Quantitative evaluation for blood-retinal barrier breakdown in experimental retinal vein occlusion produced by photodynamic thrombosis using a new photosensitizer.
Purpose. To establish a rat model of retinal vein occlusion (RVO), we applied photodynamic thrombosis using a new photosensitizer. By measuring the breakdown of the blood-retinal barrier (BRB), we evaluated the model quantitatively. We also investigated how hypertension and retinal pigment epithelium (RPE) influence the breakdown of BRB after RVO. Methods. We modified a slit lamp biomicroscope for photodynamic thrombosis. The light source was changed from white light to argon laser, which made it possible to perform fluorescein angiography (FAG) simultaneously during photodynamic thrombosis. We irradiated with a continuous diode laser to occlude three retinal veins in a rat after PAD-S31 injection. The breakdown of BRB was quantitated by measuring extravasated Evans blue dye in albino and pigmented rats. We compared hypertensive rats (SHR) to normotensive rats (WKY) and sodium iodate-treated rats to normal rats. Results. High photosensitivity of PAD-S31 made it possible to occlude any retinal veins within 120 seconds at a low dose of 10 mg/kg without retinal thermal burn at the occlusion site. Simultaneous FAG enabled us to observe the formation of thrombus during diode laser irradiation. Our measured value of intraretinal Evans blue correlated with the range of serous retinal detachment. Both albino and pigmented rats demonstrated stable and constant values of Evans blue. SHR recovered from the breakdown of BRB after venous occlusion more slowly than WKY. Sodium iodate-treated rats had smaller breakdowns of BRB and recovered earlier than normal rats. Conclusions. In this study, we established the stable and constant rat model of RVO efficiently by using a new photosensitizer. Our simultaneous FAG method was considered to have an advantage of several potential clinical applications. Our rat model of RVO allows us to study factors associated with the recovery from damage by RVO.


A 55-Year-Old Japanese Man with Multiple Sclerosis Diagnosed with Disseminated Tuberculosis Identified by Liver Function Abnormalities: A Case Report.
BACKGROUND Reactivation of latent tuberculosis infection (LTBI) is a recognized complication of immunosuppressive treatment. However, immunosuppressed patients are also at risk of hematogenous disseminated spread from a primary infection with Mycobacterium tuberculosis. This report presents the case of a 55-year-old Japanese man with a 12-year history of multiple sclerosis who was hospitalized with worsening neurological symptoms and was diagnosed with disseminated tuberculosis identified by abnormalities on liver function test results. CASE REPORT A 55-year-old Japanese man was admitted to our hospital for the treatment of multiple sclerosis with worsening symptoms. He showed mild liver dysfunction at the time of admission. A laparoscopy and biopsy were performed to identify the cause of the liver dysfunction, which was the only positive finding. The liver surface was studded with yellowish-white nodular lesions. Histological examination of a liver biopsy specimen revealed a granuloma without caseous necrosis. The patient was suspected of having tuberculosis. Although the results of an interferon-γ-releasing assay were indeterminate, asymptomatic disseminated tuberculosis was diagnosed from the serum adenosine deaminase levels, a caseating granuloma in the cervical lymph node, detection of acid-fast bacilli DNA in the cervical lymph nodes on polymerase chain reaction, and tuberculin skin test findings. Anti-tuberculosis treatment led to improvement in the liver function test findings. CONCLUSIONS This case has highlighted that tuberculosis may have an atypical presentation in the immunosuppressed patient. In addition to the reactivation of LTBI, hematogenous spread of primary tuberculosis may result in disseminated disease involving multiple organs and requiring emergency treatment.


Knowledge of Human Monkeypox Infection among Final Year Medical, Pharmacy, and Nursing Students: A Multicenter, Cross-Sectional Analysis from Pakistan.
The World Health Organization (WHO) declared the monkeypox outbreak a public health emergency in June 2022. In Pakistan, positive cases of monkeypox were reported in April 2023. Healthcare workers (HCWs) are considered as a front-line force to combat such outbreaks. A questionnaire-based cross-sectional study was conducted among 11 public sector educational institutions in Punjab, Pakistan, during May and June 2023 among final year medical, pharmacy, and nursing students concerning their knowledge of monkeypox. This included the signs/symptoms of monkeypox. Healthcare students were chosen as they are the HCWs of tomorrow. A total of 389 healthcare students participated in the study, with a mean age of 23.17 ± 1.72 years, and the majority were female. The mean knowledge score was 17.69 ± 4.55 (95% CI 17.24-18.14) out of a maximum total knowledge score of 26 (each correct answer was given a score of 1). The proportion of students with good, moderate, and poor knowledge was 21.6%, 43.2%, and 35.2%, respectively. Age (p = 0.017), gender (p < 0.001), and education (p < 0.001) had a significant impact on the knowledge score. In the multivariate linear regression model, education was the only significant factor linked to knowledge scores. Overall, the majority of future HCWs had moderate knowledge of monkeypox. Consequently, educational activities are needed to improve monkeypox-related knowledge among future HCWs. Furthermore, emerging infectious diseases should be routinely incorporated into HCW curricula.


