Psychiatric comorbidities in epilepsy.
Psychiatric disorders can be identified in 25-50% of patients with epilepsy, with higher prevalence among patients with poorly controlled seizures. These disturbances include depression, anxiety, psychotic disorders, cognitive, and personality changes occurring in the interictal or ictal/postictal states. In this chapter, we describe four areas of focus in women with epilepsy: comorbid primary psychiatric processes, integrated symptoms secondary to epilepsy, stigma and psychosocial consequences of epilepsy, and nonepileptic seizures.


Opening two benzene rings on trichalcogenasumanenes toward high performance organic optical-limiting materials.
Two flanking benzene rings on trichalcogenasumanenes (1a/1b) were opened upon oxidation, and the successive ring reconstruction afforded donor-acceptor type polycycles containing diimide units (3a/3b). 3a and 3b show excellent optical-limiting properties superior to C60.


Receptor and non-receptor mediated uptake of chylomicron remnants by the liver.
The uptake of chylomicron remnants by rodent liver is mediated by proteins residing on the microvillous surface of hepatocytes and occurs in two steps. First, initial removal of the remnants from the blood occurs through binding to the low density lipoprotein (LDL) receptor via apo E and to hepatic lipase via polar lipids and proteins on the remnant surface. Second, chylomicron remnants are taken up into the cell mainly by the LDL receptor and follow the classical receptor-mediated pathway of endocytosis. The LDL receptor-related protein (LRP), which binds weakly to chylomicron remnants via apo E, does not appear to have a significant role in the initial removal process. The remnant particles can, however, be enriched with proteoglycan-bound apo E present on hepatocytic microvilli, which increases their affinity for LRP to the extent that they are subject to endocytosis by this receptor, particularly when the LDL receptor is deficient or down-regulated. Hepatic lipase can also mediate binding of remnants to LRP, for which it has high affinity. Lipolysis of remnant lipids by hepatic lipase may promote but is not required for interaction of remnants with the endocytic receptors. Proteoglycan-bound hepatic lipase may also mediate endocytosis of chylomicron remnants independent of apo E, so that hepatic catabolism of these particles is not completely dependent upon this apoprotein. Available data from experiments in vivo thus indicate redundancy of both steps of hepatic uptake of chylomicron remnants, consistent with the centrality of this process in nutrient delivery.


Current bladder cancer tests: unnecessary or beneficial?
Bladder cancer is currently diagnosed using cystoscopy and cytology in patients with suspicious signs and symptoms. These same tests are used to monitor patients with a history of bladder cancer for recurrence. The recurrence rate for bladder cancer is high, thus necessitating long-term follow-up. Urine cytology requires an experienced cytopathologist and is costly. It has high specificity, but low sensitivity for low-grade bladder tumors. Recently many non-invasive bladder cancer tests, utilizing markers found in the urine, have been developed. The FDA has approved several of these for the use is bladder cancer diagnosis, and many others are undergoing development and investigation. An ideal bladder cancer test would be non-invasive, highly sensitive and specific, inexpensive, easy to perform, and yield highly reproducible results. Many of the tests reviewed meet some, but not all, of these criteria.


COVID-19 and Perinatal Stress Experience - a Study Conducted as Part of the COVGEN Initiative.
Introduction During the COVID-19 pandemic, stress and anxiety in the population increased due to concerns about people's own health and that of their relatives, as well as changes in everyday life due to measures taken to reduce the infection rate. Pregnant women are particularly stressed. The present study examines how the COVID-19 pandemic affects the stress experience and mental health of pregnant women and mothers of newborns and how care could be optimized. Methods As part of the international COVGEN initiative ( https://www.covgen.org ) to investigate the effects of the COVID-19 pandemic on the peripartum period, pregnant and postpartum women were asked about their experience with stress using the COPE-IS (Coronavirus Perinatal Experiences - Impact Survey) questionnaire developed for this purpose and translated from the English. In addition, demographic data, pre-existing diseases, pregnancy complications and the care situation were recorded. The questionnaire was either administered as hardcopy to inpatients at the Department of Women's Health, University Hospital Tübingen, Germany, or online. All pregnant women and mothers who were pregnant or had given birth after the official start of the COVID-19 pandemic (11 March 2020) were eligible to participate. Results Complete data sets of n = 156 pregnant women and n = 221 postpartum women were available for evaluation. The general stress level assessed with the COPE-IS was significantly increased by the COVID-19 pandemic in both, pregnant and postpartum women, with pre-existing conditions such as respiratory diseases and pregnancy-related diseases like gestational diabetes adding to the stress. The subjectively perceived quality of care/support during pregnancy also influenced the stress level. Conclusions Fears of a COVID-19 infection and changes in preventive and aftercare services were a burden for the women surveyed. Intensified care during pregnancy and puerperium could help to stabilize the mental situation and reduce stress.


Effect of anaemia on red cell metabolism in cattle.
Six enzymes: glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), glutathione reductase (GR), glutathione peroxidase (GSHPx), catalase and superoxide dismutase (SOD), and three metabolites: 2,3-diphosphoglycerate (DPG), adenosine triphosphate (ATP) and reduced glutathione (GSH), were measured in the red blood cells of cattle during experimental anaemia. There was a significant rise in the mean levels of DPG (from 0.85 to 1.69 mumol/gHb) and ATP (from 2.16 to 3.25 mumol/gHb) in the early phase of recovery. Enzyme activities also increased during the early recovery period.


Aortic distensibility after aortic root replacement assessed with cardiovascular magnetic resonance.
The changes in geometry of the aortic root during the cardiac cycle are thought to be essential for optimal valve function, both in terms of leaflet stress and dynamic behavior. Using cardiac magnetic resonance (CMR), the study aim was to determine aortic root distensibility of the homograft (group H) and the Medtronic Freestyle xenograft (group F) after aortic root replacement, from a prospective randomized trial. CMR was performed in 15 patients (six homografts, nine Freestyle) at six months and one year after surgery. Percentage change in aortic radius (PCR) and pressure strain elastic modulus (PSEM) were measured as indices of distensibility, and results related to left ventricular mass (LVM). At six months after surgery, mean PCR was 12+/-2.5 in group H and 12.9+/-6.1 in group F (p = NS), and PSEM was 428.5+/-69.8 and 493.5+/-72.7 g/cm2, respectively (p = NS). PCR was reduced to 10+/-1.7% in group H, and by 8.5+/-2.8% in group F (p = NS), while PSEM was increased to 520.5+/-87.3 and 825+/-420.4, respectively (p = NS) at the one-year follow up. Regression analysis showed a correlation between PCR and LVM (r = 0.52, p = 0.08) and LVM index (r = 0.46, p = 0.14), respectively. In addition, there was a relationship between PSEM, LVM and LVM index, suggesting that the stiffer the root wall, the higher the postoperative LVM. Up to one year after aortic root replacement, the wall of both the allogenic and xenogenic valves retained near-normal distensibility. For the first time, a correlation was demonstrated between the elastic properties of the aortic root and LVM. The longer-term behavior and clinical implications of these findings require further investigation.


Hospital management of a patient with factitial dermatitis.
A 64-year-old male with a 40-year history of dermatologic disorders is presented. The patient carries diagnoses of neurodermatitis and suspected myosis fungoides on which has been superimposed a factitial dermatitis requiring increasingly extended hospitalizations. Given a very disturbed home life and personality style, the patient showed no incentive for change, and his family and caregivers became increasingly frustrated and distraught. An operant behavioral approach proved helpful to staff in standardizing care and setting realistic expectations. This led to a reversal of a downward course and to a coordinated discharge.


Investigating the role of gap junctions in seizure wave propagation.
The effect of gap junctions as well as the biological mechanisms behind seizure wave propagation is not completely understood. In this work, we use a simple neural field model to study the possible influence of gap junctions specifically on cortical wave propagation that has been observed in vivo preceding seizure termination. We consider a voltage-based neural field model consisting of an excitatory and an inhibitory population as well as both chemical and gap junction-like synapses. We are able to approximate important properties of cortical wave propagation previously observed in vivo before seizure termination. This model adds support to existing evidence from models and clinical data suggesting a key role of gap junctions in seizure wave propagation. In particular, we found that in this model gap junction-like connectivity determines the propagation of one-bump or two-bump traveling wave solutions with features consistent with the clinical data. For sufficiently increased gap junction connectivity, wave solutions cease to exist. Moreover, gap junction connectivity needs to be sufficiently low or moderate to permit the existence of linearly stable solutions of interest.


Design of a calf muscle ergometer for the study of local muscle blood flow.
A simple ergometer is described that allows highly reproducible plethysmographic measurements of blood flow to be made within 10 seconds of ceasing rhythmic quantitated exercise of the calf muscle. A power output of some 25 Watts is developed with a mechanical efficiency of some 20%. The device seems applicable to a variety of clinical assessments.


Indoxyl glucuronide, a protein-bound uremic toxin, inhibits hypoxia-inducible factor‒dependent erythropoietin expression through activation of aryl hydrocarbon receptor.
Renal anemia is common among chronic kidney disease (CKD) patients, and is mainly caused by inadequate erythropoietin (EPO) production from kidneys due to dysfunction of intracellular hypoxia-inducible factor (HIF) signaling in renal EPO-producing cells. We have previously shown that indoxyl sulfate (IS), a representative protein-bound uremic toxin accumulated in the blood of CKD patients, inhibits hypoxia-induced HIF activation and subsequent EPO production through activation of aryl hydrocarbon receptor (AHR). In this study, we further investigated the effects of other protein-bound uremic toxins on HIF-dependent EPO expression using EPO-producing HepG2 cells. We found that indoxyl glucuronide (IG) and IS, but not p-cresyl sulfate, phenyl sulfate, 3-indoleacetic acid or hippuric acid, inhibited hypoxia mimetic cobalt chloride-induced EPO mRNA expression. Furthermore, IG at concentrations similar to the blood levels in CKD patients inhibited the transcriptional activation of HIF induced by both cobalt chloride treatment and hypoxic culture. IG also induced CYP1A1 mRNA expression and nuclear translocation of AHR protein, indicating that IG activates AHR signaling. Blockade of AHR by a pharmacological antagonist CH-223191 abolished the IG-induced inhibition of HIF activation. Collectively, this study is the first to elucidate the biological effects of IG to inhibit HIF-dependent EPO production through activation of AHR. Our data suggests that not only IS but also IG contributes to the impairment of HIF signaling in renal anemia.


