Silicon uptake and metabolism of the marine diatom Thalassiosira pseudonana: Solid-state 29Si NMR and fluorescence microscopic studies.
Uptake and metabolism of silicon by diatoms are studied by the combined use of solid-state 29Si NMR spectroscopy and confocal laser fluorescence microscopy especially with respect to the presence and nature of an intracellular silicon-storage pool. Cells of the marine diatom Thalassiosira pseudonana were synchronized by silicon starvation and frozen without any freeze-drying or chemical treatment in order to analyze integer and unmodified diatoms. The frozen samples were investigated by solid-state 29Si NMR spectroscopy to identify potential silica precursors. The developmental state of the cell culture and the formation of new siliceous girdle bands and valves were monitored by laser fluorescence microscopic studies. A comparison of fluorescence microscopic and NMR data allows the assignment of NMR spectra to the various developmental stages of the dividing diatom cells. A detailed analysis of solid-state 29Si NMR spectra suggests that the silicon-storage pool-if present-consists of four-coordinated, condensed silicon; possibly a silica sol.


The kinetic deuterium isotope effect as applied to metabolic deactivation of imatinib to the des-methyl metabolite, CGP74588.
There has recently been a burgeoning interest in impeding drug metabolism by replacing hydrogen atoms with deuterium to invoke a kinetic isotope effect. Imatinib, a front-line therapy for both chronic myeloid leukemia and of gastrointestinal stromal tumours, is often substantially metabolised via N-demethylation to the significantly less active CGP74588. Since deuterium-carbon bonds are stronger than hydrogen-carbon bonds, we hypothesised that the N-trideuteromethyl analogue of imatinib might be subject to a reduced metabolic turnover as compared to imatinib and lead to different pharmacokinetic properties, and hence improved efficacy, in vivo. Consequently, we investigated whether the N-trideuteromethyl analogue would maintain target inhibition and show a reduced propensity for N-demethylation in in vitro assays with liver microsomes and following oral administration to rats. The N-trideuteromethyl compound exhibited similar activity as a tyrosine kinase inhibitor as imatinib and similar efficacy as an antiproliferative in cellular assays. In comparison to imatinib, the trideuterated analogue also showed reduced N-demethylation upon incubation with both rat and human liver microsomes, consistent with a deuterium isotope effect. However, the reduced in vitro metabolism did not translate into increased exposure of the N-trideuteromethyl analogue following intravenous administration of the compound to rats and no significant difference was observed for the formation of the N-desmethyl metabolite from either parent drug.


Risk factors for depression among older adults living alone in Shanghai, China.
Rapid population ageing and the increasing number of older adults living alone are important phenomena. Older adults are prone to depression. This study aimed to investigate the depression of older adults living alone in Shanghai and its risk factors. The data from the survey of quality of life of community-dwelling older adults in Shanghai from April 2015 to March 2016 was used in this study. A total of 387 older adults living alone in two communities in Shanghai participated in this study. Structured questionnaires were used to collect data, including depression, the presence of chronic diseases, functional ability, self-rated health, loneliness, social support and sociodemographic variables. The prevalence of depression was 26.9%. There were significant differences across the participants of different genders, education levels, previous occupations, economic levels, residential areas, physical activity levels, chronic diseases, functional ability levels, levels of self-rated health, loneliness levels and social support levels (P < 0.05). Binary logistic regression analyses found that self-rated health (odds ratio (OR) = 5.914, P = 0.001) and loneliness (OR = 1.132, P < 0.001) were risk factors of depression. The psychological and physical health of older adults living alone need to be emphasised. Appropriate interventions could be implemented to prevent depression, so as to promote the quality of life of older adults, and achieve active ageing and healthy ageing.


Limited heterogeneity of T-cell receptor V beta gene expression in the early stage of insulitis in NOD mice.
Non-obese-diabetic (NOD) mice spontaneously develop type I diabetes. The disease starts with T cells infiltrating the islets of Langerhans. We therefore examined the T-cell receptor (TCR) V beta region repertoire in islet infiltrates from individual female NOD mice from 4 to 10 week old by polymerase chain reaction (PCR) using V beta 1-V beta 17 specific oligonucleotides. The study revealed a limited heterogeneity of TCR V beta transcripts with a predominance of V beta 1 at the onset of insulitis, i.e. at 4 weeks of age. The TCR VDJ beta sequences of the V beta 1 PCR fragments were identical in most of the individual mice. Among several different mice, similarities in the beta-junctional regions were detected. In contrast, a large heterogeneity of TCR V beta usage was found in mice with advanced insulitis, i.e., from 6 weeks of age on. Thus, these data suggest a limited heterogeneity of TCR V beta usage with a predominance of V beta 1 at the initiation phase of the disease.


Functionalized 1,3-teraryls as a new class of hepatoprotectants. Part V.
Functionalized 1,3-teraryls, synthesized through ring transformation of 6-aryl-3-carbomethoxy-4-methylthio-2H-pyran-2-one from arylketone have been screened for their hepatoprotective activity and of them have demonstrated significant protection in animal model.


Study of Electron Ionization and Fragmentation of Non-hydrated and Hydrated Tetrahydrofuran Clusters.
Mass spectroscopic investigations on tetrahydrofuran (THF, C4H8O), a common model molecule of the DNA-backbone, have been carried out. We irradiated isolated THF and (hydrated) THF clusters with low energy electrons (electron energy ~70 eV) in order to study electron ionization and ionic fragmentation. For elucidation of fragmentation pathways, deuterated TDF (C4D8O) was investigated as well. One major observation is that the cluster environment shows overall a protective behavior on THF. However, also new fragmentation channels open in the cluster. In this context, we were able to solve a discrepancy in the literature about the fragment ion peak at mass 55 u in the electron ionization mass spectrum of THF. We ascribe this ion yield to the fragmentation of ionized THF clusters. Graphical Abstract .


Adenosine A1 receptor activation mediates NMDA receptor activity in a pertussis toxin-sensitive manner during normoxia but not anoxia in turtle cortical neurons.
Adenosine is a defensive metabolite that is critical to anoxic neuronal survival in the freshwater turtle. Channel arrest of the N-methyl-d-aspartate receptor (NMDAR) is a hallmark of the turtle's remarkable anoxia tolerance and adenosine A1 receptor (A1R)-mediated depression of normoxic NMDAR activity is well documented. However, experiments examining the role of A1Rs in regulating NMDAR activity during anoxia have yielded inconsistent results. The aim of this study was to examine the role of A1Rs in the normoxic and anoxic regulation of turtle brain NMDAR activity. Whole-cell NMDAR currents were recorded for up to 2 h from turtle cortical pyramidal neurons exposed to pharmacological A1R or Gi protein modulation during normoxia (95% O(2)/5% CO2) and anoxia (95% N2/5% CO2). NMDAR currents were unchanged during normoxia and decreased 51+/-4% following anoxic exposure. Normoxic agonism of A1Rs with adenosine or N6-cyclopentyladenosine (CPA) decreased NMDAR currents 57+/-11% and 59+/-6%, respectively. The A1R antagonist 8-cyclopentyl-1,3-dimethylxanthine (DPCPX) had no effect on normoxic NMDAR currents and prevented the adenosine and CPA-mediated decreases in NMDAR activity. DPCPX partially reduced the anoxic decrease at 20 but not 40 min of treatment. The Gi protein inhibitor pertussis toxin (PTX) prevented both the CPA and anoxia-mediated decreases in NMDAR currents and calcium chelation or blockade of mitochondrial ATP-sensitive K+ channels also prevented the CPA-mediated decreases. Our results suggest that the long-term anoxic decrease in NMDAR activity is activated by a PTX-sensitive mechanism that is independent of A1R activity.


Discovery of novel, potent and bioavailable proline-urea based macrocyclic HCV NS3/4A protease inhibitors.
A novel series of P3-truncated macrocyclic HCV NS3/4A protease inhibitors containing a P2 proline-urea or carbamate scaffold was synthesized. Very potent inhibitors were obtained through the optimization of the macrocycle size, urea and proline substitution, and bioisosteric replacement of the P1 carboxylic acid moiety. Variation of the lipophilicity by introduction of small lipophilic substituents resulted in improved PK profiles, ultimately leading to compound 13Bh, an extremely potent (K(i)=0.1 nM, EC(50)=4.5 nM) and selective (CC(50) (Huh-7 cells)>50 microM) inhibitor, displaying an excellent PK profile in rats characterized by an oral bioavailability of 54% and a high liver exposure after oral administration.


Serum neurofilament light chain is more strongly associated with T2 lesion volume than with number of T2 lesions in patients with multiple sclerosis.
MR imaging provides information on the number and extend of focal lesions in multiple sclerosis (MS) patients. This study explores whether total brain T2 lesion volume or lesion number shows a better correlation with serum and cerebrospinal fluid (CSF) biomarkers of disease activity. In total, 52 patients suffering from clinically isolated syndrome (CIS)/relapsing-remitting multiple sclerosis (RRMS) were assessed including MRI markers (total brain T2 lesion volume semi-automatically outlined on 3D DIR/FLAIR sequences, number of lesions), serum and CSF biomarkers at the time of neuroimaging (neurofilament light chain (NfL), glial fibrillary acidic protein (GFAP)), and clinical parameters. After log-transformation and partial correlations adjusted for the covariates patients' age, BMI, EDSS-score and diagnosis, the Fisher's r-to-Z transformation was used to compare different correlation coefficients. The correlation between lesion volume and serum NfL (r = 0.6, p < 0.001) was stronger compared to the association between the number of T2 lesions and serum NfL (r = 0.4, p < 0.01) (z = -2.0, p < 0.05). With regard to CSF NfL, there was a moderate, positive relationship for both number of T2 lesions and lesion volume (r = 0.5 respectively, p < 0.01). We found no significant association between MRI markers and GFAP levels. Our findings suggest that there is a stronger association between serum NfL and T2 lesion volume, than there is between serum NfL and T2 lesion number. Improving robustness and accuracy of fully-automated lesion volume segmentation tools can expedite implementation into clinical routine and trials.


Determining safe teaching board heights through electromyographic analysis of the shoulder muscles.
Adjustable teaching boards have been recommended as an ergonomic practice geared at reducing the incidences of teaching-related shoulder musculoskeletal disorders among teachers. However, there are no standards guiding choice of appropriate teaching board heights. This study was aimed at evaluating shoulder muscle activities in response to five teaching board heights. Thirty participants performed writing activities on five different board heights (Diff10 = standing height minus 10% of standing height; NormH = participant's standing height; Sum10 = standing height plus 10% of standing height; Sum20 = standing height plus 20% of standing height; Sum25 = standing height plus 25% of standing height). Electromyographic analysis of the anterior deltoid (AD), upper trapezius (UT), serratus anterior (SA) and pectoralis major (PM) muscles were performed simultaneously during each task. Data were analyzed with one-way ANOVA and independent T-test at p < 0.05. Sum20 board height significantly (p < 0.001) elicited the highest activities in the AD (19.35 ± 11.26%) and UT (29.06 ± 29.53%) muscles while SA (20.37 ± 10.87%) and PM (20.86 ± 12.17%) muscles were most active at Sum25 board height. SA and PM muscles progressively increased with increasing board heights while AD and UT muscles showed alternating patterns of activation at increasing board heights. For safe ergonomic practices, recommended teaching board heights should be within the range of NormH and Sum10. Extremely high or low teaching boards have tendencies of inducing musculoskeletal shoulder discomforts.