Asteroids® and Electrocardiograms: Proof of Concept of a Simulation for Task-Switching Training.
Emergency physicians are interrupted during patient care with such tasks as reading electrocardiograms (ECGs). This phenomenon is known as task-switching which may be a teachable skill. Our objective was to evaluate the potential of a video game for simulating the cognitive demands required of task-switching. Emergency medicine residents took a pretest on ECG interpretation and then a posttest while attending to a video game, Asteroids®. The 35 residents (63%) who participated, scored worse on the ECG posttest then they did on the pretest (p<.001; effect size=1.14). There were no differences between genders or training level. Interpreting ECGs while playing the Asteroids® game significantly lowered ECG interpretation scores. This shows the potential of this activity for training residents in task-switching ability. The next phase of research will test whether ECG reading performance while task-switching improves with practice.


High-Statistics Measurement of Neutrino Quasielasticlike Scattering at 6 GeV on a Hydrocarbon Target.
We measure neutrino charged-current quasielasticlike scattering on hydrocarbon at high statistics using the wideband Neutrinos at the Main Injector beam with neutrino energy peaked at 6 GeV. The double-differential cross section is reported in terms of muon longitudinal (p_{∥}) and transverse (p_{⊥}) momentum. Cross section contours versus lepton momentum components are approximately described by a conventional generator-based simulation, however, discrepancies are observed for transverse momenta above 0.5 GeV/c for longitudinal momentum ranges 3-5 and 9-20 GeV/c. The single differential cross section versus momentum transfer squared (dσ/dQ_{QE}^{2}) is measured over a four-decade range of Q^{2} that extends to 10 GeV^{2}. The cross section turnover and falloff in the Q^{2} range 0.3-10 GeV^{2} is not fully reproduced by generator predictions that rely on dipole form factors. Our measurement probes the axial-vector content of the hadronic current and complements the electromagnetic form factor data obtained using electron-nucleon elastic scattering. These results help oscillation experiments because they probe the importance of various correlations and final-state interaction effects within the nucleus, which have different effects on the visible energy in detectors.


Economic Status and Mortality in Patients with Alzheimer's Disease in Japan: The Longevity Improvement and Fair Evidence Study.
As more countries are implementing measures to address Alzheimer's disease (AD), it is essential to update the available knowledge on the relationship between economic status and mortality in patients with AD. This study examined the influence of economic status on mortality in Japanese individuals with AD using a medical claims dataset. This was a retrospective cohort study. Medical claims data from April 2014 to March 2019 were obtained from 13 local cities participating in the Longevity Improvement and Fair Evidence study. The inclusion criteria were patients aged 65 years and older who were newly diagnosed with AD during the study period. The outcome was death during the follow-up period. We assessed economic status by household income (middle to high income and low income); data were obtained from the use of the Medical Expenditure Ceiling Application and Standard Copayment Reduction Card (fee reduction card) when receiving an AD diagnosis, as an indicator of low-income status. We performed multivariate Cox proportional hazards analyses to examine the relationship between economic status and mortality; the model was adjusted for age, sex, the Charlson comorbidity index, and antidementia drug use. We identified 39,081 newly diagnosed patients with AD from the Longevity Improvement and Fair Evidence study database (mean age, 83.6 years; female, 67.1%). Of these, 3189 individuals were identified as having a low-income status. After adjusting for possible confounders, low-income status was associated with mortality (hazard ratio, 1.95; 95% confidence interval, 1.84-2.07). Low-income status was associated with substantially poorer prognoses in new AD cases, indicating a need for a thorough examination of medical and nursing care services utilized by low-income individuals with AD and to explore improvement strategies.


Granulocytic sarcoma with expression of CD30.
A case of a young man with a spinal epidural tumour, initially diagnosed as large cell anaplastic malignant lymphoma, is reported. The tumour consisted of poorly differentiated cells showing immunoreactivity with antibodies directed against CD30 and CD45. Ten months later the patient developed acute myeloid leukaemia. The histological slides of the epidural tumour were reviewed, including additional enzymochemical and immunochemical stains. As the tumour showed immunoreactivity for myeloperoxidase and chloroacetate esterase, it was reclassified as a granulocytic sarcoma.


Structural and conformational requisites in DNA quadruplex groove binding: another piece to the puzzle.
The study of DNA G-quadruplex stabilizers has enjoyed a great momentum in the late years due to their application as anticancer agents. The recognition of the grooves of these structural motifs is expected to result in a higher degree of selectivity over other DNA structures. Therefore, to achieve an enhanced knowledge on the structural and conformational requisites for quadruplex groove recognition, distamycin A, the only compound for which a pure groove binding has been proven, has been chemically modified. Surprisingly, structural and thermodynamic studies revealed that the absence of Coulombic interactions results in an unprecedented binding position in which both the groove and the 3' end of the DNA are occupied. This further contribution adds another piece to the so far elusive puzzle of the recognition between ligands and DNA quadruplexes and will serve as a platform for a rational design of new groove binders.


A matter of font type: The effect of serifs on the evaluation of scientific abstracts.
Text-based communication is one of the substantial ways of spreading scientific information. While the content and contextual aspects of written words have been widely researched, the impact of font characteristics on text perception is an almost blank page. The following study deals with the influence of serifs on the evaluation of online-presented scientific abstracts. Yet there is only evidence for faster reading times when texts are presented in sans-serif fonts, although the opposite is stated in parts of the literature. The present work examines if the presence or absence of serifs also have an impact on the appraisal of scientific texts when all other important font characteristics do not change. For this purpose, 188 university students participated in an online experiment and rated different aspects of scientific abstracts as well as of the research outlined in the abstracts. The results show that missing serifs led to increased reading speed. However, and in contrast to the perceptual fluency hypothesis, the presence of serifs had a positive effect on all evaluation dimensions. The results of a second study with 187 participants also indicated that reading fluency counteracted the liking of texts. Implications for future studies and media production are discussed.