Reduction in locomotor ability as a cost of reproduction in gravid snakes.
Recent studies suggest that lower survival among gravid squamate reptiles may be partially the result of decreased locomotor ability during gestation. In this study, we compared the speed and endurance of female garter snakes (Thamnophis marcianus), before, during, and after pregnancy. Gravid snakes had significantly lower locomotor performance than did non-gravid females, and performance varied among stages of gestation, reaching a minimum 0-6 weeks prior to parturition. Both number of offspring and relative clutch mass were inversely correlated with locomotor performance; as females increased these traits, locomotor ability decreased. If reduced locomotor performance results in greater risk of predation and/or lowered foraging ability, then natural selection (operating via differential mortality or feeding rates of gravid females) may result in important constraints on both clutch size and relative clutch mass in squamates.


Self-Esteem and Problematic Drinking in China: A Mediated Model.
Although self-esteem is related to problematic drinking, the mechanisms by which it affects drinking remain unclear. The purpose of this study was to determine whether coping mechanisms mediate the relationship between self-esteem and problematic drinking among Chinese men and women with alcohol use disorders and to recommend appropriate interventions for drinking problems. A cross-sectional survey was conducted in Harbin, Heilongjiang Province, China. A sample of 5,689 community residents was screened, and 517 male and 172 female problematic drinkers were chosen to participate in this study. A self-esteem scale, a coping questionnaire and an alcohol use disorder identification test were completed in order to assess participants' self-esteem, coping mechanisms and alcohol use disorders, respectively. Participants' socio-demographic data were also gathered at this stage. The resulting data were examined via descriptive statistics, correlations and bootstrap analyses. Lower self-esteem levels were related to problematic drinking, and there were no gender differences in the relationship between self-esteem and problematic drinking. A relationship between low self-esteem and negative coping was observed only in men. Negative coping thus mediated the relationship between self-esteem and problematic drinking among men, but this was not the case for women. Positive coping did not mediate the relationship between self-esteem and problematic drinking among the participants, regardless of gender. Self-esteem and coping strategies are correlated among problematic drinkers. In addition, there are gender differences in the manners in which negative coping mediates the relationship between self-esteem and problematic drinking. Problematic drinking interventions directed at males should simultaneously address low self-esteem and negative coping.


Pure Progressive Ataxia and Palatal Tremor (PAPT) Associated with a New Polymerase Gamma (POLG) Mutation.
Progressive ataxia with palatal tremor (PAPT) is a syndrome caused by cerebellar and brainstem lesions involving the dentato-rubro-olivary tract and associated with hypertrophic olivary degeneration. Etiologies include acquired posterior fossa lesions (e.g. tumors, superficial siderosis, and inflammatory diseases) and genetic disorders, such as glial fibrillary acidic protein (GFAP) and polymerase gamma (POLG) mutations. We describe the case of a 52-year-old man who developed pure progressive ataxia and palatal tremor. Genetic analysis has shown that he is compound heterozygote for a known pathogenic (W748S) and a novel POLG variant (I1185N). Patients with POLG recessive mutations usually manifest a more complex clinical picture, including polyneuropathy and epilepsy; our case emphasizes the need to consider a genetic origin in a seemingly sporadic and pure PAPT.


Association of PTPN1 polymorphisms with breast cancer risk: A case-control study in Chinese females.
Breast cancer (BC) risk, development, and prognosis were closely related to obesity, diabetes mellitus, and metabolic syndrome. Protein tyrosine phosphatase, non-receptor type 1 (PTPN1) located on chromosome 20q13, could negatively regulate insulin and leptin signaling. In this study, we determined the association of PTPN1 polymorphisms with BC risk. We analyzed the distribution of 11 selected PTPN1 single nucleotide polymorphisms in Chinese female patients with BC (n = 953) and healthy controls (n = 963) based on a multicenter case-control study. The association of PTPN1 genotypes and haplotypes frequencies with BC risk were determined by logistic regression analysis. Analyses were further stratified by body mass index (BMI), waist-hip rate (WHR), diabetes mellitus history, and fasting plasma glucose level. The eQTL (expression Quantitative Trait Loci) analysis for PTPN1 was conducted by GTEx database. There were significant differences between BC cases and control groups in menopausal status, number of births, and BMI. Four single nucleotide polymorphisms (SNPs; rs3215684, rs3787345, rs718049, and rs718050) decreased overall BC risk, and other seven SNPs showed no significant association with BC risk. In multivariate analysis, BMI and rs3215684 DT + DD genotype were identified as independent risk factors for BC, and mutated genotypes of rs3215684 were correlated with increased PTPN1 expression. There are no haplotypes showed different frequencies between cases and controls. In the stratified analysis, rs2206656 showed a significant association with decreased BC risk in the subgroup of BMI ≤ 24 kg/m 2 , while rs3215684 and rs718049 showed lower BC risk in the subgroup of WHR > 0.85. Seven SNPs showed lower BC risk in the subgroup with diabetes mellitus history and/or fasting plasma glucose level ≥ 7 mM, while rs754118 decreased BC risk in the subgroup of fasting plasma glucose level < 7 mM. Our findings suggest that PTPN1 SNPs associated with BC susceptibility in Chinese females, which also suggested a novel mechanism between obesity, diabetes mellitus, and BC risk.


Risks of refractory chest pain after peroral endoscopic myotomy in achalasia-related esophageal motility disorders: short-term results from a multicenter study in Japan.
The etiology of chest pain in achalasia-related esophageal motility disorders and the frequency and risk factors of persistent chest pain after peroral endoscopic myotomy (POEM) remain unclear. A multicenter cohort study including 14 hospitals was conducted to elucidate the characteristics of patients with chest pain and the efficacy of POEM. Consecutive cases of achalasia-related esophageal motility disorders included 2294 (64.2%) and 1280 (35.8%) patients with and without chest pain, respectively. Among the 2107 patients with chest pain who underwent POEM, we observed complete remission in 1464 patients (69.5%) and nonremission in 643 patients (30.5%), including a partial response in 619 patients (29.4%) and resistance in 24 patients (1.1%). Multivariate analysis revealed that advanced age (odds ratio [OR], .28), male sex (OR, .70), prior treatment (OR, 1.39), and sigmoid type (OR, .65) were related to the prevalence of chest pain. Long disease duration (OR, .69) and esophageal dilation (OR, .79) were related to decreased severity. POEM improved patients' quality of life that was hindered by chest pain. Early onset (OR, 1.45), advanced age (OR, .58), male sex (OR, .79), prior treatment (OR, 1.37), and posterior myotomy (OR, 1.42) were associated with nonremission after POEM; high-resolution manometry (HRM) findings and myotomy length showed no statistical significance on pain etiology and persistence after POEM. The prevalence and severity of chest pain were dependent on age, sex, disease duration, prior treatment, and esophageal morphology rather than HRM findings. The efficacy of POEM is satisfactory; however, residual pain was often observed. Excessively long myotomy can be avoided, and anterior myotomy may be recommended.


Iron catalysis of lipid peroxidation in ferroptosis: Regulated enzymatic or random free radical reaction?
Duality of iron as an essential cofactor of many enzymatic metabolic processes and as a catalyst of poorly controlled redox-cycling reactions defines its possible biological beneficial and hazardous role in the body. In this review, we discuss these two "faces" of iron in a newly conceptualized program of regulated cell death, ferroptosis. Ferroptosis is a genetically programmed iron-dependent form of regulated cell death driven by enhanced lipid peroxidation and insufficient capacity of thiol-dependent mechanisms (glutathione peroxidase 4, GPX4) to eliminate hydroperoxy-lipids. We present arguments favoring the enzymatic mechanisms of ferroptotically engaged non-heme iron of 15-lipoxygenases (15-LOX) in complexes with phosphatidylethanolamine binding protein 1 (PEBP1) as a catalyst of highly selective and specific oxidation reactions of arachidonoyl- (AA) and adrenoyl-phosphatidylethanolamines (PE). We discuss possible role of iron chaperons as control mechanisms for guided iron delivery directly to their "protein clients" thus limiting non-enzymatic redox-cycling reactions. We also consider opportunities of loosely-bound iron to contribute to the production of pro-ferroptotic lipid oxidation products. Finally, we propose a two-stage iron-dependent mechanism for iron in ferroptosis by combining its catalytic role in the 15-LOX-driven production of 15-hydroperoxy-AA-PE (HOO-AA-PE) as well as possible involvement of loosely-bound iron in oxidative cleavage of HOO-AA-PE to oxidatively truncated electrophiles capable of attacking nucleophilic targets in yet to be identified proteins leading to cell demise.


A signature of 17 immune-related gene pairs predicts prognosis and immune status in HNSCC patients.
Head and neck squamous cell carcinoma (HNSCC) is an invasive malignancy with high worldwide mortality. Growing evidence has indicated a pivotal correlation between HNSCC prognosis and immune signature. This study investigated an immune-related gene pairs (IRGPs) signature to predict the prognostic value of HNSCC patients. We constructed IRGPs via integrating multiple IRG expression data sets. Moreover, we established the predictive model base on the IRGPs for HNSCC, and utilized multidimensional bioinformatics methods to validate the robustness of prognostic value of the IRGPs signature. In addition, we explored the relationship between the IRGPs model and immune status. Seventeen IRGPs signature was built as the predictive model which predicted prognosis independently and reliably for HNSCC. Compared to the high-risk group, the low-risk group demonstrated a distinctly favorable prognosis including overall survival (OS), disease-specific survival (DSS), and progression-free survival (PFS). The low-risk group showed higher-immune score and lower-tumor purity than the high-risk group. In addition, the low-risk group exhibited higher expression of Programmed cell death 1 ligand 1 (PD-L1) and Microsatellite instability (MSI) score, and lower expression of Tumor Immune Dysfunction and Exclusion (TIDE), which indicated the low-risk group was much more sensitive to immunotherapy. Lastly, the IRGs signature has achieved a higher accuracy than clinical properties for estimation of survival. The IRGPs model is an independent biomarker for estimating the prognosis, and could be also used to predict immunotherapeutic response in HNSCC patients. These findings may provide new ideas for novel biomarkers and may be helpful to formulate personalized immunotherapy strategy.


