Time course of dementia following sepsis in German health claims data.
We evaluated the short-, medium-, and long-term effects of sepsis on dementia incidence using German health claims data. A total of 161,567 patients (65 years or older) were followed from 2004 to 2015 at quarterly intervals. Time since sepsis was categorized into 0 (the effective quarter of sepsis diagnosis), 1-8, and ≥9 quarters since the latest diagnosis of sepsis, taking into account admission to intensive care unit and controlling for delirium, surgery, age, sex, and comorbidities. Incident dementia was defined for all persons who did not have a validated dementia diagnosis in 2004 and 2005 and who received a first-time, valid diagnosis between 2006 and 2015. During the quarter of sepsis diagnosis, patients not admitted to intensive care had a 3.14-fold (95% CI 2.83-3.49) increased risk, and those with intensive care stay had a 2.22-fold (95% CI: 1.83-2.70) increased risk of receiving an incident dementia diagnosis compared with patients without sepsis. The impact of sepsis on incident dementia remained in the following 2 years, remitting only thereafter. For sepsis survivors, medium-term dementia risk remains elevated, whereas long-term risk may reach the level of those without sepsis, even after controlling for delirium. These findings encourage identifying modifiable components of hospital and rehabilitation care.


Targeting genomic rearrangements in tumor cells through Cas9-mediated insertion of a suicide gene.
Specifically targeting genomic rearrangements and mutations in tumor cells remains an elusive goal in cancer therapy. Here, we used Cas9-based genome editing to introduce the gene encoding the prodrug-converting enzyme herpes simplex virus type 1 thymidine kinase (HSV1-tk) into the genomes of cancer cells carrying unique sequences resulting from genome rearrangements. Specifically, we targeted the breakpoints of TMEM135-CCDC67 and MAN2A1-FER fusions in human prostate cancer or hepatocellular carcinoma cells in vitro and in mouse xenografts. We designed one adenovirus to deliver the nickase Cas9D10A and guide RNAs targeting the breakpoint sequences, and another to deliver an EGFP-HSV1-tk construct flanked by sequences homologous to those surrounding the breakpoint. Infection with both viruses resulted in breakpoint-dependent expression of EGFP-tk and ganciclovir-mediated apoptosis. When mouse xenografts were treated with adenoviruses and ganciclovir, all animals showed decreased tumor burden and no mortality during the study. Thus, Cas9-mediated suicide-gene insertion may be a viable genotype-specific cancer therapy.


[The ontogeny of the alpha 2- and beta-adrenoreceptors in the brain following exposure to corticosterone during intrauterine development].
Ontogenesis of alpha 2- and beta-adrenoreceptors in brain cortex of rats after corticosterone injections to their mothers on days 16 and 18 days of gestation was studied using 3H-clonidine and 3H-dihydroaoprenolol binding. After corticosterone treatment the level of beta-adrenoreceptors was decreased on day 7 but increased on day 16 after birth; On the contrary, catecholamine levels and biosynthesis determined previously were increased in the first and decreased in the second age group. beta-adrenoreceptors' density and noradrenaline level during other periods of pre- and postnatal development remained unchanged. Corticosterone treatment inhibited the increase of alpha 2-adrenoreceptor density during ontogenesis. The decrease of the alpha 2-adrenoreceptors was most significant in 35- and 70-day old rates. The hormone had no effect on the adrenoreceptors' affinity to ligands. Injections of the solvent to the pregnant females which is weak stressor increasing corticosteroids levels in the blood, had a similar but less prominent effect on the ontogenesis of adrenoreceptors in the offspring brain. Thus, the effect of corticosterone on the ontogenesis of the beta-adrenoreceptors is probably associated with changes of the neurotransmitter level, and its effect on alpha 2-adrenoreceptors involves some unknown specific hormone-dependent regulator.


Psychosocial and geriatric correlates of functional status after total hip replacement.
To determine whether psychosocial factors, chronic diseases, and common geriatric problems are associated with poor physical function 3 years after primary total hip replacement (THR). We studied a sample of Medicare recipients in Ohio, Pennsylvania, and Colorado (n = 922) who underwent primary THR in 1995 (mean +/- SD age 73.1 +/- 5.6 years, 32% men). Participants completed a questionnaire regarding lifestyle factors, medical history, and quality of life approximately 3 years after the surgery. Physical function was measured using the function subscale of the Western Ontario and McMaster Universities Osteoarthritis Index. We assessed the relationship between functional outcome 3 years postsurgery and 4 predictor domains: pain or complications in the operated hip, other musculoskeletal comorbidity, medical factors (obesity, chronic medical comorbidity, rheumatoid arthritis, and such common geriatric problems as falls, poor balance, or incontinence), and psychosocial factors (mental health, regular alcohol consumption, smoking, provider role, living alone, and education). Ten percent of subjects had poor functional status. In a logistic regression model controlling for sex and age, the following factors were associated with an increased risk for poor functional status (in order of importance): pain in the back or lower extremity, severe pain in the operated hip, poor mental health, more than 1 common geriatric problem, obesity, and less than college education. Pain in the operated hip was strongly associated with poor functional status 3 years after THR. However, other factors associated with poor functional status were not related to the hip. Our results suggest that a comprehensive assessment of functional status in elderly THR patients should include assessment of common geriatric problems, mental health status, and weight.


Reference interval determination for anabasine: a biomarker of active tobacco use.
Laboratory detection of nicotine exposure is important for establishing eligibility for organ transplant and elective surgery. Nicotine testing is also used to verify compliance with nicotine replacement therapies (NRT), smoking cessation programs and for life insurance purposes. Nicotine metabolites, such as cotinine and trans-3'-hydroxycotinine, are used as biomarkers of nicotine exposure. For some clinical applications, it is important to distinguish between active use of tobacco products versus NRT. Anabasine is a tobacco alkaloid that has been used as a biomarker of active tobacco use. However, the use of anabasine as an insecticide, and its presence in consumables other than nicotine products, suggests that anabasine may not be specific to tobacco use/exposure. Here, we determine the reference interval for anabasine in the urine of nonsmokers and compare it to the range of anabasine concentrations observed in the presence or absence of nicotine metabolites.


Cloacolith in a blue-fronted amazon parrot (Amazona aestiva).
A 4-year-old blue-fronted Amazon parrot (Amazona aestiva) was admitted for vocalization secondary to constipation. Saline infusion cloacoscopy revealed the presence of a 2-cm-diameter cloacolith within the coprodeum that was obstructing the rectal opening. The cloacolith was fragmented with a pair of biopsy forceps and the pieces removed. The cloacolith was subsequently analyzed and was composed of 100% uric acid salts. The bird improved completely and was able to defecate normally after the procedure. Cloacoliths are relative uncommon cloacal conditions, and this case documents cloacoscopic findings, rectal obstruction, and confirmation of its uric acid composition by urolith analysis.


Powder diffraction: what's in a name?
The development of powder diffraction is briefly described; the extent of this development from studies of metals to protein crystal structures shows that powder diffraction is at the cutting edge of crystallography. A new name `polycrystallography' is proposed for these endeavours.


Shift in the brain network of emotional regulation in midlife women: is the menopausal transition the turning point?
The menopausal transition is marked by hormonal changes and is quite often accompanied by cognitive and emotional complaints. Recent data also suggest a heightened risk for depression. Little is known about the changes in emotional regulation that might contribute to the increased risk of depression in this population. The aim of this study was to examine the brain correlates of emotional regulation in healthy, nondepressed midlife women. Functional magnetic resonance imaging was obtained in response to a standardized emotional regulation task. Levels of congruency were set and brain activation was measured during high- and low-conflict-resolution trials. Fourteen women aged 40 to 60 years were enrolled into the study, and 11 were included in the final analyses. Activity associated with resolution of emotional conflict was observed in the dorsolateral prefrontal cortex (P < 0.05). No regions were engaged in the generation/monitoring of emotional conflict. Moreover, there was a significant deactivation of the amygdala in response to fearful faces (P < 0.05). Unlike similar studies in younger populations, these results suggest a more significant engagement of the dorsolateral prefrontal cortex and less amygdala activation in emotional regulation in midlife women. These findings are, however, consistent with previous studies in older populations. We hypothesize that a shift in emotional regulation circuitry might therefore occur in women during the menopausal transition and possibly contribute to the occurrence of mood and anxiety symptoms in women during/after this period in life.


Prevalence of symptoms of asthma, rhinitis and atopic eczema among students between 6 and 7 years of age in the city of Londrina, Brazil.
To determine the prevalence of symptoms of asthma, rhinitis and atopic eczema among students between 6 and 7 years of age in the city of Londrina, Brazil. A population-based study using the International Study of Asthma and Allergies in Childhood (ISAAC) standardized questionnaire (asthma, rhinitis and atopic eczema modules), validated for use in Brazil, in public school students between 6 and 7 years of age. Of the 3,963 questionnaires retrieved, 3,600 (90.8%) were appropriately completed and were used in the analysis. The prevalence of symptoms of asthma, rhinitis and atopic eczema in the last 12 months was 22.0%, 27.3% and 9.6%, respectively. The prevalence of physician-diagnosed asthma, rhinitis and atopic eczema was 10.4%, 23.4% and 11.4%, respectively. The prevalence of rhinoconjunctivitis and flexural eczema was 13.6% and 6.6%, respectively. Although symptoms of asthma and rhinitis were more common in males than in females, no gender difference was found regarding atopic eczema symptoms. The prevalence of symptoms of asthma, rhinitis and atopic eczema in our sample was within the range found at the facilities that participated in phases I and III of the ISAAC in Brazil. The low prevalence of physician-diagnosed asthma suggests that asthma continues to be underdiagnosed.


Letter to the editor: clarifying some aspects and the terminology of individualized human milk fortification.
This letter has been written by the components of the European Milk Bank Association (EMBA) Working Group on Human Milk Fortification in response to a recent paper published by Mathes et al. (BMC Pediatr. 2018 May 8;18(1):154) with the aim of drawing attention to the importance of the use of a metabolic marker to adapt protein intake in preterm infants. EMBA Working Group on Human Milk Fortification clarifies further the terminology and some specific aspects regarding individualized human milk fortification. There are two types of individualized human milk fortification: Adjustable human milk fortification and Targeted human milk fortification. Advantages and disadvantages of these methods are summarized.