The electrodiagnosis of the carpal tunnel syndrome.
The carpal tunnel syndrome is one of the most commonly discussed subjects among the public, as well as the medical community, these days. Even though this is a rather simple nerve compression, which can be easily corrected, the carpal tunnel syndrome can induce further complications, sometimes leading to total disability of the patient. Early, proper diagnosis and treatment is very vital. Unfortunately, the exact pathomechanism and proper diagnosis is yet to be seen. As electromyographers, the authors would like to re-visit this debatable carpal tunnel syndrome emphasizing the electrodiagnosis. Among the electrodiagnostic tests, comparison of distal motor or sensory latency of the median nerve to the ulnar nerve along with amplitude of the response, was found to be the most sensitive test. Also, the C6 cervical radiculopathy and the pronator syndrome can present similar clinical pictures and require a differentiation. After successful surgery many patients still complain of subjective residual symptoms. The electrophysiologic findings do not correspond to the clinical progress in many cases. The authors also experienced that most of the victims, following successful surgery, still would benefit from a proper exercise program including instruction for proper body mechanics before they return to their previous activities, to lessen the undesirable complications including the reflex sympathetic dystrophy.


The misuse of Kant in the debate about a market for human body parts.
Passages from the writings of Immanuel Kant concerning how a person should treat her body are often cited in the present-day debate about a market for human body parts. In this paper, I demonstrate that this has been a misuse of Kant because unlike those who cite him, Kant was not primarily concerned with prohibiting the sale of body parts. In the first section, I argue that once these particular passages are understood against the background of Kant's moral philosophy, they indicate he had much broader concerns relating to the correct moral relationship a rational person should have with her body. In the second section, I examine Stephen Munzer's unusually detailed analysis of these passages, but conclude that like those who have provided less detailed analyses, he also fails fully to understand the rationale for Kant's various prescriptions and prohibitions concerning the treatment of human body parts, and in doing so misrepresents Kant's position.


Toddler's Fractures: Time to Weight-bear With Regard to Immobilization Type and Radiographic Monitoring.
The toddler's fracture is a common pediatric nondisplaced spiral tibia fracture that is considered stable with a course of immobilization. However, there is no widely accepted type of immobilization, expected time to weight-bear, nor guidelines for radiographic monitoring. We aimed to compare immobilization type with respect to displacement and time to weight-bear, as well as determine the usefulness of follow-up radiographs. A 3-year retrospective chart review of all children aged 9 months to 4 years who had a lower leg radiograph was performed. Those who fulfilled the criteria of a nondisplaced spiral tibia fracture, without fibula or physeal injury, were included in data collection, as were subjects with a negative initial radiograph that were treated presumptively as a toddler's fracture. Subjects were compared with regard to clinical and radiographic presentation; initial and subsequent immobilization; and clinical and radiographic follow-up. There were 606 subjects with lower leg radiographs, with 192 meeting study criteria: 117 (61%) with an initially visible fracture and 75 (39%) without. Of the 75 without initially visible fractures, 70 (93%) had robust periosteal reaction on follow-up, and none were diagnosed as anything further. At final follow-up, 184 (96%) were known to be weight-bearing, with 98% of these by 4 weeks. There was an earlier return to weight-bear for those initially treated in a boot compared with short leg cast (2.5 vs. 2.8 wk, P=0.04), but there were no other differences between immobilization type. No fractures displaced at any time point, including 7 that had received no immobilization. Patients received an average of 2.5 two-radiograph series; no radiographs were noted to affect treatment decisions in follow-up. In our cohort, initial immobilization of a toddler's fracture in a boot may allow faster return to weight-bearing, but fractures were universally stable regardless of immobilization type, and nearly all regained weight-bearing by 4 weeks. This reliable healing suggests that immobilization type can be at the physician and family's discretion, and that radiographic follow-up may be unnecessary for treatment planning. Level III-this is a retrospective comparative study.


[The expression of rat brain constitutive nitric oxide synthase in NG108-15 cell].
Full length cDNA of rat brain cNOS was inserted into the polylinker area of pRC/CMV with specific orientation and an eukaryocyte expression vector pCMVcNOS was obtained. The existence of cNOS gene was demonstrated by PCR amplification, using pCMVcNOS gene as the model and primers designed in accord with the internal sequence of cNOS gene. The insertion and orientation of pCMVcNOS were further verified by enzymatic cleavage. NG108-15 cells were transfected with pCMVcNOS by calcium phosphate DNA coprecipitation and lipofectin transfection. G418 resistant monoclonal cells were selected with a culture medium containing 600 micrograms/ml G418. NOS activity of each clone was assayed by monitoring the conversion of 3H-Arginine to 3H-Citrulline. High expression cell lines were selected through measurement of the cytosol and particulate NOS activity. Out of 42 resistant monoclonal cell lines, 3 stable high expression clones have been finally selected. The increase of expressed cytosol NOS was more obvious. The result showed that the cell lines expressing cNOS at a high level had been obtained.