Mapping Pediatric Oncology Clinical Trial Collaborative Groups on the Global Stage.
The global pediatric oncology clinical research landscape, particularly in Central and South America, Africa, and Asia, which bear the highest burden of global childhood cancer cases, is less characterized in the literature. Review of how existing pediatric cancer clinical trial groups internationally have been formed and how their research goals have been pursued is critical for building global collaborative research and data-sharing efforts, in line with the WHO Global Initiative for Childhood Cancer. A narrative literature review of collaborative groups performing pediatric cancer clinical research in each continent was conducted. An inventory of research groups was assembled and reviewed by current pediatric cancer regional and continental leaders. Each group was narratively described with identification of common structural and research themes among consortia. There is wide variability in the structure, history, and goals of pediatric cancer clinical trial collaborative groups internationally. Several continental regions have longstanding endogenously-formed clinical trial groups that have developed and published numerous adapted treatment regimens to improve outcomes, whereas other regions have consortia focused on developing foundational database registry infrastructure supported by large multinational organizations or twinning relationships. There cannot be a one-size-fits-all approach to increasing collaboration between international pediatric cancer clinical trial groups, as this requires a nuanced understanding of local stakeholders and resources necessary to form partnerships. Needs assessments, performed either by local consortia or in conjunction with international partners, have generated productive clinical trial infrastructure. To achieve the goals of the Global Initiative for Childhood Cancer, global partnerships must be sufficiently granular to account for the distinct needs of each collaborating group and should incorporate grassroots approaches, robust twinning relationships, and implementation science.


The influence of plant water stress on stomatal control of gas exchange at different levels of atmospheric humidity.
Leaves of well-watered and mildly water-stressed seedlings of Betula pendula Roth. and Gmelina aroborea L. were subjected to a range of vapour pressure deficits (VPD) between 10 and 24 kPa. The stomatal conductance of birch seedlings decreased as VPD was increased and at least in mildly-stressed seedlings this response seemed to be closely linked to the water status of the air rather than to the bulk water status of the plant. Mild water stressing enhanced the degree of the stomatal humidity-response and resulted in a significant increase in the efficiency of water use at high VPD. Stomata of Gmelina were apparently insensitive to variation in VPD, but were more sensitive to a decrease in bulk leaf water status than were stomata of birch. Water use efficiency of Gmelina seedlings was comparatively high, even when VPD was high and the stomata were fully open.


Assessment of the feasibility of a rehabilitation intervention program for breast cancer survivors with cognitive complaints.
To assess the feasibility of a cognitive rehabilitation program in breast cancer survivors (BCS) with persistent post-treatment cognitive complaints. BCS with cognitive complaints, 18-months to 5-years post-treatment, were recruited for a once-weekly, five-week, group cognitive training intervention. Outcome measures included self-reported mood and cognitive function, and neurocognitive tests administered at pre-intervention, immediate-, two-month and four-month post-intervention. A sub-study in eight participants evaluated resting state quantitative electroencephalography (qEEG) changes from pre- to immediate post-intervention in relationship to post-intervention changes in cognitive complaints. Twenty-seven BCS completed the protocol and tolerated the intervention well. We observed significant reductions in total and memory-specific cognitive complaints from pre-intervention to immediate post-intervention (p = 0.031 and p = 0.009, respectively) and at four-months post-intervention (p < 0.0001 and p < 0.001, respectively). Significant improvement in neurocognitive tests were found for Symbol Digit, Stroop, and Trails A tests (df = 26, all p's <0.05). Effect sizes for changes from pre-intervention to immediate and to four-month post intervention ranged from 0.429 to 0.607, and from 0.439 to 0.741, respectively. Increase in qEEG absolute alpha power over the course of the intervention was associated with reduced complaints at immediate post-intervention (r = -0.78, p = 0.021), two-months (r range = -0.76 to -0.82, p-value range 0.004 to 0.03), and four-months (r = -0.71, p = 0.048). A five-week group cognitive training intervention is feasible and well tolerated. Cognitive complaints and neurocognitive test performances showed positive changes. qEEG may serve as a potential biomarker for improvement in self-reported complaints. A randomized clinical trial is underway to test the efficacy of the intervention.


Bacterial co-infection of the respiratory tract in ventilated children with bronchiolitis; a retrospective cohort study.
Viral bronchiolitis is the most common cause of respiratory failure requiring invasive ventilation in young children. Bacterial co-infections may complicate and prolong paediatric intensive care unit (PICU) stay. Data on prevalence, type of pathogens and its association with disease severity are limited though. These data are especially important as bacterial co-infections may be treated using antibiotics and could reduce disease severity and duration of PICU stay. We investigated prevalence of bacterial co-infection and its association with disease severity and PICU stay. Retrospective cohort study of the prevalence and type of bacterial co-infections in ventilated children performed in a 14-bed tertiary care PICU in The Netherlands. Children less than 2 years of age admitted between December 2006 and November 2014 with a diagnosis of bronchiolitis and requiring invasive mechanical ventilation were included. Tracheal aspirates (TA) and broncho-alveolar lavages (BAL) were cultured and scored based on the quantity of bacteria colony forming units (CFU) as: co-infection (TA > 10^5/BAL > 10^4 CFU), low bacterial growth (TA < 10^5/BAL < 10^4 CFU), or negative (no growth). Duration of mechanical ventilation and PICU stay were collected using medical records and compared against the presence of co-infection using univariate and multivariate analysis. Of 167 included children 63 (37.7%) had a bacterial co-infection and 67 (40.1%) low bacterial growth. Co-infections occurred within 48 h from intubation in 52 out 63 (82.5%) co-infections. H.influenza (40.0%), S.pneumoniae (27.1%), M.catarrhalis (22.4%), and S.aureus (7.1%) were the most common pathogens. PICU stay and mechanical ventilation lasted longer in children with co-infections than children with negative cultures (9.1 vs 7.7 days, p = 0.04 and 8.1vs 6.5 days, p = 0.02). In this large study, bacterial co-infections occurred in more than a third of children requiring invasive ventilation for bronchiolitis and were associated with longer PICU stay and mechanical ventilation. These findings support a clinical trial of antibiotics to test whether antibiotics can reduce duration of PICU stay.


Pulse pattern of luteinizing hormone (LH) in patients with varicocele: a preliminary report.
Serum concentrations of LH were measured every 20 minutes for 6 hours in 4 varicocele patients and 4 control subjects. In both groups, a pulsatile pattern of LH secretion was observed. Also in the two groups, LH pulse rate (2-5/6 hour) and amplitude (23.5-61%) were identical. These findings suggest that Gonadotrophin releasing hormone (Gn RH) and LH secretions have no role in the deranged sperm parameters often found in infertile men with varicocele.


MicroRNA-186-5p controls GluA2 surface expression and synaptic scaling in hippocampal neurons.
Homeostatic synaptic scaling is a negative feedback response to fluctuations in synaptic strength induced by developmental or learning-related processes, which maintains neuronal activity stable. Although several components of the synaptic scaling apparatus have been characterized, the intrinsic regulatory mechanisms promoting scaling remain largely unknown. MicroRNAs may contribute to posttranscriptional control of mRNAs implicated in different stages of synaptic scaling, but their role in these mechanisms is still undervalued. Here, we report that chronic blockade of glutamate receptors of the AMPA and NMDA types in hippocampal neurons in culture induces changes in the neuronal mRNA and miRNA transcriptomes, leading to synaptic upscaling. Specifically, we show that synaptic activity blockade persistently down-regulates miR-186-5p. Moreover, we describe a conserved miR-186-5p-binding site within the 3'UTR of the mRNA encoding the AMPA receptor GluA2 subunit, and demonstrate that GluA2 is a direct target of miR-186-5p. Overexpression of miR-186 decreased GluA2 surface levels, increased synaptic expression of GluA2-lacking AMPA receptors, and blocked synaptic scaling, whereas inhibition of miR-186-5p increased GluA2 surface levels and the amplitude and frequency of AMPA receptor-mediated currents, and mimicked excitatory synaptic scaling induced by synaptic inactivity. Our findings elucidate an activity-dependent miRNA-mediated mechanism for regulation of AMPA receptor expression.


Opioid Prescribing in the 2016 Medicare Fee-for-Service Population.
Opioid use and misuse are prevalent and remain a national crisis. This study identified beneficiary characteristics associated with filling opioid prescriptions, variation in opioid dosing, and opioid use with average daily doses (ADDs) equal to 120 morphine milligram equivalents (MMEs) or more in the 100% Medicare fee-for-service (FFS) population. In a cohort of FFS beneficiaries with 12 months of Medicare Part D coverage in 2016, we examined patient factors associated with filling an opioid prescription (n = 20,880,490) and variation in ADDs (n = 7,325,031) in a two-part model. Among those filling opioids, we also examined the probability of ADD equal to 120 MMEs or more via logistic regression. About 35% of FFS beneficiaries had one or more opioid prescription fills in 2016 and 1.5% had ADDs equal to 120 MMEs or more. Disability-eligible beneficiaries and beneficiaries with multiple chronic conditions were more likely to fill opioids, to have higher ADDs or were more likely to have ADD equal to 120 MMEs or more. Beneficiaries with chronic obstructive pulmonary disease (COPD) were more likely to fill opioids (odds ratio (OR) = 1.47, 95% confidence interval (CI) = 1.46-1.47), have higher ADDs (rate ratio = 1.06, 95% CI = 1.06-1.06) when filled and were more likely to have ADD equal to 120 MMEs or more (OR = 1.23, 95% CI = 1.21-1.24). Finally, black and Hispanic beneficiaries were less likely to fill opioids, had lower overall doses and were less likely to have ADDs equal to 120 MMEs or more compared to white beneficiaries. Several beneficiary subgroups have underappreciated risk of adverse events associated with ADD equal to 120 MMEs or more that may benefit from opioid optimization interventions that balance pain management and adverse event risk, especially beneficiaries with COPD who are at risk for respiratory depression.


