized gaussian functions, and each local unit linearly approximates the output within the partition. In this article, we propose a new on-line EMalgorithm for the NGnet, which is derived from the batch EMalgorithm (Xu, Jordan, &Hinton 1995), by introducing a discount factor. We show that the on-line EM algorithm is equivalent to the batch EM algorithm if a specific scheduling of the discount factor is employed. In addition, we show that the on-line EM algorithm can be considered as a stochastic approximation method to find the maximum likelihood estimator. A new regularization method is proposed in order to deal with a singular input distribution. In order to manage dynamic environments, where the input-output distribution of data changes over time, unit manipulation mechanisms such as unit production, unit deletion, and unit division are also introduced based on probabilistic interpretation. Experimental results show that our approach is suitable for function approximation problems in dynamic environments. We also apply our on-line EM algorithm to robot dynamics problems and compare our algorithm with the mixtures-of-experts family.


Covalent attachment of FeFe hydrogenases to carbon electrodes for direct electron transfer.
Direct electron transfer between enzymes and electrodes is now commonly achieved, but obtaining protein films that are very stable may be challenging. This is particularly crucial in the case of hydrogenases, the enzymes that catalyze the biological conversion between dihydrogen and protons, because the instability of the hydrogenase films may prevent the use of these enzymes as electrocatalysts of H(2) oxidation and production in biofuel cells and photoelectrochemical cells. Here we show that two different FeFe hydrogenases (from Chamydomonas reinhardtii and Clostridium acetobutylicum) can be covalently attached to functionalized pyrolytic graphite electrodes using peptidic coupling. In both cases, a surface patch of lysine residues makes it possible to favor an orientation that is efficient for fast, direct electron transfer. High hydrogen-oxidation current densities are maintained for up to one week, the only limitation being the intrinsic stability of the enzyme. We also show that covalent attachment has no effect on the catalytic properties of the enzyme, which means that this strategy can also used be for electrochemical studies of the catalytic mechanism.


African American patients with gout: efficacy and safety of febuxostat vs allopurinol.
African Americans are twice as likely as Caucasians to develop gout, but they are less likely to be treated with urate-lowering therapy (ULT). Furthermore, African Americans typically present with more comorbidities associated with gout, such as hypertension, obesity, and renal impairment. We determined the efficacy and safety of ULT with febuxostat or allopurinol in African American subjects with gout and associated comorbidities and in comparison to Caucasian gout subjects. This is a secondary analysis of the 6-month Phase 3 CONFIRMS trial. Eligible gouty subjects with baseline serum urate (sUA) ≥ 8.0 mg/dL were randomized 1:1:1 to receive febuxostat 40 mg, febuxostat 80 mg, or allopurinol (300 mg or 200 mg depending on renal function) daily. All subjects received gout flare prophylaxis. Primary efficacy endpoint was the proportion of subjects in each treatment group with sUA < 6.0 mg/dL at the final visit. Additional endpoints included the proportion of subjects with mild or with moderate renal impairment who achieved a target sUA < 6.0 mg/dL at final visit. Adverse events (AEs) were recorded throughout the study. Of the 2,269 subjects enrolled, 10.0% were African American and 82.1% were Caucasian. African American subjects were mostly male (89.5%), obese (BMI ≥ 30 kg/m2; 67.1%), with mean baseline sUA of 9.8 mg/dL and mean duration of gout of 10.4 years. The proportions of African American subjects with a baseline history of diabetes, renal impairment, or cardiovascular disease were significantly higher compared to Caucasians (p < 0.001). ULT with febuxostat 80 mg was superior to both febuxostat 40 mg (p < 0.001) and allopurinol (p = 0.004). Febuxostat 40 mg was comparable in efficacy to allopurinol. Significantly more African American subjects with mild or moderate renal impairment achieved sUA < 6.0 mg/dL in the febuxostat 80 group than in either the febuxostat 40 mg or allopurinol group (p < 0.05). Efficacy rates in all treatment groups regardless of renal function were comparable between African American and Caucasian subjects, as were AE rates. In African American subjects with significant comorbidities, febuxostat 80 mg is significantly more efficacious than either febuxostat 40 mg or allopurinol 200/300 mg. Febuxostat was well tolerated in this African American population.Please see related article: http://www.biomedcentral.com/1741-7015/10/15.