Biomechanical Risk Factors Associated with Running-Related Injuries: A Systematic Review.
Running is a popular form of physical activity with many health benefits. However, the incidence and prevalence of running-related injuries (RRIs) is high. Biomechanical factors may be related to the development of RRIs. This systematic review synthesizes biomechanical risk factors related to the development of RRIs in non-injured runners. PubMed, Web of Science, CINAHL, Embase, and SPORTDiscus were searched in July 2018 for original peer-reviewed prospective studies evaluating potential biomechanical factors associated with the development of RRIs. Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed. Two reviewers independently assessed articles for inclusion and methodological quality. Due to methodological heterogeneity across studies, a narrative synthesis of findings was conducted, rather than a meta-analysis. Sixteen studies, including 13 of high quality and three of moderate quality, were included. A large number of biomechanical variables were evaluated, producing inconsistent evidence overall. Limited evidence indicated greater peak hip adduction in female runners developing patellofemoral pain and iliotibial band syndrome, but not for a mixed-sex population of cross-country runners sustaining an RRI. The relationship between vertical loading rate and RRIs was inconsistent. Other kinematic, kinetic and spatiotemporal factors were only studied to a limited extent. Current prospective evidence relating biomechanical variables to RRI risk is sparse and inconsistent, with findings largely dependent on the population and injuries being studied. Future research is needed to confirm these biomechanical risk factors and determine whether modification of these variables may assist in running injury prevention and management.


Diffuse idiopathic pulmonary neuroendocrine cell hyperplasia: a systematic overview.
Diffuse idiopathic pulmonary neuroendocrine cell hyperplasia (DIPNECH) is being recognized with increasing frequency. Diagnostic and treatment information is limited. A systematic review is presented, focusing on patient demographics, clinical presentation, diagnosis, treatment options, and outcomes. A systematic electronic literature search was conducted for adult DIPNECH cases reported in the English literature during the past 6 years. Twenty-four DIPNECH cases were identified. Another case from our institution is contributed. Women represent 92% (23 of 25). Mean age at diagnosis was 58 years (range, 36-76 yr). Most were nonsmokers (16 of 24). Symptoms included cough (71%), dyspnea (63%), and wheezing (25%) occurring days to years before diagnosis. Pulmonary function testing showed obstructive ventilatory disease in 54%. Lung nodules were seen in 15 patients (63%), ground-glass attenuation in 7 patients (29%), and bronchiectasis in 5 patients (21%). Histological confirmation required surgical lung biopsy for 88%; however, transbronchial biopsies alone were diagnostic in three patients. Treatments strategies included systemic and inhaled corticosteroids, bronchodilators, and lung resection. Available follow-up data in 17 patients showed 6 clinically improved, 7 who remained stable, and 4 clinically deteriorated. The majority of patients presenting with DIPNECH are middle-aged females with symptoms of cough and dyspnea; obstructive abnormalities on pulmonary function testing; and radiographic imaging showing pulmonary nodules, ground-glass attenuation, and bronchiectasis. In general, the clinical course remains stable; however, progression to respiratory failure does occur. Long-term follow-up and treatment remains incomplete. Establishment of a national multicenter DIPNECH registry would allow formulation of optimal evidence-based guidelines for management of these patients.


Differential expression of LFA-1 molecules in non-Hodgkin's lymphoma and lymphoid leukemia.
We investigated the expression of adherence molecules lymphocyte function-associated antigens-1 alpha and -beta (LFA-1 alpha, -beta) and p150, 95 in 103 well-characterized non-Hodgkin's lymphomas (NHLs) and lymphoid leukemias (LLs). We found that NHLs and LLs differentially express LFA-1 molecules according to their lineage derivation, degree of clinical aggressiveness, and anatomic site of involvement. Specifically, (a) T-cell neoplasms nearly always express these molecules; (b) diffuse aggressive B-cell NHLs and mature LLs often lack LFA-1 alpha molecules; and (c) B-cell chronic lymphocytic leukemia (CLL) is often LFA-1 alpha-negative while B-cell small lymphocytic lymphomas (SLLs) are nearly always LFA-1 alpha-positive. Furthermore, the low expression of LFA-1 alpha in CLL is related to the low degree of homotypic lymphocyte adhesion after tumor promoter antigen stimulation that does not modulate the expression of LFA-1 alpha in vitro. The differential expression of LFA-1 by B-cell CLL and SLL and their degree of homotypic lymphocyte adhesion may account for the distinct anatomic compartmentalization and characteristic clinical behavior of these two morphologically and immunologically similar lymphoid malignancies.


Pathology of human organic mercury poisoning: Lessons from an autopsy case.
We had an opportunity to perform a general autopsy of a case with chronic organic mercury toxicosis in 2017. He had been engaged in synthesizing a variety of organic mercury compounds throughout the four years from 1966 and developed chronic organic mercury poisoning in 1969. Almost forty years on, he still remained to complain of persistent paresthesia at finger tips and tongue, and of narrowed visual field. Neurological examinations clarified a rise of two-point discrimination thresholds, a systemic increase of touch thresholds, constriction of the visual field caused by general visual depression, and sensorineural hearing loss while primary modalities of his somatic, visual, and auditory sensations were preserved. These symptoms and signs are characteristic of human organic mercury poisoning. Furthermore, he had difficulty in processing a lot of visual and auditory information at a time. His two-point discrimination thresholds and systemic elevation of touch thresholds were comparable to those of mild organic mercury poisoning cases. He had slight sensory ataxia, but not cerebellar ataxia. Brain [18F]-2-fluorodeoxyglucose positron emission tomography analysis exhibited marked hypometabolism at bilateral postcentral gyrus, striate cortex, and superior temporal gyrus, but not the cerebellum. Histopathological studies revealed considerable decrease of granular neurons and neuronal networks in bilateral primary somatosensory, visual, and auditory cortices. Those characteristic brain lesions fairly explain increase of thresholds of somatic, visual, and auditory sensations, and degradation of integrating sensory information. It is noted that damages to the peripheral nervous system and the cerebellum were not detected and that his intellectual faculties were preserved.