An early postinfection signal mediated by monoclonal anti-beta 2 microglobulin antibody is responsible for delayed production of human immunodeficiency virus type 1 in peripheral blood mononuclear cells.
We recently found (C. Devaux, J. Boucraut, G. Poirier, P. Corbeau, F. Rey, M. Benkirane, B. Perarneau, F. Kourilsky, and J.C. Chermann, submitted for publication) a latency in the human immunodeficiency virus (HIV) type 1 cytopathic effect in the human T-cell lymphotropic virus type I immortalized T-cell line MT4 that was mediated by anti-beta 2 microglobulin (beta 2m) monoclonal antibodies (MAb). Here we describe a delay in viral particle production in peripheral blood mononuclear cells (PBMC) that was mediated by three (B1-1G6, B2-62-2, and HC11-151-1) of four anti-beta 2m MAb tested, the nonefficient MAb (C21-48A) being specific for an epitope on beta 2m that was masked by association with the human leukocyte antigen class I heavy chain. Experiments were designed to determine the mechanism of interference. PBMC incubated with anti-beta 2m MAb before viral exposure were not protected from HIV infection. In addition, anti-beta 2m MAb were not efficient in preventing syncytium formation between HIV-infected PBMC and CD4-positive MT4 cells. In contrast, anti-beta 2m MAb treatment of freshly infected PBMC significantly delayed HIV production in these cells. The window of cell sensitivity to anti-beta 2m MAb treatment took place during a very early post-HIV-binding stage. The possible mechanism of anti-beta 2m MAb action is discussed.


Visuoperceptual deficits and participation in older adults after stroke.
Visuoperceptual deficits frequently occur after a stroke, but little is known about how they evolve over time. These deficits may have an impact on participation in daily activities and social roles. The aims were to (i) track changes over six months in the visual perception of older adults with persistent visuoperceptual deficits after a stroke; (ii) examine if these changes differed between participants who had and had not received rehabilitation services; and (iii) verify if participation differed between participants with and without visuoperceptual deficits. Visual perception as well as participation of 189 older adults who had had a stroke were evaluated in the first month (T1) after being discharged home from an acute care hospital (NO REHAB group) or rehabilitation unit (REHAB group). For visual perception, only participants presenting deficits at T1 were re-evaluated at three months (T2; n = 93), and those with deficits at T2 were re-evaluated at six months (T3; n = 61). A total of 57 people (30.2%) had visuoperceptual deficits six months after discharge home. Despite persistent deficits, approximately 45% of the participants in the two groups improved whereas 50% of the NO REHAB group and 24.3% of the REHAB group deteriorated. Changes in the mean scores on the MVPT-V were similar in the two groups. Participation, and especially participation in social roles, was more restricted in participants with visuoperceptual deficits (P < 0.001), independent of the severity of the stroke. Visuoperceptual deficits are common post-stroke. However, they evolve differently in different people and are associated with a reduction in participation.


Methods and utility of EEG-fMRI in epilepsy.
Brain activity data in general and more specifically in epilepsy can be represented as a matrix that includes measures of electrophysiology, anatomy and behaviour. Each of these sub-matrices has a complex interaction depending upon the brain state i.e., rest, cognition, seizures and interictal periods. This interaction presents significant challenges for interpretation but also potential for developing further insights into individual event types. Successful treatments in epilepsy hinge on unravelling these complexities, and also on the sensitivity and specificity of methods that characterize the nature and localization of underlying physiological and pathological networks. Limitations of pharmacological and surgical treatments call for refinement and elaboration of methods to improve our capability to localise the generators of seizure activity and our understanding of the neurobiology of epilepsy. Simultaneous electroencephalography and functional magnetic resonance imaging (EEG-fMRI), by potentially circumventing some of the limitations of EEG in terms of sensitivity, can allow the mapping of haemodynamic networks over the entire brain related to specific spontaneous and triggered epileptic events in humans, and thereby provide new localising information. In this work we review the published literature, and discuss the methods and utility of EEG-fMRI in localising the generators of epileptic activity. We draw on our experience and that of other groups, to summarise the spectrum of information provided by an increasing number of EEG-fMRI case-series, case studies and group studies in patients with epilepsy, for its potential role to elucidate epileptic generators and networks. We conclude that EEG-fMRI provides a multidimensional view that contributes valuable clinical information to localize the epileptic focus with potential important implications for the surgical treatment of some patients with drug-resistant epilepsy, and insights into the resting state and cognitive network dynamics.


Impact of aortopulmonary collaterals on adverse events after total cavopulmonary connection.
Effects of aortopulmonary collaterals (APCs) on outcomes after the total cavopulmonary connection (TCPC) are unclear. This study evaluated the incidence of APCs before and after TCPC and analyzed the impacts of APCs on adverse outcomes. A total of 585 patients, who underwent TCPC from 1994 to 2020 and whose preoperative angiographies were available, were included. Pre-TCPC angiograms in all patients were used for the detection of APCs, and post-TCPC angiograms were evaluated in selected patients. Late adverse events included late death, protein-losing enteropathy (PLE), and plastic bronchitis (PB). Median age at TCPC was 2.3 (1.8-3.4) years with a body weight of 12 (11-14) kg. APCs were found in 210 patients (36%) before TCPC and in 81 (14%) after TCPC. The closure of APCs was performed in 59 patients (10%) before TCPC, in 25 (4.2%) at TCPC, and in 59 (10%) after TCPC. The occurrences of APCs before and after TCPC were not associated with short-term or mid-term mortality. The APCs before TCPC were associated with chylothorax (p = 0.025), prolonged chest tube duration (p = 0.021), and PB (p = 0.008). The APCs after TCPC were associated with PLE (p < 0.001) and PB (p < 0.001). With APCs following TCPC, freedom from PLE and PB was lower than without (p < 0.001, p < 0.001). APCs before TCPC were associated with chylothorax, prolonged chest tube duration, and PB. APCs after TCPC were associated with both PLE and PB. The presence of APCs might affect the lymph drainage system and increase the incidence of chylothorax, PLE, and PB.


Reciprocal transfer of abnormalities in clonable B lymphocytes and myeloid progenitors between NZB and DBA/2 mice.
Previous studies in this laboratory revealed that NZB mice have abnormalities of myeloid progenitor populations from an early age such that they are poorly responsive to a particular type of colony-stimulating activity (CSA). In addition, these mice develop abnormally high numbers of B cells that can be cloned in semisolid agar cultures and that are atypical in resisting inhibition by anti-mu antibodies. To investigate whether these abnormalities, like other autoimmune phenomena studied previously, are transferrable with hemopoietic cell grafts, we performed reciprocal bone marrow transplants between NZB and normal DBA/2 mice. Irradiated control mice given syngeneic marrow did not change with respect to any of the parameters that were measured. In contrast, DBA/2 recipients of NZB marrow were indistinguishable from NZB mice in terms of CSA responses and incidences of anti-mu resistant B cells. The proportions but not total numbers of clonable B cells ere elevated in these mice until at least 16 wk after grafting. Conversely, NZB mice given DBA/2 cells had all of the normal characteristics of DBA/2 mice. Transplantation did not cause significant changes in hematocrits, reticulocyte counts, or spleen weights. Therefore, all elements necessary for expression of these lymphoid and nonlymphoid abnormalities of NZB mice are intrinsic to radiosensitive hemopoietic cells.


Listeria infection in transplant patients. Five cases and a review of the literature.
We report five cases of Listeria monocytogenes infection in renal transplant patients at the University of Minnesota and compare them to 15 additional patients reported on in the United States literature. All patients were noted to have fever, malaise, and nonspecific symptoms of infection. There were no consistent diagnostic laboratory findings except for positive bacteriologic studies. Successful treatment consisted of intravenous penicillin G potassium in most cases (ampicillin sodium was required in two patients). Mortality was low, with only one patient of the 20 (and no Minnesota patients) dying of listeriosis. The time interval from transplant to infection was definitively longer in the Minnesota patients; this may be due to the routine use of sulfisoxazole following renal transplantation. Listeria infection, though mild itself, may herald other infectious processes in the immunoincompetent host.


Ameliorative effect of p-coumaric acid, a common dietary phenol, on adjuvant-induced arthritis in rats.
p-Coumaric acid (3-(4-hydroxyphenyl)-2-propenoic acid), a common dietary polyphenol, is widely distributed in cereals, fruits and vegetables with antioxidant property. Numerous studies have enlightened the ability of dietary phenols to be considered as potential therapeutics against arthritis. In this study, we aimed to investigate the ameliorative effect of plant phenolic p-coumaric acid on adjuvant-induced arthritis in rats. The reference drug indomethacin was used for comparison purposes. Arthritis was induced in rats by a single intradermal injection of complete freund's adjuvant (0.1 mL) into the foot pad of right hind paw. p-Coumaric acid (100 mg/kg b wt) and indomethacin (3 mg/kg b wt) were administered intraperitoneally for 8 days from day 11 to 18 after adjuvant injection. An increase in the activities/levels of lysosomal enzymes, tissue marker enzymes, glycoproteins and paw thickness was observed in the arthritic rats, on the contrary, the body weight was found to be reduced in arthritic rats when compared to normal control rats. Administration of p-coumaric acid (100 mg/kg b wt) to the arthritic rats reverted back the altered physical and biochemical parameters to near normal levels comparable to indomethacin treatment. Histopathological evaluation of ankle joints in arthritic rats also revealed the anti-inflammatory effect of p-coumaric acid by the reduction in leukocytes infiltration. Thus, the present study clearly demonstrates the anti-inflammatory potential of the p-coumaric acid against adjuvant-induced arthritis in rats.


Quetiapine Attenuates Schizophrenia-Like Behaviors and Demyelination in a MK-801-Induced Mouse Model of Schizophrenia.
Brain demyelination is possibly one of the main pathological factors involved in schizophrenia, and targeting on myelination may be a useful strategy for schizophrenia treatment. Quetiapine, a widely used atypical antipsychotic drug for schizophrenia treatment, has been reported to have neuroprotective effects on cerebral myelination in a demyelination animal model. The objective of the present study was to evaluate the effect and underlying neuroprotective mechanism of quetiapine on the schizophrenia-like behaviors and possible cerebral demyelination induced by MK-801, an N-methyl-D-aspartate glutamate receptor antagonist. Mice were treated with chronic quetiapine (10 mg/kg/day, intraperitoneally) for 28 days. From day 22 to 28, 1 h after the administration of quetiapine, the mice were administered MK-801 (2 mg/kg/day, subcutaneously). The positive symptom of schizophrenia was measured in a locomotor activity test on day 29, the memory was evaluated by a Y-maze test on day 30, and the sensorimotor gating deficit in mice was measured by prepulse inhibition test on day 31. After the behavioral tests, the protein expression of myelin basic protein (MBP) was measured by Western Blot, and the protein expression of brain-derived neurotrophic factor (BDNF) was measured by ELISA in the frontal cortex of mice. Our results showed quetiapine attenuated schizophrenia-like behaviors including hyperactivity, memory impairment, and sensorimotor gating deficit in the MK-801 mice. In the same time, quetiapine attenuated demyelination, concurrent with attenuated BDNF decrease in the brain of MK-801-injected mice. These results suggest that the beneficial effects of quetiapine on schizophrenia might be partly related to its neuroprotective effect on brain myelin basic protein and its upregulating neuroprotective proteins such as BDNF, and indicate that modulation of cerebral demyelination could be a novel treatment target of schizophrenia.