Ketogenic diet-responsive drug-resistant epilepsy in a case of asparagine synthetase deficiency with a novel compound heterozygous missense variant.
Asparagine synthetase deficiency (ASNSD) is a rare autosomal recessive neurometabolic disorder caused by homozygous or compound heterozygous mutations in the ASNS gene. Most of the patients have early-onset intractable seizures. A 7-year-old boy was first admitted to our clinic with intractable febrile and afebrile seizures that started when he was 6 months old. He had axial hypotonia with spastic quadriparesis, mild facial dysmorphism, and acquired microcephaly at 1 year-old. Metabolic tests showed a borderline-low serum asparagine level. The electroencephalogram demonstrated epileptic discharges with a high incidence of multifocal spike-wave activity. Brain MRI showed mild cerebral atrophy. His seizures continued despite combinations of multiple antiseizure agents. Whole-exome sequencing (WES) revealed a novel compound heterozygous missense variant of the ASNS gene, and the variants were confirmed by Sanger sequencing. He was started on a ketogenic diet at five years and six months of age. In the first month of the ketogenic diet, we observed that the frequency of seizures significantly decreased. He showed a remarkable improvement in seizures and milder improvement in cognitive skills. To our knowledge, our case is the first report describing significant improvement with a ketogenic diet in intractable seizures due to ASNSD.


Biology and biomechanics of the anterior cruciate ligament.
With detailed information concerning the role of the anterior cruciate ligament in the overall kinematic response of the knee during external loading, current clinical management of the orthopedic injuries to this joint may be improved upon and refined. Among the largest challenges to be met will be elucidation of the effects of various levels and types of muscular stabilization concurrent with a precise determination of the roles of passive knee restraints.


The Alzheimer's amyloid precursor protein is produced by type I astrocytes in primary cultures of rat neuroglia.
The production of beta-amyloid precursor protein (beta APP) by primary mixed glial cell cultures from cerebral cortex of 2-3 day postnatal rats was examined by Northern and Western blotting and by immunocytochemistry. A single 3.5 kb beta APP RNA transcript was detected using probes recognizing all forms of beta APP messenger RNA. No signal was detected with a probe specific for the alternately spliced Kunitz protease inhibitor (KPI) region. In Western blot analysis of protein extracts, antisera specific for either the cytoplasmic or extracellular parts of beta APP detected several proteins ranging from 105 to 140 kDa. None of these were recognized by an antiserum specific to the KPI insert of the beta APP. Multicolor immunofluorescence showed beta APP immunoreactivity in type I (GFAP+ A2B5-) astrocytes, distributed in a fibrillar pattern like that of glial fibrillary acidic protein (GFAP). No beta APP immunoreactivity was detected in oligodendrocytes (GC+) or in A2B5+ progenitor cells. Moreover, no cultured cells showed immunostaining with an antiserum specific for the KPI sequence of beta APP. We conclude that type I astrocytes in primary culture produce amyloid precursor protein, but that oligodendrocytes and their precursors do not. Moreover, type I astrocytes produce predominantly the beta APP subtypes which lacks the KPI sequence.


Cadherin structure: a revealing zipper.
The crystal structure of the N-terminal domain of neural cadherin provides the first atomic-level picture of interacting cell-adhesion molecules, and suggests a mechanism for assembly and disassembly of intercellular adhesion zones.


The detection of differentially expressed microRNAs from the serum of ovarian cancer patients using a novel real-time PCR platform.
To determine the utility of serum miRNAs as biomarkers for epithelial ovarian cancer. Twenty-eight patients with histologically confirmed epithelial ovarian cancer were identified from a tissue and serum bank. Serum was collected prior to definitive therapy. Fifteen unmatched, healthy controls were used for comparison. Serum was obtained from all patients. RNA was extracted using a derivation of the single step Trizol method. The RNA from 9 cancer specimens was compared to 4 normal specimens with real-time PCR using the TaqMan Array Human MicroRNA panel. Twenty-one miRNAs were differentially expressed between normal and patient serum. Real-time PCR for the 21 individual miRNAs was performed on the remaining 19 cancer specimens and 11 normal specimens. Eight miRNAs of the original twenty-one were identified that were significantly differentially expressed between cancer and normal specimens using the comparative C(t) method. MiRNAs-21, 92, 93, 126 and 29a were significantly over-expressed in the serum from cancer patients compared to controls (p<.01). MiRNAs-155, 127 and 99b were significantly under-expressed (p<.01). Additionally, miRs-21, 92 and 93 were over-expressed in 3 patients with normal pre-operative CA-125. We demonstrate that the extraction of RNA and subsequent identification of miRNAs from the serum of individuals diagnosed with ovarian cancer is feasible. Real-time PCR-based microarray is a novel and practical means to performing high-throughput investigation of serum RNA samples. miRNAs-21, 92 and 93 are known oncogenes with therapeutic and biomarker potential.


A Framework for Authentic Ethical Decision Making in the Face of Grand Challenges: A Lonerganian Gradation.
This paper contributes to the contemporary business ethics narrative by proposing an approach to corporate ethical decision making (EDM) which serves as an alternative to the imposition of codes and standards to address the ethical consequences of grand challenges, like COVID-19, which are impacting today's society. Our alternative approach to EDM embraces the concept of reflexive thinking and ethical consciousness among the individual agents who collectively are the corporation and who make ethical decisions, often in isolation, removed from the collocated corporate setting. We draw on the teachings of the Canadian philosopher and theologian, Fr. Bernard Lonergan, to conceptualize an approach to EDM which focuses on the ethics of the corporate agent by nurturing the universal and invariant structure that is operational in all human beings. Embracing Lonergan's dynamic cognitive structure of human knowing, and the structure of the human good, we advance a paradigm of EDM in business which emboldens authentic ethical thought, decision making, and action commensurate with virtuous living and germane to human flourishing. Lonergan's philosophy guides us away from the imposition of over-arching corporate codes of ethics and inspires us, as individual agents, to attend to the data of our own consciousness in our ethical decision making. Such cognitional endowment leads us out of the ethics of the 'timeless present' (Islam and Greenwood in Journal of Business Ethics 170: 1-4, 2021) towards ethical authenticity in business, leaving us better placed to reflect upon and address the ethical issues emanating from grand challenges like COVID-19.


Z-scheme heterojunction ZnS/WO3 composite: Photocatalytic reduction of uranium and band gap regulation mechanism.
In the present research, ZnS/WO3 composites were prepared by coprecipitation method to construct the Z-scheme heterojunction photocatalyst with high efficiency electron separation for the photocatalytic reduction of U(VI). Compared with WO3 and ZnS, the visible light absorption, photoreduction ability and photocatalytic activity of ZnS/WO3 composites were improved. The ZnS/WO3 composites show higher photoreduction U(VI) performance under visible light irradiation with the maximum extraction capacity of U(VI) at 1.52 g g-1. The ZnS/WO3 composites exhibit high uranium reduction ability under natural light with removal efficiency reaching 93.4 %. In-situ monitoring experiments and DFT calculations were designed to explore the mechanism and pathway of photoelectron transfer in the reduction process from U(VI) to U(IV). The results show that ZnS/WO3 has an internal electric field to form a Z-scheme electron transfer, and uranium reduction is a dual-electron transfer pathway. In addition, the band gap regulation mechanism of binary composite semiconductor materials is deeply discussed.


Epidemiology and management of abdominal injuries in children.
Although more guideline-adherent care has been described in pediatric compared to adult trauma centers, we aimed to provide a more detailed characterization of management and resource utilization of children with intra-abdominal injury (IAI) within pediatric centers. Our primary objective was to describe the epidemiology, diagnostic evaluation, and management of children with IAI across U.S. children's hospitals. Our secondary objective was to describe the interhospital variation in surgical management of children with IAI. We conducted a cross-sectional study of 33 hospitals in the Pediatric Health Information System. We included children aged <18 years evaluated in the emergency department from 2010 to 2019 with IAI, as defined by ICD coding, and who underwent an abdominal computed tomography (CT). Our primary outcome was abdominal surgery. We categorized IAI by organ system and described resource utilization data. We used generalized linear regression to calculate adjusted hospital-level proportions of abdominal surgery, with a random effect for hospital. We studied 9265 children with IAI. Median (IQR) age was 9.0 (6.0-13.0) years. Abdominal surgery was performed in 16% (n = 1479) of children, with the lowest proportion of abdominal surgery observed in children aged <5 years. Liver (38.6%) and spleen (32.1%) were the most common organs injured. A total of 3.1% of children with liver injuries and 2.8% with splenic injuries underwent abdominal surgery. Although there was variation in rates of surgery across hospitals (p < 0.001), only three of 33 hospitals had rates that were statistically different from the aggregate mean of 16%. Most children with IAI are managed nonoperatively, and most children's hospitals manage children with IAI similarly. These data can be used to inform future benchmarking efforts across hospitals to assess concordance with guidelines for the management of children with IAI.


[The complex rehabilitation treatment of patients with chronic obstructive pulmonary disease using haloaerosoltherapy and blastomunil (remote results)].
The expediency of complex recovery treatment of chronic obstructive pulmonary disease patients with the usage of artificial rock salt aerosol medium (haloaerosoltherapy) and immunomodulator "Blastomunil" intake was grounded on the base of the treatment's remote results study. It was revealed that additional prescription of blastomunil resulted in the lowering the frequency of ambulant and, especially, hospital treatment, decrease of average quantity of disability days during one year after treatment and promoted the remission lengthening by 1,9-2,1 months.


Targeting phosphatidylethanolamine and phosphatidylserine for imaging apoptosis in cancer.
Both phosphatidylethanolamine (PE) and phosphatidylserine (PS) can be externalized to the outer cell membrane in apoptosis. Thus the objective was to determine whether PE-targeting 18F-duramycin and PS-targeting 18F-Zn-DPA could be used for imaging apoptosis. Duramycin and Zn-DPA were labeled with either 18F-Al or 18F-SFB. U937 cells were incubated with four different concentrations of camptothecin (CPT). For assessing the effect of incubation time on uptake, 37 MBq of radiotracer was added to cells incubated for 15, 30, 60, and 120 min at 37 °C. For blocking experiments, 150 μg duramycin and 40 μg Zn-DPA were added to cells for 15 min prior to the addition of either duramycin or Zn-DPA labeled with 18F. Apoptosis was measured by flow cytometry using an annexin-V/PI kit. Cells were co-stained with Hoechst, Cy5-duramycin, and PSVue480 (FITC-Zn-DPA) to localize fluorescent dye uptake in cells. Apoptosis in cells increased proportionally with CTP as confirmed by both flow cytometry and fluorescent staining. Both FITC-Zn-DPA and FITC-duramycin localized mainly on the cell membrane during early apoptosis and then translocated to the inside during late apoptosis. Uptake of FITC-duramycin, however, was higher than that of FITC-Zn-DPA. Cellular uptake of four different radiotracers was also proportional to the degree of apoptosis, increasing slightly over time and reaching a plateau at about 1 h. The blocking experiments demonstrated that uptake in all the control groups was predominantly non-specific, whereas the specific uptake in all the treated groups was at least 50% for both 18F labeled duramycin and Zn-DPA. Both PE-targeting 18F-duramycin and PS-targeting 18F-Zn-DPA could be considered as potential radiotracers for imaging cellular apoptosis. Advances in knowledge and implications for patient care: Cellular data support the further development of radiotracers targeting either PE or PS for imaging apoptosis, which can associate with clinical outcome for cancer patients.