Enhanced BDNF signalling following chronic hypoxia potentiates catecholamine release from cultured rat adrenal chromaffin cells.
We investigated the role of the neurotrophin BDNF signalling via the TrkB receptor in rat adrenomedullary chromaffin cells (AMCs) exposed to normoxia (Nox; 21% O2) and chronic hypoxia (CHox; 2% O2) in vitro for ∼ 48 h. TrkB receptor expression was upregulated in primary AMCs and in immortalized chromaffin (MAH) cells exposed to CHox; this effect was absent in MAH cells deficient in the transcription factor, hypoxia inducible factor (HIF)-2α. Relative to normoxic controls, activation of the TrkB receptor in chronically hypoxic AMCs led to a marked increase in membrane excitability, intracellular [Ca(2+)], and catecholamine secretion. The BDNF-induced rise of intracellular [Ca(2+)] in CHox cells was sensitive to the selective T-type Ca(2+) channel blocker TTA-P2 and tetrodotoxin (TTX), suggesting key roles of low threshold T-type Ca(2+) and voltage-gated Na(+) channels in the signalling pathway. Environmental stressors, including chronic hypoxia, enhance the ability of adrenomedullary chromaffin cells (AMCs) to secrete catecholamines; however, the underlying molecular mechanisms remain unclear. Here, we investigated the role of brain-derived neurotrophic factor (BDNF) signalling in rat AMCs exposed to chronic hypoxia. In rat adrenal glands, BDNF and its tropomyosin-related kinase B (TrkB) receptor are highly expressed in the cortex and medulla, respectively. Exposure of AMCs to chronic hypoxia (2% O2; 48 h) in vitro caused a significant increase to TrkB mRNA expression. A similar increase was observed in an immortalized chromaffin cell line (MAH cells); however, it was absent in MAH cells deficient in the transcription factor HIF-2α. A specific TrkB agonist, 7,8-dihydroxyflavone (7,8-DHF), stimulated quantal catecholamine secretion from chronically hypoxic (CHox; 2% O2) AMCs to a greater extent than normoxic (Nox; 21% O2) controls. Activation of TrkB by BDNF or 7,8-DHF increased intracellular Ca(2+) ([Ca(2+)]i), an effect that was significantly larger in CHox cells. The 7,8-DHF-induced [Ca(2+)]i rise was sensitive to the tyrosine kinase inhibitor K252a and nickel (2 mm), but not the Ca(2+) store-depleting agent cyclopiazonic acid. Blockade of T-type calcium channels with TTA-P2 (1 μm) or voltage-gated Na(+) channels with TTX inhibited BDNF-induced [Ca(2+)]i increases. BDNF also induced a dose-dependent enhancement of action potential firing in CHox cells. These data demonstrate that during chronic hypoxia, enhancement of BDNF-TrkB signalling increases voltage-dependent Ca(2+) influx and catecholamine secretion in chromaffin cells, and that T-type Ca(2+) channels play a key role in the signalling pathway.


RNA sequencing of kidney distal tubule cells reveals multiple mediators of chronic aldosterone action.
The renal aldosterone-sensitive distal tubule (ASDT) is crucial for sodium reabsorption and blood pressure regulation. The ASDT consists of the late distal convoluted tubule (DCT2), connecting tubule (CNT), and collecting duct. Due to difficulties in isolating epithelial cells from the ASDT in large quantities, few transcriptome studies have been performed on this segment. Moreover, no studies exist on isolated DCT2 and CNT cells (excluding intercalated cells), and the role of aldosterone for regulating the transcriptome of these specific cell types is largely unknown. A mouse model expressing eGFP in DCT2/CNT/initial cortical collecting duct (iCCD) principal cells was exploited to facilitate the isolation of these cells in high number and purity. Combined with deep RNA sequencing technology, a comprehensive catalog of chronic aldosterone-regulated transcripts from enriched DCT2/CNT/iCCD principal cells was generated. There were 257 significantly downregulated and 290 upregulated transcripts in response to aldosterone ( P < 0.05). The RNA sequencing confirmed aldosterone regulation of well-described aldosterone targets including Sgk1 and Tsc22d3. Changes in selected transcripts such as S100a1 and Cldn4 were confirmed by RT-qPCR. The RNA sequencing showed downregulation of Nr3c2 encoding the mineralocorticoid receptor (MR), and cell line experiments showed a parallel decrease in MR protein. Furthermore, a large number of transcripts encoding transcription factors were downregulated. An extensive mRNA transcriptome reconstruction of an enriched CNT/iCCD principal cell population was also generated. The results provided a comprehensive database of aldosterone-regulated transcripts in the ASDT, allowing development of novel hypotheses for the action of aldosterone.