Two-year results of intermittent electrical stimulation of the lower esophageal sphincter treatment of gastroesophageal reflux disease.
Lower esophageal sphincter (LES) electrical stimulation therapy (EST) has been shown to improve outcome in gastroesophageal reflux disease (GERD) patients at 1 year. The aim of this open-label extension trial (NCT01578642) was to study the 2-year safety and efficacy of LES-EST in GERD patients. GERD patients responsive partially to proton pump inhibitors (PPI) with off-PPI GERD health-related quality of life (HRQL) of ≥20, 24-hour esophageal pH ≤4.0 for >5% of the time, hiatal hernia ≤3 cm, and esophagitis LA grade C or lower participated in this trial. Bipolar stitch electrodes and a pulse generator (EndoStim BV, The Hague, The Netherlands) were implanted laparoscopically. LES-EST at 20 Hz, 215 μs, 3-8 mAmp was delivered over 30-minute sessions, 6-12 sessions per day, starting on day 1 after implantation. Patients were evaluated using GERD-HRQL, symptom diaries, Short Form-12, and esophageal pH testing at regular intervals. Stimulation sessions were optimized based on residual symptoms and esophageal pH at follow-up. Twenty-five patients (mean age [SD] = 52 [12] years; 14 men) were implanted successfully; 23 patients participated in the 2-year extension trial, and 21 completed their 2-year evaluation. At 2 years, there was improvement in their median GERD-HRQL on LES-EST compared with both their on-PPI (9 vs 0; P = .001) and off-PPI (23.5 vs. 0; P < .001) baseline scores. Median 24-hour distal esophageal acid exposure improved from 10% at baseline to 4% (per-protocol analysis; P < .001) at 2 years with 71% demonstrating either normalization or a ≥50% decrease in their distal esophageal acid exposure. All except 5 patients (16/21) reported complete cessation of PPI use; only 2 patients were using a PPI regularly (≥50% of days). There was significant improvement in sleep quality and daily symptoms of heartburn and regurgitation on LES-EST. At baseline, 92% of the subjects (22/24) reported that they were "unsatisfied" with their condition off-PPI and 71% (17/24) on-PPI compared with 0% (0/21) "unsatisfied" at the 24-month visits on LES-EST. There were no device- or therapy-related serious adverse events and no untoward sensation or dysphagia reported with LES-EST. LES-EST is safe and effective for treating patients with GERD over a period of 2 years. LES-EST resulted in a significant and sustained improvement in GERD symptoms, and esophageal acid exposure and eliminated PPI use in majority of patients (16 of 21). Further, LES-EST was not associated with any gastrointestinal side effects or adverse events.


Mitogen-activated protein kinase kinase kinase 1 protects against nickel-induced acute lung injury.
Nickel compounds are environmental and occupational hazards that pose serious health problems and are causative factors of acute lung injury. The c-jun N-terminal kinases (JNKs) are regulated through a mitogen-activated protein (MAP) 3 kinase-MAP2 kinase cascade and have been implicated in nickel toxicity. In this study, we used genetically modified cells and mice to investigate the involvement of two upstream MAP3Ks, MAP3K1 and 2, in nickel-induced JNK activation and acute lung injury. In mouse embryonic fibroblasts, levels of JNK activation and cytotoxicity induced by nickel were similar in the Map3k2-null and wild-type cells but were much lower in the Map3k1/Map3k2 double-null cells. Conversely, the levels of JNK activation and cytotoxicity were unexpectedly much higher in the Map3k1-null cells. In adult mouse tissue, MAP3K1 was widely distributed but was abundantly expressed in the bronchiole epithelium of the lung. Accordingly, MAP3K1 ablation in mice resulted in severe nickel-induced acute lung injury and reduced survival. Based on these findings, we propose a role for MAP3K1 in reducing JNK activation and protecting the mice from nickel-induced acute lung injury.


Amorphous Chloride Solid Electrolytes with High Li-Ion Conductivity for Stable Cycling of All-Solid-State High-Nickel Cathodes.
Solid electrolytes (SEs) are central components that enable high-performance, all-solid-state lithium batteries (ASSLBs). Amorphous SEs hold great potential for ASSLBs because their grain-boundary-free characteristics facilitate intact solid-solid contact and uniform Li-ion conduction for high-performance cathodes. However, amorphous oxide SEs with limited ionic conductivities and glassy sulfide SEs with narrow electrochemical windows cannot sustain high-nickel cathodes. Herein, we report a class of amorphous Li-Ta-Cl-based chloride SEs possessing high Li-ion conductivity (up to 7.16 mS cm-1) and low Young's modulus (approximately 3 GPa) to enable excellent Li-ion conduction and intact physical contact among rigid components in ASSLBs. We reveal that the amorphous Li-Ta-Cl matrix is composed of LiCl43-, LiCl54-, LiCl65- polyhedra, and TaCl6- octahedra via machine-learning simulation, solid-state 7Li nuclear magnetic resonance, and X-ray absorption analysis. Attractively, our amorphous chloride SEs exhibit excellent compatibility with high-nickel cathodes. We demonstrate that ASSLBs comprising amorphous chloride SEs and high-nickel single-crystal cathodes (LiNi0.88Co0.07Mn0.05O2) exhibit ∼99% capacity retention after 800 cycles at ∼3 C under 1 mA h cm-2 and ∼80% capacity retention after 75 cycles at 0.2 C under a high areal capacity of 5 mA h cm-2. Most importantly, a stable operation of up to 9800 cycles with a capacity retention of ∼77% at a high rate of 3.4 C can be achieved in a freezing environment of -10 °C. Our amorphous chloride SEs will pave the way to realize high-performance high-nickel cathodes for high-energy-density ASSLBs.


Move over LOCF: principled methods for handling missing data in sleep disorder trials.
Missing data, e.g. patient attrition, are endemic in sleep disorder clinical trials. Common approaches for dealing with this situation include complete-case analysis (CCA) and last observation carried forward (LOCF). Although these methods are simple to implement, they are deeply flawed in that they may introduce bias and underestimate uncertainty, leading to erroneous conclusions. There are alternative principled approaches, however, that are available in statistical software namely mixed-effects models and multiple imputation. In this paper we introduce terminology used to describe different assumptions about missing data. We emphasize that understanding reasons for missingness is a critical step in the analysis process. We describe and implement both linear mixed-effects models and an inclusive multiple imputation strategy for handling missing data in a randomized trial examining sleep outcomes. These principled strategies are compared with "complete-case analysis" and LOCF. These analyses illustrate that methodologies for accommodating missing data can produce different results in both direction and strength of treatment effects. Our goal is for this paper to serve as a guide to sleep disorder clinical trial researchers on how to utilize principled methods for incomplete data in their trial analyses.


Novel Production of Bovine Papillomavirus Pseudovirions in Tobacco Plants.
Vaccine efficacy requires the production of neutralising antibodies which offer protection against the native virus. The current gold standard for determining the presence of neutralising antibodies is the pseudovirion-based neutralisation assay (PBNA). PBNAs utilise pseudovirions (PsVs), structures which mimic native virus capsids, but contain non-viral nucleic material. PsVs are currently produced in expensive cell culture systems, which limits their production, yet plant expression systems may offer cheaper, safer alternatives. Our aim was to determine whether plants could be used for the production of functional PsVs of bovine papillomavirus 1 (BPV1), an important causative agent of economically damaging bovine papillomas in cattle and equine sarcoids in horses and wild equids. BPV1 capsid proteins, L1 and L2, and a self-replicating reporter plasmid were transiently expressed in Nicotiana benthamiana to produce virus-like particles (VLPs) and PsVs. Strategies to enhance particle yields were investigated and optimised protocols were established. The PsVs' ability to infect mammalian cells and express their encapsidated reporter genes in vitro was confirmed, and their functionality as reagents in PBNAs was demonstrated through their neutralisation by several different antibodies. This is the first report of BPV PsVs expressed in plants and demonstrates the potential for the development of therapeutic veterinary vaccines in planta.


Characterization of Vibrio parahaemolyticus manganese-resistant mutants in reference to the function of the ferric uptake regulatory protein.
In many bacteria, the ferric uptake regulatory protein (Fur) has a central role in the negative regulation of genes affected by iron limitation. In this study, Vibrio parahaemolyticus strains carrying mutations in the fur gene encoding Fur were isolated by the manganese selection method to assess the function of Fur in connection with alternations in the coordinate expression of the siderophore vibrioferrin (VF) and iron-repressible outer membrane proteins (IROMPs). Ten out of 25 manganese-resistant mutants constitutively produced VF and expressed at least two IROMPs irrespective of the iron concentration in the medium. PCR-direct DNA sequencing of the fur genes in these mutants identified four different point mutations causing amino acid changes. Moreover, a fur overexpressing plasmid was constructed to prepare antiserum against V. parahaemolyticus Fur. Western blotting with this antiserum revealed that the intracellular abundance of the wild-type Fur was not significantly affected by the iron concentrations in the growth medium, and that the Fur proteins of the mutant strains occurred at substantially smaller amounts and/or migrated more rapidly in sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the wild-type Fur. These data afford an additional insight into the structure-function relationship of Fur and imply its involvement in the iron acquisition systems of V. parahaemolyticus, although it is yet unknown whether its action on the target genes is direct or indirect.


Role of phospholipid transfer protein and prebeta-high density lipoproteins in maintaining cholesterol efflux from Fu5AH cells to plasma from insulin-resistant subjects.
Plasma phospholipid transfer protein (PLTP) enhances the generation of prebeta-high density lipoproteins (HDL) that may act as initial acceptors of cellular cholesterol, and are likely to play an important role in the antiatherogenic process of reverse cholesterol transport. We examined the interrelationships between insulin resistance, the ability of plasma to stimulate cellular cholesterol efflux, HDL cholesterol, plasma PLTP activity and prebeta-HDL in 12 non-diabetic, non-smoking, normotriglyceridaemic men. Cholesterol efflux from Fu5AH cells to plasma, plasma lipoproteins, PLTP activity and prebeta-HDL formation as measured in incubated plasma were determined after a 12-h fast. Insulin sensitivity was assessed by a euglycaemic, hyperinsulinaemic clamp (M-value). HDL cholesterol was positively correlated with the M-value (r=0.65, p< 0.05), whereas plasma PLTP activity (r= -0.59, p <0.05) and prebeta-HDL in incubated plasma (r= -0.66, p<0.05) were negatively correlated with the M-value. Thus, the lower the insulin sensitivity, the lower was HDL cholesterol and the higher were plasma PLTP activity and prebeta-HDL. Cellular cholesterol efflux tended to be correlated with HDL cholesterol (r=0.55, p < 0.10) as well as with plasma PLTP activity (r=0.56, p<0.10) and was positively correlated with prebeta-HDL in incubated plasma (r=0.74, p<0.01). No positive correlation between the M-value and cellular cholesterol efflux was found (r= -0.34, ns). These preliminary results support the hypothesis that, despite a lower HDL cholesterol, the ability of plasma from insulin-resistant subjects to promote cellular cholesterol efflux is not impaired, as a consequence of a higher plasma PLTP activity and enhanced prebeta-HDL formation.