[Analgesia and sedation during placement of peripherally inserted central catheters in neonates].
This study aimed to characterize the analgesia and sedation strategies in neonates having a peripherally inserted central catheter (PICC) placed, and to relate it to the number of venipunctures, duration of procedure and catheter tip position. This was a cross-sectional study with prospective data collection, conducted in a neonatal intensive care unit of a private hospital in the city of São Paulo, during the period from August 31, 2010 to July 1, 2011, which evaluated 254 PICC insertions. The adoption of analgesic or sedative strategies occurred in 88 (34.6%) catheter placements and was not related to the number of venipunctures, duration of procedure or catheter tip position. Intravenous administration of midazolam, in 47 (18.5%), and fentanyl, in 19 (7.3%), catheter insertions were the most frequent strategies. Wider adoption of analgesic strategies is recommended before, during and after the procedure.


Early development of the optic chiasm in the gray short-tailed opossum, Monodelphis domestica.
We have studied the early development of the uncrossed retinofugal projection in the gray short-tailed opossum. Axons that form the adult uncrossed retinofugal projection arise from the temporal crescent of the retina and reach the optic chiasm on postnatal day 7. The sites at which the uncrossed fibres segregate from the crossed fibres and the pattern of this segregation are very different from those seen in eutherian mammals. In the opossum, the uncrossed fibres segregate from the crossed fibres within the juxtachiasmatic part of the optic nerve before they have encountered either the fibres of the other eye or midline structures of the ventral diencephalon. The uncrossed fibres turn perpendicular to the axis of the nerve and grow dorsoventrally through the crossed projection to gather as a discrete bundle at the ventral edge of the nerve. The abrupt divergence of the uncrossed fibres occurs at a border between two glial cell types: the interfascicular glia that characterise the main part of the optic nerve and the radial glia of the juxtachiasmatic part of the nerve. At the ventral part of the nerve, the bundle of uncrossed fibres turns caudally across the axis of the nerve and enters the ipsilateral optic tract. When retinofugal fibres encounter the border between the interfascicular and radial glia, a very specific axonal reorganisation occurs in marsupials, and this is strikingly different from the axonal reorganisation that occurs at the same site in eutherians, where essentially all retinofugal fibres reorganise, not just the uncrossed component. We believe this to be an important example of an identified cellular element that has quite distinct axon-guidance properties in different species.


AN INQUIRY INTO SOME MECHANICAL FACTORS IN THE PRODUCTION OF LYMPHOCYTOSIS.
1. The lymph of the thoracic duct furnishes to the blood a larger proportion than is usually supposed of the lymphocytes in circulation. Gross variations in its output of such cells must affect very considerably the blood picture. 2. The quantity of lymphocytes supplied through the thoracic duct of the healthy dog remains practically constant from hour to hour, if the physiological conditions are not notably changed. Transient change in physiological conditions may alter the output of cells, but with the disappearance of this change the output tends to resume its previous rate. These facts indicate that the tissues producing lymphocytes are "set" at a rate of activity definite in the individual. 3. Muscular activity (struggle) produces a prompt increase in the output of lymphocytes through the thoracic duct. (a) This is assured by the presence of an increased number of cells per cubic millimeter of lymph, combined with an increase in the amount of lymph voided. (b) The lymphocyte-output may be tripled or quadrupled during a long-continued struggle. (c) Following prolonged struggle the output of lymphocytes is for a short time less than previous to the exertion. 4. The increased lymph-flow caused by a lymphagogue of the second class (glucose) brings with it increased output of lymphocytes through the thoracic duct. (a) The individual cubic millimeters of lymph are often poor in cells, during the rapid lymph-flow, yet the total number of elements transported is large. (b) The results with glucose support the theory of Ehrlich, that a rapidly appearing lymphocytosis may be produced through the flushing effect of increased lymph-flow. 5. A comparison of the effects of struggle with those of glucose demonstrates that in the former some factor besides increase in lymph-flow per se (Ehrlich) works to cause the large output of lymphocytes. The nature of this factor has not yet been determined. 6. The variations caused by muscular exertion and by increased lymph-flow in the number of lymphocytes coursing through the thoracic duct are so pronounced as to suggest that the total number of lymphocytes in circulation must be considerably influenced by them. Clinical findings by other observers indicate that this is true; and the clinical findings themselves become much simpler of interpretation. 7. The results in general prove the existence, reserved from circulation, of a large fund of lymphocytes, which is quickly yielded to the blood under certain physiological conditions.