A Novel Tomato Fusarium Wilt Tolerance Gene.
The reduced mycorrhizal colonization (rmc) tomato mutant is unable to form mycorrhiza and is more susceptible to Fusarium wilt compared with its wild-type isogenic line 76R. The rmc mutant has a chromosomal deletion affecting five genes, one of which is similar to CYCLOPS. Loss of this gene is responsible for non-mycorrhizality in rmc but not enhanced Fusarium wilt susceptibility. Here, we describe assessment of a second gene in the rmc deletion, designated Solyc08g075770 that is expressed in roots. Sequence analyses show that Solyc08g075770 encodes a small transmembrane protein with putative phosphorylation and glycosylation sites. It is predicted to be localized in the plasma membrane and may function in transmembrane ion transport and/or as a cell surface receptor. Complementation and knock-out strategies were used to test its function. Some putative CRISPR/Cas-9 knock-out transgenic events exhibited Fusarium wilt susceptibility like rmc and some putative complementation lines were 76R-like, suggesting that the tomato Solyc08g075770 functions in Fusarium wilt tolerance. This is the first study to demonstrate that Solyc08g075770 is the contributor to the Tfw locus, conferring tolerance to Fusarium wilt in 76R which was lost in rmc.


Pain related-visual imagery is associated with distress in chronic pain sufferers.
Chronic pain patients often describe their pain in ways that suggest vivid mental images, with some reporting images relating to their pain. Despite these clinical observations, there are few studies describing the nature and consequences of these images. This study examined whether mental imagery of pain is associated with levels of reported distress, cognitions, disability or pain severity. In a postal survey, 83 adult chronic pain patients indicated whether or not they experienced mental images of their pain. They also completed standardized measures of depression, anxiety, cognitions, disability and pain severity. Those reporting images were compared to those not reporting images on their responses to the other variables. People with pain imagery reported significantly higher levels of anxiety, depression and catastrophizing than patients who did not report such images. No differences were found on measures of physical disability. Mental images of pain appear to be associated with higher levels of anxiety, depression and catastrophizing. It is possible that these images play a role in maintaining such difficulties. For these patients, imagery may provide a route via which clinicians can work with patients to help them reinterpret or respond more flexibly to their pain.


Interferon-gamma drives programmed death-ligand 1 expression on islet β cells to limit T cell function during autoimmune diabetes.
Type 1 diabetes is caused by autoreactive T cell-mediated β cell destruction. Even though co-inhibitory receptor programmed death-1 (PD-1) restrains autoimmunity, the expression and regulation of its cognate ligands on β cell remains unknown. Here, we interrogated β cell-intrinsic programmed death ligand-1 (PD-L1) expression in mouse and human islets. We measured a significant increase in the level of PD-L1 surface expression and the frequency of PD-L1+ β cells as non-obese diabetic (NOD) mice aged and developed diabetes. Increased β cell PD-L1 expression was dependent on T cell infiltration, as β cells from Rag1-deficient mice lacked PD-L1. Using Rag1-deficient NOD mouse islets, we determined that IFN-γ promotes β cell PD-L1 expression. We performed analogous experiments using human samples, and found a significant increase in β cell PD-L1 expression in type 1 diabetic samples compared to type 2 diabetic, autoantibody positive, and non-diabetic samples. Among type 1 diabetic samples, β cell PD-L1 expression correlated with insulitis. In vitro experiments with human islets from non-diabetic individuals showed that IFN-γ promoted β cell PD-L1 expression. These results suggest that insulin-producing β cells respond to pancreatic inflammation and IFN-γ production by upregulating PD-L1 expression to limit self-reactive T cells.


Understanding patient needs without understanding the patient: the need for complementary use of professional interpreters in end-of-life care.
High-quality doctor-patient communication in end-of-life care results in better quality of life for patients. In linguistically and culturally diverse societies, language discordant consultations become daily practice, leading to difficulties in eliciting patient preferences toward end-of-life care. Although family members invariably act as informal interpreters, this may cause some ethical dilemmas. We present a case of a palliative patient whose son acted as an interpreter. This case generated a triple- layered ethical dilemma: (i) how to safeguard patient autonomy against paternalistic interventions by family members, (ii) how to respect the relational context in which patient autonomy can be realized, and (iii) how to respect the ethno-cultural values of the patient and his family. These issues are being discussed and reflected upon within the framework shared decision making involving informal- and professional interpreters. The complementary use of professional interpreters next to family members acting as informal interpreters is recommended.


Nutrient and trace element dynamics in blended topsoils containing spent foundry sand and compost.
Spent foundry sand (SFS) could be used with compost to produce blended synthetic topsoil material but many environmental agencies restrict such use out of concern for leaching and plant uptake of contaminants in SFS. A 3 yr field experiment assessed the potential for contaminant leaching and plant uptake from natural soils and blended synthetic topsoils made with three SFSs and three composts. Plots were instrumented with lysimeters and planted with perennial ryegrass (Lolium perenne L.). Growth was better on blended soils than natural soils and tissue content of most nutrients and trace elements remained within normal ranges in plants grown on both blended and natural soils, though concentrations of N, P, K, S, B, Cu, Mo, and Zn were larger from some of the blended soils. Blended soils leached more P and K than natural soils and blended soil made with low C/N ratio compost leached more NH(4) than the natural soil. All blended soils leached more Cu, and some leached more As, Mo, and Zn than natural soils. With the exception of As in leachates from one blended soil no trace elements exceeded drinking water standards. None of the 13 polyaromatic hydrocarbons or 11 phenolics analyzed was detected in the leachates. We conclude use of SFS in blended soils presents low risk from trace metal or organic contaminants in SFS. Blended soils with SFS and compost perform well as topsoil substitutes and the greatest environmental risk is N and P leaching from use of low C/N ratio or immature composts.