Experimental demonstration of the hybrid opto-electronic correlator for target recognition.
Optical target recognition using correlators is an important technique for fast verification and identification of images. The hybrid opto-electronic correlator (HOC) recently proposed by us bypasses the need for nonlinear materials such as photorefractive polymer films by using detectors instead, and the phase information is yet conserved by the interference of plane waves with the images. In this paper, we demonstrate experimentally the basic working principle of the HOC architecture using currently available technologies. For matched reference and query images, the output signal shows a sharp peak, indicating a match is found. For an unmatched case, a much lower peak value is observed, indicating no match. We also demonstrate the dependence of the output signal on the phases of the interfering plane waves and describe a technique using an interferometer and a servo for optimizing the output signal. As such, the work reported here paves the way for further development of the HOC for practical applications.


Slow initial β-lactam infusion and oral paracetamol to treat childhood bacterial meningitis: a randomised, controlled trial.
New antimicrobials or adjunctive treatments have not substantially reduced mortality from acute childhood bacterial meningitis. Paracetamol seems to have beneficial effects in bacteraemic adults and some experts recommend initial slow β-lactam infusion. We investigated whether these treatments had benefits in children with bacterial meningitis. We did a prospective, double-blind, single-centre study with a two-by-two factorial design in Luanda, Angola. 723 participants aged 2 months to 13 years were randomly assigned two 12 h intravenous infusions, without loading doses, of 125 mg/kg bodyweight cefotaxime (total dose 250 mg/kg) given over 24 h, or 250 mg/kg bodyweight cefotaxime given as four boluses, one every 6 h over 24 h. Patients also received oral paracetamol at an initial dose of 30 mg/kg then 20 mg/kg every 6 h for 48 h or placebo. Two primary endpoints, death or severe neurological sequelae and deafness, were analysed by intention to treat. The study was registered as ISRCTN62824827. 183 patients were assigned cefotaxime infusion plus paracetamol and 180 patients to each of the other three treatment groups. Causative agents were identified in 63% of cases and were mostly Haemophilus influenzae type b, Streptococcus pneumoniae, or Neisseria meningitidis. Death or severe neurological sequelae were seen in 340 (47%) of 723 children and deafness in 45 (12%) of 374 tested, both distributed similarly across treatment groups. In a predefined subgroup analysis of death or any sequelae, by causative agent, a benefit was seen in favour of infusion over bolus in children with pneumococcal meningitis (infusion plus placebo, odds ratio 0·18, 95% CI 0·03-0·90, p=0·04). A similar effect was seen for children receiving cefotaxime infusion plus paracetamol, but the difference was not significant (OR 0·22, 95% CI 0·04-1·09, p=0·06). A post-hoc analysis suggested that cefotaxime infusion plus paracetamol lowered mortality at least during the first 3 days, irrespective of cause. Although no tested regimen improved the final outcomes of these very ill children, studies of longer courses of β-lactam infusion plus paracetamol seem warranted. The Päivikki and Sakari Sohlberg, the Sigrid Jusélius, and the Paediatric Research Foundations, and the daily newspaper Helsingin Sanomat.


Neurological diseases in Peru: a systematic analysis of the global burden disease study.
Disease burden indicators assess the impact of disease on a population. They integrate mortality and disability in a single indicator. This allows setting priorities for health services and focusing resources. To analyze the burden of neurological diseases in Peru from 1990-2015. A descriptive study that used the epidemiological data published by the Institute for Health Metrics and Evaluation of Global Burden of Diseases from 1990 to 2015. Disease burden was measured using disability-adjusted life years (DALY) and their corresponding 95% uncertainty intervals (UIs), which results from the addition of the years of life lost (YLL) and years lived with disability (YLD). The burden of neurological diseases in Peru were 9.06 and 10.65%, in 1990 and 2015, respectively. In 2015, the main causes were migraine, cerebrovascular disease (CVD), neonatal encephalopathy (NE), and Alzheimer's disease and other dementias (ADD). This last group and nervous system cancer (NSC) increased 157 and 183% of DALY compared to 1990, respectively. Young population (25 to 44 years old) and older (>85 years old) were the age groups with the highest DALY. The neurological diseases produced 11.06 and 10.02% of the national YLL (CVD as the leading cause) and YLD (migraine as the main cause), respectively. The burden of disease (BD) increased by 1.6% from 1990 to 2015. The main causes were migraine, CVD, and NE. ADD and NSC doubled the DALY in this period. These diseases represent a significant cause of disability attributable to the increase in the life expectancy of our population among other factors. Priority actions should be taken to prevent and treat these causes.


Differential requirements for the activation domain and FOG-interaction surface of GATA-1 in megakaryocyte gene expression and development.
GATA1 is mutated in patients with 2 different disorders. First, individuals with a GATA1 mutation that blocks the interaction between GATA-1 and its cofactor Friend of GATA-1 (FOG-1) suffer from dyserythropoietic anemia and thrombocytopenia. Second, children with Down syndrome who develop acute megakaryoblastic leukemia harbor mutations in GATA1 that lead to the exclusive expression of a shorter isoform named GATA-1s. To determine the effect of these patient-specific mutations on GATA-1 function, we first compared the gene expression profile between wild-type and GATA-1-deficient megakaryocytes. Next, we introduced either GATA-1s or a FOG-binding mutant (V205G) into GATA-1-deficient megakaryocytes and assessed the effect on differentiation and gene expression. Whereas GATA-1-deficient megakaryocytes failed to undergo terminal differentiation and proliferated excessively in vitro, GATA-1s-expressing cells displayed proplatelet formation and other features of terminal maturation, but continued to proliferate aberrantly. In contrast, megakaryocytes that expressed V205G GATA-1 exhibited reduced proliferation, but failed to undergo maturation. Examination of the expression of megakaryocyte-specific genes in the various rescued cells correlated with the observed phenotypic differences. These studies show that GATA-1 is required for both normal regulation of proliferation and terminal maturation of megakaryocytes, and further, that these functions can be uncoupled by mutations in GATA1.


Junctional ectopic tachycardia following repair of congenital heart defects-experience in multimodal management from a West African Centre.
Postoperative junctional ectopic tachycardia (JET) is a rare and transient phenomenon occurring after repair of congenital heart defects. Report on this arrhythmia in the subregion is rare. We set out to determine the incidence of this arrhythmia and review the treatment and outcomes of treatment in our centre. Retrospective search of the records of all patients aged 18 years and below admitted into the intensive care unit (ICU) following repair or palliation of a congenital heart defect over 5 years, from January 1, 2006 to December 31, 2010. A review of clinical notes, operative records, anaesthetic charts, cardiopulmonary bypass (CPB) records, nursing observation charts, electrocardiograms (ECGs) and out-patient follow-up records was undertaken. 510 children under 18 years were enlisted. 7 cases of postoperative JET were recorded, (1.37%). 184 (36.1%) of these were performed under CPB. All JET cases were from cases done under CPB, 3.8%. Median age was 3 years and median weight 11.3 kg. No patient was febrile at diagnosis. 4 patients had amiodarone administration, 5 had magnesium sulphate infusion, 2 patients had direct current shock (DCS) whilst 3 patients had all three therapeutic modalities. All patients had control of the arrhythmia with conversion to sinus rhythm and no recurrence. We report a JET incidence of 1.37% among children undergoing CPB for repair of congenital heart defects. We demonstrate the therapeutic effectiveness of amiodarone, magnesium sulphate infusions and DCS alone or in combination in the management of JET on various substrates with good outcome.


Baseline frontostriatal-limbic connectivity predicts reward-based memory formation.
Reward mediates the acquisition and long-term retention of procedural skills in humans. Yet, learning under rewarded conditions is highly variable across individuals and the mechanisms that determine interindividual variability in rewarded learning are not known. We postulated that baseline functional connectivity in a large-scale frontostriatal-limbic network could predict subsequent interindividual variability in rewarded learning. Resting-state functional MRI was acquired in two groups of subjects (n = 30) who then trained on a visuomotor procedural learning task with or without reward feedback. We then tested whether baseline functional connectivity within the frontostriatal-limbic network predicted memory strength measured immediately, 24 h and 1 month after training in both groups. We found that connectivity in the frontostriatal-limbic network predicted interindividual variability in the rewarded but not in the unrewarded learning group. Prediction was strongest for long-term memory. Similar links between connectivity and reward-based memory were absent in two control networks, a fronto-parieto-temporal language network and the dorsal attention network. The results indicate that baseline functional connectivity within the frontostriatal-limbic network successfully predicts long-term retention of rewarded learning.


The Dutch registry for facioscapulohumeral muscular dystrophy: Cohort profile and longitudinal patient reported outcomes.
Facioscapulohumeral dystrophy (FSHD) is the second most prevalent inherited muscular disorder and currently lacks a pharmaceutical treatment. The Dutch FSHD Registry was initiated in 2015 as a result of an international collaboration on trial readiness. This paper presents the cohort profile and six years of follow-up data of the registered FSHD patients. At the time of self-registration and every six months thereafter, participants were invited to complete a digital survey of patient and disease characteristics and the Dutch versions of the Checklist Individual Strength (CIS20R), the Individualised Neuromuscular Quality of Life Questionnaire (INQoL), the Beck Depression Index - Primary Care and the McGill Pain Questionnaire. From March 2015 to March 2021, 373 participants completed at least one survey. At baseline, fatigue and muscle weakness were the most frequently reported symptoms (median CIS20R sumscore 77 [IQR 60-92], median INQoL Fatigue score 58 [IQR 42-68] and median INQoL weakness score 58 [IQR 42-68]). Pain was experienced most often in the head and shoulder region (193, 52%). Nineteen of the 23 (sub)sections of questionnaires showed no significant changes over time. We conclude that the Dutch FSHD Registry was successfully set up, enabling collection of longitudinal data and facilitating recruitment in several studies.