Factors related to delays in obtaining contraception among pregnancy-capable adults in New York state during the COVID-19 pandemic: The CAP study.
To investigate factors associated with delays to obtaining contraception during the COVID-19 pandemic among pregnancy-capable adults in New York State. We administered a cross-sectional survey in June-July 2020 to female/transgender male New York State residents aged 18-44 years (n = 1,525). This analysis focused on respondents who were not pregnant and sought contraception (n = 953). We conducted bivariate and multinomial logistic regression analyses to assess sociodemographic, social, and health characteristics, by the outcome of delays to obtaining birth control (delayed due to COVID-19, delayed due to other reasons, no delay). We also analyzed a sub-sample who reported COVID-19 as a reason for delays (n = 317) and report the frequencies of type of contraceptive methods/procedures delayed and availability of telemedicine visits. Half of respondents had no contraceptive delays, 39% reported delays due to COVID-19, and 11% reported delays due to reasons other than COVID-19. In adjusted analyses, those who missed a rent/mortgage payment during the pandemic (aOR: 2.23; CI: 1.55, 3.22), participated in a supplemental government program in 2019 (aOR: 1.88; CI: 1.36, 2.60), and themselves/household member had COVID-19 (aOR: 1.48; CI: 1.04, 2.12) were more likely to report delays to contraception due to COVID-19 (versus no delays). In the sub-sample, 63% reported available virtual contraceptive visits, 28% unavailable, and 9% not sure. The most frequently (42%) reported delays were new prescriptions for the pill, patch, or ring. Reducing financial barriers that help individuals maintain their housing and living necessities, and promoting telemedicine visits, may help increase access to contraception.


Plant immunity in soybean: progress, strategies, and perspectives.
Soybean (Glycine max) is one of the most important commercial crops worldwide. Soybean hosts diverse microbes, including pathogens that may cause diseases and symbionts that contribute to nitrogen fixation. Study on soybean-microbe interactions to understand pathogenesis, immunity, and symbiosis represents an important research direction toward plant protection in soybean. In terms of immune mechanisms, current research in soybean lags far behind that in the model plants Arabidopsis and rice. In this review, we summarized the shared and unique mechanisms involved in the two-tiered plant immunity and the virulence function of pathogen effectors between soybean and Arabidopsis, providing a molecular roadmap for future research on soybean immunity. We also discussed disease resistance engineering and future perspectives in soybean.


Haematological indices as surrogate markers of factors affecting mortality after hip fracture.
This study investigated the relationship between different components of the full blood count, such as haemoglobin (Hb), total white cell count, total lymphocyte count on admission and total neutrophil count as possible surrogate markers of conditions that lead to increased mortality in hip-fracture patients. A total of 791 patients were studied with 81.2% being females. The 1-year mortality was 26.4%(32.9% in males compared with 24.9% in females). A significant increase in mortality was seen in anaemic patients, especially with Hb 80–100 g l(-1) (1-year mortality was 49.2%, p-value < 0.001) and patients with lymphocyte count ≤ 1.1 × 10(9) l(-1) (33.2% mortality, p-value < 0.0001). There was no relationship between total white cell count or neutrophil count and mortality. This information clearly suggests that Hb and total lymphocyte counts may be surrogate markers for increased mortality after a hip fracture and can be used for audit purposes to adjust for different case mixes between groups.


A Paratesticular Serous Borderline Tumor in a Pediatric Patient With Proteus Syndrome.
Proteus syndrome is a rare disorder of asymmetric overgrowth of various tissues of the body and is associated with specific tumors appearing before the second decade. Although there have been reports of lesions of the genitourinary tract associated with Proteus syndrome, a case of serous borderline tumor of the paratestis has not been previously recorded. We report the first such case in a 20-month-old child who presented with a left-sided testicular mass that was found on histology to be a serous borderline tumor of the paratestis. Surgical management included a left inguinal radical orchiectomy and surveillance follow-up.


Insulin down-regulates cardioprotective SUR2A in the heart-derived H9c2 cells: A possible explanation for some adverse effects of insulin therapy.
Some recent studies associated insulin therapy with negative cardiovascular events and shorter lifespan. SUR2A, a KATP channel subunit, regulate cardioprotection and cardiac ageing. Here, we have tested whether glucose and insulin regulate expression of SUR2A/KATP channel subunits and resistance to metabolic stress in heart H9c2 cells. Absence of glucose in culture media decreased SUR2A mRNA, while mRNAs of Kir6.2, Kir6.1, SUR1 and IES SUR2B were increased. 2-deoxyglucose (50 mM) decreased mRNAs of SUR2A, SUR2B and SUR1, did not affect IES SUR2A and IES SUR2B mRNAs and increased Kir6.2 mRNA. No glucose and 2-deoxyglucose (50 mM) decreased resistance to an inhibitor of oxidative phosphorylation, DNP (10 mM). 50 mM glucose did not alter KATP channel subunits nor cellular resistance to DNP (10 mM). Insulin (20 ng/ml) in both physiological and high glucose (50 mM) down-regulated SUR2A while upregulating Kir6.1 and Kir6.2 (in high glucose only). Insulin (20 ng/ml) in physiological and high glucose decreased cell survival in DNP (10 mM). As opposed to Kir6.2, infection with SUR2A resulted in titre-dependent cytoprotection. We conclude that insulin decreases resistance to metabolic stress in H9c2 cells by decreasing SUR2A expression. Lower cardiac SUR2A levels underlie increased myocardial susceptibility to metabolic stress and shorter lifespan.


Pelvic stability during simulated total hip arthroplasty motions: Comparing different hip positioners.
Total hip arthroplasty (THA) requires forceful maneuvers that can cause the pelvis to shift from its original position. Various methods for stabilizing the pelvis in the lateral decubitus position exist, but there is limited data quantifying the relative stability of each hip positioner. We sought to quantify the pelvic movement that occurred in four commercially available hip positioners during surgeon induced motion of the hip. An infrared marker was attached to the ilium of a cadaver secured in the lateral decubitus position. Four commercially available hip positioners were used for positioning: Beanbag, Pegboard, Stulberg, and ExactFit. Rotation and translation was captured using an infrared marker and camera system while the hip was moved through six motions (Flexion, Extension, Internal Rotation, External Rotation, Push, and Pull). The Beanbag had the greatest amount of rotation and translation of the pelvis, with maximum hip rotation of 41.5°. The Stulberg and Pegboard positioners showed intermediate stability, with a maximum rotation of 7.8° and 17.1°, respectively. The ExactFit hip positioner resulted in the least amount of motion of the pelvis, with a maximum rotation of the pelvis of up to 3.2°. Of the simulated motions performed, internal rotation and flexion of the hip led to the greatest changes in pelvic rotation and translation. The ExactFit positioner was associated with the smallest amount of pelvic motion during simulated motions of hip arthroplasty, followed by the Stulberg, Pegboard, and Beanbag positioners. Further studies are required to correlate this information with clinical outcomes following total hip arthroplasty.


Clinical and electrophysiologic effects of amiodarone in patients with atrial fibrillation complicating the Wolff-Parkinson-White syndrome.
The clinical and electrophysiologic effects of oral amiodarone were evaluated in 10 patients with a history of spontaneous atrial fibrillation and a rapid ventricular response, complicating the Wolff-Parkinson-White syndrome. Five patients developed ventricular fibrillation during an episode of atrial fibrillation. Seven patients underwent electrophysiology study and arrhythmia induction, both before and during chronic amiodarone treatment. During a mean 30 months of follow-up (range 8 to 59 months), no recurrences of atrial fibrillation or ventricular fibrillation were noted. One patient had recurrent supraventricular tachycardia and underwent surgery. One had serious and five had minor side effects. Arrhythmia suppression was associated with prolongation of the anterograde accessory pathway (+38%, p less than 0.01) and atrial (+34%, p less than 0.01) effective refractory periods. Amiodarone also slowed the ventricular response to induced atrial fibrillation, prolonging the mean (+90%, p less than 0.01) and minimum (+104%, p = 0.01) R-R intervals. Thus, amiodarone is a safe and effective alternative for arrhythmia prevention if surgery is inadvisable or is not desired by the patient.


Analysis and occurrence of endocrine-disrupting compounds and estrogenic activity in the surface waters of Central Spain.
Endocrine-disrupting compounds (EDCs) are chemical compounds with the ability to alter the hormonal systems of organisms. Such compounds are used in several industrial and domestic activities and reach the aquatic environment via wastewater discharge. The aim of this study is to assess the occurrence of 30 EDCs and related compounds in the surface waters of central Spain and to determine the overall estrogenic activity of environmental samples. This study analyzed a large number of EDCs and other emergent or suspected compounds with endocrine-disrupting activity. The results have shown the presence of 19 EDCs at concentrations ranging from 2 to 5928 ng L(-1). Organophosphorus-based flame retardants, alkylphenolic compounds and anticorrosives were found at the highest concentrations. Furthermore, although insufficient data are available to calculate an average over time, these preliminary results show the need to monitor the waters in both rivers studied. Alkylphenolic compounds, particularly nonylphenol, were the main contributors to overall estrogenicity. A higher concentration of the compounds studied was detected in the river Jarama, although the estrogenicity expressed as estradiol equivalents (EEQs) was higher in the river Manzanares due to a higher concentration of nonylphenol. However, the total estrogenicity did not exceed 1 ng L(-1) (EEQ), which is the level that may cause estrogenic effects in aquatic organisms, in any of the samples. In conclusion, the potential estrogenic risk in both rivers is low, although organophosphorus-based flame retardants may increase this risk as they were found at high levels in all samples. Unfortunately, these compounds could not be taken into account when calculating the estrogenic activity due to the lack of activity data for them. For future investigations, it will be important to assess the estrogenicity provided by these flame retardants. Due to the significant concentrations of EDCs detected in both rivers, further studies in this region are required.


Rab13 Sustains Breast Cancer Stem Cells by Supporting Tumor-Stroma Cross-talk.
Cancer stem cells (CSC) are supported by the tumor microenvironment, and non-CSCs can regain CSC phenotypes in certain niches, leading to limited clinical benefits of CSC-targeted therapy. A better understanding of the mechanisms governing the orchestration of the CSC niche could help improve the therapeutic targeting of CSCs. Here, we report that Rab13, a small GTPase, is highly expressed in breast CSCs (BCSC). Rab13 depletion suppressed breast cancer cell stemness, tumorigenesis, and chemoresistance by reducing tumor-stroma cross-talk. Accordingly, Rab13 controlled the membrane translocation of C-X-C chemokine receptor type 1/2 (CXCR1/2), allowing tumor cells to interact with tumor-associated macrophages and cancer-associated fibroblasts to establish a supportive BCSC niche. Targeting the Rab13-mediated BCSC niche with bardoxolone-methyl (C-28 methyl ester of 2-cyano-3, 12-dioxoolen-1, 9-dien-28-oic acid; CDDO-Me) prevented BCSC stemness in vitro and in vivo. These findings highlight the novel regulatory mechanism of Rab13 in BCSC, with important implications for the development of therapeutic strategies for disrupting the BCSC niche. Targeting Rab13 perturbs formation of the breast cancer stem cell niche by inhibiting cross-talk between cancer cells and the tumor microenvironment, providing a therapeutic opportunity for niche-targeted breast cancer treatment.