"Getting my Independence Back": Economic solvency program ideas from residents of a women's shelter.
Intimate partner violence (IPV) is a serious public health issue that affects many women all over the world. One of the most consistent risk factors for IPV is poverty. On the other hand, IPV is also a risk factor for continued poverty, which puts many women in a cycle of violence and poverty that is difficult to escape. Interventions are needed to help women improve their economic solvency and decrease their poverty-based risk for IPV. The current study is a qualitative study to examine how women who have experienced IPV would plan and execute an intervention to improve economic solvency. The participants were 21 diverse women living in a women's shelter. The major themes that came from the women's suggestions were as follows: addressing mental health, accountability, financial management classes, job skills and education, and other resources. Implications of this study include the need for more comprehensive programs, a general desire amongst the women for mentoring and accountability, and the need to address knowledge deficits about resources. This information can help resource providers to help build interventions to improve economic solvency in women who have experienced IPV and improve their overall safety.


The terminal period of cancer: definition and descriptive epidemiology.
There is considerable current interest in the care of the terminally ill. This study provides information on the incidence, prevalence and duration of terminal cancer, and examines patient characteristics and patterns of care during the terminal care period (TCP). The data were derived from a retrospective study of 144 cancer deaths, utilizing information from physicians, medical records, and interviews with surviving relatives. It was estimated that 90% of all cancer deaths had a TCP, of average duration 94 days. Better functional status was associated with a longer TCP, and with spending a greater proportion of the TCP at home. Several areas of concern were noted in patterns of care, including use of a tumor-directed therapy during the TCP (in 18% of patients with a TCP) and the lack of face-to-face contact with the physician outside institutions, particularly as death approached.


Use of a Smartphone App to Explore Potential Underuse of Prophylactic Aspirin for Preeclampsia.
Preeclampsia is a leading preventable cause of maternal morbidity and mortality. Initiation of low-dose aspirin (LDASA) treatment at or before 16 weeks' gestation may prevent preeclampsia onset for patients with specific risk factors. To assess potential underuse of LDASA and reasons for underuse using data from a prenatal care smartphone app. In this prospective cohort study, English-speaking pregnant patients aged 18 years or older from the UPMC health care system received an invitation to use the MyHealthyPregnancy app at their first prenatal appointment. Use of the app was voluntary. The study took place between September 23, 2019, and August 31, 2020, as part of a quality-improvement initiative. The app offered educational information, monitoring tools, and routine screenings tailored to patient-entered gestational age and demographic and clinical characteristics. App-based questions included LDASA eligibility based on US Preventive Services Task Force criteria for preeclampsia risk and a monthly prompt about LDASA recommendations from the patient's health care practitioner. The primary outcomes were the receipt of LDASA recommendations from a practitioner and adherence to any such recommendation, as self-reported on the app. Patients' medical records were examined to cross-reference their self-reports of an LDASA recommendation. Multivariable logistic regression was used to model patient-perceived recommendation as a function of factors associated with preeclampsia. The patient cohort consisted of 2563 patient participants (2036 [79%] White; mean [SD] age, 30 [5.2] years) with 2567 pregnancies; 1882 pregnancies (73.3%) were among women with private or employer-based insurance, and 1246 (48.5%) were among nulliparous patients. At least 1 factor associated with high risk for preeclampsia was reported in 316 pregnancies (12.3%), and 2 or more factors associated with moderate risk were reported in 1051 (40.9%). Of the 1015 pregnancies for which patients answered voluntary questions about aspirin use, 124 (12.2%) met at least 1 criterion for highest risk of preeclampsia. In 57 (46.0%) of these pregnancies, the patient indicated that their practitioner recommended LDASA; after examination of the medical records, 90 pregnancies (72.6%) had evidence of an LDASA recommendation and 34 (27.4%) did not. Of the 90 pregnancies with a documented LDASA recommendation, 33 patients (36.7%) were unaware of it. Prior preeclampsia (28 weeks' gestation: odds ratio, 20.1; 95% CI, 11.0-36.9) and chronic hypertension (28 weeks' gestation: odds ratio, 17.4; 95% CI, 6.3-48.2) were the primary high-risk factors associated with recommendation of LDASA. In this cohort study, only 46.0% of prenatal care app users who met the criteria for highest preeclampsia risk reported receiving an LDASA recommendation from their practitioner, and medical records suggested that there may have been frequent miscommunication between patients and practitioners about LDASA use. Digital tools such as the MyHealthyPregnancy app might offer an opportunity to improve identification of patients at risk for preeclampsia and communication with these patients about aspirin use.


Current use of high-resolution mass spectrometry in the environmental sciences.
During the last two decades, mass spectrometry (MS) has been increasingly used in the environmental sciences with the objective of investigating the presence of organic pollutants. MS has been widely coupled with chromatographic techniques, both gas chromatography (GC) and liquid chromatography (LC), because of their complementary nature when facing a broad range of organic pollutants of different polarity and volatility. A clear trend has been observed, from the very popular GC-MS with a single quadrupole mass analyser, to tandem mass spectrometry (MS-MS) and, more recently, high-resolution mass spectrometry (HRMS). For years GC has been coupled to HR magnetic sector instruments, mostly for dioxin analysis, although in the last ten years there has been growing interest in HRMS with time-of-flight (TOF) and Orbitrap mass analyzers, especially in LC-MS analysis. The increasing interest in the use of HRMS in the environmental sciences is because of its suitability for both targeted and untargeted analysis, owing to its sensitivity in full-scan acquisition mode and high mass accuracy. With the same instrument one can perform a variety of tasks: pre- and post-target analysis, retrospective analysis, discovery of metabolite and transformation products, and non-target analysis. All these functions are relevant to the environmental sciences, in which the analyst encounters thousands of different organic contaminants. Thus, wide-scope screening of environmental samples is one of the main applications of HRMS. This paper is a critical review of current use of HRMS in the environmental sciences. Needless to say, it is not the intention of the authors to summarise all contributions of HRMS in this field, as in classic descriptive reviews, but to give an overview of the main characteristics of HRMS, its strong potential in environmental mass spectrometry and the trends observed over the last few years. Most of the literature has been acquired since 2005, coinciding with the growth and popularity of HRMS in this field, with a few exceptions that deserve to be mentioned because of their relevance.


CD4⁺ CD45RA⁻ FoxP3high activated regulatory T cells are functionally impaired and related to residual insulin-secreting capacity in patients with type 1 diabetes.
Accumulating lines of evidence have suggested that regulatory T cells (T(regs)) play a central role in T cell-mediated immune response and the development of type 1A and fulminant type 1 diabetes. CD4(+) forkhead box protein 3 (FoxP3)(+) T cells are composed of three phenotypically and functionally distinct subpopulations; CD45RA(+) FoxP3(low) resting T(regs) (r-T(regs)), CD45RA(-) FoxP3(high) activated T(regs) (a-T(regs)) and CD45RA(-) FoxP3(low) non-suppressive T cells (non-T(regs)). We aimed to clarify the frequency of these three subpopulations in CD4(+) FoxP3(+) T cells and the function of a-T(regs) with reference to subtypes of type 1 diabetes. We examined 20 patients with type 1A diabetes, 15 patients with fulminant type 1 diabetes, 20 patients with type 2 diabetes and 30 healthy control subjects. A flow cytometric analysis in the peripheral blood was performed for the frequency analysis. The suppressive function of a-T(regs) was assessed by their ability to suppress the proliferation of responder cells in a 1/2:1 co-culture. A flow cytometric analysis in the peripheral blood demonstrated that the frequency of a-T(regs) was significantly higher in type 1A diabetes, but not in fulminant type 1 diabetes, than the controls. Further, the proportion of a-T(regs) among CD4(+) FoxP3(+) T cells was significantly higher in patients with type 1A diabetes with detectable C-peptide but not in patients with type 1A diabetes without it and with fulminant type 1 diabetes. A proliferation suppression assay showed that a-T(regs) were functionally impaired both in fulminant type 1 diabetes and in type 1A diabetes. In conclusion, a-T(regs) were functionally impaired, related to residual insulin-secreting capacity and may be associated with the development of type 1 diabetes.


Effects of bioirrigation of non-biting midges (Diptera: Chironomidae) on lake sediment respiration.
Bioirrigation or the transport of fluids into the sediment matrix due to the activities of organisms such as bloodworms (larvae of Diptera, Chironomidae), has substantial impacts on sediment respiration in lakes. However, previous quantifications of bioirrigation impacts of Chironomidae have been limited by technical challenges such as the difficulty to separate faunal and bacterial respiration. This paper describes a novel method based on the bioreactive tracer resazurin for measuring respiration in-situ in non-sealed systems with constant oxygen supply. Applying this new method in microcosm experiments revealed that bioirrigation enhanced sediment respiration by up to 2.5 times. The new method is yielding lower oxygen consumption than previously reported, as it is only sensitive to aerobic heterotrophous respiration and not to other processes causing oxygen decrease. Hence it decouples the quantification of respiration of animals and inorganic oxygen consumption from microbe respiration in sediment.


Breast cancer survivorship and sexual dysfunction: a population-based cohort study.
Breast cancer is the most common non-skin cancer in women and an increasing number of people are living as breast cancer survivors. While the prognosis of breast cancer continues to improve, the rates of sexual dysfunction and the risk related to cancer treatments have not been well characterized in a population-based study. We identified a cohort of 19,709 breast cancer survivors diagnosed between 1997 and 2017 from the Utah Cancer Registry, and 93,389 cancer-free women who were matched by age and birth state from the Utah Population Database. Sexual dysfunction diagnoses were identified through ICD-9 and ICD-10 codes from electronic medical records and statewide healthcare facilities data. Cox proportional hazard models were used to estimate hazard ratios for risk of sexual dysfunction. Breast cancer survivors were at higher risk of sexual dysfunction diagnosis (9.1% versus 6.9%, HR 1.60, 95% CI 1.51-1.70) compared to the general population. This risk increased 2.05-fold within 1 to 5 years after cancer diagnosis (95% CI 1.89-2.22) and 3.05-fold in individuals diagnosed with cancer at < 50 years of age (95% CI 2.65-3.51). Cancer treatments including endocrine therapy, chemotherapy and radiation therapy were associated with an increased risk of sexual dysfunction among breast cancer survivors. Risk of sexual dysfunction in breast cancer survivors is higher than in the general population, but may be underdiagnosed in the clinical setting. Health care professionals should be encouraged to address the topic of sexual health early on in the treatment of breast cancer, and routinely screen patients for symptoms of sexual dysfunction.