Mast cells are not required for anaphylatoxin-induced ileal smooth muscle contraction.
The complement-derived anaphylatoxin peptides, C3a and C5a, have long been considered to manifest their spasmogenic activities primarily through stimulation of mast cells. Although mast cells represent the major non-circulating repository for histamine, these cells also elaborate a number of additional, highly potent spasmogenic mediators derived from arachidonic acid. The same lipid mediators can be released by many other cell types. As a result, evaluation of the role of mast cells in anaphylatoxin-dependent responses cannot be based exclusively upon an analysis of the mediators released. We evaluated the role of mast cells in anaphylatoxin-induced ileal smooth muscle contractions by testing isolated segments of ileal tissues derived from genetically mast cell-deficient mice and their congenic normal (+/+) littermates. Isolated tissues from either congenic normal (+/+) or mast cell-deficient Sl/Sld mice responded similarly to acetylcholine, histamine, serotonin, prostaglandin E2, and the thromboxane A2 analog, U-46619. At 1 microgram/ml, histamine induced contractions of greater magnitude in tissues from mast cell-deficient animals; however, this mediator also desensitized the tissues to repeat challenge with histamine at the same concentration. C5a at 1 nM resulted in contractions equivalent to approximately 50% of the maximal KCl response; normal and mast cell-deficient tissues responded in a similar manner. C5a also released histamine from the normal mouse ileum, in addition to causing contraction of the tissues. C3a at 200 nM also produced similar contractile responses in both +/+ and S1/S1d tissues. These studies show that the anaphylatoxin peptides C3a and C5a are capable of contracting smooth muscle-containing tissues by a mechanism completely independent of mast cells. In addition, we also demonstrated that mast cell degranulation does not necessarily provoke ileal contraction. Thus compound 48/80, a mast cell degranulating agent unrelated to the anaphylatoxins, did not induce contractions in ileal tissues, even when used at concentrations as high as 100 micrograms/ml. Compound 48/80 did release histamine from the +/+ ileum, however, indicating that the agent was able to cause degranulation of ileal mast cells. Taken together, these data indicate that spasmogenic responses to anaphylatoxins (and possibly other agents) that are associated with mast cell degranulation need not necessarily require mast cell mediator release for their expression.


Coordination among the rearfoot, midfoot, and forefoot during walking.
Examining coordination between segments is essential for prevention and treatment of injuries. However, traditional methods such as ratio, cross-correlation technique, and angle-time plot may not provide a complete understanding of intersegmental coordination. The present study aimed to quantify the coordination among the rearfoot, midfoot, and forefoot segments during walking. Twenty healthy young men walked barefoot on a treadmill. Reflective markers were fixed to their right shank and foot based on the Leardini foot model. Three-dimensional joint angles were calculated at the distal segment, and were expressed relative to the adjacent proximal segment. The coupling angle representing intersegmental coordination was calculated by using the modified vector coding technique, and categorized into the following four coordination patterns: in-phase with proximal dominancy, in-phase with distal dominancy, tanti-phase with proximal dominancy, and anti-phase with distal dominancy. The results showed that the midfoot was dominantly everted compared with the rearfoot and forefoot during the early stance (i.e., the rearfoot-midfoot coordination and midfoot-forefoot coordination were mainly in-phase with distal and proximal dominancy, respectively). This result may suggest that the midfoot plays a more significant role than the rearfoot and forefoot during early stance. The results of the present study can help in understanding the interaction of the intersegmental foot kinematic time series during walking. The results could be used as data to distinguish the presence of injuries or abnormal inter-segmental foot motions such as pes planus. Additionally, these data might be used in the future in a comparison with data on foot deformities.


Genome sequencing and conservation genomics in the Scandinavian wolverine population.
Genetic approaches have proved valuable to the study and conservation of endangered populations, especially for monitoring programs, and there is potential for further developments in this direction by extending analyses to the genomic level. We assembled the genome of the wolverine (Gulo gulo), a mustelid that in Scandinavia has recently recovered from a significant population decline, and obtained a 2.42 Gb draft sequence representing >85% of the genome and including >21,000 protein-coding genes. We then performed whole-genome resequencing of 10 Scandinavian wolverines for population genomic and demographic analyses. Genetic diversity was among the lowest detected in a red-listed population (mean genome-wide nucleotide diversity of 0.05%). Results of the demographic analyses indicated a long-term decline of the effective population size (Ne ) from 10,000 well before the last glaciation to <500 after this period. Current Ne appeared even lower. The genome-wide FIS level was 0.089 (possibly signaling inbreeding), but this effect was not observed when analyzing a set of highly variable SNP markers, illustrating that such markers can give a biased picture of the overall character of genetic diversity. We found significant population structure, which has implications for population connectivity and conservation. We used an integrated microfluidic circuit chip technology to develop an SNP-array consisting of 96 highly informative markers that, together with a multiplex pre-amplification step, was successfully applied to low-quality DNA from scat samples. Our findings will inform management, conservation, and genetic monitoring of wolverines and serve as a genomic roadmap that can be applied to other endangered species. The approach used here can be generally utilized in other systems, but we acknowledge the trade-off between investing in genomic resources and direct conservation actions.


Outcomes of combined Furlow palatoplasty and sphincter pharyngoplasty for velopharyngeal insufficiency.
To compare surgical outcomes between pharyngeal flap, sphincter pharyngoplasty, and combined Furlow palatoplasty and sphincter pharyngoplasty in the management of pediatric velopharyngeal insufficiency. Case series with chart review. Tertiary care pediatric hospital. After exclusion of children with velocardiofacial syndrome, 96 patients who underwent surgical intervention between 2008 and 2012 were identified. Surgical interventions were categorized as pharyngeal flap, sphincter pharyngoplasty, and combined Furlow palatoplasty and sphincter pharyngoplasty. Main outcome measures included perceptual speech analyses, complications, and surgical revision rates. Of the 96 reviewed patients, 38 (39.6%) underwent pharyngeal flap, 20 (20.8%) sphincter pharyngoplasty, and 38 (39.6%) combined Furlow palatoplasty and sphincter pharyngoplasty. Choice of surgical intervention was based on patient characteristics, observed palatal length, and formal speech assessments. There were no differences in patient demographics or preoperative perceptual speech analysis scores among the 3 surgical groups. The mean speech improvement was significantly greater in both the pharyngeal flap (P = .031) and combined procedure (P = .013) compared with sphincter pharyngoplasty alone, but no differences were observed between the pharyngeal flap and combined procedure (P = .797). There were no differences in complications among the 3 surgical interventions (P = .220). The combined procedure required significantly less surgical revisions than the pharyngeal flap (P = .019). Combined Furlow palatoplasty and sphincter pharyngoplasty is an effective procedure for the management of pediatric velopharyngeal insufficiency and may result in superior speech outcomes and lower revision rates than sphincter pharyngoplasty and pharyngeal flap, respectively.


Characterisation of full-length mitochondrial copies and partial nuclear copies (numts) of the cytochrome b and cytochrome c oxidase subunit I genes of Toxoplasma gondii, Neospora caninum, Hammondia heydorni and Hammondia triffittae (Apicomplexa: Sarcocystidae).
Genomic DNA was extracted from three oocyst isolates of Hammondia triffittae from foxes and two oocyst isolates of Hammondia heydorni from dogs, as well as from cell culture-derived tachyzoites of Toxoplasma gondii (RH strain) and Neospora caninum (NC-Liverpool strain), and examined by PCR with primers targeting the cytochrome b (cytb) and the cytochrome c oxidase subunit I (cox1) genes in order to characterise both genes and, if possible, the remainder of the mitochondrial genome of these species. Several primers were designed and used in various combinations to amplify regions within and between both genes and to determine gene order. When certain forward primers targeting cytb were used in combination with certain reverse primers targeting cox1, two overlapping sequences were obtained for each species and isolate studied, which showed that a full-length copy of cytb was followed 36-37 bp downstream by a full-length copy of cox1, and these sequences are believed to represent the true mitochondrial genes and the gene order in the mitochondrial genome of the four species examined. The cytb of T. gondii, N. caninum, H. heydorni and H. triffittae comprised a total of 1,080 bp (359 amino acids) and used ATG and TAA as start and stop codon, respectively. The cox1 of these species also used TAA as stop codon, whereas the most likely start codon was ATG, resulting in a gene comprising 1,491 bp (496 amino acids). Pair-wise sequence comparisons based on either cytb or cox1 clearly separated T. gondii from N. caninum and both of these species from the two Hammondia species, whereas the latter two species were 100 % identical at cytb and shared 99.3 % identity at cox1. Phylogenetic analyses using the maximum-likelihood method confirmed these findings and placed T. gondii in a clade separate from the three other species and all four Toxoplasmatinae in a sister clade to Eimeria spp. PCR with other primers and/or primer pairs than those used to obtain the full-length mitochondrial genes yielded several types of about 1-1.5 kb long sequences, which comprised stretches of the primer-targeted genes at both ends and an intervening non-coding sequence of various length and composition. Thus, portions of cytb could be found both upstream and downstream from portions of cox1 and portions of the same gene could be found adjacent to each other (cytb→cox1; cox1→cytb; cytb→cytb; cox1→cox1). Sequence comparisons revealed that some of these gene fragments were truncated genes, whereas others included the putative start or stop codon of the full-length mitochondrial genes. From the nature of the gene fragments and/or their flanking sequences, they are assumed to be located on the chromosomes of the nuclear genome and to represent nuclear mitochondrial DNA segments (numts) or pseudogenes. In the four species examined, there were no nucleotide differences between the full-length mitochondrial copies of cytb and cox1 and their various incomplete nuclear counterparts. With a few exceptions, identical numt types and closely similar flanking sequences were obtained for all four species, which would indicate that the original transfer of these mitochondrial genes to the nuclear genome and/or the majority of any subsequent rearrangements of these gene fragments within the nuclear genome happened before the four species diverged. Yet, there were species-specific differences in the nucleotide composition of the nuclear gene fragments, identical to the differences in the mitochondrial genes, which would indicate that the incomplete nuclear copies of cytb and cox1 have been continuously updated during evolution to conform to their mitochondrial parent genes. The PCR-based findings of numts were further supported by Basic Local Alignment Search Tool (BLAST) searches against genome sequences of T. gondii and N. caninum using the concatenated mitochondrial cytb/cox1 sequences as queries. These searches revealed the presence of numerous numts of eighth distinct types in both species, with each one having a fixed starting and end point with respect to the nucleotide positions in the full-length mitochondrial genes. Four numt types were completely homologous between both species, whereas four other types differed with respect to their end point and/or the absence/presence of a 96-bp deletion. Each starting and end point was associated with a unique 100-200-bp long flanking sequence, which further revealed the presence of numts. For both species, the numt types and their various arrangements with respect to each other were identical or similar to those obtained by PCR in all four species examined. None of the identified numts covered a full-length gene, but together, the various numts covered the entire mitochondrial cytb and cox1 genes in an overlapping manner. In addition, they were fairly closely spaced on the chromosomes, and these features may explain why the nuclear copies were preferentially amplified to the exclusion of the true mitochondrial genes with most primers and primer pairs used in the present study. The possibility of a similar high prevalence of numts occurring in the nuclear genome of dinoflagellates is discussed.