Fruquintinib versus placebo in patients with refractory metastatic colorectal cancer (FRESCO-2): an international, multicentre, randomised, double-blind, phase 3 study.
There is a paucity of effective systemic therapy options for patients with advanced, chemotherapy-refractory colorectal cancer. We aimed to evaluate the efficacy and safety of fruquintinib, a highly selective and potent oral inhibitor of vascular endothelial growth factor receptors (VEGFRs) 1, 2, and 3, in patients with heavily pretreated metastatic colorectal cancer. We conducted an international, randomised, double-blind, placebo-controlled, phase 3 study (FRESCO-2) at 124 hospitals and cancer centres across 14 countries. We included patients aged 18 years or older (≥20 years in Japan) with histologically or cytologically documented metastatic colorectal adenocarcinoma who had received all current standard approved cytotoxic and targeted therapies and progressed on or were intolerant to trifluridine-tipiracil or regorafenib, or both. Eligible patients were randomly assigned (2:1) to receive fruquintinib (5 mg capsule) or matched placebo orally once daily on days 1-21 in 28-day cycles, plus best supportive care. Stratification factors were previous trifluridine-tipiracil or regorafenib, or both, RAS mutation status, and duration of metastatic disease. Patients, investigators, study site personnel, and sponsors, except for selected sponsor pharmacovigilance personnel, were masked to study group assignments. The primary endpoint was overall survival, defined as the time from randomisation to death from any cause. A non-binding futility analysis was done when approximately one-third of the expected overall survival events had occurred. Final analysis occurred after 480 overall survival events. This study is registered with ClinicalTrials.gov, NCT04322539, and EudraCT, 2020-000158-88, and is ongoing but not recruiting. Between Aug 12, 2020, and Dec 2, 2021, 934 patients were assessed for eligibility and 691 were enrolled and randomly assigned to receive fruquintinib (n=461) or placebo (n=230). Patients had received a median of 4 lines (IQR 3-6) of previous systemic therapy for metastatic disease, and 502 (73%) of 691 patients had received more than 3 lines. Median overall survival was 7·4 months (95% CI 6·7-8·2) in the fruquintinib group versus 4·8 months (4·0-5·8) in the placebo group (hazard ratio 0·66, 95% CI 0·55-0·80; p<0·0001). Grade 3 or worse adverse events occurred in 286 (63%) of 456 patients who received fruquintinib and 116 (50%) of 230 who received placebo; the most common grade 3 or worse adverse events in the fruquintinib group included hypertension (n=62 [14%]), asthenia (n=35 [8%]), and hand-foot syndrome (n=29 [6%]). There was one treatment-related death in each group (intestinal perforation in the fruquintinib group and cardiac arrest in the placebo group). Fruquintinib treatment resulted in a significant and clinically meaningful benefit in overall survival compared with placebo in patients with refractory metastatic colorectal cancer. These data support the use of fruquintinib as a global treatment option for patients with refractory metastatic colorectal cancer. Ongoing analysis of the quality of life data will further establish the clinical benefit of fruquintinib in this patient population. HUTCHMED.


Associations of adolescent lifestyle behaviors with body mass index within a nationally representative sample of US adolescents: a quantile regression analysis.
The purpose of this study was to examine the associations between lifestyle behavior variables such as physical activity, television watching, computer use, and school night sleep duration with body mass index percentile (BMI%) using quantile regression within a representative sample of adolescents who completed the 2017 US National Youth Risk Behavior Survey. The study design was a cross-sectional study. A multistage cluster sampling procedure obtained a representative sample of US adolescents. The number of sampled adolescents submitting questionnaires with BMI% data was 13,146. To examine the associations between lifestyle behaviors and BMI%, simultaneous quantile regression was used. When relationships were modeled at every 10th percentile, more precise parameter estimates were observed at higher percentiles. Across the interquartile range, physical activity associated with lower BMI% at the 50th and 75th percentiles (brange = -2.27% to -5.24%, P < 0.05), television watching associated with higher BMI% at the 25th to 75th percentiles (brange = 2.29%-4.16%, P < 0.05), sleep durations less than 8 h per school night associated with higher BMI% at the 25th and 50th percentile (brange = 2.81%-8.26%, P < 0.05), and 10 or more hours of school night sleep associated with higher BMI% at the 50th and 75th percentile (brange = 3.43%-7.53%, P < 0.05). Higher levels of physical activity associated with lower BMI% and longer time watching television, school night sleep durations less than 8 h, and school night sleep durations of 10 h or more at higher quantiles associated with higher BMI% in adolescents. Estimates of association were more precise within higher quantiles.


Prognostic implications of different clinical profiles in hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is a myocardial genetic disease relatively common in the general population with heterogenous clinical presentation, natural history and prognosis. About 60% of HCM patients have a stable clinical course, while others may experience a variety of HCM-related complications which follows relatively independent pathways, and that can be distinguished in different subgroups. These subgroups are represented by patients with left ventricular outflow tract obstruction; patients with end-stage disease and reduced or preserved systolic function; patients with apical hypertrophy; patients with apical aneurysm; patients with atrial fibrillation, patients at high risk of sudden death and patients with preclinical HCM. The purpose of this review was to describe each of these clinical profiles with its prognostic implications.


Patterns in Patient Encounters and Emergency Department Capacity in California, 2011-2021.
The health care system has undergone major changes in the past decade, and emergency department (ED) crowding has worsened over time; however, the most recent patterns in ED capacity and use in California have yet to be studied. To analyze patterns in ED capacity and utilization in California hospitals from 2011 to 2021. This retrospective cohort study used data from the California Department of Health Care Access and Information and the US Census Bureau to analyze ED facility characteristics from more than 400 general acute care hospitals with more than 320 EDs in California as well as patients who presented to those EDs between January 1, 2011, and December 31, 2021. Linear patterns (measured by percentage change) in total annual ED capacity (volume of hospital beds, EDs, ED treatment stations, and trauma centers) and ED use (ED visits by disposition and acuity) were assessed as primary outcomes. Patterns in ambulance diversion hours and the number of patients who left the ED without being seen were also examined as secondary outcomes. Visit acuity was categorized into 5 levels by increasing severity (minor, low to moderate, moderate, severe without threat, and severe with threat) based on California Department of Health Care Access and Information descriptions corresponding to Current Procedural Terminology codes. In the prepandemic period (2011-2019), the total population of California increased from 37 638 369 to 39 512 223 (5.0%; 95% CI, 4.1%-5.8%), then decreased to 39 237 836 in 2021 (0.7%; 95% CI, -3.9% to 2.5%). Over the entire study period (2011-2021), the total California population increased by 4.2% (95% CI, 3.3%-5.2%). From 2011 to 2019, the annual number of ED visits increased from 12 054 885 to 14 876 653 (23.4%; 95% CI, 20.0%-26.8%) before decreasing to 12 944 692 in 2021 (-13.0%; 95% CI, -33.1% to 7.1%); from 2011 to 2021, total ED visits increased by 7.4% (95% CI, 5.6%-9.1%). From 2011 to 2021, the total number of EDs decreased from 339 to 326 (-3.8%; 95% CI, -4.4% to -3.2%) and the total number of hospital beds decreased from 75 940 to 74 052 (-2.5%; 95% CI, -3.3% to -1.6%), while the number of ED treatment stations in these fewer EDs increased from 7159 to 8667 (21.1%; 95% CI, 19.7%-22.4%). The number of visits rated as severe with threat also increased, from 2 011 637 in 2011 to 3 375 539 in 2021 (67.8%; 95% CI, 59.7%-75.9%), while visits rated as minor decreased from 913 712 to 336 071 (-63.2%; 95% CI, -75.2% to -51.2%) over the same period. In this cohort study, multiple measures of ED capacity did not proportionally increase with the increasing demand for services; however, the COVID-19 pandemic appears to have substantially affected some of these patterns. These findings may be helpful to policy makers and health care stakeholders when planning resource allocation of limited health care resources.


TRAF4-mediated nonproteolytic ubiquitination of androgen receptor promotes castration-resistant prostate cancer.
Castration-resistant prostate cancer (CRPC) poses a major clinical challenge with the androgen receptor (AR) remaining to be a critical oncogenic player. Several lines of evidence indicate that AR induces a distinct transcriptional program after androgen deprivation in CRPCs. However, the mechanism triggering AR binding to a distinct set of genomic loci in CRPC and how it promotes CRPC development remain unclear. We demonstrate here that atypical ubiquitination of AR mediated by an E3 ubiquitin ligase TRAF4 plays an important role in this process. TRAF4 is highly expressed in CRPCs and promotes CRPC development. It mediates K27-linked ubiquitination at the C-terminal tail of AR and increases its association with the pioneer factor FOXA1. Consequently, AR binds to a distinct set of genomic loci enriched with FOXA1- and HOXB13-binding motifs to drive different transcriptional programs including an olfactory transduction pathway. Through the surprising upregulation of olfactory receptor gene transcription, TRAF4 increases intracellular cAMP levels and boosts E2F transcription factor activity to promote cell proliferation under androgen deprivation conditions. Altogether, these findings reveal a posttranslational mechanism driving AR-regulated transcriptional reprogramming to provide survival advantages for prostate cancer cells under castration conditions.


Endothelin inhibits adenylate cyclase and stimulates phosphoinositide hydrolysis in adult cardiac myocytes.
We have assessed the effects of endothelin-1 (ET-1) on transmembrane signaling in adult rat ventricular myocytes. ET-1 stimulates phosphoinositide hydrolysis with an EC50 of 0.3-0.8 nM. This stimulation is linear for up to 30 min in the presence of a protease inhibitor, is additive with the effects of other stimulators of phosphoinositide hydrolysis, is not inhibited by the Ca2+ entry blocker, nifedipine, and is insensitive to pertussis toxin. ET-1 also reduces cyclic AMP production in myocytes in response to isoproterenol and forskolin (EC50, 1 nM). This cyclic AMP-lowering effect of ET-1 is sensitive to pertussis toxin, can be demonstrated directly in assays of adenylate cyclase activity of myocyte membranes, and seems to be mediated by Gi. These data indicate that the effects of endothelin on adult cardiac myocytes involve multiple signaling pathways, including enhanced activity of the inositol phosphate pathway and a decrease in cyclic AMP-mediated responses, neither of which seems likely to account for the positive contractile effects of endothelin.