Survey of endogenous virus and TVB* receptor status of commercial chicken stocks supplying specific-pathogen-free eggs.
Endogenous avian leukosis virus (ALVE) and the ALVE receptor (TVB*S1) status of six commercial chicken lines supplying specific-pathogen-free eggs were analyzed. All commercial chicken lines are certified free of the avian leukosis virus (ALV) by screening for expression of the p27 protein using the standard enzyme-linked immunosorbent assay. The commercial chicken lines A, E, and F contained replication competent ALVE inserts. Line A was fixed for ALVE21, and lines E and F were segregating for ALVE10. In addition, ALVE1 was detected in all the chicken lines. Chicken lines B, D, and F were essentially fixed for the TVB*S1 allele that confers susceptibility to ALVE, whereas lines A, C, B, and E were resistant, containing either the TVB*S3 or TVB*R alleles. The results show that lines selected to be ALV p27 negative give rise to two different genotypes. One genotype lacks the TVB*S1 receptor for ALVE. Chicken lines with the TVB*S1 negative genotype can retain replication competent endogenous virus inserts such as ALVE2, 10, or 21 and still display the p27 negative phenotype. These replication competent ALVE viruses are phenotypically p27 negative in the absence of the TVB*S1 receptor because their chromosomal integration sites restrict transcription and subsequent production of the p27 protein and virus particles to levels below the detection limit. If the TVB*S1 receptor is present, the limited production of ALVE virus particles reinfects and integrates into more productive chromosomal locations in the cell. Increased production of infective virus particles and detectable levels of p27 follow this reinfection and integration into more active regions of the cells genome. The other genotype observed in the commercial lines retains the ALVE receptor (TVB*S1) but either lacks replication competent inserts or expresses the envelope encoded protein from defective inserts such as ALVE3 or ALVE6. In this phenotype, the env-coded glycoprotein encoded by the defective inserts binds to the TVB*S1 receptor and blocks the reinfection of the replication competent ALVE virus. This receptor interference stops reinfection and subsequent production of detectable virus particles and the p27 protein. Mixtures of different p27 negative phenotypes can result in the p27 positive phenotype and ALVE virus production. For example, mixtures of ALVE receptor positive (TVB*S1) but ALVE negative (p27 negative and envelope negative) chick embryo fibroblasts (CEFs) with fibroblasts that are receptor negative but ALVE positive could generate cells expressing high levels of p27 and ALVE virus. In this situation, the undetectable levels of ALVE virus from the receptor negative CEFs would infect and integrate into the receptor positive CEFs and produce detectable levels of ALVE virus. The implications of these findings for vaccine manufacturers and regulatory agencies are discussed.


The role of an integrated back stability program in patients with chronic low back pain.
A controlled clinical trial. To assess the effects of an integrated back stability (IBS) programme on a chronic low back pain (CLBP) population in a time restricted private clinic environment. Studies assessing stability training CLBP have reported inconsistent results. Methods used within trials vary, with some authors focusing on muscle isolation and others using whole body movements. IBS uses an exercise progression beginning with posturally based exercise and progressing from muscle isolation through to complex movements. Fifty-nine chronic low back patients were divided into control (n=32) and intervention (n=27) groups. Participants in the intervention group were prescribed a 6 week individualized exercise programme in three stages. In stage 1, exercises addressed posture and movement dysfunction and activated the core stabilizing muscles. In stage 2, 'back fitness' was enhanced using progressive exercise principles. Stage 3 emphasized technique specific actions. Participants in the control group received a backcare advice leaflet only. Pre- and post-test scores were analysed for each of the outcome measures within the control group using a Wilcoxin signed ranks test. At an alpha level of p<or=0.0071, no differences were observed. For the intervention group, a Mann-Whitney U-test showed significant differences between groups in the Roland and Morris Disability Questionaire (RMDQ), short form McGill Pain Questionnaire (SF-MPQ), and the Tampa Scale of Kinesiophobia (TSK) (p<or=0.0071). Patient satisfaction was assessed by questionnaire, 89% of patients considering their level of pain and functional impairment acceptable following the programme. IBS significantly reduced pain and disability in the subject group studied. Patients reported a positive experience of the programme.


Causes of Death and Pathological Findings in Stranded Harbour Porpoises (Phocoena phocoena) from Swedish Waters.
Harbour porpoises (Phocoena phocoena) are useful indicators of the health of their wild populations and marine ecosystems, yet their elusive nature makes studying them in their natural environment challenging. Stranded porpoises provide an excellent source of data to study the health and biology of these animals and identify causes of death, diseases and other threats. The aim of this study was to document pathology, and where possible, cause of death in porpoises from Swedish waters. Post-mortem examinations were performed on 128 stranded porpoises collected from 2006 to 2020. Overall, bycatch including definitive and probable cases was the most common cause of death (31.4%), followed by disease (21.3%), predominantly pneumonia. In adults, infectious disease was the most common cause of death. Bacteria with zoonotic potential such as Erysipelothrix rhusiopathiae and Brucella sp. were documented for the first time in porpoises from Swedish waters, as was the porpoise-adapted group B Salmonella enterica ST416/ST417. Three of four deaths from non-infectious diseases involved parturition complications. Four cases of suspected predation were documented, but further analyses are required to confirm these findings. Our results are consistent with those from other regions in Europe and serve as a reference for future monitoring for changing patterns of health and disease of porpoises and their environments.


MOF-derived CeO2/Co3O4-Fe2O3@CC nanocomposites as highly sensitive electrochemical sensor for bisphenol A detection.
A novel CeO2/Co3O4-Fe2O3@CC electrode derived from CeCo-MOFs was developed for detecting the endocrine disruptor bisphenol A (BPA). Firstly, bimetallic CeCo-MOFs were prepared by hydrothermal method, and obtained material was calcined to form metal oxides after doping Fe element. The results suggested that hydrophilic carbon cloth (CC) modified with CeO2/Co3O4-Fe2O3 had good conductivity and high electrocatalytic activity. By the analyses of cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), the introduction of Fe increased the current response and conductivity of the sensor, greatly increasing the effective active area of the electrode. Significantly, electrochemical test proves that the prepared CeO2/Co3O4-Fe2O3@CC had excellent electrochemical response to BPA with a low detection limit of 8.7 nM, an excellent sensitivity of 20.489 μA/μM·cm2, a linear range of 0.5-30 μM, and strong selectivity. In addition, the CeO2/Co3O4-Fe2O3@CC sensor had a high recovery rate for the detection of BPA in real tap water, lake water, soil eluent, seawater, and PET bottle samples, which showed its potential in practical applications. To sum up, the CeO2/Co3O4-Fe2O3@CC sensor prepared in this work had excellent sensing performance, good stability and selectivity for BPA, which can be well used for the detection of BPA.


Nailfold videocapillaroscopy in patients with systemic lupus erythematosus.
Nailfold videocapillaroscopy was performed in 21 controls (C) and 21 patients (P) with systemic lupus erythematosus (SLE) classified according to the American College of Rheumatology, with, at least, 1 year of diagnosed disease and having low activity (MEX-SLEDAI) and sequel (SLICC) indexes at the time of the examination, paired by sex and age. Red blood cell velocity (RBCV, mm/s) at rest and after the release of 60s arterial occlusion (RBCVmax, mm/s), time to reach it (TRBCVmax, s), functional capillary density (FCD, number of capillaries /mm2), afferent, apical and efferent capillary diameters (microm) (DAF, DAP and DEF, respectively) were obtained from videotapes analyzed by the CapImage software. The results did not show any significant difference between the groups that were analyzed, suggesting that morphological (capillary diameters) and functional (RBCV, RBCVmax, TRBCVmax and FCD) parameters are not affected by SLE when low activity and sequel indexes of the disease are present.


Characterization and prognostic implication of delayed complete response in AL amyloidosis.
Little is known on continued response following completion of therapy in light chain (AL) amyloidosis. We studied 373 AL amyloidosis patients who achieved complete response (CR) or very good partial response (VGPR) to first-line therapy. By end of therapy (EOT), 46% of patients achieved a CR and 54% a VGPR. With no further therapy, 17.5% of patients were upstaged from VGPR to CR (delayed CR), with a median of 9 months. Compared with CR and VGPR at EOT, patients with a delayed CR were characterized by higher proportion of t(11;14) and lower rate of trisomies. Autologous stem cell transplant was more frequent in the delayed CR group. Patients with a delayed CR were characterized by minimal residual disease negativity and organ response rates similar to patients with CR at EOT and higher than patients achieving VGPR at EOT. Patients with a delayed CR had a longer PFS/OS compared to patients with CR or VGPR by EOT (median PFS 149 vs 92 vs 52 months, P < .001; 10-year OS 87% vs 71% vs 56%, P < .001). This study characterizes delayed CR in AL amyloidosis, highlights its prognostic impact which is at least similar to those who achieved CR at EOT, and underlines another aspect of response monitoring.


Tailoring the pH dependence of human non-pancreatic secretory phospholipase A2 by engineering surface charges.
Human non-pancreatic secretory phospholipase A2 (hnpsPLA2) catalyzes the sn-2 acyl hydrolysis of phospholipids. It was reported that hnpsPLA2 is involved in various diseases like inflammation, cancer, and so on. This enzyme also exhibits anti-bacterial and anti-virus activities. It is active over a broad pH range, with higher activity at alkaline conditions. In order to make it suitable as a potential bactericide, high activity at neutral pH is preferable. We have tried to tailor the pH dependence of hnpsPLA2 activity by replacing its surface charged residues. Three surface charge replacements, Arg42Glu, Arg100Glu, and Glu89Lys, showed increased activities at neutral pH, which are 2.3, 2.8, and 2.3 times that of the wild-type enzyme at pH 7. Both the positive-to-negative and negative-to-positive mutations lowered the optimum enzymatic reaction pH of hnpsPLA2, indicating that the enzyme pH profile depends on a delicate balance of charged residues. The activity changes are in good agreement with the recently proposed calcium-coordinated catalytic triad mechanism. This study also provides a general means of enhancing hnpsPLA2 activity at low pH.