Social Participation in Schools: A Survey of Occupational Therapy Practitioners.
We sought to identify strategies and practices that school-based occupational therapy practitioners use in addressing social participation, their perceptions of competence, and factors influencing their strategies and practices. Surveys were sent to 500 randomly selected members of the American Occupational Therapy Association's Early Intervention and School Special Interest Section; 112 usable surveys were received from 36 states. Respondents reported focusing on internal client factors when addressing social participation. Fewer than half (46.5%) indicated that they understood their role (mean [M] = 4.23, standard deviation [SD] = 1.22), and 57.1% desired greater understanding or ability (M = 4.64, SD = 1.29). Differences were found based on years of experience, service delivery model used, and services provided by diagnosis category. School-based occupational therapy practitioners may need to expand their current practices in addressing student social participation. Findings may be used to develop interventions to address this area of practice.


Phosphorylation-dependent regulation of cytosolic localization and oncogenic function of Skp2 by Akt/PKB.
Skp2 is an F-box protein that forms the SCF complex with Skp1 and Cullin-1 to constitute an E3 ligase for ubiquitylation. Ubiquitylation and degradation of the p27 are critical for Skp2-mediated entry to the cell cycle, and overexpression and cytosolic accumulation of Skp2 have been clearly associated with tumorigenesis, although the functional significance of the latter is still unknown. Here we show that Akt/protein kinase B (PKB) interacts with and directly phosphorylates Skp2. We find that Skp2 phosphorylation by Akt triggers SCF complex formation and E3 ligase activity. A phosphorylation-defective Skp2 mutant is drastically impaired in its ability to promote cell proliferation and tumorigenesis. Furthermore, we show that Akt-mediated phosphorylation triggers 14-3-3beta-dependent Skp2 relocalization to the cytosol, and we attribute a specific role to cytosolic Skp2 in the positive regulation of cell migration. Finally, we demonstrate that high levels of activation of Akt correlate with the cytosolic accumulation of Skp2 in human cancer specimens. Our results therefore define a novel proto-oncogenic Akt/PKB-dependent signalling pathway.


Abrupt burial imparts persistent changes to the bacterial diversity of turbidite-associated sediment profiles.
The emplacement of subaqueous gravity-driven sediment flows imposes a significant physical and geochemical impact on underlying sediment and microbial communities. Although previous studies have established lasting mineralogical and biological signatures of turbidite deposition, the response of bacteria and archaea within and beneath debris flows remains poorly constrained. Both bacterial cells associated with the underlying sediment and those attached to allochthonous material must respond to substantially altered environmental conditions and selective pressures. As a consequence, turbidites and underlying sediments provide an exceptional opportunity to examine (i) the microbial community response to rapid sedimentation and (ii) the preservation and identification of displaced micro-organisms. We collected Illumina MiSeq sequence libraries across turbidite boundaries at ~26 cm sediment depth in La Jolla Canyon off the coast of California, and at ~50 cm depth in meromictic Twin Lake, Hennepin County, MN. 16S rRNA gene signatures of relict and active bacterial populations exhibit persistent differences attributable to turbidite deposition. In particular, both the marine and lacustrine turbidite boundaries are sharply demarcated by the abundance and diversity of Chloroflexi, suggesting a characteristic sensitivity to sediment disturbance history or to differences in organic substrates across turbidite profiles. Variations in the abundance of putative dissimilatory sulfate-reducing Deltaproteobacteria across the buried La Jolla Canyon sediment-water interface reflect turbidite-induced changes to the geochemical environment. Species-level distinctions within the Deltaproteobacteria clearly conform to the sedimentological boundary, suggesting a continuing impact of genetic inheritance distinguishable from broader trends attributable to selective pressure. Abrupt, <1-cm scale changes in bacterial diversity across the Twin Lake turbidite contact are consistent with previous studies showing that relict DNA signatures attributable to sediment transport may be more easily preserved in low-energy, anoxic environments. This work raises the possibility that deep subsurface microbial communities may inherit variations in microbial diversity from sediment flow and deformation events.


