The Central Role of Protein Kinase C Epsilon in Cyanide Cardiotoxicity and Its Treatment.
In adult mouse myocytes, brief exposure to sodium cyanide (CN) in the presence of glucose does not decrease ATP levels, yet produces profound reduction in contractility, intracellular Ca2+ concentration ([Ca2+]i) transient and L-type Ca2+ current (ICa) amplitudes. We analyzed proteomes from myocytes exposed to CN, focusing on ionic currents associated with excitation-contraction coupling. CN induced phosphorylation of α1c subunit of L-type Ca2+ channel and α2 subunit of Na+-K+-ATPase. Methylene blue (MB), a CN antidote that we previously reported to ameliorate CN-induced reduction in contraction, [Ca2+]i transient and ICa amplitudes, was able to reverse this phosphorylation. CN decreased Na+-K+-ATPase current contributed by α2 but not α1 subunit, an effect that was also counteracted by MB. Peptide consensus sequences suggested CN-induced phosphorylation was mediated by protein kinase C epsilon (PKCε). Indeed, CN stimulated PKC kinase activity and induced PKCε membrane translocation, effects that were prevented by MB. Pretreatment with myristoylated PKCε translocation activator or inhibitor peptides mimicked and inhibited the effects of CN on ICa and myocyte contraction, respectively. We conclude that CN activates PKCε, which phosphorylates L-type Ca2+ channel and Na+-K+-ATPase, resulting in depressed cardiac contractility. We hypothesize that this inhibition of ion fluxes represents a novel mechanism by which the cardiomyocyte reduces its ATP demand (decreased ion fluxes and contractility), diminishes ATP turnover and preserves cell viability. However, this cellular protective effect translates into life-threatening cardiogenic shock in vivo, thereby creating a profound disconnect between survival mechanisms at the cardiomyocyte level from those at the level of the whole organism.


Geminivirus C5 proteins mediate formation of virus complexes at plasmodesmata for viral intercellular movement.
Movement proteins (MPs) encoded by plant viruses deliver viral genomes to plasmodesmata (PD) to ensure intracellular and intercellular transport. However, how the MPs encoded by monopartite geminiviruses are targeted to PD is obscure. Here, we demonstrate that the C5 protein of tomato yellow leaf curl virus (TYLCV) anchors to PD during the viral infection following trafficking from the nucleus along microfilaments in Nicotiana benthamiana. C5 could move between cells and partially complement the traffic of a movement-deficient turnip mosaic virus (TuMV) mutant (TuMV-GFP-P3N-PIPO-m1) into adjacent cells. The TYLCV-C5 null mutant (TYLCV-mC5) attenuates viral pathogenicity and decreases viral DNA and protein accumulation, and ectopic overexpression of C5 enhances viral DNA accumulation. Interaction assays between TYLCV-C5 and the other eight viral proteins described in TYLCV reveal that C5 associates with C2 in the nucleus and with V2 in the cytoplasm and at PD. The V2 protein is mainly localized in the nucleus and cytoplasmic granules when expressed alone; in contrast, V2 forms small punctate granules at PD when co-expressed with C5 or in TYLCV-infected cells. The interaction of V2 and C5 also facilitates their nuclear export. Furthermore, C5-mediated PD localization of V2 is conserved in two other geminiviruses. Therefore, this study solves a long-sought-after functional connection between PD and the geminivirus movement and improves our understanding of geminivirus-encoded MPs and their potential cellular and molecular mechanisms.


Diversity versus disparity and the radiation of modern cetaceans.
Modern whales are frequently described as an adaptive radiation spurred by either the evolution of various key innovations (such as baleen or echolocation) or ecological opportunity following the demise of archaic whales. Recent analyses of diversification rate shifts on molecular phylogenies raise doubts about this interpretation since they find no evidence of increased speciation rates during the early evolution of modern taxa. However, one of the central predictions of ecological adaptive radiation is rapid phenotypic diversification, and the tempo of phenotypic evolution has yet to be quantified in cetaceans. Using a time-calibrated molecular phylogeny of extant cetaceans and a morphological dataset on size, we find evidence that cetacean lineages partitioned size niches early in the evolutionary history of neocetes and that changes in cetacean size are consistent with shifts in dietary strategy. We conclude that the signature of adaptive radiations may be retained within morphological traits even after equilibrium diversity has been reached and high extinction or fluctuations in net diversification have erased any signature of an early burst of diversification in the structure of the phylogeny.


The utilitarian argument for medical confidentiality: a pilot study of patients' views.
To develop and pilot a questionnaire based assessment of the importance patients place on medical confidentiality, whether they support disclosure of confidential information to protect third parties, and whether they consider that this would impair full disclosure in medical consultations. Questionnaire administered to 30 consecutive patients attending a GP surgery. Overall patients valued confidentiality, felt that other patients might be deterred from seeking treatment if it were not guaranteed, but did not think that they would withhold information for this reason themselves. When presented with brief details of five clinical situations in which a breach of confidentiality might be considered, a clear majority of subjects believed that doctors should disclose information in two of the situations, but subjects were not confident that doctors would do so. In three situations, about half felt that disclosure was justified--these included the only scenario in which disclosure was clearly mandated by statute. There was little change in patients' general attitude to confidentiality after considering the scenarios. However, the views expressed were often inconsistent with responses to the clinical scenarios, suggesting that complex opinions were not accurately reflected in the responses. The format of the questionnaire has been amended, and the study will be repeated with other groups of patients.


The plasma membrane redox system is impaired by amyloid β-peptide and in the hippocampus and cerebral cortex of 3xTgAD mice.
Membrane-associated oxidative stress has been implicated in the synaptic dysfunction and neuronal degeneration that occurs in Alzheimer's disease (AD), but the underlying mechanisms are unknown. Enzymes of the plasma membrane redox system (PMRS) provide electrons for energy metabolism and recycling of antioxidants. Here, we show that activities of several PMRS enzymes are selectively decreased in plasma membranes from the hippocampus and cerebral cortex of 3xTgAD mice, an animal model of AD. Our results that indicate the decreased PMRS enzyme activities are associated with decreased levels of coenzyme Q(10) and increased levels of oxidative stress markers. Neurons overexpressing the PMRS enzymes (NQO1 or cytochrome b5 reductase) exhibit increased resistance to amyloid β-peptide (Aβ). If and to what extent Aβ is the cause of the impaired PMRS enzymes in the 3xTgAD mice is unknown. Because these mice also express mutant tau and presenilin-1, it is possible that one or more of the PMRS could be adversely affected by these mutations. Nevertheless, the results of our cell culture studies clearly show that exposure of neurons to Aβ1-42 is sufficient to impair PMRS enzymes. The impairment of the PMRS in an animal model of AD, and the ability of PMRS enzyme activities to protect neurons against Aβ-toxicity, suggest enhancement PMRS function as a novel approach for protecting neurons against oxidative damage in AD and related disorders.


Involvement of MAPKs and NF-kappaB in LPS-induced VCAM-1 expression in human tracheal smooth muscle cells.
Lipopolysaccharide (LPS) has been shown to induce the expression of adhesion molecules on airway epithelial and smooth cells and contributes to inflammatory responses. Here, the roles of mitogen-activated protein kinases (MAPKs) and nuclear factor-kappaB (NF-kappaB) pathways for LPS-induced vascular cell adhesion molecule (VCAM)-1 expression were investigated in HTSMCs. LPS-induced expression of VCAM-1 protein and mRNA in a time-dependent manner, was significantly inhibited by inhibitors of MEK1/2 (U0126), p38 (SB202190), and c-Jun-N-terminal kinase (JNK; SP600125). The involvement of p42/p44 MAPK and p38 in these responses was further confirmed by that transfection with small interference RNAs (siRNA) direct against MEK, p42, and p38 significantly attenuated LPS-induced VCAM-1 expression. Consistently, LPS-stimulated phosphorylation of p42/p44 MAPK and p38 was attenuated by pretreatment with U0126 or SB202190, and transfection with these siRNAs, respectively. In addition, LPS-induced VCAM-1 expression was significantly blocked by a specific NF-kappaB inhibitor helenalin. LPS-stimulated translocation of NF-kappaB into the nucleus and degradation of IkappaB-alpha was blocked by helenalin, U0126, SB202190, or SP600125. Moreover, the resultant enhancement of VCAM-1 expression increased the adhesion of polymorphonuclear cells to monolayer of HTSMCs which was blocked by pretreatment with helenalin, U0126, or SP600125 prior to LPS exposure. Taken together, these results suggest that in HTSMCs, activation of p42/p44 MAPK, p38, and JNK pathways, at least in part, mediated through NF-kappaB, is essential for LPS-induced VCAM-1 gene expression. These results provide new insight into the mechanisms of LPS action that bacterial toxins may promote inflammatory responses in the airway disease.


From incarceration to community care: structured assessment of correctional adaptation.
In this study, the authors extend their evaluation of a structured instrument for assessing the persistence of attitudes and behaviors developed by mentally ill offenders during periods of incarceration (Structured Assessment of Correctional Adaptation; SACA) and seek to demonstrate further the clinical significance of the construct of correctional adaptation. The subjects, patients at a state psychiatric center, were administered the SACA, along with the Brief Psychiatric Rating Scale the Psychopathy Checklist: Screening Version and Working Alliance Inventory. Chart review captured relevant demographic, diagnostic, and correctional history variables. Results were analyzed comparing patients with and without incarceration histories. The SACA total score demonstrated strong interrater reliability and association with criminal history indices of validity. Patients with histories of incarceration were significantly more likely to score higher on the Correctional Adaptation measure and lower on the Bond subscale of working alliance. Controlling for symptom severity and psychopathy did not alter the negative relationship between correctional adaptation score and the Working Alliance Inventory.


A randomised crossover trial comparing volume assured and pressure preset noninvasive ventilation in stable hypercapnic COPD.
Recent randomised controlled trials suggest non-invasive ventilation may offer benefit in the long-term management of ventilatory failure in stable COPD. The best mode of ventilation is unknown and newer volume assured modes may offer advantages by optimising ventilation overnight when treatment is delivered. This study compares volume assured with pressure preset non-invasive ventilation. Randomised crossover trial including twenty five subjects previously established on long-term non-invasive ventilation to manage COPD with chronic ventilatory failure. Two 8-week treatment periods of volume assured and pressure preset non-invasive ventilation. The primary outcomes were daytime arterial blood gas tensions and mean nocturnal oxygen saturation. Secondary outcomes included lung function, exercise capacity, mean nocturnal transcutaneous carbon dioxide, health status and compliance. No significant differences were seen in primary or secondary outcomes following 8 weeks of treatment when comparing volume assured and pressure preset ventilation. Primary outcomes assessed: mean (standard deviation) PaO(2) 7.8 (1.2) vs 8.1(1) kPa, PaCO(2) 6.7 (1.1) vs 6.3 (1.2) kPa and mean nocturnal oxygenation 90 (4) vs 91 (3)% volume assured versus pressure preset, respectively. Volume assured and pressure preset non-invasive ventilation appear equally effective in the long-term management of ventilatory failure associated with stable COPD.