Glucose activation of liver glycogen synthase. Insulin-mediated restoration of glucose effect in diabetic rats is blocked by protein synthesis inhibitor.
The loss of glucose regulation of glycogen synthase in perfused livers from diabetic rats was associated with a substantial reduction in synthase phosphatase activity. Treatment of diabetic rats with insulin alone resulted in total restoration of the glucose effect and synthase phosphatase activity, while simultaneous treatment with cycloheximide severely reduced the hormonal effect. Although treatment of normal rats with cycloheximide had no effect on glucose activation of synthase, it did result in severe depletion of liver glycogen, increased liver glycogen phosphorylase activity, and elevation of liver adenosine 3',5'-monophosphate (cyclic AMP), but without elevation of liver protein kinase activity. Simultaneous treatment of alloxan-diabetic rats with insulin and cycloheximide resulted in reduction of total liver glycogen, increased phosphorylase activity, a reduction in the ability of insulin to lower hepatic cyclic AMP, and a further reduction of protein kinase activity. In summary, the effect of insulin treatment of diabetic rats to restore glucose regulation of hepatic glycogen synthase probably involves synthesis of new protein, and the data remain consistent with the hypothesis that the defect may be due to a diabetes-related deficiency in a specific synthase phosphatase and/or alteration of the synthase molecule itself.


Prevalence of visual impairment in adults with intellectual disabilities in the Netherlands: cross-sectional study.
To obtain the first representative and valid population-based prevalence figures on visual impairment and blindness in adults with intellectual disabilities (ID) and to identify risk groups. Cross-sectional survey. An age-Down's syndrome-stratified random sample of 1,598 persons from a base population of 9,012 adult users of ID services with mild to profound intellectual disabilities was screened. Participants underwent protocollised on-site screening of visual functions. Results were related to degree of ID, occurrence of Down's syndrome (DS) and age. Prevalences of visual impairment and blindness in the study population and in subgroups and weighted prevalences in the total Dutch population using ID services. Prevalences of visual impairment ranged from 2.2% (95% confidence interval (CI), 0.5-6.4) in young adults with mild ID and no Down's syndrome to 66.7% (95% CI, 41.0-86.7) in older adults with profound ID and Down's syndrome; prevalences of blindness ranged from 0.7% (95% CI, 0.1-4.1) to 38.9% (95% CI, 28.1-50.3). Weighted prevalences of visual impairment and blindness in the total Dutch population of adult users of intellectual disability services are 13.8% (95% CI, 9.3-18.4) and 5.0% (95% CI, 3.8-6.2), respectively. Prior to this study, visual impairment or blindness had remained undiagnosed in 106/261 (40.6%) persons. As compared to published figures for the general Dutch population aged 55 years and over (visual impairment 1.4%, blindness 0.5%), prevalences of visual impairment and blindness are higher in all subgroups with intellectual disabilities, including the young and mildly handicapped group. The diagnosis is too often missed. All persons with severe or profound intellectual disabilities, and all older adults with Down's syndrome, should be considered visually impaired until proved otherwise.


Transgenesis and imaging in birds, and available transgenic reporter lines.
Avian embryos are important model organism to study higher vertebrate development. Easy accessibility to developing avian embryos enables a variety of experimental applications to understand specific functions of molecules, tissue-tissue interactions, and cell lineages. The whole-mount ex ovo culture technique for avian embryos permits time-lapse imaging analysis for a better understanding of cell behaviors underlying tissue morphogenesis in physiological conditions. To study mechanisms of blood vessel formation and remodeling in developing embryos by using a time-lapse imaging approach, a transgenic quail model, Tg(tie1:H2B-eYFP), was generated. From a cell behavior perspective, Tg(tie1:H2B-eYFP) quail embryos are a suitable model to shed light on how the structure and pattern of blood vessels are established in higher vertebrates. In this manuscript, we give an overview on the biological and technological background of the transgenic quail model and describe procedures for the ex ovo culture of quail embryos and time-lapse imaging analysis.


Liver-Targeted AAV8 Gene Therapy Ameliorates Skeletal and Cardiovascular Pathology in a Mucopolysaccharidosis IVA Murine Model.
Mucopolysaccharidosis type IVA (MPS IVA) is due to the deficiency of GALNS (N-acetylgalactosamine 6-sulfate sulfatase) and is characterized by systemic skeletal dysplasia. We have evaluated adeno-associated virus 8 (AAV8) vectors expressing different forms of human GALNS under a liver-specific promoter. The vectors were delivered intravenously into 4-week-old MPS IVA knockout (KO) and immune tolerant (MTOL) mice at a dose of 5 × 1013 genome copies (GC)/kg. These mice were monitored for 12 weeks post-injection. GALNS enzyme activity was elevated significantly in plasma of all treated mice at 2 weeks post-injection. The activity observed was 4- to 19-fold higher than that in wild-type mice and was maintained throughout the monitoring period. Treatment with AAV vectors resulted in a reduction of keratan sulfate (KS) levels in plasma to normal levels 2 weeks post-injection, which were maintained until necropsy. Both vectors reduced the storage in articular cartilage, ligaments, and meniscus surrounding articular cartilage and growth plate region as well as heart muscle and valves. Our results suggest that the continuous presence of high levels of circulating enzyme increases the penetration into bone and heart and reduces the KS level, thereby improving storage in these regions. The current data support a strategy for developing a novel treatment to address the bone and heart disease in MPS IVA using AAV gene therapy.


Peripheral brain-derived neurotrophic factor (BDNF) as a biomarker in bipolar disorder: a meta-analysis of 52 studies.
The neurotrophic hypothesis postulates that mood disorders such as bipolar disorder (BD) are associated with a lower expression of brain-derived neurotrophic factor (BDNF). However, its role in peripheral blood as a biomarker of disease activity and of stage for BD, transcending pathophysiology, is still disputed. In the last few years an increasing number of clinical studies assessing BDNF in serum and plasma have been published. Therefore, it is now possible to analyse the association between BDNF levels and the severity of affective symptoms in BD as well as the effects of acute drug treatment of mood episodes on BDNF levels. We conducted a systematic review and meta-analysis of all studies on serum and plasma BDNF levels in bipolar disorder. Through a series of meta-analyses including a total of 52 studies with 6,481 participants, we show that, compared to healthy controls, peripheral BDNF levels are reduced to the same extent in manic (Hedges' g = -0.57, P = 0.010) and depressive (Hedges' g = -0.93, P = 0.001) episodes, while BDNF levels are not significantly altered in euthymia. In meta-regression analyses, BDNF levels additionally negatively correlate with the severity of both manic and depressive symptoms. We found no evidence for a significant impact of illness duration on BDNF levels. In addition, in plasma, but not serum, peripheral BDNF levels increase after the successful treatment of an acute mania episode, but not of a depressive one. In summary, our data suggest that peripheral BDNF levels, more clearly in plasma than in serum, is a potential biomarker of disease activity in BD, but not a biomarker of stage. We suggest that peripheral BDNF may, in future, be used as a part of a blood protein composite measure to assess disease activity in BD.


One-hour blood xylose test: a reliable index of small bowel function.
A retrospective analysis of oral D-xylose tolerance tests in 435 pediatric patients was performed. A significant difference was found between 126 normal subjects and 47 untreated children with celiac disease one hour after load. The one-hour value was found to be more reliable than was fecal fat analysis in screening children for celiac disease. Sex, age, weight, and body surface did not influence the results of the one-hour value in the control subjects. Forty-eight children with cystic fibrosis had one-hour xylose levels within the normal range, but the means at 90, 120, and 180 minutes after load exceeded significantly (P less than 0.01) those of the controls. In a nonceliac group of 63 children with abnormally low xylose levels, almost all were found to have clinical conditions compatible with upper small bowel mucosal impairment. This study shows that a single estimation of xylose in blood one hour after load is a reliable index of small bowel mucosal function. It is a good screening test for celiac disease and small bowel disorders producing malabsorption in children.


Retroperitoneal lymph node dissection after chemotherapy for germ cell tumours: is a full bilateral template always necessary?
To evaluate the long-term oncological outcome in selected patients treated for nonseminomatous germ cell tumours (NSGCT) with a retroperitoneal lymph node dissection after chemotherapy (pcRPLND) and not using the full bilateral template. From 1988 to 2005, 102 patients with retroperitoneal stage II NSGCT, who at initial presentation had metastases in the primary retroperitoneal site only, had pcRPLND within a restricted template, whether computed tomography showed complete or incomplete remission. In all, 78 patients had a unilateral template dissection and 24 an open modified template dissection. Of the 102 patients, 30 had stage IIC, 63 IIB, and nine tumour marker-positive stage IIA disease. Active tumour was found in two RPLND specimens; mature teratoma in 46 of the 102 patients, and necrosis/fibrosis in 54. Antegrade ejaculation was preserved in all 78 patients who had unilateral RPLND and in 18 of 24 who had a modified RPLND (overall antegrade ejaculation rate 94%). During a median follow-up of 102 months there were only three recurrences, two outside the retroperitoneum and one in the retroperitoneum. Only the latter was retrocaval, above the level of the inferior mesenteric artery within the boundaries of a full bilateral RPLND. This patient had had left unilateral template dissection after chemotherapy for IIC disease which was restricted to the para-aortic nodes. In a selected group of patients with stage II NSGCT, pcRPLND within the modified template might be oncologically efficient and allow ejaculation to be preserved.


Pulsed field gel electrophoresis and random amplified polymorphic DNA molecular characterization of Ralstonia pickettii isolates from patients with nosocomial central venous catheter related bacteremia.
Over a period of 18 months 3 clusters of central venous catheter-related Ralstonia pickettii bacteremia occurred in 3 different units of the same hospital. In order to investigate the relatedness of the clinical isolates we studied 15 strains using pulsed-field gel electrophoresis (PFGE) and randomly amplified polymorphic DNA (RAPD) techniques. The combined analysis of the results obtained by these two methods led us to conclude that all the patients except one were infected by a single clone comprising two variants circulating in the units. Only one case was due to a different strain, probably originating outside the ward. PFGE and RAPD appear to be discriminatory techniques to study the clonal relationship among the isolates and can represent a good tool to perform the epidemiological investigation of an outbreak.


Predictor variables in competency to stand trial decisions.
Clinicians possess significant discretion in competency to stand trial assessment. Therefore, it is paramount to explore the contribution of individual variables to ensure that the decision-making process is devoid of bias and solely relates to the legal criterion. To test for the possibility of bias in clinical decision-making, we examined the predictive efficiency of clinical, criminological, and sociodemographic variables in a sample of 468 criminal defendants referred for competency evaluations. Only clinical diagnostic variables and employment status were significant predictors. This finding supports the idea that examiner decisions of competency appear to be unbiased and relate primarily to a defendant's functional ability.