A spiking neuron as information bottleneck.
Neurons receive thousands of presynaptic input spike trains while emitting a single output spike train. This drastic dimensionality reduction suggests considering a neuron as a bottleneck for information transmission. Extending recent results, we propose a simple learning rule for the weights of spiking neurons derived from the information bottleneck (IB) framework that minimizes the loss of relevant information transmitted in the output spike train. In the IB framework, relevance of information is defined with respect to contextual information, the latter entering the proposed learning rule as a "third" factor besides pre- and postsynaptic activities. This renders the theoretically motivated learning rule a plausible model for experimentally observed synaptic plasticity phenomena involving three factors. Furthermore, we show that the proposed IB learning rule allows spiking neurons to learn a predictive code, that is, to extract those parts of their input that are predictive for future input.


Expansion and countrywide dissemination of ST11, ST15 and ST147 ciprofloxacin-resistant CTX-M-15-type beta-lactamase-producing Klebsiella pneumoniae epidemic clones in Hungary in 2005--the new 'MRSAs'?
To investigate the molecular epidemiology of ciprofloxacin-resistant CTX-M-15-producing Klebsiella pneumoniae epidemic clones (ECs) isolated from six nosocomial outbreaks and sporadic cases during 2005 in Hungary. Two hundred and eighty-one extended-spectrum beta-lactamase (ESBL)-producing K. pneumoniae clinical isolates collected from 41 centres were submitted to the National ESBL Reference Laboratory for further investigations. Of the 281 strains, 75 isolates proved to be SHV producers, whereas 6 isolates were ciprofloxacin-susceptible CTX-M-type ESBL producers. One hundred and ninety-six ciprofloxacin-resistant CTX-M-type beta-lactamase-producing isolates collected from 35 centres were subjected to macrorestriction profile analysis. Furthermore, molecular typing was performed by PCR and sequencing of several antibiotic resistance genes, plasmid profile analysis, transfer of resistance determinants and multilocus sequence typing (MLST). PFGE revealed the existence of three genetic clusters defined as ECs, where 129 isolates belonged to the previously described Hungarian EC (HEC), 46 isolates to epidemic clone II (EC II) and 21 isolates to epidemic clone III (EC III), respectively. All isolates harboured plasmids ranging from 2.0 to 230 kb. PstI digestion of plasmid DNA from transconjugants/transformants revealed diverse restriction patterns from distinct ECs. Sequence analysis of beta-lactamase genes from 19 selected isolates detected bla(CTX-M-15) and bla(OXA-1) in strains from all three ECs and bla(TEM-1) in EC III isolates located on large plasmids. ISEcpI associated with CTX-M-15 was detected only on a 50 kb non-conjugative plasmid from EC III. MLST identified three allelic profiles: ST 15 (HEC), ST 11 (EC III) and the novel ST 147 (EC II), which correspond to the PFGE clusters, respectively. In 2005, 97% of all CTX-M-producing K. pneumoniae isolates detected across Hungary were highly ciprofloxacin-resistant CTX-M-15 producers and represented just three stable genetic clones.


Human ribosomes from cells with reduced dyskerin levels are intrinsically altered in translation.
Dyskerin is a pseudouridine (ψ) synthase involved in fundamental cellular processes including uridine modification in rRNA and small nuclear RNA and telomere stabilization. Dyskerin functions are altered in X-linked dyskeratosis congenita (X-DC) and cancer. Dyskerin's role in rRNA pseudouridylation has been suggested to underlie the alterations in mRNA translation described in cells lacking dyskerin function, although relevant direct evidences are currently lacking. Our purpose was to establish definitely whether defective dyskerin function might determine an intrinsic ribosomal defect leading to an altered synthetic activity. Therefore, ribosomes from dyskerin-depleted human cells were purified and 1) added to a controlled reticulocyte cell-free system devoid of ribosomes to study mRNA translation; 2) analyzed for protein contamination and composition by mass spectrometry, 3) analyzed for global pseudouridylation levels. Ribosomes purified from dyskerin-depleted cells showed altered translational fidelity and internal ribosome entry site (IRES)-mediated translation. These ribosomes displayed reduced uridine modification, whereas they were not different in terms of protein contamination or ribosomal protein composition with respect to ribosomes from matched control cells with full dyskerin activity. In conclusion, lack of dyskerin function in human cells induces a defect in rRNA uridine modification, which is sufficient to alter ribosome activity.