Dairy products and the risk of stroke and coronary heart disease: the Rotterdam Study.
We examined whether consumption of total dairy and dairy subgroups was related to incident stroke and coronary heart disease (CHD) in a general older Dutch population. The study involved 4,235 participants of the Rotterdam Study aged 55 and over who were free of cardiovascular disease (CVD) and diabetes at baseline (1990-1993). Multivariable Cox proportional hazards models were used to calculate hazard ratios (HRs) for the intake of total dairy and dairy subgroups in relation to incident CVD events. Median intake of total dairy was 397 g/day, which mainly comprised low-fat dairy products (median intake of 247 g/day). During a median follow-up time of 17.3 years, 564 strokes (182 fatal) and 567 CHD events (350 fatal) occurred. Total dairy, milk, low-fat dairy, and fermented dairy were not significantly related to incident stroke or fatal stroke (p > 0.2 for upper vs. lower intake categories). High-fat dairy was significantly inversely related to fatal stroke (HR of 0.88 per 100 g/day; 95% CI 0.79, 0.99), but not to incident stroke (HR of 0.96 per 100 g/day; 95% CI 0.90, 1.02). Total dairy or dairy subgroups were not significantly related to incident CHD or fatal CHD (HRs between 0.98 and 1.05 per 100 g/day, all p > 0.35). In this long-term follow-up study of older Dutch subjects, total dairy consumption or the intake of specific dairy products was not related to the occurrence of CVD events. The observed inverse association between high-fat dairy and fatal stroke warrants confirmation in other studies.


Cell-bound complement activation products as lupus biomarkers: diagnosis, monitoring and stratification.
Cell-bound complement activation products (CB-CAPs) were first reported in 2004, since which time multiple laboratories have demonstrated their value as biomarkers for diagnosis, monitoring, and stratification of patients with systemic lupus erythematosus. Areas covered: This review summarizes the highlights of these 14 years of CB-CAPs discovery and validation, concluding with a view toward their future potential for precision medicine. Expert commentary: The practice of medicine is both art and science and each physician can be considered both artist and scientist with a variable blend of the two skill sets. There is arguably no disease that presents a greater challenge, nor a greater opportunity, for implementation of precision medicine, as does lupus. The physician who is presented with diagnosis and/or management of a patient suspected of having lupus will need to augment artistic skills with scientific guidance, and that science will be delivered in the form of biomarkers. Ultimately, we will likely have a 'lupus liquid biopsy' that will be 100% sensitive and 100% specific for a diagnosis of lupus. This will undoubtedly be a panel of biomarkers rather than an individual laboratory test. Such a liquid biopsy could transform lupus diagnosis to an entirely scientific process.


The hdrRM operon of Streptococcus mutans encodes a novel regulatory system for coordinated competence development and bacteriocin production.
The Streptococcus mutans hdrRM operon encodes a novel two-gene regulatory system induced by high cell density. Previous studies identified hdrM as the only known negative regulator of competence development in S. mutans. In the present study, we demonstrated that the HdrRM system bypasses the prototypical competence gene regulators ComC and ComDE in the transcriptional regulation of the competence-specific sigma factor comX and the late competence genes. Similarly, the HdrRM system can abrogate the requirement for ComE to produce the bacteriocin mutacin IV. To further probe the regulatory mechanism of hdrRM, we created an hdrR overexpression strain and showed that it could reproduce each of the hdrM competence and mutacin phenotypes, indicating that HdrM acts as a negative regulator of HdrR activity. Using a mutacin IV-luciferase reporter, we also demonstrated that the hdrRM system utilizes the same promoter elements recognized by ComE and thus appears to comprise a novel regulatory pathway parallel to ComCDE.


Multiplex real-time RT-PCR for prospective evaluation of WT1 and fusion gene transcripts in newly diagnosed de novo acute myeloid leukemia.
Prognostic assessment is crucial for the management of AML. Although the use of karyotype analysis for risk-stratification is widely accepted, prognosis of AML remains ambiguous, particularly for patients categorized into the intermediate cytogenetic risk group and additional markers are required for an accurate prediction of outcome. For this study, we used multiplex real-time RT-PCR, which can simultaneously quantify WT1 and 10 distinct fusion gene transcripts, to prospectively evaluate the pre-treatment bone marrow findings of 53 de novo AML patients. Five patients with normal karyotype or insufficient metaphases detected by conventional karyotype analysis proved to have AML1-MTG8, CBFbeta-MYH11 or PML-RARalpha fusion transcripts. WT1 overexpression was observed in 92% of the patients, and the levels were significantly higher in the cytogenetic favorable risk group, especially patients with PML-RARalpha. WT1 levels also correlated with the percentage of blasts in bone marrow, especially in cases of core-binding factor leukemia. There was no association between initial WT1 levels and outcome in terms of event-free survival or overall survival. These results suggest that multiplex real-time RT-PCR is rapid and useful for the precise cytogenetic stratification of AML, and that WT1 levels at presentation correlate with several biologic features of leukemia, but have no prognostic significance.


Physiological studies of cortical spreading depression.
Cortical spreading depression (CSD) produces propagating waves of transient neuronal hyperexcitability followed by depression. CSD is initiated by K+ release following neuronal firing or electrical, mechanical or chemical stimuli. A triphasic (30-50 s) cortical potential transient accompanies localized transmembrane redistributions of K+, glutamate, Ca2+, Na+, Cl- and H+. Accumulated K+ in the restricted interstitial space can cause both further neuronal depolarisation and inward movement of K+ into astrocytes that buffers this increased extracellular K+ concentration ([K+])o. However, astrocyte interconnections may then propagate the CSD wave by K+ liberation through an opening of remote K+ channels by volume, Ca2+ or N-methyl-D-aspartate receptor activation. Changes in cerebral blood volume and in apparent water diffusion co-efficient (ADC) accompanying CSD were first studied using magnetic resonance imaging (MRI) in whole lissencephalic brains. Diffusion-weighted echoplanar imaging in gyrencephalic brains went on to demonstrate CSD features that paralleled classical migraine aura. The ADC activity persisted minutes/hours post KCl stimulus. Pixelwise analyses distinguished single primary events and multiple, spatially restricted, slower propagating, secondary events whose detailed features varied with the nature of the originating stimulus. These ADC changes varied reciprocally with T2*-weighted (i.e. referring to spin-spin relaxation times) waveforms reflecting local blood flow. There followed prolonged decreases in cerebral blood flow culminating in late cerebrovascular changes blocked by the antimigraine agent sumatriptan. CSD phenomena have possible translational significance for human migraine aura and other cerebral pathologies such as the peri-infarct depolarisation events that follow ischaemia and brain injury.


The organization and regulation of the macrophage actin skeleton.
To move, leukocytes extend portions of their cortical cytoplasm as pseudopods. These pseudopods are filled with a three-dimensional actin filament skeleton, the reversible assembly of which in response to receptor stimulation is thought to play a major role in providing the mechanical force for these protrusive movements. The organization of this actin skeleton occurs at different levels within the cell, and a number of macrophage proteins have been isolated and shown to affect the architecture, assembly, stability, and length of actin filaments in vitro. The architecture of cytoplasmic actin is regulated by proteins that cross-link filaments in higher-order structures. Actin-binding protein plays a major role in defining network structure by cross-linking actin filaments into orthogonal networks. Gelsolin may have a central role in regulating network structure. It binds to the sides of actin filaments and severs them, and binds the "barbed" filament end, thereby blocking monomer addition at this end. Gelsolin is activated to bind actin filaments by microM calcium. Dissociation of gelsolin bound on filament ends occurs in the presence of the polyphosphoinositides, PIP and PIP2. Calcium and PIP2 have been shown to be intracellular messengers of cell stimulation.


Human schwann cells in tissue culture: histochemical and ultrastructural studies.
Schwann cells cultures were established from 40 human peripheral nerves that underwent biopsy according to an explant-reexplantation technique that markedly reduces non-Schwann cells and achieves cultures greatly enriched in Schwann cells suitable for a wide range of studies. Described are the growth characteristics and histochemical, ultrastructural, and ultrastructural-cytochemical patterns of normal human cultured Schwann cells from 12 patients with demonstrable peripheral nerve abnormality. These findings serve as a basis of comparison when seeking abnormalities of Schwann cells grown from peripheral nerves of patients with putative dysschwannian neuropathies.


[The effect of trematodes on the cellular immunity of the dragonfly Aeschna grandis (Odonata) larvae].
The high percent of the infestation by the trematodes of the families Plagiorchidae and Prosthogonimidae is recorded in the dragonfly Aeschna grandis larvae of the last stage. About 30% of the trematode cysts were melanized. It is established, that the parasitising of the trematodes do not effects on the process of the incapsulation of foreign bodies, but it suppresses partly the formation of oxygen free radicals and phenoloxidase activity in the haemocytes of dragonfly larvae.


Effects of improved complementary feeding and improved water, sanitation and hygiene on early child development among HIV-exposed children: substudy of a cluster randomised trial in rural Zimbabwe.
HIV-exposed uninfected children may be at risk of poor neurodevelopment. We aimed to test the impact of improved infant and young child feeding (IYCF) and improved water, sanitation and hygiene (WASH) on early child development (ECD) outcomes. Sanitation Hygiene Infant Nutrition Efficacy was a cluster randomised 2×2 factorial trial in rural Zimbabwe ClinicalTrials.gov NCT01824940). Pregnant women were eligible if they lived in study clusters allocated to standard-of-care (SOC; 52 clusters); IYCF (20 g small-quantity lipid-based nutrient supplement/day from 6 to 18 months, complementary feeding counselling; 53 clusters); WASH (pit latrine, 2 hand-washing stations, liquid soap, chlorine, play space, hygiene counselling; 53 clusters) or IYCF +WASH (53 clusters). Participants and fieldworkers were not blinded. ECD was assessed at 24 months using the Malawi Developmental Assessment Tool (MDAT; assessing motor, cognitive, language and social skills); MacArthur Bates Communication Development Inventory (assessing vocabulary and grammar); A-not-B test (assessing object permanence) and a self-control task. Intention-to-treat analyses were stratified by maternal HIV status. Compared with SOC, children randomised to combined IYCF +WASH had higher total MDAT scores (mean difference +4.6; 95% CI 1.9 to 7.2) and MacArthur Bates vocabulary scores (+8.5 words; 95% CI 3.7 to 13.3), but there was no evidence of effects from IYCF or WASH alone. There was no evidence that that any intervention impacted object permanence or self-control. Combining IYCF and WASH interventions significantly improved motor, language and cognitive development in HIV-exposed children. NCT01824940.