Identification and replication of the interplay of four genetic high-risk variants for urinary bladder cancer.
Little is known whether genetic variants identified in genome-wide association studies interact to increase bladder cancer risk. Recently, we identified two- and three-variant combinations associated with a particular increase of bladder cancer risk in a urinary bladder cancer case-control series (Leibniz Research Centre for Working Environment and Human Factors at TU Dortmund (IfADo), 1501 cases, 1565 controls). In an independent case-control series (Nijmegen Bladder Cancer Study, NBCS, 1468 cases, 1720 controls) we confirmed these two- and three-variant combinations. Pooled analysis of the two studies as discovery group (IfADo-NBCS) resulted in sufficient statistical power to test up to four-variant combinations by a logistic regression approach. The New England and Spanish Bladder Cancer Studies (2080 cases and 2167 controls) were used as a replication series. Twelve previously identified risk variants were considered. The strongest four-variant combination was obtained in never smokers. The combination of rs1014971[AA] near apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3A (APOBEC3A) and chromobox homolog 6 (CBX6), solute carrier family 1s4 (urea transporter), member 1 (Kidd blood group) (SLC14A1) exon single nucleotide polymorphism (SNP) rs1058396[AG, GG], UDP glucuronosyltransferase 1 family, polypeptide A complex locus (UGT1A) intron SNP rs11892031[AA] and rs8102137[CC, CT] near cyclin E1 (CCNE1) resulted in an unadjusted odds ratio (OR) of 2.59 (95% CI = 1.93-3.47; P = 1.87 × 10-10), while the individual variant ORs ranged only between 1.11 and 1.30. The combination replicated in the New England and Spanish Bladder Cancer Studies (ORunadjusted = 1.60, 95% CI = 1.10-2.33; P = 0.013). The four-variant combination is relatively frequent, with 25% in never smoking cases and 11% in never smoking controls (total study group: 19% cases, 14% controls). In conclusion, we show that four high-risk variants can statistically interact to confer increased bladder cancer risk particularly in never smokers.


Effect of interphase and interpulse delay in high-frequency irreversible electroporation pulses on cell survival, membrane permeabilization and electrode material release.
To achieve high efficiency of electroporation and to minimize unwanted side effects, the electric field parameters must be optimized. Recently, it was suggested that biphasic high-frequency irreversible electroporation (H-FIRE) pulses reduce muscle contractions. However, it was also shown for sub-microsecond biphasic pulses that the opposite polarity phase of the pulse cancels the effect of the first phase if the interphase delay is short enough. We investigated the effect of interphase and interpulse delay (ranging from 0.5 to 10,000 µs) of 1 µs biphasic H-FIRE pulses on cell membrane permeabilization, on survival of four mammalian cell lines and determined metal release from aluminum, platinum and stainless steel electrodes. Biphasic H-FIRE pulses were compared to 8 × 100 µs monophasic pulses. We show that a longer interphase and interpulse delay results in lower cell survival, while the effects on cell membrane permeabilization are ambiguous. The cancellation effect was observed only for the survival of one cell line. Application of biphasic H-FIRE pulses results in lower metal release from electrodes but the interphase and interpulse delay does not have a large effect. The electrode material, however, importantly influences metal release - the lowest release was measured from platinum and the highest from aluminum electrodes.


The efficacy of wet-cupping in the treatment of tension and migraine headache.
Wet-cupping is an ancient medical technique still used in several contemporary societies, but little empirical study has been devoted to test its efficacy to treat tension and migraine headache. Using a pre-post research design, 70 patients with chronic tension or migraine headache were treated with wet-cupping. Three primary outcome measures were considered at the baseline and 3 months following treatment: headache severity, days of headache per month, and use of medication. Results suggest that, compared to the baseline, mean headache severity decreased by 66% following wet-cupping treatment. Treated patients also experienced the equivalent of 12.6 fewer days of headache per month. We conclude that wet-cupping leads to clinical relevant benefits for primary care patients with headache. Possible mechanisms of wet-cupping's efficacy, as well as directions for future research are discussed.