COVID-19, traffic demand, and activity restriction in China: A national assessment.
The global COVID pandemic of 2020, affected travel patterns across the world. The level of impact was influenced not only by the virus itself, but also by the nature, extent, and duration of governmental restriction on commerce and personal activity to limit its spread. This paper focuses on the interaction between COVID-19 transmission and traffic volume and further explores the impact of traffic control policies on the interaction. Roadway traffic volume was used to quantify and assess the Chinese response to the pandemic; specifically, the relationship between government restrictions, travel activity, and COVID-19 progression across 29 provinces. Space and time distributions of traffic volume across China during the first half of 2020, were used to quantity the response and recovery of travel during the critical initial onset period of the virus. Most revealing of these trends were the impact of the Chinese restriction policies on both travel and the virus as well as the relationship of traffic trends during the closure period with the speed and extent of the recovery "bounce" across individual provinces based on location, economic activity, and restriction policy. These suggest that the most significant and rapid declines in traffic volume during the restriction period resulted in the most pronounced returns to normal (or more) demand levels. Based on these trends a Susceptible Infection Recovery model was created to simulate a range of outbreak and restriction policies to examine the relationship between COVID-19 spread and traffic volume in China.


Deviations between transpalpebral tonometry using TGDc-01 and Goldmann applanation tonometry depending on the IOP level.
Recent comparisons between transpalpebral tonometry using TGDc-01 and Goldmann applanation tonometry were performed in populations with IOPs between 10 and 20 mmHg. The purpose of this study was to evaluate device deviations depending on different IOP levels (range 5-40 mmHg). A total of 68 eyes of 68 patients were included and assigned to four IOP levels according to an initial applanation tonometry assessment: level I, <10 mmHg (n=8); level II, 10-19 mmHg (n=20); level III, 20-29 mmHg (n=20); and level IV, > or =30 mmHg (n=20). Two independent and randomized observers performed three replicate measurements per eye-observer 1 using TGDc-01 tonometry, and observer 2 using Goldmann applanation tonometry. Intraindividual deviations between measurement results were investigated concerning clinical relevance by medians and quartiles, concerning statistical significance by pairwise sign tests; p values <0.05 indicate local statistical significance. In patients with initial IOP > or =20 mmHg, TGDc-01-based tonometry significantly underestimated the IOP as based on Goldmann applanation tonometry (p<0.001). This effect increased with increasing IOP: IOP level III median difference (TGDc-01 - Goldmann) -1.3 mmHg (interquartile range, -2.5, -0.4), IOP level IV median difference -2.7 mmHg (-3.7, -1.0). In patients with initial IOP <10 mmHg, an at least gradual underestimation by TGDc-01 tonometry (p=0.219; median difference, -0.6, -1.6, 0) was observed. A total 18% of patients showed device deviations > +/-3 mmHg, and even 35% of those patients with initial IOP > or =30 mmHg. TGDc-01-based tonometry demonstrated an increasing underestimation of IOP with increasing IOP levels when compared with the current standard method of Goldmann applanation tonometry.


Controlling a virtual forehand prosthesis using an adaptive and affective Human-Machine Interface.
This paper presents the design of an adaptable Human-Machine Interface (HMI) for controlling virtual forearm prosthesis. Direct physical performance measures (obtained score and completion time) for the requested tasks were calculated. Furthermore, bioelectric signals from the forehead were recorded using one pair of electrodes placed on the frontal region of the subject head to extract the mental (affective) measures while performing the tasks. By employing the proposed algorithm and above measures, the proposed HMI can adapt itself to the subject's mental states, thus improving the usability of the interface. The quantitative results from 15 subjects show that the proposed HMI achieved better physical performance measures in comparison to a conventional non-adaptive myoelectric controller (p < 0.001).


Cell-mediated immunity in anorexia nervosa augmented lymphocyte transformation response to concanavalin A and lack of increased risk of infection.
Cell-mediated immunity (CMI) was studied in 33 consecutively admitted patients with anorexia nervosa (AN) who had a mean weight loss of 30%. Clinical infections were not diagnosed in the patients. The acute phase reactant, C-reactive protein, was slightly elevated in only one patient, but was within the normal range in the remainder. The absolute number of T-lymphocytes in peripheral blood was significantly increased (p < 0.01) in the AN-patients compared to the control group while the number of granulocytes was reduced (p < 0.01). However, a significant negative correlation between the relative weight loss and the numbers of both total and T-lymphocytes in the AN-patients was established (p < 0.05). Mitogen stimulation of lymphocytes with phytohemagglutinin (PHA), poke-weed-mitogen (PWM) and concanavalin A (Con A) showed no reduction in the proliferative response in AN-patients compared to the control group. On the contrary, Con A transformation was significantly higher (p < 0.03) in the AN-patients. The lymphocyte response to PWM (p < 0.02) was, however, diminished in the patients with the most advanced weight loss compared to those with the least weight loss.


Relationship of embryotoxicity to genotoxicity of lead nitrate in mice.
Frequency of sister chromatid exchanges (SCEs), nucleolar organizing regions (NORs) and chromosomal aberrations were analysed, in maternal bone marrow and fetal liver and/or lung cells of ICR Swiss Webster mice, following maternal exposure to lead nitrate on gestational day 9. The number of implantations and morphological changes in day 18 fetuses, following the treatment, were also noted. Chemical analysis of lead in maternal and fetal tissues showed that it is readily transferred across the placenta. Lead caused a moderate, but statistically significant, increase in the frequency of SCEs in maternal bone marrow cells and significant reduction in NORs at the 2 highest dose levels (150 and 200 mg/kg b.w.). Lead treated animals showed several specific chromosomal aberrations, mostly deletions in maternal bone marrow and fetal cells. Aneuploidy was found to be frequently associated with the lowest dose levels of lead nitrate (100 mg/kg). Maternal treatment with lead nitrate also significantly increased embryonic resorptions and reduced placental weights. The results suggest that the embryotoxic effects of lead might be associated with the chromosomal changes.


Hsp27 protects adenocarcinoma cells from UV-induced apoptosis by Akt and p21-dependent pathways of survival.
Transcriptional activation of p53 target genes, due to DNA damage, causes either apoptosis or survival by cell cycle arrest and DNA repair. However, the regulators of the choice between cell death and survival signaling have not been completely elucidated. Here, we report that human adenocarcinoma cells (MCF-7) survive UV-induced DNA damage by heat shock protein 27 (Hsp27)-assisted Akt/p21 phosphorylation/translocation. Protein levels of the p53 target genes, such as p21, Bcl-2, p38MAPK, and Akt, showed a positive correlation to Hsp27 level during 48 hours postirradiation, whereas p53 expression increased initially but started decreasing after 12 hours. Hsp27 prevented the G(1)-S phase cell cycle arrest, observed after 8 hours of post-UV irradiation, and PARP-1 cleavage was inhibited. Conversely, silencing Hsp27 enhanced G(1)-S arrest and cell death. Moreover, use of either Hsp27 or Akt small interference RNA reduced p21 phosphorylation and enhanced its retention in nuclei even after 48 hours postirradiation, resulting in enhanced cell death. Our results showed that Hsp27 expression and its direct chaperoning interaction increases Akt stability, and p21 phosphorylation and nuclear-to-cytoplasm translocation, both essential effects for the survival of UV-induced DNA-damaged cells. We conclude that the role of Hsp27 in cancer is not only for enhanced p53 proteolysis per se, rather it is also a critical determinant in p21 phosphorylation and translocation.


Disinhibition as a mechanism for visual cortex neurons to tune to cross-shaped figures.
A discrete simulation model of a receptive field selectively responding to cross-shaped figures, as seen in 40% of primary visual cortex neurons in the cat, was studied. The model was based on disinhibition of end-stop inhibition in the receptive field by the lateral disinhibition zone. These experiments showed that this mechanism can produce selective or, conversely, invariant tuning to the shape and orientation of cross-shaped figures and could underlie the high sensitivity of neurons to second-order image features.


Visualization of Transcription in early mouse embryos.
Transcriptional activity during early mammalian embryogenesis was examined in 2 cell, 4--8 cell and late morula/early blastocyst mouse embryos. Zona pellucida-free embryos were obtained after pronase digestion followed by a PMSF wash to inhibit proteolytic activity. The embryos were then lysed with NP40 detergent and spread for electron microscopy according to Miller and Bakken (1972). pre-mRNA transcription was observed at all stages. Comparison of growing RNP chain lengths revealed statistically significant differences in the distribution of shorter fibrils between the successive developmental stages examined. The number of shorter fibrils was lower in the 4--8 cell stage than in either of the two other stages. Transcription complexes of ribosomal type were detected only in 4--8 cell and morula/blastocyst embryos. Structures resembling replication loops were observed within chromatin from all stages. Similar loop-like structures as well as individually transcribed RNP fibrils were also occasionally found on fibres emerging from mitotic chromosomes. The results are discussed in the context of recent findings concerning genetic expression in early mouse embryos.


N-acetylcysteine attenuates PM2.5-induced apoptosis by ROS-mediated Nrf2 pathway in human embryonic stem cells.
While the effects of fine particulate matter (PM2.5) on embryonic toxicity are widely accepted, its exact mechanisms have not yet been fully elucidated, which partially attribute to lack of ideal research model. Embryonic stem cells (ESCs) have the capacity to differentiate into all cell types of three germ layers. Thus, they are ideal resources for the reproductive toxicity assessment in vitro. In the present study, we investigated the effects of PM2.5 exposure on the oxidative stress and apoptosis of human ESCs (hESCs) and its possible mechanism. Our results showed that strong cytotoxicity high reactive oxygen species (ROS) level and fragmentation of nuclei were observed in the PM2.5-treated hESCs. Meanwhile, up-regulation of apoptosis as well as down-regulation of Nrf2 signaling pathway were also observed after PM2.5 treatment. However, we did not detect significant expression change or phosphorylation of Akt and Erk in PM2.5-treated hESCs. Interestingly, scavenging of PM2.5-induced ROS by N-acetylcysteine (NAC) could block cell apoptosis and rescue the activity of Nrf2 signaling pathway. In conclusion, we demonstrate that PM2.5 is toxic to hESCs by inhibition of ROS-mediated Nrf2 pathway activity. Our findings suggest activation of Nrf2 pathway will help develop new strategies for the prevention and treatment of PM2.5-associated disease.