[Studies on plasma insulin-like growth factor (IGF)-I levels in normal subjects and in patients with various endocrine and metabolic diseases using radioimmunoassay for synthetic IGF-I (26-46)].
Determination of plasma IGF-I concentrations is not easily accessible to clinical use at present because of extremely limited supply of purified natural IGF-I essential for its assay system. Thus, an alternative method has recently been introduced by the development of a specific radioimmunoassay (RIA) for IGF-I (26-46). We examined the specificity and sensitivity of this assay system, and then investigated the changes in plasma concentrations of IGF-I in normal children, adults and in patients with various endocrine and metabolic diseases. Each plasma sample was subjected to acid-ethanol treatment before assay to separate IGF-I from its binding protein. The recovery rate of known amount of IGF-I (26-46) added to untreated plasma sample was more than 90%. The coefficients of variation of intra- and interassay were 9.0% and 13.6%, respectively. This assay system was able to detect IGF-I as low as 10 pg/tube. When plasma sample of a patient with active acromegaly was applied to Sephadex G-75 column, immunoreactive IGF-I was eluted at the position of 7,000 molecular weight. An inhibition curve of plasma extract from an acromegalic patient was parallel to that of IGF-I (26-46), indicating that the RIA could detect IGF-I. There was no remarkable difference between IGF-I values of plasma and serum from the same individual. The value of IGF-I concentration of cord plasma was considerably low (144 +/- 6.7 pg/ml, M +/- SEM) as compared with that of sera of 49 normal children aged 7-12 12 years (320 +/- 14.3 pg/ml). The highest value (460 +/- 54 pg/ml) was attained at the age of 13 years, followed by gradual decrease toward adult age. Plasma IGF-I concentration of normal adults between 20 and 69 years of age was 290 +/- 10 pg/ml. When plasma IGF-I values of adult males and females were separately plotted against age group of each decade, the value declined gradually with age in males while in females there was a remarkable increase in plasma IGF-I concentration at 4th and 5th decades, suggesting the effect of hormonal change at menopause on plasma IGF-I levels. There was a good correlation between disorders of GH secretion and plasma IGF-I concentrations. In 10 cases of active acromegaly the level was 506 +/- 67 pg/ml (285-970 pg/ml). On the other hand in 20 patients with pituitary dwarfism it was only 180 +/- 15 pg/ml.(ABSTRACT TRUNCATED AT 400 WORDS)


A baseline controlled examination of a 5-day intensive treatment for pediatric obsessive-compulsive disorder.
This study extends support for a 5-day intensive exposure and response prevention (ERP) treatment protocol for pediatric obsessive compulsive disorder (OCD). Twenty-two children with OCD received ERP treatment twice daily for 5 days. The treatment also emphasized teaching children and parents how to conduct ERP independently after they returned home. Symptoms were assessed at four time-points: Baseline, 4 weeks later at pre-treatment, one week after the intensive treatment 5-day treatment, and at 3 month follow-up. Changes on the primary outcome measure, clinician severity ratings on the Anxiety Disorders Interview Schedule for Children, and secondary measures, indicated that OCD symptoms remained stable from the evaluation to baseline and improved significantly from baseline to follow-up. Moreover, parental accommodation of OCD decreased significantly from baseline to post-treatment and from post-treatment to follow-up. These data suggest that the 5-day intervention demonstrates efficacy in reducing OCD symptoms and may initiate change in parent accommodation that continues to improve after the family returns home.


Cdk1 promotes cytokinesis in fission yeast through activation of the septation initiation network.
In Schizosaccharomyces pombe, late mitotic events are coordinated with cytokinesis by the septation initiation network (SIN), an essential spindle pole body (SPB)-associated kinase cascade, which controls the formation, maintenance, and constriction of the cytokinetic ring. It is not fully understood how SIN initiation is temporally regulated, but it depends on the activation of the GTPase Spg1, which is inhibited during interphase by the essential bipartite GTPase-activating protein Byr4-Cdc16. Cells are particularly sensitive to the modulation of Byr4, which undergoes cell cycle-dependent phosphorylation presumed to regulate its function. Polo-like kinase, which promotes SIN activation, is partially responsible for Byr4 phosphorylation. Here we show that Byr4 is also controlled by cyclin-dependent kinase (Cdk1)-mediated phosphorylation. A Cdk1 nonphosphorylatable Byr4 phosphomutant displays severe cell division defects, including the formation of elongated, multinucleate cells, failure to maintain the cytokinetic ring, and compromised SPB association of the SIN kinase Cdc7. Our analyses show that Cdk1-mediated phosphoregulation of Byr4 facilitates complete removal of Byr4 from metaphase SPBs in concert with Plo1, revealing an unexpected role for Cdk1 in promoting cytokinesis through activation of the SIN pathway.


Association between the degree of obstructive sleep apnea and the severity of COVID-19: An explorative retrospective cross-sectional study.
Obstructive sleep apnea (OSA) on its own, as well as its risk factors, have been found to be associated with the outcome of Coronavirus disease 2019 (COVID-19). However, the association between the degree of OSA and COVID-19 severity is unclear. Therefore, the aim of the study was to evaluate whether or not parameters to clinically evaluate OSA severity and the type of OSA treatment are associated with COVID-19 severity. Patient data from OSA patients diagnosed with COVID-19 were reviewed from outpatients from the Isala Hospital and patients admitted to the Isala Hospital, starting from March until December 2020. Baseline patient data, sleep study parameters, OSA treatment information and hospital admission data were collected. Apnea hypopnea index (AHI), low oxyhemoglobin desaturation (LSAT), oxygen desaturation index (ODI), respiratory disturbance index (RDI), and the type of OSA treatment were regarded as the independent variables. COVID-19 severity-based on hospital or intensive care unit (ICU) admission, the number of days of hospitalization, and number of intubation and mechanical ventilation days-were regarded as the outcome variables. Multinomial regression analysis, binary logistic regression analysis, and zero-inflated negative binomial regression analysis were used to assess the association between the parameters to clinically evaluate OSA severity and COVID-19 severity. A total of 137 patients were included. Only LSAT was found to be significantly associated with the COVID-19 severity (p<0.05) when COVID-19 severity was dichotomized as non-hospitalized or hospitalized and ICU admission or death. Therefore, our findings showed that LSAT seems to be a significant risk factor for COVID-19 severity. However, the degree of OSA-based on AHI, ODI, and RDI-and OSA treatment were not found to be risk factors for COVID-19 severity when looking at hospital or ICU admission, the number of days of hospitalization, and number of intubation and mechanical ventilation days.


PIP2 is a necessary cofactor for ion channel function: how and why?
Phosphatidylinositol 4,5-bisphosphate (PIP2) is a minority phospholipid of the inner leaflet of plasma membranes. Many plasma membrane ion channels and ion transporters require PIP2 to function and can be turned off by signaling pathways that deplete PIP2. This review discusses the dependence of ion channels on phosphoinositides and considers possible mechanisms by which PIP2 and analogues regulate ion channel activity.


School life satisfaction and peer connectedness of intellectually gifted adolescents in France: Is there a labeling effect?
Intellectual giftedness is commonly associated with a high level of intellectual functioning, an identification process whereby individuals are labeled as gifted, and adjustments in schools such as grade skipping. During adolescence, all these factors are prone to reduce peer connectedness and school life satisfaction. The aim of the present study was to disentangle the effects of these factors in a sample of 492 sixth and 10th graders. We identified three subsamples based on different characteristics associated with giftedness: students previously identified as gifted (n = 66), students who scored in the top 10% on a general intelligence test (n = 49), and students who had skipped a grade (n = 57). Comparative analysis showed that none of these subsamples differed from their respective control groups on school life satisfaction. Students labeled as gifted reported a lower level of peer connectedness, and the latter's contribution to school life satisfaction was significantly stronger within this subsample. These results underscore the importance of social integration for adolescents identified as intellectually gifted and exclude grade skipping as a risk factor. Moreover, high intellectual level does not seem to impact either school life satisfaction or peer connectedness.


Structural and biochemical determinants of ligand binding by the c-di-GMP riboswitch .
The bacterial second messenger c-di-GMP is used in many species to control essential processes that allow the organism to adapt to its environment. The c-di-GMP riboswitch (GEMM) is an important downstream target in this signaling pathway and alters gene expression in response to changing concentrations of c-di-GMP. The riboswitch selectively recognizes its second messenger ligand primarily through contacts with two critical nucleotides. However, these two nucleotides are not the most highly conserved residues within the riboswitch sequence. Instead, nucleotides that stack with c-di-GMP and that form tertiary RNA contacts are the most invariant. Biochemical and structural evidence reveals that the most common natural variants are able to make alternative pairing interactions with both guanine bases of the ligand. Additionally, a high-resolution (2.3 A) crystal structure of the native complex reveals that a single metal coordinates the c-di-GMP backbone. Evidence is also provided that after transcription of the first nucleotide on the 3'-side of the P1 helix, which is predicted to be the molecular switch, the aptamer is functional for ligand binding. Although large energetic effects occur when several residues in the RNA are altered, mutations at the most conserved positions, rather than at positions that base pair with c-di-GMP, have the most detrimental effects on binding. Many mutants retain sufficient c-di-GMP affinity for the RNA to remain biologically relevant, which suggests that this motif is quite resilient to mutation.


[Treatment of fresh fibular capsular ligament lesion].
340 patients with ruptures in the region of the fibular ligaments at the ankle joint could be followed up clinically and by x-ray after an average period of 3.4 years following trauma and treatment. Significantly better treatment results were seen in the group of 246 patients who had primarily been treated by surgery (77% very good to good) than in the group of 94 patients who had been treated conservatively (41.5% very good to good). On the basis of our own follow-up results, the international literature and with particular reference to the experiences collected in sports orthopedics and sports traumatology, it continues to seem imperative to arrive at an individual indication in each case as to whether a rupture of the lateral ligaments at the ankle joint should be treated by surgery or not; the results we have reviewed point to a generally more favorable outcome of surgical treatment than of the conservative approach.