Role of Bcl-2 family members in invertebrates.
Proteins belonging to the Bcl-2 family function as regulators of 'life-or-death' decisions in response to various intrinsic and extrinsic stimuli. In mammals, cell death is controlled by pro- and anti-apoptotic members of the Bcl-2 family, which function upstream of the caspase cascade. Structural and functional homologues of the Bcl-2 family proteins also exist in lower eukaryotes, such as nematodes and flies. In nematodes, an anti-apoptotic Bcl-2 family protein, CED-9, functions as a potent cell death inhibitor, and a BH3-only protein, EGL-1, acts as an inhibitor of CED-9 to facilitate the spatio-temporal regulation of programmed cell death. On the other hand, the Drosophila genome encodes two Bcl-2 family proteins, Drob-1/Debcl/dBorg-1/dBok and Buffy/dBorg-2, both of which structurally belong to the pro-apoptotic group, despite abundant similarities in the cell death mechanisms between flies and vertebrates. Drob-1 acts as a pro-apoptotic factor in vitro and in vivo, and Buffy/dBorg-2 exhibits a weak anti-apoptotic function. The ancestral role of the Bcl-2 family protein may be pro-apoptotic, and the evolution of the functions of this family of proteins may be closely linked with the contribution of mitochondria to the cell death pathway.


Spatiotemporal Proteomic Analysis of Stress Granule Disassembly Using APEX Reveals Regulation by SUMOylation and Links to ALS Pathogenesis.
Stress granules (SGs) are cytoplasmic assemblies of proteins and non-translating mRNAs. Whereas much has been learned about SG formation, a major gap remains in understanding the compositional changes SGs undergo during normal disassembly and under disease conditions. Here, we address this gap by proteomic dissection of the SG temporal disassembly sequence using multi-bait APEX proximity proteomics. We discover 109 novel SG proteins and characterize distinct SG substructures. We reveal dozens of disassembly-engaged proteins (DEPs), some of which play functional roles in SG disassembly, including small ubiquitin-like modifier (SUMO) conjugating enzymes. We further demonstrate that SUMOylation regulates SG disassembly and SG formation. Parallel proteomics with amyotrophic lateral sclerosis (ALS)-associated C9ORF72 dipeptides uncovered attenuated DEP recruitment during SG disassembly and impaired SUMOylation. Accordingly, SUMO activity ameliorated C9ORF72-ALS-related neurodegeneration in Drosophila. By dissecting the SG spatiotemporal proteomic landscape, we provide an in-depth resource for future work on SG function and reveal basic and disease-relevant mechanisms of SG disassembly.


Augmented nuclease activity during cellular senescence in vitro.
The molecular correlates of the limited proliferative potential of normal human diploid fibroblasts and extensive single-strand breaks in the genomic DNA of these cells were examined by transfection analyses in which DNA replication could be uncoupled from DNA damage and repair. Both supercoiled (fmI), and restriction endonuclease-cleaved, linear (fmIII) molecules of a well-defined bacterial plasmid DNA, pBR322, were transfected into, and subsequently recovered from, early and late passage fibroblasts. Southern blot analysis revealed that fmI DNA was converted by random nicks into fmII DNA slightly more rapidly in late passage cells compared with cells at early passage. Similarly, fmII and fmIII DNAs also sustained multiple random nicks and no appreciable net religation of free ends of fmIII DNA could be detected at either passage. In addition, the efficiency of in vitro ligation of fmIII DNA recovered from late passage cells was also reduced, compared with that from early passage cells, as determined by Southern blotting. These data suggest that in the absence of DNA replication, a putative nuclease activity may contribute to DNA damage observed in senescent cells, which, in turn, may be causally related to their limited replicative potential.


Ultrasound monitoring of testis and prostate maturation in hypogonadotropic hypogonadic males during gonadotropin-releasing hormone treatment.
The effects of the administration of gonadotropin-releasing hormone (GnRH) on the increase of testis and prostate volume was monitored by ultrasound in six patients affected by idiopathic hypogonadotropic hypogonadism. A significant increase of testis volume was observed after 90 and 180 days (6.65 versus 3.32 mL, 99.1% net increase and 8.47 mL, 176.8% increase, respectively) of pulsatile GnRH treatment. A similar increase of prostate volume was observed at day 90 (12.67 versus 7.78 mL, 70.3% net increase) and day 180 (14.70 mL, 97.7% increase). The ultrasound monitoring of the modifications of testis and prostate volume may represent a biological assay of the effects of GnRH treatment and offer additional data on the response of target organs to the hormonal treatment.


The effect of different exercise programs on sarcopenia criteria in older people: A systematic review of systematic reviews with meta-analysis.
To compare the results of different modalities of physical exercises on the sarcopenia diagnostic criteria in older people. Systematic review of systematic reviews. Search strategy included older people and sarcopenia MeSh, performed at mainly databases. Selected studies include older adults, submitted to physical training (Intervention Group: IG) compared to control groups (CG). Quantitative analyses with the inverse variance statistic method (random effects). The effect measures mean difference. Heterogeneity measured with Q-Test. 494 systematic reviews found. After screening, 5 were included (48 papers. n=3,877). Mean age: 74.02±6.1. 73.44% female. Mean interventions duration: 17.38 weeks (average: 2.56 weekly sessions). AMSTAR and PRISMA showed high methodological quality. Meta-analyses compared results of resistance training interventions (RTA) with other than resistance (Non-resistance Training interventions: NRTA). Handgrip strength, skeletal muscle mass (SMM) and gait speed showed statistically significant differences (SSD) favorable to IG. In chair stand test, RTA showed SSD favorable to the IG, and NRTA to CG. The timed-up-and-go do not showed SSD. The SMM and strength showed better results in RTA, although the confidence intervals (CI) overlap. Both valences can be trained with similar volume and training intensity, which can modify muscle volume and strength. Physical performance obtained better results in NRTA, even with CI overlap. For severely sarcopenic, training including more than one valence may be best. In sarcopenia diagnosis most studies do not take into account the consensuses of standardization, making hard the larger groups analysis. Being part of any training program can be beneficial for sarcopenia in older people, with resistance training better for strength and SMM, and mixed modalities for physical performance.


Correction: Fast Tac Metabolizers at Risk-It is Time for a C/D Ratio Calculation. J. Clin. Med. 2019, 8, 587.
The authors wish to make the following corrections to this paper [...].


Investigating sources of inefficiency in residential mental health facilities.
Data Envelopment Analysis (DEA) was used to measure efficiency of residential mental health facilities. The sample consisted of 50 half-way houses, 8 nursing homes, and 32 sheltered homes. In total, 68 facilities belonged to the public sector and were 22 supervised by private non-profit organizations. Variables chosen to characterize production were: structure size (m2), staff, salaries and operational costs, and the output measure was patient numbers. An input oriented DEA model, allowing for variable returns to scale, was applied and units were ranked according to a benchmarking approach. Mean efficiency, for the whole sample, was 73.2% and 18 best practice units were found, on average, 33.1% over-efficient. The other 72 were under-performing, with 54 appearing more than 20% inefficient. The mean efficiency scores for public and private non-profit units were 68.8 and 86.6%, respectively, and significantly different (p < 0.001). Results suggest that efficiency improvements are possible with better use of resources but more research employing various data sets is required.


Amyloid beta-protein precursor accumulates in dystrophic neurites of senile plaques in Alzheimer-type dementia.
We raised two rabbit antisera against synthetic peptides corresponding to the carboxyl- and amino-terminal regions of the predicted amyloid beta-protein precursor (APP). Both antisera recognized the same 106-135 kDa proteins of human brain extract by immunoblot analysis. Immunocytochemical studies showed that these antisera both reacted with the same dystrophic neurites within the senile plaques of Alzheimer brains. These results indicated that APP accumulated in the dystrophic neurites of the senile plaques.


[PTEN and NBS1 gene mutations in familial breast cancer and early-onset breast cancer from Hunan Province in China].
To investigate the profile and potential significance of PTEN and NBS1 mutations among patients with familial or at early onset breast cancer in Hunan province. A total of 131 breast cancer patients with familial history or suffered from breast cancer at the age of less than 35 years old were included in this study. A comprehensive phosphatase and tensin homolog (PTEN) and nibrin (NBS1) mutation analysis was performed through denaturing high performance liquid chromatography (DHPLC) and subsequent DNA direct sequencing. Among 131 patients, a reported mutation IVS4+109insTCTTA in PTEN gene were identified in two patients. The mutation frequency of IVS4+109insTCTTA was 1.15%. Two mutations in PTEN gene, 225 A>C (Thr 160 Pro) and IVS5+13T>C, was firstly discovered. Another reported missense mutation was rs121909229 G>A (Arg 130 Gln). Three mutations were detected in NBS1 gene, of which IVS6+43A>G and IVS6+127A>G were firstly discovered and another reported synonymous mutations was rs1805794 G>C (Glu 185 Gln). The novel mutations in PTEN and NBS1 might be specific to the familial and early-onset breast cancer of Chinese Hunan population.


Nodular secondary syphilis mimicking cutaneous lymphoreticular process.
We hereby report a case presenting a rare form of secondary syphilis with an unusual nodular eruption accompanied by pruritus and generalized lymphadenopathy, bearing a striking resemblance to lymphoma. The histologic picture was also misleading and was compatible with lymphocytoma cutis. The diagnosis was eventually made after results of serologic tests for syphilis were found to be positive, with an extremely high titer of VDRL, and dark-field microscopy had demonstrated spirochetes in a nodular lesion. Secondary syphilis should thus be considered in the differential diagnosis of nodular lesions resembling lymphoreticular disease. It is also suggested that syphilis be added to the list of diagnoses belonging to the category of pseudolymphoma.


Experiences with the Milan Automated Neurobehavioral System (MANS) in occupational neurotoxic exposure.
It is known that in neurobehavioral toxicology, the data obtained by means of psychological testing can acquire sense only when testing conditions and procedures, among others, are kept under rigorous control. This demand, together with the diffusion of multicentered studies aimed at finding mutual agreement on the parameters to be measured and the interpretation criteria to be adopted, has determined our decision to convert six out of the seven tests of the WHO-NCTB into a computer-aided system (MANS). The choice of one of the two systems depends on the characteristics of the testing situation and on the different requirements of clinical, epidemiological and experimental studies. The validation of MANS, as to its reliability and sensitivity, is ongoing but some data have already been collected which show a high correlation of the computer-administered test results with the paper and pencil form as well as a good degree of discriminating power in different testing situations. The experience, made in the past three years, seems to show that the goal of having a simple, easy to handle and reliable instrument to be used and, if necessary easily adapted to a variety of situations and cultures, has been met. Further work is to be carried out to confirm its validity in the early detection of neurobehavioral impairments due to neurotoxic agents.