CXCL8, CCL2, and CMV Seropositivity as New Prognostic Factors for a Severe COVID-19 Course.
The exact pathophysiology of severe COVID-19 is not entirely elucidated, but it has been established that hyperinflammatory responses and cytokine storms play important roles. The aim of this study was to examine CMV status, select chemokines, and complement components in COVID-19, and how concentrations of given molecules differ over time at both molecular and proteomic levels. A total of 210 COVID-19 patients (50 ICU and 160 non-ICU patients) and 80 healthy controls were enrolled in this study. Concentrations of select chemokines (CXCL8, CXCL10, CCL2, CCL3, CCR1) and complement factors (C2, C9, CFD, C4BPA, C5AR1, CR1) were examined at mRNA and protein levels with regard to a COVID-19 course (ICU vs. non-ICU group) and CMV status at different time intervals. We detected several significant differences in chemokines and complement profiles between ICU and non-ICU groups. Pro-inflammatory chemokines and the complement system appeared to greatly contribute to the pathogenesis and development of severe COVID-19. Higher concentrations of CXCL8 and CCL2 in the plasma, with reduced mRNA expression presumably through negative feedback mechanisms, as well as CMV-positive status, correlated with more severe courses of COVID-19. Therefore, CXCL8, CCL2, and CMV seropositivity should be considered as new prognostic factors for severe COVID-19 courses. However, more in-depth research is needed.


"Effect of resveratrol on oxidative and inflammatory stress in liver and spleen of streptozotocin-induced type 1 diabetic rats".
"It has been well known that both oxidative stress and inflammatory activity play crucial roles in the pathogenesis of type 1 diabetes mellitus (T1DM). Resveratrol (RSV), a naturally occurring polyphenol found in grapes and red wine, has recently been shown to exert potent anti-diabetic, anti-oxidative and anti-inflammatory actions. In the present study, we investigated the effect of RSV on oxidative stress and inflammatory response in the liver and spleen of streptozotocin (STZ)-induced type 1 diabetic animal models. Male Long-Evans rats were injected with 65 mg/kg STZ to induce diabetes for 2 weeks, and subsequently administrated with the dosage of 0.1 or 1 mg/kg/day RSV for 7 consecutive days. Hepatic and splenic tissues were dissected for evaluation of oxidative and inflammatory stress. Oxidative stress was assessed by quantification of oxidative indicators including superoxide anion content, lipid and protein oxidative products, as well as manganese superoxide dismutase (Mn-SOD) and nitro-tyrosine protein expression levels. Inflammatory stress was evaluated by the levels of nuclear factor κB (NF-κB) and the proinflammatory cytokine tumor necrosis factorα (TNF-α), interleukin 1 β (IL-1 β ) and IL-6. The experimental results indicated that RSV significantly decreased oxidative stress (superoxide anion content, protein carbonyl level and Mn-SOD expression) in both tissues and hepatic inflammation (NF- κB and IL-1 β ), but implicated proinflammatory potential of RSV in diabetic spleen (TNF-α and IL-6). The results of this study suggest that RSV may serve as a potent antioxidant, but RSV possesses a proinflammatory potential in certain circumstances in diabetes."


[A decade of checkpoint inhibitors: current standard of care and future trends].
Therapy with checkpoint inhibitors still revolutionizes the therapeutical landscape in oncology. Since the first approval of a checkpoint inhibitor for the therapy of malignant melanoma 2011, many other approvals in the field of hematology and oncology followed. Besides monotherapy, a rapidly increasing number of trials is investigating checkpoint inhibitors in different combination therapies for advanced disease. Cumulating evidence suggests checkpoint blockade also as promising option for (neo)-adjuvant treatment. Here we review the different treatment strategies of mono- and combination-therapies. Additionally, important biomarkers for the treatment with checkpoint inhibitors are discussed.


An investigation on water quality of Darlik Dam drinking water using satellite images.
Darlik Dam supplies 15% of the water demand of Istanbul Metropolitan City of Turkey. Water quality (WQ) in the Darlik Dam was investigated from Landsat 5 TM satellite images of the years 2004, 2005, and 2006 in order to determine land use/land cover changes in the watershed of the dam that may deteriorate its WQ. The images were geometrically and atmospherically corrected for WQ analysis. Next, an investigation was made by multiple regression analysis between the unitless planetary reflectance values of the first four bands of the June 2005 Landsat TM image of the dam and WQ parameters, such as chlorophyll-a, total dissolved matter, turbidity, total phosphorous, and total nitrogen, measured at satellite image acquisition time at seven stations in the dam. Finally, WQ in the dam was studied from satellite images of the years 2004, 2005, and 2006 by pattern recognition techniques in order to determine possible water pollution in the dam. This study was compared to a previous study done by the authors in the Küçükçekmece water reservoir, also in Istanbul City.


HER2 status in gastric and gastroesophageal junction cancer assessed by local and central laboratories: Chinese results of the HER-EAGLE study.
Trastuzumab has been approved for human epidermal growth factor receptor 2 (HER2)-positive advanced gastric and gastroesophageal junction cancers (GC and GJC) in combination with chemotherapy. The aim of this HER2 early/advanced gastric epidemiology (HER-EAGLE) study was to evaluate the frequency of HER2 over-expression and to evaluate agreement on HER2 status assessment in GC and GJC patients in local laboratories versus a central laboratory in China. Tumor samples from 734 GC or GJC patients who were enrolled at 11 different hospitals in China were examined. HER2 status was assessed by immunohistochemistry (IHC), and followed by dual-color silver-enhanced in Situ hybridization (DSISH) in IHC 2+ cases. Clinicopathologic characteristics were collected from all of the patients. HER2-positive tumors were identified in 12.0% (88/734) of the GC and GJC cases. There were significantly higher rates of HER2 positivity in patients with GJC (GJC: 18.1%, GC: 9.7%, P=0.002), and intestinal-type cancers using the Lauren classification (intestinal: 23.6%, diffuse/mixed: 4.3%, P<0.0001). No significant difference in HER2 positivity was identified between resection and biopsy samples, or between early and advanced disease stages. The agreement between local laboratories and the central laboratory on HER2 status scoring was good (kappa=0.86). The main reason of HER2 status discordance between local and the central laboratories was IHC result mis-interpretation in local laboratories. These results suggest that IHC followed by DSISH testing is an accurate and cost-effective procedure in China.


Efficient polyhydroxybutyrate production using acetate by engineered Halomonas sp. JJY01 harboring acetyl-CoA acetyltransferase.
Polyhydroxybutyrate (PHB) is a well-known biodegradable bioplastic synthesized by microorganisms and can be produced from volatile fatty acids (VFAs). Among VFAs acetate can be utilized by Halomonas sp. YLGW01 for growth and PHB production. In this study, Halomonas sp. JJY01 was developed through introducing acetyl-CoA acetyltransferase (atoAD) with LacIq-Ptrc promoter into Halomonas sp. YLGW01. The effect of expression of atoAD on acetate was investigated by comparison with acetate consumption and PHB production. Shake-flask study showed that Halomonas sp. JJY01 increased acetate consumption rate, PHB yield and PHB production (0.27 g/L/h, 0.075 g/g, 0.72 g/L) compared to the wild type strain (0.17 g/L/h, 0.016 g/g, 0.11 g/L). In 10 L fermenter scale fed-batch fermentation, the growth of Halomonas sp. JJY01 resulted in higher acetate consumption rate, PHB yield and PHB titer (0.55 g/L/h, 0.091 g/g, 4.6 g/L) than wild type strain (0.35 g/L/h, 0.067 h/h, 2.9 g/L). These findings demonstrate enhanced acetate utilization and PHB production through the introduction of atoAD in Halomonas strains.


Lipoprotein(a) particle concentration and lipoprotein(a) cholesterol assays yield discordant classification of patients into four physiologically discrete groups.
There is little known about the relative predictive value of different lipoprotein(a) [Lp(a)] assays in clinical use, although each has been shown to predict similar incremental risk over conventional clinical and lipid risk factors. Thus, we examined the classification behavior of two commonly used Lp(a) assays and their associations with other lipid parameters. Serum lipid and Lp(a) concentrations were measured in 144 primary and secondary prevention patients. Lp(a) cholesterol [Lp(a)-C] was measured with the Vertical Auto Profile (upper limit of normal, 10 mg/dL). Lp(a) particle concentrations [Lp(a)-P] were measured with an isoform-independent molar assay (upper limit of normal, 70 nmol/L). The subjects were divided into the following four groups on the basis of their Lp(a)-C and Lp(a)-P levels: normal Lp(a)-P and Lp(a)-C; high Lp(a)-P and normal Lp(a)-C; normal Lp(a)-P and high Lp(a)-C; and high Lp(a)-P and Lp(a)-C. The proportion of subjects with values above the upper limit of normal was similar with both assays (P = .15). However, the Lp(a)-C and Lp(a)-P assays discordantly classified 23% of the study's subjects. In addition, the four Lp(a)-defined groups displayed differences in their relationships with other lipoproteins. The two groups with elevated Lp(a)-C showed significant associations with higher high-density lipoprotein cholesterol, apolipoprotein AI, and high-density lipoprotein cholesterol/apolipoprotein AI ratios. Triglycerides were also noted to be above normal in discordant and normal within concordant Lp(a) groups. Finally, the amount of cholesterol per Lp(a) particle [Lp(a)-C/Lp(a)-P] varied widely across the four groups. These findings suggest that the four Lp(a)-defined groups are physiologically discrete. Further investigation is warranted to assess which parameters among Lp(a)-P, Lp(a)-C, and Lp(a)-C/Lp(a)-P can be used to more accurately characterize Lp(a)-associated cardiovascular risk.