Nephrogenic diabetes insipidus in a patient with L1 syndrome: a new report of a contiguous gene deletion syndrome including L1CAM and AVPR2.
We report on an infant boy with congenital hydrocephalus due to L1 syndrome and polyuria due to diabetes insipidus. We initially believed his excessive urine loss was from central diabetes insipidus and that the cerebral malformation caused a secondary insufficient pituitary vasopressin release. However, he failed to respond to treatment with a vasopressin analogue, which pointed to nephrogenic diabetes insipidus (NDI). L1 syndrome and X-linked NDI are distinct clinical disorders caused by mutations in the L1CAM and AVPR2 genes, respectively, located in adjacent positions in Xq28. In this boy we found a deletion of 61,577 basepairs encompassing the entire L1CAM and AVPR2 genes and extending into intron 7 of the ARHGAP4 gene. To our knowledge this is the first description of a patient with a deletion of these three genes. He is the second patient to be described with L1 syndrome and NDI. During follow-up he manifested complications from the hydrocephalus and NDI including global developmental delay and growth failure with low IGF-1 and hypothyroidism.


Screening of athletic knee injuries.
The screening of athletic knee injuries requires a great deal of judgment and experience on the part of the physician. There are many gray areas with differences of opinion regarding methods of treatment by experts in the field. There is, however, unanimity of opinion regarding the necessity for immediate careful and thorough evaluation of injuries to the athlete's knee as well as prompt, appropriate treatment in order to prevent severe disability in the future.


Is myeloablative dose intensity necessary in allogeneic hematopoietic cell transplantation for lymphomas?
The advent of novel immunotherapy and tyrosine kinase inhibitors has ushered a new era in the treatment of Hodgkin and non-Hodgkin lymphomas. Allogeneic hematopoietic cell transplantation remains, however, a vital component in the management and potential cure of lymphomas, especially in the relapsed setting. Considering the biological and clinical heterogeneity of various subtypes of lymphomas, the optimal intensity of conditioning regimens remains controversial. Reduced intensity conditioning regimens have broadened applicability of the procedure to older and frail patients. Observational studies suggest that although reduced intensity allografting is associated with higher risk of relapse, overall survival is comparable and in some cases even better, than observed with myeloablative regimens. Here, we review the available published data pertaining to allogeneic hematopoietic cell transplantation using reduced intensity or myeloablative conditioning for various lymphoma histologies. Owing to the lack of randomized prospective trials, recommendations are mainly based on registry and single-institution studies. Special emphasis must be given to implementing strategies to prevent relapse when using reduced intensity regimens. Identifying particular patients who may benefit from myeloablative regimens in lymphomas remains to be better defined.


Transactivation from Gal4-VP16 transgenic insertions for tissue-specific cell labeling and ablation in zebrafish.
Prior studies with transgenic zebrafish confirmed the functionality of the transcription factor Gal4 to drive expression of other genes under the regulation of upstream activator sequences (UAS). However, widespread application of this powerful binary system has been limited, in part, by relatively inefficient techniques for establishing transgenic zebrafish and by the inadequacy of Gal4 to effect high levels of expression from UAS-regulated genes. We have used the Tol2 transposition system to distribute a self-reporting gene/enhancer trap vector efficiently throughout the zebrafish genome. The vector uses the potent, hybrid transcription factor Gal4-VP16 to activate expression from a UAS:eGFP reporter cassette. In a pilot screen, stable transgenic lines were established that express eGFP in reproducible patterns encompassing a wide variety of tissues, including the brain, spinal cord, retina, notochord, cranial skeleton and muscle, and can transactivate other UAS-regulated genes. We demonstrate the utility of this approach to track Gal4-VP16 expressing migratory cells in UAS:Kaede transgenic fish, and to induce tissue-specific cell death using a bacterial nitroreductase gene under UAS control. The Tol2-mediated gene/enhancer trapping system together with UAS transgenic lines provides valuable tools for regulated gene expression and for targeted labeling and ablation of specific cell types and tissues during early zebrafish development.


Anti-Müllerian Hormone as a marker of ovarian reserve in relation to cardio-metabolic health: a narrative review.
The final hallmark of diminishing ovarian reserve is menopause, a state known to be inextricably linked to the deterioration of female cardiovascular health. The menopausal transition is associated with an increased risk of future cardiovascular morbidity and mortality, irrespective of chronological age. The aim of this narrative review is to identify studies investigating the association between Anti-Müllerian Hormone (AMH), a marker of ovarian reserve status, and factors of cardio-metabolic risk. Both for regularly cycling women and women with polycystic ovary syndrome (PCOS), current reports are conflicting and heterogeneous, with some indicating presence and others absence of a correlation between AMH and cardio-metabolic risk factors. The occurrence of hypertensive complications in pregnancy, known to increase the risk of later cardiovascular sequelae, is associated with reduced AMH levels in various study populations. Further research remains a prerequisite in order to further elucidate a possible common mechanism for ovarian and cardiovascular decline. More knowledge of the temporal or causal association between ovarian and cardiovascular decline may enable timely identification of women with increased risk of cardiovascular disease or early onset ovarian aging. Following this, AMH may in the future play a role beyond the scope of female reproduction.


Gallstone prevalence and risk factors in patients with ulcerative colitis in Korean population.
This study aims to evaluate the prevalence, risk factors, and relative risk of gallstones and associated disease in patients with ulcerative colitis (UC). Medical records of 311 patients diagnosed with UC between January 2004 and February 2015 were reviewed retrospectively. To assess relative risk, 622 patients matched by age, sex, and body mass index were included as a control group. Gallstones were detected in 8% (25/311) of UC patients and in 3.9% (24/622) of the control group. Prevalence was significantly higher in the UC group (odds ratio [OR], 2.178; P = .007). Mean age of gallstone patients was 57.1 ± 17.8 years in the UC group, and mean disease duration of UC was 67.2 ± 38.8 months. The male-to-female ratio of gallstone patients in the UC group was 2.13:1. Mean interval from diagnosis of UC to detection of gallstones was 17.8 ± 30 months. Six UC patients with gallstones underwent cholecystectomy or endoscopic retrograde cholangiography for symptomatic disease and complications. In univariate analysis, diabetes, hypertension, age ≥65 years, and history of more than 3 admissions were significantly associated with gallstone in UC patients. In multivariate analysis, age ≥65 years (OR, 2.655; P = .033) and hospitalization ≥3 times (OR, 4.1; P = .001) were statistically significant risk factors for gallstones in UC patients. This study shows that UC patients have a significantly higher risk of gallstones compared to the general population (OR, 2.178; P = .007), especially those who are older, with a history of multiple admissions.


A somatic-mutational process recurrently duplicates germline susceptibility loci and tissue-specific super-enhancers in breast cancers.
Somatic rearrangements contribute to the mutagenized landscape of cancer genomes. Here, we systematically interrogated rearrangements in 560 breast cancers by using a piecewise constant fitting approach. We identified 33 hotspots of large (>100 kb) tandem duplications, a mutational signature associated with homologous-recombination-repair deficiency. Notably, these tandem-duplication hotspots were enriched in breast cancer germline susceptibility loci (odds ratio (OR) = 4.28) and breast-specific 'super-enhancer' regulatory elements (OR = 3.54). These hotspots may be sites of selective susceptibility to double-strand-break damage due to high transcriptional activity or, through incrementally increasing copy number, may be sites of secondary selective pressure. The transcriptomic consequences ranged from strong individual oncogene effects to weak but quantifiable multigene expression effects. We thus present a somatic-rearrangement mutational process affecting coding sequences and noncoding regulatory elements and contributing a continuum of driver consequences, from modest to strong effects, thereby supporting a polygenic model of cancer development.


Prevalence of cardiovascular and renal complications in older adults with normal or impaired glucose tolerance or NIDDM. A population-based study.
To determine the prevalence of CVD and renal disease by glucose tolerance status in a population-based study. The study consisted of 991 men and 1243 women, aged 50-89 yr, living in the upper-middle class community of Rancho Bernardo, CA. All participants received a standard OGTT between 1984 and 1987. A total of 159 men and 158 women had NIDDM, whereas 237 men and 348 women had IGT. Age-adjusted prevalence of complications generally was highest in those who were previously diagnosed with NIDDM. A significant correlation was observed between increases in the prevalence of complications and increases in glucose intolerance for microproteinuria, macroproteinuria, and myocardial infarction (borderline), as well as for stroke and claudication in men and myocardial infarction and stroke in women. In men with IGT or NIDDM, the age-adjusted prevalence of proteinuria and CVD was highest when other complications were present whereas in women such a prevalence was highest when other complications were absent. In men with diabetes, microproteinuria was associated with duration of disease. Prevalent CVD and renal disease were increased in adults with NIDDM and elevated in adults with IGT compared to those with normal glucose tolerance. The prevalence of complications in NIDDM subjects in this population-based study was substantially lower than that reported for clinic-based diabetic populations.


Neovascularized bone grafts: experimental investigation.
Vascularized bone grafts are standardized procedures in reconstructive surgery but there are some disadvantages: donor site morbidity, limited number of "natural" donor sites, and complex technique. In this study, we test the possibility of creating a "neovascularized" bone graft utilizing a vascular implantation procedure in a rabbit model. Sixteen New Zealand adult white rabbits were used. In each animal, two iliac crest bone grafts (7 x 7 x 10 mm) were harvested. Vascular implantation of the right superficial femoral vessels was performed in one of the two grafts, which was wrapped in a silicone envelope to avoid neovascularization from the surrounding tissues and positioned in a subcutaneous pocket in the right medial thigh. On the left side, the bone block, wrapped in the silicone envelope, was buried subcutaneously without vascular implantation. The operated animals were divided into two groups: Group I included eight rabbits explanted 4 weeks postoperatively and Group II included eight rabbits explanted 8 weeks postoperatively. Tetracycline injection was performed 72 hours preexplantation to evaluate new bone formation. Selective colloidal ink injection in the axial artery was performed to investigate the neovascularization before inclusion in poly-methyl-methacrylate (PMMA). Histological examination was performed in all explanted specimens comparatively. Histological examination 8 weeks after surgery showed a marked neovascularization, with normal bone cells. Tetracycline labeling showed new bone formation with a normal pattern. In all nonvascularized specimens, no viable cells or neovascularization and no bone formation were found. The vascular implantation procedure can induce a good neovascularization with new bone formation in a small bone graft. The possibility of neovascularization induction by the simple vascular implantation procedure has several clinical implications in reconstructive surgery.