Alignment of liquid crystals with patterned isotropic surfaces.
The molecules of a nematic liquid crystal exposed to an isotropic surface adopt a mean tilt relative to the normal but have no in-plane alignment-that is, they are free to have any azimuthal orientation in the surface plane. Pursuing the theoretical suggestion by Meyer that, in spite of this azimuthal degeneracy, spatially inhomogeneous isotropic surfaces combine with liquid crystal elastic anisotropy to produce alignment, we show that a boundary line between two isotropic regions that differ in mean tilt does indeed align the liquid crystal. The boundaries on a patterned surface of distinct isotropic regions thus act as a system of lines that the molecular orientation locally follows. This enables the development of liquid crystal alignment surfaces based on printing or lithographic patterning.


Retention of black and white participants in the selenium and vitamin E cancer prevention trial (SWOG-coordinated intergroup study S0000).
Disproportionally low retention of minority populations can adversely affect the generalizability of clinical research trials. We determine the overall retention rates for White and Black participants from the Selenium and Vitamin E Cancer Prevention Trial (SELECT) and explore participant and site characteristics associated with retention failure (study disengagement) for these groups. A secondary analysis of 28,118 White (age ≥55), and 4,322 Black (age ≥ 50) SELECT participants used multivariate Cox regression to estimate overall retention rates and to calculate HRs and 95% confidence intervals (CI). Blacks had higher age-adjusted risk of disengagement than Whites (HR, 1.92; 95% CI, 1.77-2.08). Among Black participants, those ages 50 to 54 were at three times the risk of disengagement than those ≥65 years of age (HR, 3.61; 95% CI, 2.41-5.41). Blacks age ≥65 had 1.6 times the risk of disengagement than Whites age ≥65 (HR, 1.60; 95% CI, 1.38-1.87). By 6 years after randomization, 84% of Whites and 69% of Blacks remained engaged in the study. Current smoking status was an independent risk factor for study disengagement for both White and Black participants. For both groups, sites whose staffs missed SELECT training sessions or who received SELECT Retention and Adherence grants were associated with increased and decreased disengagement risks, respectively. SELECT retention was disproportionately lower for Blacks than for Whites. The observed difference in retention rates for Blacks and Whites and factors identified by race for study disengagement in SELECT may inform retention efforts for future long-term, cancer prevention trials.


TIEG1/KLF10 modulates Runx2 expression and activity in osteoblasts.
Deletion of TIEG1/KLF10 in mice results in a gender specific osteopenic skeletal phenotype with significant defects in both cortical and trabecular bone, which are observed only in female animals. Calvarial osteoblasts isolated from TIEG1 knockout (KO) mice display reduced expression levels of multiple bone related genes, including Runx2, and exhibit significant delays in their mineralization rates relative to wildtype controls. These data suggest that TIEG1 plays an important role in regulating Runx2 expression in bone and that decreased Runx2 expression in TIEG1 KO mice is in part responsible for the observed osteopenic phenotype. In this manuscript, data is presented demonstrating that over-expression of TIEG1 results in increased expression of Runx2 while repression of TIEG1 results in suppression of Runx2. Transient transfection and chromatin immunoprecipitation assays reveal that TIEG1 directly binds to and activates the Runx2 promoter. The zinc finger containing domain of TIEG1 is necessary for this regulation supporting that activation occurs through direct DNA binding. A role for the ubiquitin/proteasome pathway in fine tuning the regulation of Runx2 expression by TIEG1 is also implicated in this study. Additionally, the regulation of Runx2 expression by cytokines such as TGFβ1 and BMP2 is shown to be inhibited in the absence of TIEG1. Co-immunoprecipitation and co-localization assays indicate that TIEG1 protein associates with Runx2 protein resulting in co-activation of Runx2 transcriptional activity. Lastly, Runx2 adenoviral infection of TIEG1 KO calvarial osteoblasts leads to increased expression of Runx2 and enhancement of their ability to differentiate and mineralize in culture. Taken together, these data implicate an important role for TIEG1 in regulating the expression and activity of Runx2 in osteoblasts and suggest that decreased expression of Runx2 in TIEG1 KO mice contributes to the observed osteopenic bone phenotype.