High-Sensitivity 3D ZIF-8/PDA Photonic Crystal-Based Biosensor for Blood Component Recognition.
Optical biosensors are sensitive devices used in bioanalytics detection. Analysis of blood constituents is very important for the detection of major diseases and also performs a significant role in the diagnosis of diabetes, various cancers, and cardiovascular disorders. In this work, a three-dimensional photonic crystal-based biosensor composed of zeolitic imidazolate framework-8 (ZIF-8) nanoarrays are placed on polydopamine (PDA) coated on a silicon substrate. This sensor is designed, simulated, and evaluated for various blood components in the wavelength range of 1.1 to 1.5 μm by the finite-difference time-domain (FDTD) method. The proposed biosensor was used for 10 types of blood components such as biotin-streptavidin, bovine serum albumin (BSA), cytop, glucose (40 mg/100 mL), hemoglobin, blood plasma, Sylgard184, white blood compounds, urethane dimethacrylate, and polyacrylamide. The FDTD technique was used for the performance analysis of the biosensor. The design parameters of the radius, the lattice constant, the thickness of the ZIF-8 arrays, and the PDA layer thickness are chosen to optimize the photonic crystal structure. This study indicates that the thickness of the PDA is the most important parameter for peak wavelength value in comparison to the other physical parameters. The factors for optimizing the photonic crystal-based biosensors such as the peak wavelength value (PWV), sensitivity, full width at half-maximum (FWHM), and figure of merit (FOM) are significant in comparison with pertinent works in this field, which evaluated 171 nm/RIU, 7.62 nm, and 22.5 RIU-1, respectively. A change of 0.01 nm in the refractive index of the constituents of the blood leads to a shift of 80 nm in the maximum peak wavelength, therefore acting as a functional biosensor with a high detection limit of 0.004 RIU.


[The main caregiver of oncologic terminally ill patient, repercussions of this task].
to identify the profile of the caregivers of terminally ill patient, to inquire the activities, changes and difficulties. The researchers applied a questionnaire. From all the 53 caregivers, 48 were female. 92% of the caregivers were relative to the patient. Despite of the financial problems, their relationship between caregiver and patient improved. The hygiene cares, feeding, medication were responsibility of the caregiver. The activities developed increased their amount of job, negative physical and psychical repercussions appeared. 60% of the caregivers answered that the most convenient place of the patient's death was the hospital, 40% chose home. There is a distinction between the routine of care and the death of the patient; the caregivers in fact take care of life.


[Significance of inflammatory diseases of otorhinolaryngological organs in etiology and pathogenesis of myopia in the Air Forces flying personnel].
Complex ophthalmologic examination (visorefracto- and accommodometry) and study of disease structure were conducted in 1189 persons of flying staff. In the main group (persons with myopia) LOR-diseases were noticed 3.6 times more often than in those without myopia. In pilots with myopic refraction chronic inflammatory diseases of palatine tonsils: chronic compensated (35.5%) and decompensated (27%) tonsillitis occur 7-12 times more often than in control group. Investigation of visual function has revealed that in the group of persons without the history of chronic tonsillitis the least degree of myopia, the greatest visual acuity without correction, the least value of progress gradient and minimal decrease in indices of relative and absolute accommodation were observed. In persons after tonsillectomy the mentioned parameters of visual functions are close to the values obtained in pilots without the history of chronic tonsillitis; only accommodative functions were practically twice decreased. Chronic tonsillitis causes the greater degree of myopia and gradient of its progress and respectively more lower visual acuity and significant decrease in accommodation indices (P < 0.05). Also it has been detected that if myopia was diagnosed in a year or more after tonsillectomy then myopia and degree of visual function disorders respectively were minimal. If the operation was performed in a year or more after myopia detection the disorders in visual functions were more expressed and myopia reached significantly higher values (P < 0.05).


A five-year follow-up study on the pathological changes of gastric mucosa after H. pylori eradication.
To investigate the relationship between H. pylori infection, gastric cancer and other gastric diseases through the changes in gastric mucosa and the status of different gastric diseases within 5 years after H. pylori eradication in H. pylori-positive subjects in a high incidence region of gastric cancer. One thousand and six adults were selected from the general population in Yantai, Shandong province, a high incidence region for gastric cancer in China. Gastroscopy and Campylobacter-like organism (CLO) testing were performed on all subjects. Biopsy samples from the gastric antrum and body were obtained for histology and assessment of H. pylori infection. All H. pylori-positive subjects were then randomly divided into two groups: treatment group receiving Omeprazole Amoxicillin Clarythromycin (OAC) triple therapy and placebo as controls. These subjects were endoscopically followed up in the second and fifth year. We compared the endoscopic appearance and histology of the biopsy specimens from the same site obtained at the first and last visits. All 552 H. pylori-positive subjects were randomly and evenly divided into treatment group or control group. During the five-year follow-up, the numbers of patients who continued to be negative or positive for H. pylori were 161 and 198, respectively. Statistical analysis revealed that: (1) At the initial visit, there were no significant differences in the severity and activity of inflammation, atrophy and intestinal metaplasia between the biopsy specimens from the antrum and body respectively in both groups. (2) The severity and activity of inflammation in both the antrum and body were markedly reduced after H. pylori eradication (P = 0.000). (3) Within five years after H. pylori eradication, intestinal metaplasia in the antrum either regressed or had no progression, while the proportion of intestinal metaplasia in the H. pylori-positive group increased significantly (P = 0.032). (4) After H. pylori eradication, the atrophy in both the antrum and body had no significant regression. The P value was 0.223 and 0.402, respectively. H. pylori eradication results in remarkable reduction in the severity and activity of chronic gastritis, marked resolution of intestinal metaplasia in the antrum. On the other hand, continuous H. pylori infection leads to progressive aggravation of atrophy and intestinal metaplasia.


The course and branching pattern of pudendal nerve in fetus.
The pudendal nerve is a considerably large branch of the sacral plexus. There are many articles in the literature concerning the pudendal nerve in adults, but as far as we know, there is none on the branching pattern and variations in pudendal nerve anatomy in fetus. This study investigates the pudendal nerve trunking with respect to the piriformis muscle in 25 formalin-fixed fetuses (50 sides of pelves, 15 females, 10 males), ranging from 20 to 37 weeks of gestation. We investigate pudendal nerve trunking in four types: Type I-a is defined as single-trunk with the inferior rectal nerve branching proximal to the dorsal nerve of penis/clitoris (38%), Type I-b is also single-trunk with the dorsal nerve of penis/clitoris branching proximal to the inferior rectal nerve (24%), Type II is double-trunk with medial trunk as an inferior rectal nerve (34%), and Type III is triple-trunk (4%). We measured the average diameter of the main trunk of pudendal nerve in Type I-a and I-b groups to be 0.98 +/- 0.33 mm. We also measured the average length of the pudendal nerve trunks before the dorsal nerve of penis/clitoris branch to be 7.35 +/- 3.50 mm. There was no significant statistical difference in the average length, diameter, number of trunks, and pudendal nerve variations between male and female and also right and left sides of the pelves. This first and detailed fetal study of pudendal nerve trunking with respect to the piriformis muscle would be useful for educational anatomy dissections and anatomical landmark definitions for relevant clinical procedures.


Endotoxin shock: thermoregulatory mechanisms.
To clarify mechanisms of hypothermia in lipopolysaccharide (LPS) shock, four experiments were conducted in 72 chronically instrumented Wistar rats. They were intended to accomplish the following: experiment 1, determine the dose of intravenous Escherichia coli LPS that induces a body temperature (Tb) fall at a minimal mortality [the dose chosen (0.5 mg/kg) was then used in experiments 2-4]; experiment 2, identify the time course of the arterial blood pressure (BP) fall (shock) during the response to LPS; experiment 3, measure threshold Tb values for skin vasodilation and activation of metabolic heat production (M) during the LPS shock; and experiment 4, ascertain behavioral thermoregulation in LPS shock. For experiments 1-3, rats were kept in restrainers; ambient temperature (Ta) was 26 degrees C. In experiment 4, rats freely moved in a thermogradient (18-33 degrees C). Variables monitored were colonic (Tc) and tail skin (Tsk) temperatures (experiment 1); BP (experiment 2); hypothalamic temperature (Thy), M (from oxygen consumption), and Tsk (experiment 3); and preferred Ta (Tpr) and abdominal temperature (experiment 4). In experiment 1, LPS induced no Tc changes at 0 mg/kg, a biphasic fever (no mortality) at 0.05 mg/kg, a biphasic hypothermia (42% mortality) at 0.5 mg/kg, and a rapid fall of Tc (100% mortality) at 5 mg/kg. LPS-induced (0.5 mg/kg) hypotension (experiment 2) occurred simultaneously with the first hypothermic phase; both Tc and BP reached their nadirs (-0.8 +/- 0.1 degrees C and -34 +/- 12 mmHg) at approximately 1.5 h post-LPS. The major autonomic mechanism of the shock hypothermia was a shift in the threshold Thy for M from 37.9 +/- 0.3 to 36.0 +/- 0.3 degrees C (experiment 3; P < 0.05). In experiment 4, rats selected Tpr below 25 degrees C (vs. 28-30 degrees C in control; P < 0.05) throughout the duration of the shock; their Tb dropped to 36.2 +/- 0.3 degrees C (P < 0.05). In sum, the LPS shock-associated hypothermia involves a decrease in the threshold Tb for M, the resultant widening of the interthreshold zone, and cold-seeking behavior.


Neoplasms of the canine urinary bladder.
Evaluation of histologic data obtained from 90 breeds of dogs with 3,837 neoplasms collected over 10 years (1961-1971) revealed 21 cases of urinary bladder neoplasmia in 14 breeds of dogs. There was no breed predispostion. The mean age of dogs with urinary bladder neoplasia was 9.5 years; 6 were males, 13 were females, and 2 were spayed females.


Reproduction in women reporting chronic musculoskeletal disorders.
The present population based study assessed aspects of female health as a part of a national cardiovascular screening program in 40-42 year old women in the Sor-Trondelag county. Fertility, events related to the menstrual cycle, reproduction, and the use of contraceptive methods in women reporting musculoskeletal diseases (MSD) were compared to women without these disorders (references). A higher proportion of women with MSD reported subfertility. No significant difference in mean age of menarche or menstrual cycling pattern was found, however women reporting MSD had significantly more often metrorrhagia than references. Mother's age at birth of the first and last child differed from references only in two of the patient groups, and interval between pregnancies was longer in women with MSD. Women with MSD more often chose sterilisation as a contraceptive method. Reproduction was altered in women with MSD. It remains to elucidate the interrelationship between rheumatic disorders and gynecological dysfunction.