Surgical treatment of farmyard pox. Orf, milker's nodules, bovine papular stomatitis pox.
Superficial epidermal surgical removal of localized bullous lesions of viral origin is recommended. An example is presented in which the patient had what we have termed "farmyard pox." This is our generic label for the clinically indistinguishable parapox viral infections acquired from farm animals, which include orf, milker's nodules, and the pox of bovine papular stomatitis. The surgery is simple and rapid, and completely removes the lesions. This eliminates the possibility of enlargement and contagion, and also promotes rapid healing.


Verapamil reduces glucose and fatty acid metabolism of beating and nonbeating rat heart cells.
The effects of various dosages of verapamil on glucose or fatty acid utilization by beating or nonbeating rat heart myocytes in tissue culture were determined. Myocytes were incubated with verapamil and either D-6-14C glucose or 1-14C palmitic acid as substrate. After incubation the subsequently generated 14CO2 was captured with hyamine hydroxide and the equivalent oxygen values were calculated. Low doses of verapamil (50 ng/ml) treatment produced a 52% reduction in myocyte glucose utilization and a 16% reduction in fatty acid utilization that appeared to be independent of its effect on myocyte contractile rate since these effects were evident in both beating and nonbeating myocytes. In addition, verapamil treatment caused differences in the myocyte handling of substrate. Verapamil (50 ng/ml) lowered cellular accumulation of glucose by 21% compared to controls. Contrary to glucose, myocyte concentrations of palmitic acid were significantly increased by 117% relative to controls in verapamil treated cultures. These results suggest that verapamil may have a direct effect on basal heart cell metabolism in a way that is unrelated to myocyte contractile activity. In addition, verapamil may interfere with glucose membrane transport.


Endoscopic Full-thickness Resection with the Full-Thickness Resection Device (FTRD) for "difficult to resect" colonic lesions. A single-center experience.
Aim of our observational and retrospective study is to compare efficacy and indications of endoscopic full-thickness resection device (FTRD) with the over-the-scope (OVESCO) clip closure for en bloc resection of colorectal lesions (including adenomas, early carcinomas, inflammatory polyps and neuroendocrine tumors). This article collected 36 cases of colorectal neoplasms from a single Italian referral center per colorectal disease treatment. Primary endpoints included en bloc resection, R0 resection and an early discharge of the patient. Secondary endpoints included procedure-related adverse events. Mean procedure time± standard deviation (SD) was 19.6±22.1 minutes and mean hospital stay (± SD) was 2.2±1.1 days. Overall, an en bloc resection was achieved in 34 cases (94.4%), with an R0 resection rate of 91.6%. Among the three not R0 patients, further additional treatments were needed. Along the same line of other already published articles, the main current indications of EFTR by FTRD-OVESCO are limited to superficial or low-risk malignancy lesions (eg, adenomas, early cancers or subepithelial tumors), not suitable to conventional endoscopic resection or in patients with a severe surgical risk. Both en bloc resection rate and complication rate are aligned with other authors' data. EFTR by FTRD system represents an effective and safe options whenever a recurrent lesion in a challenging environment occurres (eg, recent scar, low rectum or beyond a large colonic bend). Procedure-related adverse events are potentially severe, so that this novel technique should be performed by "expert hands". Difficult polypectomies, Early carcinomas, Endoscopic Full-Thickness Resection (EFTR), Full-Thickness Resection Device (FTRD) by Over-The-Scope (OVESCO) clip closure, Literature overview, Single center experience.


The role of methylation, DNA polymorphisms and microRNAs on HLA-G expression in human embryonic stem cells.
The human leukocyte antigen (HLA)-G gene seems to play a pivotal role in maternal tolerance to the fetus. Little is known about HLA-G expression and its molecular control during in vivo human embryogenesis. Human embryonic stem cells (hESC) provide an interesting in vitro model to study early human development. Different studies reported discrepant findings on whether HLA-G mRNA and protein are present or absent in hESC. Several lines of evidence indicate that promoter CpG methylation and 3' untranslated region (3'UTR) polymorphisms may influence HLA-G expression. We investigated how HLA-G expression is linked to the patterns of promoter methylation and explored the role of the 3'UTR polymorphic sites and their binding microRNAs on the post-transcriptional regulation of HLA-G in eight hESC lines. We showed that, while the gross expression levels of HLA-G are controlled by promoter methylation, the genetic constitution of the HLA-G 3'UTR, more specifically the 14bp insertion in combination with the +3187A/A and +3142G/G SNP, plays a major role in HLA-G mRNA regulation in hESC. Our findings provide a solid first step towards future work using hESC as tools for the study of early human developmental processes in normal and pregnancy-related disorders such as preeclampsia.


[Transrenal ureteric embolisation. Experimental and clinical results (author's transl)].
Transrenal ureteric embolisation with the tissue adhesive butyl 2-cyano-acrylate mixed with lipiodol and tantalum powder produces rapid and effective occlusion of the ureter. This can be combined with the use of a Gianturco spiral. In an experimental series of eight dogs observed for 54 days, the adhesive disappeared partly in three animals and totally in a further three. Fibrotic ureteric stenosis was observed in three animals. Clinical results were rather better because of the presence of external urinary drainage. Twenty-one ureters in eighteen patients were occluded either unilaterally or bilaterally, and sometimes combined with percutaneous contralateral renal embolisation. After one to four months, the ureter was still occluded in three out of six cases; after five to 17 months, three out of five cases were no longer totally occluded, but in two cases they were still blocked. The procedure is new and suitable for the treatment of otherwise untreatable conditions, such as extensive urinary fistulae, bladder tenesmus and haematuria due to extensive tumours of the minor pelvis.


Acute rhinosinusitis in adults.
Rhinosinusitis is one of the most common conditions for which patients seek medical care. Subtypes of rhinosinusitis include acute, subacute, recurrent acute, and chronic. Acute rhinosinusitis is further specified as bacterial or viral. Most cases of acute rhinosinusitis are caused by viral infections associated with the common cold. Symptomatic treatment with analgesics, decongestants, and saline nasal irrigation is appropriate in patients who present with nonsevere symptoms (e.g., mild pain, temperature less than 101°F [38.3°C]). Narrow-spectrum antibiotics, such as amoxicillin or trimethoprim/sulfamethoxazole, are recommended in patients with symptoms or signs of acute rhinosinusitis that do not improve after seven days, or that worsen at any time. Limited evidence supports the use of intranasal corticosteroids in patients with acute rhinosinusitis. Radiographic imaging is not recommended in the evaluation of uncomplicated acute rhinosinusitis. Computed tomography of the sinuses should not be used for routine evaluation, although it may be used to define anatomic abnormalities and evaluate patients with suspected complications of acute bacterial rhinosinusitis. Rare complications of acute bacterial rhinosinusitis include orbital, intracranial, and bony involvement. If symptoms persist or progress after maximal medical therapy, and if computed tomography shows evidence of sinus disease, referral to an otolaryngologist is warranted.


[Structure of lymphatic vessels in the human thighs].
Peculiarities of construction of muscular coat of lymphatics of human legs (long, polylymphangion muscular bands, longitudinally oriented, localized in deep layers of lymphatics walls and forming branches into adjacent valves of lymphangions) are the base for grouped contractions of lymphangions in them. Predominance of these muscular fascicles in muscular coat of lymphatics of thigh provide combined action of group of adjacent lymphangions versus gravitation force in the walking man.


Marketing the rural hospital-based laboratory: building a customer-centered outreach program.
Rural hospital laboratories, which operate 24 hours/day, 7 days/week to meet inpatient and emergency-room care requirements, are situated uniquely to provide outpatient laboratory services for their communities. Laboratory managers looking to augment current services should consider implementing an outreach program. This article will explore the marketing aspects involved with developing a customer-centered outreach program, including finding a market niche, developing a business plan using elements of the consumption chain, addressing customer service issues, and business plan testing and validation.


Characterization of plasmids carrying oqxAB in bla(CTX-M)-negative Escherichia coli isolates from food-producing animals.
To study the characteristics of plasmids harboring oqxAB among bla(CTX-M)-negative Escherichia coli isolates and search for oqxAB-harboring plasmids similar to plasmids carrying oqxAB-bla(CTX-M) reported previously, conjugation experiment was performed for 115 randomly selected oqxAB-positive but bla(CTX-M)-negative E. coli isolates from diseased animals in Guangdong, China. S1 nuclease pulsed-field gel electrophoresis (PFGE) and southern blotting experiments were performed to investigate the location of oqxAB and other resistance genes. The EcoRI digestion profiles of the plasmids with oqxAB were also analyzed. The clonal relatedness of donor isolates was investigated by PFGE. In this study, 32 oqxAB transconjugants were successfully obtained and most transconjugants showed multidrug resistances. Eleven replicon combination types were found in these transconjugants. floR and oqxAB were found on the same plasmids in all nine transconjugants resistant to florfenicol. The sequences between floR and oqxAB were identical in most transconjugants and the two genes were both linked with tnp in insertion sequences. Nine F18:A-:B1 plasmids with only oqxAB shared identical EcoRI digestion profiles and the profiles were also identical with that of a plasmid carrying oqxAB-bla(CTX-M) found previously. Co-transfer of plasmids carrying oqxAB and fosA3, respectively, was also observed in one isolate. This study demonstrates the dissemination of oqxAB among bla(CTX-M)-negative E. coli isolates was mainly mediated by identical F18:A-:B1 plasmids. A novel arrangement of regions between floR and oqxAB might play an important role in the dissemination of floR-oqxAB. This is the first description of the genetic environment of the relationship between oqxAB and floR in E. coli.


Humans perceive flicker artifacts at 500 Hz.
Humans perceive a stable average intensity image without flicker artifacts when a television or monitor updates at a sufficiently fast rate. This rate, known as the critical flicker fusion rate, has been studied for both spatially uniform lights, and spatio-temporal displays. These studies have included both stabilized and unstablized retinal images, and report the maximum observable rate as 50-90 Hz. A separate line of research has reported that fast eye movements known as saccades allow simple modulated LEDs to be observed at very high rates. Here we show that humans perceive visual flicker artifacts at rates over 500 Hz when a display includes high frequency spatial edges. This rate is many times higher than previously reported. As a result, modern display designs which use complex spatio-temporal coding need to update much faster than conventional TVs, which traditionally presented a simple sequence of natural images.