Discrete Fourier Transform as applicable technique in electrochemical detection of hydrazine using multi-walled carbon nanotube/polyacrylonitrile ceramic fiber as working electrode.
Effect of "Discrete Fourier Transform" (DFT) is studied for electrochemical detection of some electroactive species using multi-walled carbon nanotube/polyacrylonitrile ceramic fiber as ultra micro electrode. Based on DFT theory, two independent phases i.e. the imaginary and real phases are evaluated during the oxidation/reduction of the quasi-reversible or irreversible electroactive species, revealing the independent components of imaginary (IImaginary) and real (IReal) currents. The results show that, in different electrochemical modes such as cyclic voltammetry (CV), the contribution of DFT to the electrochemical signals significantly improves the detection limit of the electrochemical technique. More sensitive signals are obtained at high scan rates according to the combination of electrochemical techniques with the DFT theory. The reliability of DFT algorithm was evaluated for rapid determination of trace amount of hydrazine (N2H4) at a scan rate up to 800 V s(-1). In this study, the amounts of phase and amplitude were estimated to 1.69 and 31.57, respectively. The detection limit of hydrazine was 4.13×10(-9) M. The application of this technique was also evaluated for determination of hydrazine in different industrial wastewater samples.


Therapeutic window of globular adiponectin against cerebral ischemia in diabetic mice: the role of dynamic alteration of adiponectin/adiponectin receptor expression.
Recent studies have demonstrated that adiponectin (APN) attenuates cerebral ischemic/reperfusion via globular adiponectin (gAD). However, the therapeutic role of gAD in cerebral ischemic injury in type 1 diabetes mellitus (T1DM) remains unclear. Our results showed that gAD improved neurological scores and reduced the infarct volumes in the 8-week T1DM (T1DM-8W) mice, but not in the 2-week T1DM (T1DM-2W) mice. Moreover, the ischemic penumbra APN levels increased and peaked in T1DM-2W mice, and reduced to normal in T1DM-8W mice, while the APN receptor 1 (AdipoR1) expression change was the opposite. Administration of rosiglitazone in T1DM-2W mice up-regulated the expression of AdipoR1 and restored the neuroprotection of gAD, while intracerebroventricular injection of AdipoR1 small interfering RNA (siRNA) in T1DM-8W mice reversed it. Furthermore, the expression of p-PERK, p-IRE1 and GRP78 were increased whereas the expressions of CHOP and cleaved caspase-12 as well as the number of apoptotic neurons were decreased after gAD treatment in T1DM-8W mice. These beneficial effects of gAD were reversed by pretreatment with AdipoR1 siRNA. These results demonstrated a dynamic dysfunction of APN/AdipoR1 accompanying T1DM progression. Interventions bolstering AdipoR1 expression during early stages and gAD supplementation during advanced stages may potentially reduce the cerebral ischemic injury in diabetic patients.


The ability of single site, single depth sacral lateral branch blocks to anesthetize the sacroiliac joint complex.
To determine the physiologic effectiveness of single site, single depth sacral lateral branch injections. Randomized, controlled, and double-blinded study. Outpatient pain management center. Fifteen asymptomatic volunteers. The dorsal sacroiliac ligament was probed and the sacroiliac joint was injected with contrast medium until capsular distension occurred. The presence or absence of pain with each maneuver was noted. Under double-blind conditions, subjects returned 1 week later for L5 dorsal ramus and S1-4 lateral branch injections; 10 subjects received 4% lidocaine (active) injections while five subjects received saline (control) injections. After 30 minutes, subjects had repeat ligamentous probing and capsular distension of the same sacroiliac joint that was previously tested. The presence or absence of pain with each maneuver was noted. In a parallel anatomic study, S1 and S2 lateral branch injections with green dye were performed on two nonembalmed cadavers. Dissection was undertaken to quantify the degree of staining of these target lateral branch nerves. Presence or absence of pain for ligamentous probing and sacroiliac joint capsular distension. Forty percent had no discomfort upon repeat ligamentous probing after active lateral branch injections while 100% retained pain upon repeat ligamentous probing with control lateral branch injections. Forty percent of the active group and 20% of the control group did not feel repeat capsular distension of the sacroiliac joint after the lateral branch injections. In the anatomic study, 11 lateral branch nerves were isolated while staining occurred in only four cases or 36%. Anatomic limitations exist with single site, single depth sacral lateral branch injections rendering them physiologically ineffective on a consistent basis.


Pneumatosis cystoides intestinalis associated with perforated chronic duodenal ulcer and Meckel's diverticulum.
A case of pneumatosis cystoides intestinalis (PCI) associated with perforated duodenal ulcer and Meckel's diverticulum is presented. The patient was managed with direct suturing of the perforation and reinforcement with an omental patch. Meckel's diverticulum was excised. PCI was followed up and disappeared postoperatively in the 4th week.


Volatile Sulfur Compounds Produced by the Anaerobic Bacteria Porphyromonas spp. Isolated from the Oral Cavities of Dogs.
Porphyromonas spp. are oral anaerobic Gram-negative bacteria that form black-pigmented colonies on blood agar and produce volatile sulfur compounds (VSCs), such as hydrogen sulfide (H2S), methyl mercaptan (CH3SH), and dimethyl sulfide ((CH3)2S), which cause halitosis and the destruction of periodontal tissues. P. gulae is considered the main pathogen involved in periodontal disease in dogs. However, the characteristics of the VSCs produced by P. gulae are unknown. In the present study, VSCs were measured in 26 isolates of P. gulae and some isolates of the other Porphyromonas spp. obtained from the oral cavities of dogs with periodontal disease using an in vitro assay with an Oral ChromaTM gas chromatograph. The results demonstrated that P. gulae was able to produce large amounts of H2S and CH3SH, and the dominant product was CH3SH (CH3SH/H2S was approximately 2.2). Other Porphyromonas spp. that were also obtained from the oral cavities of dogs with periodontal disease indicated similar levels of production of H2S and CH3SH to those of P. gulae. It is strongly suggested that the high levels of H2S and CH3SH produced by P. gulae and other Porphyromonas spp. contribute to halitosis and the destruction of periodontal tissues during the progression of periodontal disease in dogs.


Bystander LECT2 amyloidosis in tumor nephrectomy.
Pathologic evaluation of the non-neoplastic renal parenchyma in tumor nephrectomy specimens is critical and can detect both renal-limited and systemic pathologies. We report the case of a 69-year-old Punjabi male who underwent cytoreductive nephrectomy for advanced renal cell carcinoma after immunotherapy. We detected clinically unexpected leukocyte chemotactic factor 2 (LECT2) amyloidosis during pathologic analysis of the surrounding non-neoplastic renal parenchyma, which was confirmed by mass spectrometry. LECT2 amyloidosis occurs predominantly in Hispanic patients and has only rarely been described in Punjabi patients. This case highlights the importance of careful pathologic evaluation of the non-neoplastic renal parenchyma of nephrectomy specimens and raises awareness that LECT2 amyloidosis can occur outside of the typical demographic of Hispanic patients.


Direct immunoenzyme double staining applicable for monoclonal antibodies.
A novel method for immunoenzymatic double staining was developed, using primary antibodies directly labeled with either horseradish peroxidase or alkaline phosphatase. The enzyme-antibody conjugates were applied simultaneously on sections of human tonsil. Intracytoplasmic antigens like immunoglobulins and light chains could easily be detected simultaneously in the same tissue section. With antibodies against cell surface antigens like IgM and T cell antigens areas containing B and T cells could be clearly distinguished. This method opens the possibility to perform double staining with two monoclonal antibodies.


Racial differences in drug response: isoproterenol effects on heart rate following intravenous metoprolol.
Healthy young black men and white men received single intravenous doses of metoprolol (0.07 mg/kg) or participated in an isoproterenol sensitivity study before and after metoprolol (0.07 mg/kg followed by 50 micrograms/min) in a randomized, crossed-over fashion. Noncompartmental pharmacokinetic parameters were calculated. The dose of isoproterenol versus change in heart rate response curves were constructed, and comparisons of dose ratio, ED50, Emax, and Ka, with the apparent association constant for metoprolol binding to beta 1-receptors, were made. There were no pharmacokinetic differences observed between the groups. The predicted Emax for the black group was 52.7 +/- 8.7 beats/min at a metoprolol concentration of 29.8 +/- 6.1 ng/ml, which was higher (p less than 0.05) than that in the white group, i.e., 43.7 +/- 7.3 beats/min at a concentration of 27.6 +/- 9.1 ng/ml. There were no differences in dose ratio, ED50, or Ka. The racial differences in beta 1-receptor responses to exogenous isoproterenol following metoprolol can simply be explained by an increase in beta 1-receptor activity in the black subjects, assuming homogeneity in cardiac beta 2-receptor responses.


Post-partum pyogenic abscess containing Ascaris lumbricoides.
We report an unusual case of multiple pyogenic liver abscesses containing Ascariasis lumbricoides in a 35-year-old post-partum female who had delivered 1 month back. Open drainage of liver abscess along with liver worm was done. Patient did well post-operatively.


Yonetani-Theorell analysis of hepatic carnitine palmitoyltransferase-I inhibition indicates two distinct inhibitory binding sites.
Analysis of inhibitor studies indicates that carnitine palmitoyltransferase-I has two separate sites for inhibitor binding. One site is located on the cytoplasmic side of the mitochondrial outer membrane. Malonyl-CoA, the most important physiological inhibitor of carnitine palmitoyltransferase-I, binds primarily to this site, but it can also bind to another site. A second inhibitory site is located at the active site of carnitine palmitolytransferase-I. Coenzyme A, a product/inhibitor of carnitine palmitoyltransferase-I binds primarily at this site and can inhibit carnitine palmitoyltransferase-I at physiological concentrations, but can also attenuate malonyl-CoA inhibition. Analogs of malonyl-CoA and other simpler compounds containing two carbonyl groups but no coenzyme A moiety inhibit only at the cytoplasmic site, indicating that this site has an absolute requirement for two carbonyl groups but has no absolute requirement for a coenzyme A moiety. Inhibitors acting through the active site included the active-site-directed inhibitor (+)-hemipalmitoylcarnitinium. These studies support the existence of two regulatory binding sites for the control of hepatic fatty acid oxidation: (a) the active site, for regulation by the inhibitory binding of coenzyme A and acetyl-CoA, and (b) a separate regulatory malonyl-CoA-binding site that is physical separated from the active site.


Benzodiazepine enhancement of gamma-aminobutyric acid-mediated chloride ion flux in rat brain synaptoneurosomes.
Benzodiazepine agonists such as Ro 11-6896 [B10(+)], diazepam, clonazepam, and flurazepam were found to enhance muscimol-stimulated 36Cl- uptake into rat cerebral cortical synaptoneurosomes. The rank order of potentiation was B10(+) greater than diazepam greater than clonazepam greater than flurazepam. These benzodiazepines had no effect on 36Cl-uptake in the absence of muscimol. Further, the inactive enantiomer, Ro 11-6893 [B10(-)], and the peripheral benzodiazepine receptor ligand Ro 5-4864 did not potentiate muscimol-stimulated 36Cl- uptake at concentrations up to 10 microM. In contrast, the benzodiazepine receptor inverse agonists ethyl-beta-carboline-3-carboxylate and 6,7-dimethoxy-4-ethyl-beta- carboline-3-carboxylic acid methyl ester inhibited muscimol stimulated 36Cl- uptake. Benzodiazepines and beta-carbolines altered the apparent K0.5 of muscimol-stimulated 36Cl- uptake, without affecting the Vmax. The effects of both benzodiazepine receptor agonists and inverse agonists were reversed by the benzodiazepine antagonists Ro 15-1788 and CGS-8216. These data further confirm that central benzodiazepine receptors modulate the capacity of gamma-aminobutyric acid receptor agonists to enhance chloride transport and provide a biochemical technique for studying benzodiazepine receptor function in vitro.