Armenian hamster female protein: a pentraxin under complex regulation.
The serum of Armenian hamster (Cricetulus migratorius) contains a protein homologous to female protein (FP) that has been characterized in the Syrian (golden) hamster. Of unknown function, FP belongs to a family of proteins (called pentraxins) that have a common ancestral gene and are widely expressed in nature. Whereas serum concentration of FP in Syrian hamsters (SFP) is many fold greater (200- to 300-fold) in females vs. males, Armenian hamster FP (AFP) is only moderately elevated (approximately 3-fold) in female vs. males and only for the fall-winter months of the year. In the Armenian hamster testosterone administration to females or castration of males has no effect on AFP serum levels, whereas in Syrian hamster these treatments change SFP serum concentration to that characteristic of the opposite sex. Some sex steroid control of hepatic AFP synthesis is evident, however, as serum levels decrease after exogenous estrogen treatment. In contrast to Syrian hamster FP, normal levels of AFP are dependent on an intact hypophysial-pituitary axis and also are influenced by the season of the year. As an acute-phase protein, AFP responds in a typical fashion, with increasing serum levels detected in both sexes in contrast to the divergent sex-limited response in Syrian hamsters. Although AFP and SFP are similar structurally, morphologically, and antigenetically and share common binding specificities, the regulation of FP synthesis in Armenian hamster is very different from that previously found in Syrian hamster.


Thy1.2 YFP-16 transgenic mouse labels a subset of large-diameter sensory neurons that lack TRPV1 expression.
The Thy1.2 YFP-16 mouse expresses yellow fluorescent protein (YFP) in specific subsets of peripheral and central neurons. The original characterization of this model suggested that YFP was expressed in all sensory neurons, and this model has been subsequently used to study sensory nerve structure and function. Here, we have characterized the expression of YFP in the sensory ganglia (DRG, trigeminal and vagal) of the Thy1.2 YFP-16 mouse, using biochemical, functional and anatomical analyses. Despite previous reports, we found that YFP was only expressed in approximately half of DRG and trigeminal neurons and less than 10% of vagal neurons. YFP-expression was only found in medium and large-diameter neurons that expressed neurofilament but not TRPV1. YFP-expressing neurons failed to respond to selective agonists for TRPV1, P2X(2/3 and TRPM8 channels in Ca2+ imaging assays. Confocal analysis of glabrous skin, hairy skin of the back and ear and skeletal muscle indicated that YFP was expressed in some peripheral terminals with structures consistent with their presumed non-nociceptive nature. In summary, the Thy1.2 YFP-16 mouse expresses robust YFP expression in only a subset of sensory neurons. But this mouse model is not suitable for the study of nociceptive nerves or the function of such nerves in pain and neuropathies.


Light adaptation mechanisms in the eye of the fiddler crab Afruca tangeri.
A great diversity of adaptations is found among animals with compound eyes and even closely related taxa can show variation in their light-adaptation strategies. A prime example of a visual system evolved to function in specific light environments is the fiddler crab, used widely as a model to research aspects of crustacean vision and neural pathways. However, questions remain regarding how their eyes respond to the changes in brightness spanning many orders of magnitude, associated with their habitat and ecology. The fiddler crab Afruca tangeri forages at low tide on tropical and semi-tropical mudflats, under bright sunlight and on moonless nights, suggesting that their eyes undergo effective light adaptation. Using synchrotron X-ray tomography, light and transmission electron microscopy and in vivo ophthalmoscopy, we describe the ultrastructural changes in the eye between day and night. Dark adaptation at dusk triggered extensive widening of the rhabdoms and crystalline cone tips. This doubled the ommatidial acceptance angles and increased microvillar surface area for light capture in the rhabdom, theoretically boosting optical sensitivity 7.4 times. During daytime, only partial dark-adaptation was achieved and rhabdoms remained narrow, indicating strong circadian control on the process. Bright light did not evoke changes in screening pigment distributions, suggesting a structural inability to adapt rapidly to the light level fluctuations frequently experienced when entering their burrow to escape predators. This should enable fiddler crabs to shelter for several minutes without undergoing significant dark-adaptation, their vision remaining effectively adapted for predator detection when surfacing again in bright light.


Gene expression profile of ovalbumin-induced lung inflammation in a murine model of asthma.
Asthma is a chronic inflammatory disease that is associated with airway hyperresponsiveness, tissue remodeling, and airway obstruction, and that involves coordinate expression of multiple inflammatory genes in the lungs. To evaluate the gene expression pattern in a mouse model of asthma and assess the effect of a new drug, R142571, on the gene expression profile. Lung tissue from ovalbumin-sensitized mice was used to examine gene expression on the CodeLink oligonucleotide mouse 20 K bioarray platform. Data were validated for some genes by semiquantitative reverse-transcriptase polymerase chain reaction. Of the 19,736 genes represented on the microarray, expression of 378 genes was differentially regulated (215 upregulated and 163 downregulated), with at least a 2-fold change in expression (P <.05). The differentially regulated transcripts included genes known to be involved in several different biological processes, including signaling, DNA-dependent transcriptional regulation, immune response, proteolysis, and peptidolysis. Cluster analysis of the differentially regulated genes showed that at least 16 were downregulated by R142571 treatment at both of the doses used (1 and 10 mg/kg). In addition, 46 and 29 genes were downregulated at doses of 10 mg/kg and 1 mg/kg, respectively, as compared to the animals treated with vehicle. The cytokine expression pattern in our data, suggests that the murine model exhibits a predominantlyT helper 2-type response, as observed in asthmatic human subjects. Based on this study, we suggest that this mouse model would be an appropriate system for screening new drug molecules for treatment of atopic asthma.


Pediatric Primary Cutaneous Marginal Zone Lymphoma Treated With Doxycycline.
Primary cutaneous marginal zone lymphoma (PCMZL) is a cutaneous B-cell lymphoma that rarely occurs in children. We present a 13-year-old boy with multiple asymptomatic erythematous papules and nodules on the trunk and arms that were confirmed on biopsy to be PCMZL. He was treated with doxycycline and intralesional triamcinolone with improvement of lesions. This case supports the use of doxycycline for the treatment of pediatric PCMZL in patients with widespread involvement despite negative Borrelia serology. Multiple low-risk treatment modalities may be used in conjunction to clear disease in pediatric patients.


[Outpatient care of patients with ovarian cancer after primary treatment and the prevention of recurrence].
According to the scheme worked out by the staff of the 1st oncogynecologic department of the Institute, all patients with ovarian cancer in whom primary treatment was effective should be followed up. Clinical and cytologic examination should be carried out every 2 months during the first 1.5-2 years when recurrence is most likely to develop. Thereafter, cytologic follow-up (every 3 to 4 months) should be continued. Second-look operations should be carried out on a wider scale in suspicious cases. Prevention of recurrence should be assured by adequate primary treatment of ovarian cancer, particularly, at early stage and regular posttreatment check-ups.


Analysis of sodium current fluctuations in the cut-open squid giant axon.
Patch pipettes were used to record the current arising from small populations of sodium channels in voltage-clamped cut-open squid axons. The current fluctuations associated with the time-variant sodium conductance were analyzed with nonstationary statistical techniques in order to obtain an estimate for the conductance of a single sodium channel. The results presented support the notion that the open sodium channel in the squid axon has only one value of conductance, 3.5 pS.


Adaptive evolution and antiviral activity of the conserved mammalian cytidine deaminase APOBEC3H.
The APOBEC3 genes encode cytidine deaminases that act as components of an intrinsic immune defense that have potent activity against a variety of retroelements. This family of genes has undergone a rapid expansion from one or two genes in nonprimate mammals to at least seven members in primates. Here we describe the evolution and function of an uncharacterized antiviral effector, APOBEC3H, which represents the most evolutionarily divergent APOBEC3 gene found in primates. We found that APOBEC3H has undergone significant adaptive evolution in primates. Consistent with our previous findings implicating adaptively evolving APOBEC3 genes as antiviral effectors, APOBEC3H from Old World monkeys (OWMs) has efficient antiviral activity against primate lentiviruses, is sensitive to inactivation by the simian immunodeficiency virus Vif protein, and is capable of hypermutating retroviral genomes. In contrast, human APOBEC3H is inherently poorly expressed in primate cells and is ineffective at inhibiting retroviral replication. Both OWM and human APOBEC3H proteins can be expressed in bacteria, where they display significant DNA mutator activity. Thus, humans have retained an APOBEC3H gene that encodes a functional, but poorly expressed, cytidine deaminase with no apparent antiviral activity. The consequences of the lack of antiviral activity of human APOBEC3H are likely to be relevant to the current-day abilities of humans to combat retroviral challenges.


Does walkable neighbourhood design influence the association between objective crime and walking?
Few studies have investigated associations between objectively measured crime and walking, and findings are mixed. One explanation for null or counterintuitive findings emerges from criminology studies, which indicate that the permeable street layouts and non-residential land uses that underpin walkable neighbourhoods are also associated with more crime. This study examined associations between objective crime and walking, controlling for the characteristics of walkable neighbourhoods. A population representative sample of adults (25-65 years) (n = 3,487) completed the Western Australian Health and Wellbeing Survey (2006-2008) demographic and walking frequency items. Objective environmental measures were generated for each participant's 400 m and 1600 m neighbourhood areas, including burglary, personal crime (i.e., crimes committed against people) in public space, residential density, street connectivity and local destinations. Log-linear negative binomial regression models were used to examine associations between crime and walking frequency/week, with progressive adjustment for residential density, street connectivity and local destinations. Burglary and personal crime occurring within a participant's 400 m and 1600 m neighbourhoods were positively and significantly associated with walking frequency. For example, for every additional 10 crimes against the person/year within 400 m of a participant's home, walking frequency increased by 8% (relative change = 1.077, p = 0.017). Associations remained constant after controlling for residential density and street connectivity, but attenuated after adjusting for local destinations (e.g., for personal crime in 400 m: relative change = 1.054, p = 0.104). This pattern of attenuation was evident across both crime categories and both neighbourhood sizes. The observed positive associations between objective crime and walking appear to be a function of living in a more walkable environment, as the presence of destinations has the capacity to both promote walking and attract crime. This study provides a plausible explanation for some mixed findings emerging from studies examining crime as a barrier to walking. In some settings, the hypothesised deterrent effect of crime on walking may be insufficient to outweigh the positive impacts of living in a more walkable environment.


Combination of RGB and multispectral imagery for discrimination of cabernet sauvignon grapevine elements.
This paper proposes a sequential masking algorithm based on the K-means method that combines RGB and multispectral imagery for discrimination of Cabernet Sauvignon grapevine elements in unstructured natural environments, without placing any screen behind the canopy and without any previous preparation of the vineyard. In this way, image pixels are classified into five clusters corresponding to leaves, stems, branches, fruit and background. A custom-made sensory rig that integrates a CCD camera and a servo-controlled filter wheel has been specially designed and manufactured for the acquisition of images during the experimental stage. The proposed algorithm is extremely simple, efficient, and provides a satisfactory rate of classification success. All these features turn out the proposed algorithm into an appropriate candidate to be employed in numerous tasks of the precision viticulture, such as yield estimation, water and nutrients needs estimation, spraying and harvesting.