Evolutionary legacy of a forest plantation tree species (Pinus armandii): Implications for widespread afforestation.
Many natural systems are subject to profound and persistent anthropogenic influence. Human-induced gene movement through afforestation and the selective transportation of genotypes might enhance the potential for intraspecific hybridization, which could lead to outbreeding depression. However, the evolutionary legacy of afforestation on the spatial genetic structure of forest tree species has barely been investigated. To do this properly, the effects of anthropogenic and natural processes must be examined simultaneously. A multidisciplinary approach, integrating phylogeography, population genetics, species distribution modeling, and niche divergence would permit evaluation of potential anthropogenic impacts, such as mass planting near-native material. Here, these approaches were applied to Pinus armandii, a Chinese endemic coniferous tree species, that has been mass planted across its native range. Population genetic analyses showed that natural populations of P. armandii comprised three lineages that diverged around the late Miocene, during a period of massive uplifts of the Hengduan Mountains, and intensification of Asian Summer Monsoon. Only limited gene flow was detected between lineages, indicating that each largely maintained is genetic integrity. Moreover, most or all planted populations were found to have been sourced within the same region, minimizing disruption of large-scale spatial genetic structure within P. armandii. This might be because each of the three lineages had a distinct climatic niche, according to ecological niche modeling and niche divergence tests. The current study provides empirical genetic and ecological evidence for the site-species matching principle in forestry and will be useful to manage restoration efforts by identifying suitable areas and climates for introducing and planting new forests. Our results also highlight the urgent need to evaluate the genetic impacts of large-scale afforestation in other native tree species.


Impact of Fgf10 deficiency on pulmonary vasculature formation in a mouse model of bronchopulmonary dysplasia.
Bronchopulmonary dysplasia (BPD), characterized by alveoli simplification and dysmorphic pulmonary microvasculature, is a chronic lung disease affecting prematurely born infants. Pulmonary hypertension (PH) is an important BPD feature associated with morbidity and mortality. In human BPD, inflammation leads to decreased fibroblast growth factor 10 (FGF10) expression but the impact on the vasculature is so far unknown. We used lungs from Fgf10+/- versus Fgf10+/+ pups to investigate the effect of Fgf10 deficiency on vascular development in normoxia (NOX) and hyperoxia (HOX, BPD mouse model). To assess the role of fibroblast growth factor receptor 2b (Fgfr2b) ligands independently of early developmentaldefects, we used an inducible double transgenic system in mice allowing inhibition of Fgfr2b ligands activity. Using vascular morphometry, we quantified the pathological changes. Finally, we evaluated changes in FGF10, surfactant protein C (SFTPC), platelet endothelial cell adhesion molecule (PECAM) and alpha-smooth muscle actin 2 (α-SMA) expression in human lung samples from patients suffering from BPD. In NOX, no major difference in the lung vasculature between Fgf10+/- and control pups was detected. In HOX, a greater loss of blood vessels in Fgf10+/- lungs is associated with an increase of poorly muscularized vessels. Fgfr2b ligands inhibition postnatally in HOX is sufficient to decrease the number of blood vessels while increasing the level of muscularization, suggesting a PH phenotype. BPD lungs exhibited decreased FGF10, SFTPC and PECAM but increased α-SMA. Fgf10 deficiency-associated vascular defects are enhanced in HOX and could represent an additional cause of morbidity in human patients with BPD.


Defects in Efflux (oprM), β-Lactamase (ampC), and Lipopolysaccharide Transport (lptE) Genes Mediate Antibiotic Hypersusceptibility of Pseudomonas aeruginosa Strain Z61.
Antibiotic hypersensitive bacterial mutants (e.g., Escherichia coliimp) are used to investigate intrinsic resistance and are exploited in antibacterial discovery to track weak antibacterial activity of novel inhibitor compounds. Pseudomonas aeruginosa Z61 is one such drug-hypersusceptible strain generated by chemical mutagenesis, although the genetic basis for hypersusceptibility is not fully understood. Genome sequencing of Z61 revealed nonsynonymous single-nucleotide polymorphisms in 153 genes relative to its parent strain, and three candidate mutations (in oprM, ampC, and lptE) predicted to mediate hypersusceptibility were characterized. The contribution of these mutations was confirmed by genomic restoration of the wild-type sequences, individually or in combination, in the Z61 background. Introduction of the lptE mutation or genetic inactivation of oprM and ampC genes alone or together in the parent strain recapitulated drug sensitivities. This showed that disruption of oprM (which encodes a major outer membrane efflux pump channel) increased susceptibility to pump substrate antibiotics, that inactivation of the inducible β-lactamase gene ampC contributed to β-lactam susceptibility, and that mutation of the lipopolysaccharide transporter gene lptE strongly altered the outer membrane permeability barrier, causing susceptibility to large antibiotics such as rifampin and also to β-lactams.


Utilization of occupational therapy services and relation to survival in people taking dementia-specific medication in Austria-A retrospective population-based study with a 13-year observation period.
Evidence-based treatment of dementia includes pharmacological and non-pharmacological methods of which psycho-social interventions are an important component, commonly administered by occupational therapists. The aim of this study was to investigate the utilization of occupational therapy (OT) services and its association with survival in people taking dementia-specific medication in a population-based Austrian dataset compared to a two times as large control group without dementia-specific medication. A retrospective study with a 13-year observation period (2003-2016) was conducted on real-world data. Two stratifications were done and we used descriptive statistics, Chi-squared/Fisher's Exact Tests and survival analyses including three Cox models. Data from 286,553 participants were analysed. Only 4.5% (n = 12,950) received OT services. In the dementia-medication group (n = 111,033), participants who received OT services (3.6%; n = 4032) had significantly more comorbidities (4.7%) compared to those without OT (3.5%; p < 0.001) and were also more likely to be male (4 vs. 3.5%; p < 0.001). While persons taking dementia-specific medication showed a slightly reduced survival with OT (p < 0.001) compared to those without, the result in the control group without dementia-specific medication showed a slightly better result of the participants who received OT (p < 0.001). The reduced survival in the dementia-medication group with OT is likely to be related to the higher number of comorbidities in this group. People receiving dementia-specific medication were more likely to receive OT if they had additional comorbidities, however our analysis showed that utilization of OT services in Austria was very low indicating an overall insufficient accessibility of OT services for patients who needed it.


Studies of the cardiac-like action potential in crayfish giant axons induced by platinized tungsten metal electrodes.
A lightly platinized tungsten (Pt-W) wire electrode, axially inserted into a crayfish giant axon, causes the development of cardiac-like action potentials with durations of up to 4 s. The plateau in membrane potential typically occurs within 10 min of the start of action potential elongation. The effect occurs without passing current through the Pt-W electrode and is temporally related to a dramatic decrease in intracellular pH (pHi). Such an effect cannot be induced by a decrease in pHi produced by equilibrating the axon with HCO3(-)-CO2 solution (pH6), and NH4Cl rebound or direct intracellular injection of PO4(3-) buffer (pH 4 X 5). Action potential elongation is accompanied by a block of delayed rectification and the possibility that inward rectification also develops cannot be ruled out. Plateau generation requires Na+ and Ca2+ inward currents as demonstrated by abolition of the plateau by [Na+]o or [Ca2+]o depletion or treatment with tetrodotoxin (TTX) or verapamil. The block of outward rectification by Pt-W requires external Na+ or Ca2+. Action potential elongation produced by 3,4-diaminopyridine is not sensitive to verapamil and the waveform is different from that produced by Pt-W. The data support the possibility that different classes of excitable membranes have similar channel populations and that the functional differences between them reside in the inhibitory or masking influences that are present in the microenvironments of the various membrane channels.


Research utilization and evidence-based practice in occupational therapy: a scoping study.
Many articles have been written on the barriers to and facilitators of the use of evidence in practice in nursing and medicine, but to date no extensive review has been published of the literature on evidence-based practice (EBP) supports in occupational therapy. This article presents the results of a scoping review that examined factors that support the integration of research into practice. A review of 69 articles revealed four themes: (1) attitudes toward, perceptions of, confidence in, and use of research and EBP; (2) factors that support the use of research in practice; (3) effects of interventions targeting changes in knowledge, attitudes, skills, behaviors, and evidence-based practices; and (4) identification of the processes involved in the acquisition of EBP skills and their application in clinical practice. A process that integrates client-centered practice, structured reflection, case application, and peer consultations within a scholarship of practice model facilitates occupational therapists' evaluation and integration of research evidence.


[Role of antiphospholipid antibodies in occlusion of retinal vessels in various vascular eye diseases].
A strong association between the presence of antiphospholipid antibodies (aPLab) and arterial and venous thrombosis was observed during the past decade. aPLab represent a heterogeneous group of immunoglobulins that include the lupus anticoagulant (LA), anticardiolipin antibodies (aCL), and antibodies responsible for false-positive test for syphilis. aPLab react with negatively charged, rarely with neutral phospholipids and/or phospholipid-binding proteins. aPLab are associated with vascular occlusion in autoimmune diseases, e.g. systemic lupus erythematosus (SLE). The role of aCL in ocular vascular occlusive disease involving retinal vessels is unclear. The aim of this study was to determine the relationship between aPLab and occlusive retinal disease. A relationship between aCL (IgG), LA, and thrombotic retinal events has been detected in SLE patients, but no association between aPLab and other occlusive retinal diseases (central retinal vein thrombosis, isolated retinal vasculitis, optic nerve vasculitis). In our study the incidence of aCL IgG was higher in patients with Behcet disease with non-occlusive thrombosis than in patients with retinal occlusive events or in patients without occlusions or thrombosis. It seems that different mechanisms are responsible for these conditions.