Effect of amiloride on sodium and water reabsorption in the rabbit gall-bladder.
The effects of the Na+-channel-blocking diuretic agent amiloride were assessed in the rabbit gall-bladder epithelium, a low-resistance epithelium with an isosmotic, coupled NaCl transport mechanism. Amiloride caused a rapid, reversible, and dose-dependent decrease in fluid absorption when applied from the mucosal side in concentrations between 8.8 X 10(-5) and 1.76 X 10(-3) M. These concentrations were without effect from the serosal side, suggesting an action of amiloride in the luminal cell membrane as in high-resistance epithelia. Amiloride did not affect the epithelial resistance or the passive serosa-to-mucosa Na+ flux, while net Na+ and water reabsorption were inhibited in parallel. Thus, amiloride did not affect the paracellular tight junction pathway, but inhibited a transcellular, coupled salt and water transport mechanism. The kinetics of the amiloride effect were of a Michaelis-Menten type. The dose of amiloride giving 50% inhibition of fluid absorption (ID50) was 4 X 10(-4) M, a value about three orders of magnitude higher than in high-resistance, Na+-retaining epithelia. The percentage inhibitory effect at each concentration of amiloride increased with increasing rate of spontaneous (control) fluid transport, reaching maximal responses fitting a Michaelis-Menten kinetic with an ID50 of 1.5 X 10(-4) M. No effects of changing the extracellular Na+ concentration between 51 and 145 mequiv/l on the maximal inhibitory effect of amiloride on Na+ and water reabsorption were observed. This suggests a non-competitive type of action of amiloride on a Na+-dependent isosmotic fluid transport mechanism. Removal of mucosal Ca2+ did not alter the effect of amiloride. The implications of these findings are discussed in relation to concepts concerning the mechanism of isosmotic salt and water transport. The data are compatible with the concept that amiloride interferes with a Na+-dependent formation and transcellular transport of isosmotic fluid volumes in a sequestered compartment in the epithelial cells.


Neurobehavioral profiles in individuals with hyperimmunoglobulin E Syndrome (HIES) and brain white matter hyperintensities.
Individuals with hyperimmunoglobulin E Syndrome (HIES) have central nervous system abnormalities, including focal white matter hyperintensities (WMH), or unidentified bright objects. This cross-sectional study aimed to describe the cognitive and emotional functioning and quality of life of people with HIES. We also sought to explore the relationship between cognitive functioning and WMHs in this population. Twenty-nine individuals (13 males) with autosomal-dominant HIES (mean age = 35.1 years, range 16-55) were administered a comprehensive psychological assessment as part of a natural history protocol. The assessment included measures of global cognitive functioning (Wechsler Adult Intelligence Scale-III), memory (California Verbal Learning Test-II, Wechsler Memory Scale-III), executive skills (Delis Kaplan Executive Function System), and attention (Test of Everyday Attention). Emotional symptoms and quality of life also were assessed. All mean cognitive scores were within normal limits. Mean scores on memory and executive functioning measures were significantly lower than Full Scale IQ scores (ps < .05). Substantial percentages of patients self-reported executive skills to be in the clinical range. Patients with fewer (1-20) versus more (21+) WMHs scored significantly better on measures of global cognitive skills, visual-perceptual skills, and working memory. Mean scores on emotional symptom and quality of life measures were in the average range and unrelated to WMHs. Global cognitive functioning was average to high average in our sample of individuals with HIES. However, focal brain lesions were associated with lower scores in specific domains. Emotional functioning and quality of life are within normal limits in this sample.


[Advantages of cryosurgical treatment of malignant skin transformations in the region of the nose and auricle (author's transl)].
Since the beginning of 1974 70 basal cell carcinomas of the face were treated by cryosurgery at the ENT clinic in Göttingen. The cryosurgical removal of basalomas on a cartilagenous base (nose, auricle) avoids hospitalization of the patient and relieves the surgeon of a time-consuming plastic operation. Scar formation after cryonecrosis is, in general, excellent and cosmetically unobtrusive. All patients must undergo strict follow-up examinations since for the surgeon there is no histologic safeguarding by an "excision into healthy tissue".


Intracellular labeling of T-cells with superparamagnetic contrast agents.
Isolated rat T-cells have been labeled intracellularly, using endocytosis uptake of two superparamagnetic contrast agents, AquaMag100 and BMS180549, which are both iron-oxide particles coated with dextran. No deterioration of cell proliferation response to mitogen stimulation was observed after labeling with either superparamagnetic contrast agent. AquaMag100 particles show aggregation and co-precipitation in culture media for T-cells. BMS180549 particles not only produce no observable aggregation or co-precipitation, but also have a higher efficiency for labeling T-cells than AquaMag100. The efficiency of cell labeling was determined by measuring the decrease in the spin-spin relaxation time of the water proton in cell samples containing 1 x 10(7) labeled T-cells/milliliter of 2% w/w gelatin. After optimization of the labeling procedures, a shortening of the spin-spin relaxation time by a factor of approximately 7 to 10 has been demonstrated. Under the present experimental conditions, the up-regulation of low density lipoprotein receptor does not increase the labeling efficiency by endocytosis. Our results suggest that intracellular labeling of specific cell types can be achieved with good efficiency and the labeled cells can be detected by magnetic resonance imaging in rat testicles in vivo.


Calorimetric effects accompanying ion adsorption at the charged metal oxide/electrolyte interfaces: effects of the correlations between the binding-to-surface energies of various surface complexes.
As has been shown in our earlier publications, a theoretical description of ion adsorption at the real, energetically heterogeneous oxide/electrolyte interfaces, involves necessary assumption about the correlations between the adsorption energies of the various surface complexes formed at this interface. So far, only two extreme models have been considered in such theoretical descriptions: one model assuming that high correlations exist, and the other assuming that practically no correlations exist. The purpose of this paper is to develop yet another description based on a model assuming that some partial correlations exist between the adsorption energies of the various surface complexes. The other purpose of this publication is to test these three models by analysing some experimental data reported so far in literature. Such data sets must include necessary information about the enthalpic effects accompanying ion adsorption. This is because enthalpic effects are known to be more sensitive to the mechanistic models underlying a theoretical description of adsorption systems. With such requirements in mind, we have selected three adsorption systems: Al2O3/NaCl, TiO2/NaCl and silica/NaCl, for our analysis. Our theoretical-numerical analysis of the behaviour of these adsorption systems suggests that either none or partial correlations exist between the adsorption energies of the various surface complexes. However, that analysis also shows, that the present accuracy of the experimental data does not allow us to draw more elaborate conclusions.


Psychometric features of Neuropathic Pain Symptom Inventory in Iranian people with Parkinson's disease.
Neuropathic pain is a type of pain reported in people with Parkinson's disease. There are various scales to evaluate the characteristics of this kind of pain. The purpose of this study was to investigate the psychometric properties of the Neuropathic Pain Symptom Inventory (NPSI), a specific scale that measures neuropathic pain in Iranian people with Parkinson's disease. Four hundred forty-seven individuals with Parkinson's disease were recruited in the study. Acceptability, internal consistency (Cronbach's alpha), and test-retest reliability (intraclass correlation coefficient, ICC) of NPSI were calculated. Dimensionality was examined through exploratory factor analysis. For convergent validity, correlations of NPSI with Douleur Neuropathic 4, Brief Pain Inventory, King's Pain Parkinson disease Scale, and Visual Analog Scale-Pain were used. Discriminative validity and sensitivity to change between On- and Off- medication states were analyzed. A marked floor effect was observed for this scale (64.2%). Cronbach's alpha and ICC were 0.90 and 0.87, respectively. Items of NPSI were placed in 4 factors. A moderate to the strong association (rs = 0.55 to 0.85) between NPSI and other scales was obtained. The results of discriminative validity and sensitivity to change indicate the ability of NPSI to show differences between medication states. The results of this study suggest that NPSI has acceptable reliability, validity, and sensitivity to change, indicating that this scale is suitable for measuring neuropathic pain in Iranian people with Parkinson's disease.


Upregulation of water channel aquaporin-4 in experimental autoimmune encephalomyeritis.
Aquaporin-4 (AQP4) is a water channel protein that plays an important role in water movement in the central nervous system (CNS). Recently, presence of anti-AQP4 antibody has been reported in the sera from patients with neuromyelitis optica. AQP4 is therefore a possible target for inflammatory mechanisms in CNS. In the present investigation, we performed semi-quantitative analysis of AQP4-mRNA in brain and spinal cord from mice affected with experimental autoimmune encephalomyelitis (EAE) using real-time PCR. AQP4-mRNA expression was increased in EAE; reaching a peak in the spinal cord at 14 days, and in the brain at 21 days after first inoculation. Immunohistochemical analysis showed that AQP4 is expressed on astrocytes, indicating that the increase in AQP4 expression may correlate with astrocytic activation. This is the first study to demonstrate upregulation of AQP4 in EAE. The upregulation of AQP4 could be involved in the development of inflammation in the acute phase of EAE.


The carboxyl terminal mutational hotspot of the ciliary disease protein RPGRORF15 (retinitis pigmentosa GTPase regulator) is glutamylated in vivo.
Mutations inRPGR(ORF15)(retinitis pigmentosa GTPase regulator) are a major cause of inherited retinal degenerative diseases. RPGR(ORF15)(1152 residues) is a ciliary protein involved in regulating the composition and function of photoreceptor cilia. The mutational hotspot in RPGR(ORF15)is an unusual C-terminal domain encoded by exon ORF15, which is rich in polyglutamates and glycine residues (Glu-Gly domain) followed by a short stretch of basic amino acid residues (RPGR(C2)domain; residues 1072-1152). However, the properties of the ORF15-encoded domain and its involvement in the pathogenesis of the disease are unclear. Here we show that RPGR(ORF15)is glutamylated at the C-terminus, as determined by binding to GT335, which recognizes glutamylated substrates. This reactivity is lost in two mouse mutants ofRpgr, which do not express RPGR(ORF15)due to disease-causing mutations in exon ORF15. Our results indicate that RPGR(ORF15)is posttranslationally glutamylated in the Glu-Gly domain and that the GT335 antibody predominantly recognizes RPGR(ORF15)in photoreceptor cilia.