Automated quantitative fluorescent image analysis of cervical cytology.
Cervical cytology from 122 patients with histories of one or more abnormal Pap smears were studied using slide-based automated quantitative fluorescence image analysis (QFIA) in order to determine the usefulness of the QFIA technique in detecting neoplasias. The purpose of the study was to evaluate the accuracy and validity of QFIA as a cytological method for the early detection of cervical cancer, based on comparisons with the conventional Pap smear and histopathology. The results of the analysis demonstrated that the sensitivity of the automated QFIA cytology was 100% for high-grade neoplasms (CIN III and invasive carcinoma), compared with 82% for the Pap smear. Evaluation of lower grade neoplasms was also performed. Initial analyses of the data indicate that QFIA, which uses objective biochemical criteria to identify cells with increased nuclear nucleic acid content, is a more sensitive detector of cervical neoplasia of all grades than is conventional Pap cytology.


Emergency medical care in Turkey: current status and future directions.
We outline recent developments in the delivery of emergency medicine and prehospital care in the Republic of Turkey. Residency training in emergency medicine is in its infancy in Turkey, and the presence of specialists will help define the future of emergency medical care in this country.


Phospholipase A2-induced hypotension in the rat and its pharmacological modulation.
1. An increase in circulating levels of extracellular group II phospholipase A2 (PLA2) has been detected in patients with septic shock as well as in rats, rabbits and human volunteers following intravenous endotoxin administration. 2. Group II PLA2 from Naja mocambique mocambique (NajaPLA2) snake venom (1-10 micrograms i.v.) induced a dose-dependent hypotension and leucopenia in the rat similar to that induced by intravenous administration of endotoxin. 3. NajaPLA2 did not aggregate rat platelet, but did aggregate purified rat neutrophils in vitro. However intravenous PLA2 caused rat platelets to aggregate in vivo. 4. The hypotensive effect was reduced either by inactivating NajaPLA2 in vitro with para-bromophenacyl bromide or by infusing in vivo polyclonal rabbit antiPLA2 antisera. Neither the hypotension nor the leucopenia was affected by several agonists and inhibitors. 5. The leukotriene D4 antagonist L-649,923 produced a dose-related (5-20 mg/kg i.v.) inhibition of NajaPLA2-induced hypotension while the dual inhibitor of lipo- and cyclooxygenase BW755c (5-50 mg/kg i.v.) was ineffective. 6. In rats rendered leucopenic with methotrexate the L-649,923 was ineffective implying that the L-649,923 effect could be partially mediated through its action on neutrophils.


Management of High-Risk Breast Lesions.
High-risk breast lesions (HRLs) are a group of heterogeneous lesions that can be associated with a synchronous or adjacent breast cancer and that confer an elevated lifetime risk of breast cancer. Management of HRLs after core needle biopsy may include close imaging and clinical follow-up or excisional biopsy to evaluate for cancer. This article reviews histologic features and clinical presentation of each of the HRLs, current evidence with regard to management, and guidelines from the American Society of Breast Surgeons and National Comprehensive Cancer Network. In addition, imaging surveillance and risk-reduction strategies for women with HRLs are discussed.


Association between corneal hysteresis and central corneal thickness in glaucomatous and non-glaucomatous eyes.
We aimed to determine corneal hysteresis values (CH) using the ocular response analyser (ORA) in non-glaucomatous and glaucomatous eyes and their relationship with central corneal thickness (CCT). Corneal hysteresis, intraocular pressure (IOP) as measured by Goldmann applanation tonometry (GAT) and CCT were prospectively evaluated in 74 non-glaucoma subjects with IOP < 21 mmHg and in 108 patients with treated primary open-angle glaucoma (POAG). One eye in each subject was randomly selected for inclusion in the analysis. Mean (+/- standard deviation [SD]) age was 59.2 +/- 14.2 years in the non-glaucoma group and 62.4 +/- 9.8 years in the glaucoma group. Mean (+/- SD) GAT IOP was 15.7 +/- 2.65 mmHg and 16.38 +/- 2.73 mmHg in the non-glaucoma and glaucoma groups, respectively. There was no statistically significant difference between the two groups in mean age (p = 0.396) or mean GAT IOP (p = 0.098). Mean (+/- SD) CH was 10.97 +/- 1.59 mmHg in the non-glaucoma and 8.95 +/- 1.27 mmHg in the glaucoma groups, respectively. The difference in mean CH between the two groups was statistically significant (p < 0.0001). There was a strong positive correlation between CH and CCT in the non-glaucoma group (r = 0.743) and a significantly (p = 0.001) weaker correlation (r = 0.426) in the glaucoma group. Corneal hysteresis was significantly lower in eyes with treated POAG than in non-glaucomatous eyes. The corneal biomechanical response was strongly associated with CCT in non-glaucoma subjects, but only moderately so in glaucoma patients. It can be assumed that diverse structural factors, in addition to thickness, determine the differences in the corneal biomechanical profile between non-glaucomatous and glaucomatous eyes. Corneal hysteresis could be a useful tool in the diagnosis of glaucoma.


The supporting cast. Strong, focused committees and task forces are essential to overall governance excellence.
The best committees improve board decision-making and efficiency.


Connective tissue diseases and autoimmune thyroid disorders in the first trimester of pregnancy.
To investigate the rates and coexistence of autoimmune thyroid and connective tissue diseases (CTD) during the first trimester of pregnancy and their influence on pregnancy outcome. A cohort study of 150 women with CTD diagnosed during first trimester of pregnancy and 150 negative controls. Screening of CTD by a self-reported questionnaire, rheumatic and thyroid autoantibody detection, clinical rheumatological evaluation and obstetric outcomes. Out of 3852 women screened, 61 (1.6%) were diagnosed with undefined connective tissue disease (UCTD), 28 (0.7%) with major CTD (six rheumatoid arthritis, five systemic lupus erythematosus, eight Sjogren syndrome, five anti-phospholipid syndrome, two systemic sclerosis, one mixed CTD and one monoarticular arthritis) and 61 (1.6%) had insufficient criteria for a diagnosis of a rheumatic disease. The overall prevalence of either thyroid peroxidase (TPO-a) or thyroglobulin (TG-a) autoantibodies detection was 8% (12/150) among controls, 62.3% (38/61) among UCTD and 60.7% (17/28) in women with a major CTD (p<.001 compared to controls for both comparisons). After adjustment for confounders, overall CTDs (major or undefined) (OR=3.54, 95% CI; 1.61-7.78) and TPO-a plus TG-a positivity (OR=2.78, 95% CI;1.29-5.98) were independently associated with increased risks of moderate-severe complications of pregnancy (miscarriage, fetal growth restriction, preeclampsia, delivery before 34 weeks). Rheumatic and thyroid autoantibodies during pregnancy are closely associated. Thyroid antibodies could add to the risk of adverse pregnancy outcome associated with connective tissue diseases.


Interlaboratory studies in clinical chemistry--the Canadian experience.
This report summarizes the results of eleven cross-Canada interlaboratory surveys between 1974 and 1985, for a small number of analytes. Statistical analysis of data for precision and accuracy by comparison to approved reference method values are presented.


Prevalence and distribution of carcinoma in patients with solitary and multiple thyroid nodules on sonography.
Controversy remains as to the optimal management of patients with multiple thyroid nodules. The objective of this study was to determine the prevalence, distribution, and sonographic features of thyroid cancer in patients with solitary and multiple thyroid nodules. We describe a retrospective observational cohort study that was carried out from 1995 to 2003. The study was conducted in a tertiary care hospital. Patients with one or more thyroid nodules larger than 10 mm in diameter who had ultrasound-guided fine needle aspiration (FNA) were included in the study. The main outcome measures were prevalence and distribution of thyroid cancer and the predictive value of demographic and sonographic features. A total of 1985 patients underwent FNA of 3483 nodules. The prevalence of thyroid cancer was similar between patients with a solitary nodule (175 of 1181 patients, 14.8%) and patients with multiple nodules (120 of 804, 14.9%) (P = 0.95, chi(2)). A solitary nodule had a higher likelihood of malignancy than a nonsolitary nodule (P < 0.01). In patients with multiple nodules larger than 10 mm, cancer was multifocal in 46%, and 72% of cancers occurred in the largest nodule. Multiple logistic regression analysis of statistically significant features demonstrates that the combination of patient gender (P < 0.02), whether a nodule is solitary vs. one of multiple (P < 0.002), nodule composition (P < 0.01), and presence of calcifications (P < 0.001) can be used to assign risk of cancer to each individual nodule. Risk ranges from a 48% likelihood of malignancy in a solitary solid nodule with punctate calcifications in a man to less than 3% in a noncalcified predominantly cystic nodule in a woman. In a patient with one or more thyroid nodules larger than 10 mm in diameter, the likelihood of thyroid cancer per patient is independent of the number of nodules, whereas the likelihood per nodule decreases as the number of nodules increases. For exclusion of cancer in a thyroid with multiple nodules larger than 10 mm, up to four nodules should be considered for FNA. Sonographic characteristics can be used to prioritize nodules for FNA based on their individual risk of cancer.


STR typing of skin swabs from individuals after an allogeneic hematopoietic stem cell transplantation.
One of the pre-requisites for forensic DNA analysis is the fact that all nucleated cells of a person carry the same genetic information. However, this is not the case for individuals who have received an allogeneic hematopoietic stem cell or bone marrow transplantation, as all new cells formed by the bone marrow no longer show the genetic information of the recipient but that of the donor, while all other cells still carry the original information before transplantation. Thus, STR typing of a blood sample after successful transplantation yields a DNA profile that differs from the recipient's original profile and corresponds to the donor genotype instead. Evidence from a routine case suggests that transplanted individuals may show donor alleles in skin swabs, as well. In order to examine this issue more closely, various skin swabs from 28 patients who have received an allogeneic hematopoietic stem cell transplantation were examined in this study. Swabs from the right and left palm, the back of the hand, one of the two upper arms, and the neck were collected from each person. Ninety-one of the 140 resulting swabs delivered useful results. All of those samples showed mixtures of recipient and donor DNA with different mixture ratios and the proportions of donor and recipient alleles revealed inter- and intra-individual differences. Those results were discussed with respect to graft versus host disease.