Needle type and the risk of post-lumbar puncture headache in the outpatient neurology clinic.
Post-lumbar puncture headaches (PLPHs) are a common complication of diagnostic lumbar punctures (LPs) caused by a persistent leak of spinal fluid from the dural puncture site. We conducted a prospective study to determine risk factors associated with PLPHs in the neurology outpatient setting. Clinical information from all diagnostic LPs performed at the Johns Hopkins Lumbar Puncture Clinic between September 2008 and June 2009 was reviewed. As standard of care, each patient was contacted by telephone by the attending physician within 2-5 days of having an LP to ascertain health status and the presence of PLPH. We performed multiple logistic regression analysis to evaluate the association between PLPH and needle type (traditional Quincke cutting needle 20 and 22 gauge, 20Q and 22Q, and Sprotte non-traumatic gauge 22 needle, 22S) adjusting for important variables such as traumatic LPs, number of attempts, positioning and volume of CSF drawn. The prevalence of PLPH was 32% with the popular gauge 20Q and 22Q needles compared to 19% with the 22S non-traumatic needle. Compared to the 20Q needle, the non-traumatic 22S needle was associated with 69% decreased odds of PLPH (adjusted OR: 0.31, 95% CI 0.12-0.82). In subset analysis, the odds of PLPH increased 4-fold when the 22Q needle was used compared to the 22S needle (adjusted OR=3.99, 95% CI 1.32-12.0). Our outpatient findings support the American Academy of Neurology recommendations to use smaller non-traumatic needles to reduce the risk of PLPH.


Extension of life span by down-regulation of enzymes catalyzing tryptophan conversion into kynurenine: Possible implications for mechanisms of aging.
The end products of catabolism of tryptophan (Trp), an essential amino acid, are known to affect mechanism(s) of aging, a neurodegenerative condition. This review focuses on the possible role of the initial step of Trp catabolism, kynurenine (Kyn) formation from Trp, in aging mechanism(s). Rate-limiting enzymes of Trp conversion into Kyn are tryptophan 2,3-dioxygenase 2 (TDO) or indoleamine 2,3-dioxygenase (IDO). Aging is associated with up-regulated production of cortisol, an activator of TDO, and pro-inflammatory cytokines, inducers of IDO. The other rate-limiting enzyme of Kyn formation from Trp is ATP-binding cassette (ABC) transporter that regulates Trp availability as a substrate for TDO. Inhibitors of TDO (alpha-methyl tryptophan) and ABC transporter (5-methyltryptophan) extended life span of wild-type Drosophila. Life span prolongation was observed in TDO knockdown of Caenorhabditis elegans and in TDO or ABC transporter-deficient Drosophila mutants. Down-regulation of enzymes catalyzing Kyn conversion into kynurenic acid (KYNA) and 3-hydroxykynurenine decreases life span. Considering that down-regulation of Methuselah (MTH) gene prolonged life span, aging-accelerating effect of KYNA, a GPR35/MTH agonist, might depend on MTH gene activation. Mice treated with TDO inhibitor, benserazide, an ingredient of anti-Parkinson medication carbidopa, and TDO-deficient Drosophila mutants were resistant to inducement of aging-associated Metabolic Syndrome by high-sugar or high-fat diets. Up-regulation of Kyn formation was associated with accelerated aging and increased mortality in human subjects. Trp-Kyn pathway is evolutionary conserved (from yeasts, through insects, worms, vertebrates to humans). Further studies might explore possible antiaging effect of down-regulation of Kyn formation from Trp by dietary, pharmacological, and genetic interventions.


Quantifying changes in the thiol redox proteome upon oxidative stress in vivo.
Antimicrobial levels of reactive oxygen species (ROS) are produced by the mammalian host defense to kill invading bacteria and limit bacterial colonization. One main in vivo target of ROS is the thiol group of proteins. We have developed a quantitative thiol trapping technique termed OxICAT to identify physiologically important target proteins of hydrogen peroxide (H(2)O(2)) and hypochlorite (NaOCl) stress in vivo. OxICAT allows the precise quantification of oxidative thiol modifications in hundreds of different proteins in a single experiment. It also identifies the affected proteins and defines their redox-sensitive cysteine(s). Using this technique, we identified a group of Escherichia coli proteins with significantly (30-90%) oxidatively modified thiol groups, which appear to be specifically sensitive to either H(2)O(2) or NaOCl stress. These results indicate that individual oxidants target distinct proteins in vivo. Conditionally essential E. coli genes encode one-third of redox-sensitive proteins, a finding that might explain the bacteriostatic effect of oxidative stress treatment. We identified a select group of redox-regulated proteins, which protect E. coli against oxidative stress conditions. These experiments illustrate that OxICAT, which can be used in a variety of different cell types and organisms, is a powerful tool to identify, quantify, and monitor oxidative thiol modifications in vivo.


Fungicide effect on nitrogenase activity in methylotrophic bacteria.
The study comprised tests on the effect of fungicides: Oxafun T, Funaben T and Baytan Universal on the nitrogenase activity of methylotrophic bacteria, selected from the rhizosphere and non-rhizosphere soil of spring barley. The obtained results indicated that the field dose of fungicides - 0.5 ppm had no effect on all the examined strains of methylotrophic bacteria. The dose of 50 ppm, however, had varied effects, suppressing or stimulating, depending on the particular strain. The highest concentration of fungicides applied, i.e. 100 ppm in all cases suppressed the nitrogenase activity of the strains under examination.


Evaluation of a New Personalized Health Dashboard in Preventive Child Health Care: Protocol for a Mixed Methods Feasibility Randomized Controlled Trial.
A new dashboard, the 360ºCHILD-profile, was developed to adopt personalized health care within preventive child health care. On this profile, holistic health data are visualized in a single image to provide parents, adolescents, and caregivers direct access to a manageable résumé of a child's medical record. Theoretical ordering, conforming to "International Classification of Functioning, Disability and Health for Children and Youth", guides clinical reasoning toward the biopsychosocial concept of health. It is yet unknown if and how this promising tool functions in practice, and a variety of feasibility questions must be addressed. This paper describes the design and methods of a feasibility randomized controlled trial (RCT), with the aim of evaluating the RCT's feasibility (recruitment, response, measure completion, and intervention allocation) and 360ºCHILD-profile's feasibility (usability and potential effectiveness). A pragmatic mixed methods study design was chosen, starting with an RCT to measure feasibility and health literacy in 2 parallel groups (1:1). Qualitative research will then be used to understand and explain quantitative findings and to explore the stakeholders' perspectives on the potential of the 360ºCHILD-profile. Participants will include child health care professionals (n≥30), parents (n≥30), and caregivers (n≥10) of children who experience developmental problems (age 0-16 years). Children will only be able to participate if they are older than 11 years (adolescents, n≥10). The 2 groups included in the study will receive standard care. The experimental group will additionally receive personalized 360ºCHILD-profiles. After an intervention period of 6 months, quantitative outcomes will be measured, analyzed (descriptive feasibility statistics and preliminary between-group differences) and used to purposively sample for semistructured interviews. Study results will provide knowledge for building theory on the 360ºCHILD-profile and designing future (effect) studies. Netherlands Trial Register NTR6909; https://www.trialregister.nl/trial/6731. DERR1-10.2196/21942.


Comprehensive peptidomic and glycomic evaluation reveals that sweet whey permeate from colostrum is a source of milk protein-derived peptides and oligosaccharides.
Whey permeate is a co-product obtained when cheese whey is passed through an ultrafiltration membrane to concentrate whey proteins. Whey proteins are retained by the membrane, whereas the low-molecular weight compounds such as lactose, salts, oligosaccharides and peptides pass through the membrane yielding whey permeate. Research shows that bovine milk from healthy cows contains hundreds of naturally occurring peptides - many of which are homologous with known antimicrobial and immunomodulatory peptides - and nearly 50 oligosaccharide compositions (not including structural isomers). As these endogenous peptides and oligosaccharides have low-molecular weight and whey permeate is currently an under-utilized product stream of the dairy industry, we hypothesized that whey permeate may serve as an inexpensive source of naturally occurring functional peptides and oligosaccharides. Laboratory fractionation of endogenous peptides and oligosaccharides from bovine colostrum sweet whey was expanded to pilot-scale. The membrane fractionation methodology used was similar to the methods commonly used industrially to produce whey protein concentrate and whey permeate. Pilot-scale fractionation was compared to laboratory-scale fractionation with regard to the identified peptides and oligosaccharide compositions. Results were interpreted on the basis of whether industrial whey permeate could eventually serve as a source of functional peptides and oligosaccharides. The majority (96%) of peptide sequences and the majority (96%) of oligosaccharide compositions found in the laboratory-scale process were mirrored in the pilot-scale process. Moreover, the pilot-scale process recovered an additional 33 peptides and 1 oligosaccharide not identified from the laboratory-scale extraction. Both laboratory- and pilot-scale processes yielded peptides deriving primarily from the protein β-casein. The similarity of the laboratory-and pilot-scale's resulting peptide and oligosaccharide profiles demonstrates that whey permeate can serve as an industrial-scale source of bovine milk peptides and oligosaccharides.


Impact of hepatitis C virus infection on kidney transplant outcome.
One hundred and forty kidney transplant recipients were evaluated to study the impact of hepatitis C virus (HCV) infection on patient and graft outcome. There .were 98 males arid 42 females with a mean age of 32.1 +/- 13 years. The duration of follow-up ranged from 6-60 months with a mean period of 27.8 +/- 18.2 months. Seventy-four (53%) patients had received cadaveric kidneys while 66 (47%) received living donor grafts. Anti-HCV reactivity was tested using second generation enzyme-linked immunosorbent assay and positivity was confirmed by recombinant immunoblot assay. HCV infection was diagnosed in 29 cases (20.7%) while HBsAg was found in nine (6.4%) and concomitant anti-HCV and HBsAg positivity was observed in two patients (1.4%). Seventeen of 29 (58.6%) patients with anti-HCV reactivity showed elevated ALT levels as against 17 of 111 (17.3%) anti-HCV non-reactive patients (P< 0.001). There was no association between the sex of the patient, source of the graft, and anti-HCV reactivity. Serum creatinine values were higher in the anti-HCV positive group, but this did not rank to statistical significance. We observed a significantly higher graft loss among the anti-HCV reactive group (27.6% versus 1.8%, P< 0.003). Thirteen anti-HCV reactive patients were subjected to 18 liver biopsies; the commonest lesion observed was chronic active hepatitis, which was progressive in two patients subjected to re-biopsy. We conclude that HCV infection is a serious health problem among kidney transplant recipients and it significantly affects the graft outcome.