[Dental feats 1. An implant-supported orbital prosthesis after ablative tumor surgery].
Extraoral implants are used with increasing frequency, in order to provide retention for facial prostheses. Facial prostheses can replace lost extraoral hard and soft tissues using acrylic or silicone materials. The surgeon, maxillofacial prosthodontist and dental technician are facing particular treatment considerations and practical problems. Some of these are demonstrated and discussed in this article by means of a case-report. A patient received an orbital prosthesis which obtains its retention from implants, placed in the lateral orbital rim.


A comparison of the inhibitory effects of disodium pyrophosphate and disodium ethane-hydroxy-1, 1-diphosphonate on simple calcergy.
Both disodium pyrophosphate and disodium ethane-hydroxy-1, 1-diphosphate inhibit the reaction of simple calcergy induced in the mouse by lead acetate. The pyrophosphate is effective only by a direct interaction with the lead salt and the disphosphonate is effective only before the occurrence of histochemically demonstrable calcification.


Homoeologous gene silencing in tissue cultured wheat callus.
In contrast to diploids, most polyploid plant species, which include the hexaploid bread wheat, possess an additional layer of epigenetic complexity. Several studies have demonstrated that polyploids are affected by homoeologous gene silencing, a process in which sub-genomic genomic copies are selectively transcriptionally inactivated. This form of silencing can be tissue specific and may be linked to developmental or stress responses. Evidence was sought as to whether the frequency of homoeologous silencing in in vitro cultured wheat callus differ from that in differentiated organs, given that disorganized cells are associated with a globally lower level of DNA methylation. Using a reverse transcription PCR (RT-PCR) single strand conformation polymorphism (SSCP) platform to detect the pattern of expression of 20 homoeologous sets of single-copy genes known to be affected by this form of silencing in the root and/or leaf, we observed no silencing in any of the wheat callus tissue tested. Our results suggest that much of the homoeologous silencing observed in differentiated tissues is probably under epigenetic control, rather than being linked to genomic instability arising from allopolyploidization. This study reinforces the notion of plasticity in the wheat epi-genome.


Isolation of rat immunoglobulin class switch variants of rat-mouse hybridomas by enzyme-linked immunosorbent assay and sequential sublining.
The use of sequential sublining in combination with highly specific and sensitive enzyme-linked immunosorbent assays for the isolation of spontaneous rat Ig heavy chain class switch variants is described. These methods allowed us to isolate switch variants from mouse-rat hybridoma lines secreting monoclonal rat antibodies. Switch variants from IgM to IgG2a, from IgG2a or IgG2b to IgE and from IgE to IgA were obtained. Members of the BA1.2 family, which consists of IgG2b, IgE and IgA antibodies are shown to exhibit identical rhamnose-inhibitable binding to the O18A antigen of Escherichia coli and to the paratope-associated anti-idiotypic antibody BA114.


Structure, pharmacology and function of GABA-A receptors in cochlear outer hair cells.
There is evidence that the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) is released from some efferent olivocochlear nerve endings terminating at outer hair cells (OHCs). Using monoclonal antibodies against postsynaptic GABAA receptor from bovine cerebral cortex we confirm the presence of GABA and benzodiazepine bindings sites of alpha- and beta-subunits of GABAA receptors at the basal pole of isolated OHCs. Whole-cell recording with viable OHCs revealed that the application of 10(-3)-10(-8) M GABA to the cell surface was followed by a concentration-dependent hyperpolarization of the outer cell membrane. Hyperpolarization was increased in the presence of 2.5 x 10(-5) M chlorazepate, a benzodiazepine derivative. Electrophysiological effects caused by GABA alone or in combination with chlorazepate were specifically inhibited by 10(-6) M of the GABA-receptor antagonist picrotoxin. Moreover, 10(-5)-10(-7) M GABA caused reversible slow elongation of the cylindrical hair cell body in OHCs examined. These neurotransmitter-induced motile responses were specifically blocked by 10(-4) M picrotoxin. The results suggest that a subpopulation of OHCs express alpha- and beta-subunits of GABAA receptors which both form a GABA/benzodiazepine-receptor complex at the basal pole of isolated OHCs. These receptors are thought to allow GABA which is released from efferent auditory nerve terminals to bind to the cell surface of OHCs, resulting in GABAA-receptor activation. This probably gates a GABAA-receptor-associated chloride channel in the postsynaptic OHC membrane, allowing hyperpolarization and elongation of the cell.


The identification and characterisation of a new deltamethrin resistance-associated gene, UBL40, in the diamondback moth, Plutella xylostella (L.).
Differential expression of ubiquitin was previously reported between Plutella xylostella strains that are resistant or susceptible to the pesticide deltamethrin (DM). This finding hinted at the potential involvement of ubiquitin in deltamethrin resistance, a theory that demanded further testing. Real-time PCR analyses revealed that one of the ubiquitin genes, UBL40, was overexpressed in the deltamethrin-resistant strain during the fourth instar. To investigate the functional relationship between this gene and deltamethrin resistance, RNA interference (RNAi) and cell transfection were utilised. UBL40 knockdown was observed to significantly reduce the level of resistance in RNAi-treated larvae after 48 h. Conversely, overexpression of UBL40 in Drosophila Kc cells conferred a degree of protection against deltamethrin. These results represent the first evidence that UBL40 plays a role in the regulation of deltamethrin resistance in P. xylostella.


Global warming not so harmful for all plants - response of holomycotrophic orchid species for the future climate change.
Current and expected changes in global climate are major threat for biological diversity affecting individuals, communities and ecosystems. However, there is no general trend in the plants response to the climate change. The aim of present study was to evaluate impact of the future climate changes on the distribution of holomycotrophic orchid species using ecological niche modeling approach. Three different scenarios of future climate changes were tested to obtain the most comprehensive insight in the possible habitat loss of 16 holomycotrophic orchids. The extinction of Cephalanthera austiniae was predicted in all analyses. The coverage of suitable niches of Pogoniopsis schenckii will decrease to 1-30% of its current extent. The reduction of at least 50% of climatic niche of Erythrorchis cassythoides and Limodorum abortivum will be observed. In turn, the coverage of suitable niches of Hexalectris spicata, Uleiorchis ulaei and Wullschlaegelia calcarata may be even 16-74 times larger than in the present time. The conducted niche modeling and analysis of the similarity of their climatic tolerance showed instead that the future modification of the coverage of their suitable niches will not be unified and the future climate changes may be not so harmful for holomycotrophic orchids as expected.


Differential tyrosine phosphorylation of leukemic cells during apoptosis as a result of treatment with imatinib mesylate.
Bcr-Abl fusion tyrosine kinase contributes to leukemic transformation. Imatinib mesylate inhibits Bcr-Abl tyrosine kinase, resulting in a blockage of tyrosine phosphorylation in its downstream pathways. We analyzed the alteration of tyrosine phosphorylation, on BCR/ABL+ chronic myelogenous leukemia cells, after treatment with imatinib mesylate. Data were collected using a two-dimensional gel electrophoresis followed by Western blot and mass spectrometry. The inhibition of Bcr-Abl tyrosine kinase by 2.5 microM imatinib mesylate caused both cell cycle arrest in the G0/G1 phase and increased the portion of apoptotic cells. As a result, the population of leukemic cells decreased by 30% and 70% compared to controls at 24 and 72 h, respectively. Furthermore, treatment with imatinib mesylate altered tyrosine phosphorylation of 24 protein spots as the incubation time proceeded from 0 to 24 and 72 h. Ten of the 24 protein spots are visible at all three times. Four are detectable at both the 0 and 24 h points in time. Eight were detectable only at time 0.


On the hepatotoxicity of 1,1,2,2-tetrachloroethane.
Intoxication of male and female mice with a single dose (300 or 600 mg/kg) of 1,1,2,2-tetrachloroethane (TTCE) resulted in significant decreases in cytochrome P-450 (to 58-73% of the control) and NADPH-cytochrome (P-450) c-reductase (to 29-35% of the control) in hepatic microsomes. This was accompanied by an alteration of mixed function monooxygenases stemming from the marked reduction (to 20-64% of the control) of several oxidative activities to selected substrates towards different P-450 isozymes (classes IA1, IA2, IIB1, IIE1 and IIIA). As phase II markers, epoxide hydrolase (approximately 35% loss), UDP-glucuronosyl transferase (approximately 42% loss) and to a lesser extent glutathione S-transferase (approximately 17% loss) were all affected. Also, the activity of delta-aminolevulinic (ALA) synthetase was decreased (approximately 57% of the control). On the contrary, heme oxygenase activity was increased (up to 35%) at the maximal dose tested. The decrease of P-450-function may be explained in terms of an alteration in the rate of heme biosynthesis and degradation, provoking a loss of heme content (approximately 33%) as well as of the direct inactivation of both P-450 and reductase. Because of increasing evidence on the involvement of free radical intermediates in the case of toxicity of haloalkanes, electron spin resonance spectroscopy (ESR) spin-trapping in vivo techniques were used to characterize the possible free radical species involved in the observed liver damage. The results obtained with the spin-trap N-benzylidene-2-methylpropylamine N-oxide (phenyl t-butylnitrone, PBN) provide evidence for the formation and trapping of the CHCl2CHCl free radicals. The detection of conjugated diene signals by means of second-derivative spectrophotometry, have enabled us to show that in vivo lipid peroxidation may be one of the main mechanisms responsible for TTCE hepatotoxicity.


Apparent relationships between toxins elaborated by the cyanobacterium Trichodesmium erythraeum and those present in the flesh of the narrow-barred Spanish mackerel Scomberomorus commersoni.
The marine cyanobacterium Trichodesmium erythraeum contains toxic water-soluble material that produces signs in mice similar to those produced by water-soluble extracts of the flesh of a specimen of pelagic fish Scomberomorus commersoni from a batch that had been implicated in a poisoning resembling ciguatera. Extracts of water-soluble material from both the cyanobacterium and the fish contained toxins that were chromatographically indistinguishable. A peptide and an alkaloid were detected in partially purified extracts of the water-soluble material. In addition to this material toxic lipid-soluble material was present in some batches of T. erythraeum. Elution of this material with 9:1 chloroform:methanol using column chromatography produced material that was chromatographically indistinguishable from ciguatoxin-like material from S. commersoni and produced signs in mice similar to those produced by this material. Elution of the lipid-soluble material with 97:3 chloroform:methanol yielded a toxin resembling in its chromatographic and toxic properties a scaritoxin-like substance from S. commersoni. Other toxins with Rf values lying between that of the ciguatoxin-like material and that of the scaritoxin-like material were also detected in extracts of T. erythraeum. It is postulated that T. erythraeum is the progenitor of major toxins carried by some ciguateric fish and that water-soluble toxins released into the ambient sea water by T. erythraeum may constitute a health hazard for humans.