Alternative splicing and its role in pathologies of the endocrine system.
Alternative splicing of pre-mRNA is a process in which noncoding regions of primary transcript are removed and coding regions are joined in different manners to produce mRNA molecules of different sequences. Alternative splicing affects nearly all human genes and is a key source of diversity of proteins coded by a relatively small number of genes. Since alternative splicing is of crucial importance for the proper functioning of cells, including those involved in hormonal signalling, aberrations of alternative splicing can lead to disruption of cellular mechanisms and in consequence result in serious endocrine pathologies. Disturbances of alternative splicing include mutations of consensus splice regulatory sites and improprieties in the action of splicing factors, the proteins involved in regulating the process. In consequence of disturbed alternative splicing, improperly spliced mRNA and protein isoforms can be produced which can lead to disruption of function of their wild type counterparts. This review aims to discuss the role of alternative splicing in pathologies of the endocrine system and gives examples that highlight the importance of this process in the proper functioning of hormones, hormone receptors and other factors involved in hormonal regulation. The examples given include endocrine-related tumours (pituitary tumours, cancers of the thyroid, prostate, ovary and breast, and insulinoma), isolated growth hormone deficiency, and Frasier syndrome. Non-endocrine pathologies in which aberrant alternative splicing of transcripts of genes involved in hormonal signalling have been detected are also described. Finally, we discuss future perspectives on the possible usage of alternative splicing in diagnostics and therapy.


Transobturator tape compared with tension-free vaginal tape for the treatment of stress urinary incontinence: a randomized controlled trial.
To compare the safety and efficacy of the transobturator tape to tension-free vaginal tape (TVT) in the treatment of stress urinary incontinence in patients with and without concurrent pelvic organ prolapse. One-hundred seventy women with urodynamic stress incontinence, including those with and those without pelvic organ prolapse, from three academic medical centers were randomized to receive TVT or transobturator tape. Subjects with detrusor overactivity or previous sling surgery were excluded. The primary outcome was the presence or absence of abnormal bladder function, a composite outcome defined as the presence of any the following: incontinence symptoms of any type, a positive cough stress test, or retreatment for stress incontinence or postoperative urinary retention assessed 1 year after surgery. This study is a noninferiority study design. Of 180 women who enrolled in the study, 170 underwent surgery and 168 returned for follow-up, with a mean follow-up of 18.2+/-6 months. Mean operating time, length of stay, and postoperative pain scores were similar between the two groups. Bladder perforations occurred more frequently in the TVT group (7% compared with 0%, P=.02); otherwise, the incidence of perioperative complications was similar. Abnormal bladder function occurred in 46.6% of TVT patients and 42.7% of transobturator tape patients, with a mean absolute difference of 3.9% favoring transobturator tape (95% confidence interval -11.0% to 18.6%.). The P value for the one-sided noninferiority test was .006, indicating that transobturator tape was not inferior to TVT. The transobturator tape is not inferior to TVT for the treatment of stress urinary incontinence and results in fewer bladder perforations. ClinicalTrials.gov, www.clinicaltrials.gov, NCT00475839.


Commercial insulin immunoassays fail to detect commonly prescribed insulin analogues.
Blood insulin and C-peptide are key investigations in the differential diagnosis of hypoglycaemia. Analogues of insulin have modified primary-sequences compared to native human insulin, as such may not cross react with insulin assays. This has important implications in detecting surreptitious or malicious insulin administration. The aim of this study is to assess the cross-reactivity of all insulins currently listed in the British National Formulary (BNF65, 2013) in clinical insulin assays currently used in UK clinical laboratories. Sample sets were prepared for all 15 exogenous insulin classes listed in the BNF, at concentrations of 1000 pmol/L and 300 pmol/L, using pooled human serum. Samples were sent blinded to 5 participating analytical laboratories to cover analysis on the 10 major clinical insulin assays used in the UK. The ability of insulin assays to detect exogenous insulin preparations was highly variable and ranged from 0% to >140% for a single exogenous insulin. Four assays were highly specific for the human insulin sequence and had no cross-reactivity with any synthetic analogue insulin. Two detected all insulin types (human sequence, animal and synthetic analogue), with the remaining having variable cross-reactivity. The cross-reactivity of the 15 exogenous insulin preparations is highly variable in the assays used in clinical laboratories around the UK. It is important that laboratories and clinicians are aware of the limitations of their local assays to avoid missing the important diagnosis of hypoglycaemia secondary to excessive exogenous insulin. Where necessary, samples should be referred to specialist centres for insulin analysis and ideally by a validated and fully-quantitative mass spectrometry-based method.


Explicit non-Markovian susceptible-infected-susceptible mean-field epidemic threshold for Weibull and Gamma infections but Poisson curings.
Although non-Markovian processes are considerably more complicated to analyze, real-world epidemics are likely non-Markovian, because the infection time is not always exponentially distributed. Here, we present analytic expressions of the epidemic threshold in a Weibull and a Gamma SIS epidemic on any network, where the infection time is Weibull, respectively, Gamma, but the recovery time is exponential. The theory is compared with precise simulations. The mean-field non-Markovian epidemic thresholds, both for a Weibull and Gamma infection time, are physically similar and interpreted via the occurrence time of an infection during a healthy period of each node in the graph. Our theory couples the type of a viral item, specified by a shape parameter of the Weibull or Gamma distribution, to its corresponding network-wide endemic spreading power, which is specified by the mean-field non-Markovian epidemic threshold in any network.


["Hypertensive heart" symptoms in the elderly].
Arterial hypertension is a leading pathology in the elderly. This pathology become extremely important in a spread atherosclerotic process commonly observed in the geriatric patients. While the aspects of left ventricle in arterial hypertensive patients are rather well known, this pathology needs further investigations connected with the studies on age-related changes in cardiovascular system. Particular attention should be paid to a symptomatic of "hypertensive heart" in the elderly patients.


Defect induced ferromagnetism in Mn3Ga.
Ni-substituted Mn3Ga displays a weak ferromagnetism embedded in an antiferromagnetic phase. Upon field cooling, the alloy exhibits exchange bias and an open hysteresis loop, signifying kinetic arrest at room temperature. For the first time, a kinetic arrest is seen in a compound due to the first order transition of an embedded defect phase. A systematic study of crystal structure, local structure, and magnetic properties of Mn3-xNixGa (x = 0, 0.25) alloys reveal the origin of ferromagnetism
in Mn2.75Ni0.25Ga is due to the segregation of a Heusler-type environment around Ni in the cubic Mn3Ga matrix. Upon temper annealing at 400◦C, these local structural defects around the Ni phase separate into a modulated ferromagnetic Ni-Mn-Ga Heusler phase. A strong interaction between the antiferromagnetic host and the
ferromagnetic defect phase gives rise to exchange bias. The first-order transition of the defect phase seems to be responsible for the observed kinetic arrest in Mn2.75Ni0.25Ga.


Evaluation of tumor size response metrics to predict survival in oncology clinical trials.
Model-based drug development in oncology is still lagging despite a good momentum in the clinical pharmacology and pharmacometry community in the past few years. The failure rate of late-stage oncology studies is one of the highest across therapeutic areas. The modeling of the relationship between longitudinal tumor size and overall survival has been proposed to enhance learning in early clinical studies, to predict overall survival, and to simulate clinical trials. This approach has the potential to support proof of concept, early clinical decisions, and design of late-stage trials, but it is not yet widely integrated into the oncology drug development process. In this article, we review the state of these modeling efforts and discuss several key applications of these models. We conclude by suggesting a few paths forward.


Precipitate production by some Malassezia species on Dixon's agar.
The production of a precipitate by Malassezia species on Dixon's agar was observed. Malassezia furfur (n = 12), M. obtusa (n = 2) and M. slooffiae (n = 3) were precipitate negative, while M. sympodialis (n = 32) and M. globosa (n = 6) were precipitate positive. This test may be useful in differentiating Malassezia species.


Antiphospholipid antibodies: an epiphenomenon in Tourette syndrome.
Antiphospholipids (aPLAs) have been previously identified in children with Tourette syndrome (TS), which has led to the speculation that these antibodies might have a pathophysiologic role in this disorder. Therefore, 21 healthy children and adolescents with TS, whose ages ranged from 7 to 17 years, underwent laboratory studies designed to diagnose the lupus anticoagulant, anticardiolipin (aCL) antibodies [immunoglobulin (Ig) G, IgA, and IgM], and antinuclear antibodies. Although five subjects had at least one value that differed from accepted laboratory standards, the changes were marginal in four of them. Lupus anticoagulant was identified in one patient, based on a minimal requirement of a prolonged dilute Russell viper venom time, clotting studies that did not correct after mixture with normal plasma, and an abnormal platelet neutralization procedure. A prolonged (but correctable) activated partial thromboplastin time was found in one individual, and aCL IgG was marginally increased in three subjects. Two (10%) of a control population of 20 same-age children also had low positive aCL IgG levels. There were no differences in tics (onset, type, frequency, severity, and family history) or comorbid features between children with normal or "abnormal" laboratory study results. Our data suggest that the presence of aPLAs in TS represents an epiphenomenon rather than a pathophysiologic mechanism.


[Hemodynamic changes in healthy persons and in patients with arterial hypertension during hyperventilation].
Hemodynamic changes in response to the hyperventilation test were examined in normal males and females of varying age, and in patients with first- and second-stage essential hypertension, renal arterial hypertension and hypertensive neurocirculatory dystonia. Normal subjects showed increased heart rate (HR) and cardiac index (CI), and lowered specific peripheral resistance in response to hyperventilation, the magnitude of HR and CI changes declining with age. The increase in HR and CI during the test was somewhat more pronounced in patients with essential hypertension as compared to normal subjects, whereas in hypertensive neurocirculatory dystonia hemodynamic shifts were similar to those of normal subjects.


Stabilization of acyclic water tetramer in a copper(II) malonate framework structure.
Copper(II) complex [Cu(dpq)(mal)(H(2)O)]·3H(2)O (1) (dpq = dipyrido-[3,2-d:2',3'-f]-quinoxaline, mal = malonato) was synthesized and characterized by elemental analysis, infrared spectroscopy, thermogravimetric analysis and single-crystal X-ray crystallography. The single-crystal X-ray structure of 1 reveals a square pyramidal structure, with the dipyrido-[3,2-d:2',3'-f]-quinoxaline and malonato at the equatorial positions and a water molecule at the axial position. The molecule acts as a building block generating a supramolecular three-dimensional metal-organic framework (MOF) encapsulating metal linked acyclic water tetramer. The H-bonding capacity of malonato and the π-π stacking interactions of dipyrido-[3,2-d:2',3'-f]-quinoxaline further reinforce the framework. The copper(II) bound hydroxyl group is demonstrated to mediate hydrolytic cleavage of plasmid pBR322 DNA under dark conditions.