Pharmacokinetics, analgesic effect, and tolerability of a single preprocedural dose of intranasal fentanyl in patients undergoing drain removal after breast reduction or augmentation surgery: A prospective, randomized, double-blind, placebo-controlled study.
Although acetaminophen is used to reduce pain after breast reduction or augmentation surgery, pain during the removal of the surgical drains is typically not specifically treated. Intranasally administered fentanyl may be suitable for pain control during removal of drains. The reported therapeutic window of fentanyl is between 0.2 and 1.2 ng/mL. The aim of this study was to evaluate the analgesic effect, tolerability, and pharmacokinetics of a single preprocedural dose of intranasal fentanyl administered before removal of surgical drains in patients who had undergone breast reduction or augmentation surgery. This was a randomized, double-blind, prospective study in healthy women (American Society of Anesthesiologists physical status I or II) between the ages of 18 and 65 years who were scheduled to undergo removal of surgical drains 1 to 4 days after breast reduction or augmentation surgery. A single dose of fentanyl nasal spray 0.05 mg/0.1 mL or placebo (preserved normal saline) 0.1 mL was administered 10 minutes before removal of drains. Because drain removal is generally carried out without specific analgesia, no rescue medication was provided. Pain intensity was measured on a visual analog scale (VAS) from 0 = no pain at all to 100 = worst pain possible. Pain intensity was evaluated immediately before administration of study medication (t = 0), at the time of drain removal (t = 10), and at 15, 20, 25, 40, and 70 minutes after administration of study medication. Safety measures included oxygen saturation, respiratory rate, heart rate, and blood pressure. Local and systemic adverse events were elicited by direct questioning throughout the study. Blood samples for pharmacokinetic analysis were collected at baseline and at 5, 10, 15, 30, 60, and 120 minutes after administration of study medication. The population pharmacokinetic parameters of fentanyl were calculated according to a 1-compartment open model with an iterative 2-stage Bayesian fitting procedure. Thirty-six women were randomized to treatment, and 33 completed the study. Their mean (SD) age was 39.2 (13.0) years, and their mean weight was 68.9 (10.7) kg. Mean VAS scores at baseline were 14.8 (17.8) for the fentanyl group and 6.0 (9.7) for the placebo group (P = NS); at the time of drain removal, the corresponding VAS scores were 31.0 (20.6) and 33.8 (25.7) (P = NS). Analysis of a random-effects model with mean VAS scores as a function of time as the dependent variable indicated a significant difference in mean VAS scores between the fentanyl and placebo groups (P = 0.006). The overall incidence of adverse events was 39.4% (13/33). Among the 17 patients in the fentanyl group, 8 reported > or =1 adverse event; among the 16 patients in the placebo group, 9 reported > or =1 adverse event. A mean estimated C(max) of 0.184 (0.069) ng/mL was reached at 13.76 (3.56) minutes after administration of intranasal fentanyl. The mean measured C(max) was 0.22 (0.088) ng/mL. In these women who had undergone breast reduction or augmentation surgery, a single preprocedural dose of intranasal fentanyl was significantly more effective than placebo in reducing pain intensity over the hour after removal of surgical drains. However, there was no significant difference in pain intensity between fentanyl at the time of drain removal and placebo. Intranasal fentanyl was generally well tolerated. At the dose used (0.05 mg), plasma fentanyl concentrations were below the reported therapeutic window.


Assessing the ecological impact of banana farms on water quality using aquatic macroinvertebrate community composition.
In Costa Rica, considerable effort goes to conservation and protection of biodiversity, while at the same time agricultural pesticide use is among the highest in the world. Several protected areas, some being wetlands or marine reserves, are situated downstream large-scale banana farms, with an average of 57 pesticide applications per year. The banana industry is increasingly aware of the need to reduce their negative environmental impact, but few ecological field studies have been made to evaluate the efficiency of proposed mitigation strategies. This study compared the composition of benthic macroinvertebrate communities up- and downstream effluent water from banana farms in order to assess whether benthic invertebrate community structure can be used to detect environmental impact of banana farming, and thereby usable to assess improvements in management practises. Aquatic invertebrate samples were collected at 13 sites, using kick-net sampling, both up- and downstream banana farms in fast flowing streams in the Caribbean zone of Costa Rica. In total, 2888 invertebrate specimens were collected, belonging to 15 orders and 48 families or taxa. The change in community composition was analysed using multivariate statistics. Additionally, a biodiversity index and the Biological Monitoring Working Party (BMWP) score system was applied along with a number of community composition descriptors. Multivariate analyses indicated that surface waters immediately up- and downstream large-scale banana farms have different macroinvertebrate community compositions with the most evident differences being higher dominance by a single taxa and a much higher total abundance, mostly of that same taxon. Assessment of macroinvertebrate community composition thus appears to be a viable approach to detect negative impact from chemical-intensive agriculture and could become an effective means to monitor the efficacy of changes/proposed improvements in farming practises in Costa Rica and similar systems.


Accuracy and precision of five analog radioimmunoassays for free thyroxin compared.
We compared the precision of free thyroxin (FT4) measurements by kits involving analog RIA and the use of antibody-coated tubes (Becton Dickinson and Coat-A-Count), magnetic separation (Amerlex Magnetic and Corning Magic), or centrifugation of antibodies linked to solid beads (Amerlex). Results of kits with magnetic separation were the most reproducible. Amerlex, Amerlex Magnetic, and Becton Dickinson kits gave values comparing best with those obtained by direct equilibrium dialysis. Coat-A-Count and Corning Magic results differed significantly from dialysis values, both for patients' samples and kit standards. The kits had equal diagnostic efficiency in patients with suspected thyroid disease. On measurement of FT4 some patients were reclassified from "subclinical thyroid disease" to "overt disease." Most patients with triiodothyronine thyrotoxicosis had increased FT4. Several kit values were low for pregnant women and patients with nonthyroidal illness but the Amerlex and Amerlex Magnetic assays had fewer low results. The Amerlex Magnetic FT4 assay gave the best precision, agreement with the reference method, and diagnostic efficiency.


Outcome of robotic surgery for endometrial cancer as a function of patient age.
This study aimed to evaluate and compare robot-assisted surgical staging on clinical outcomes, including quality of life and survival, as a function of patient age. Evaluation and comparison of perioperative morbidity, survival, and postoperative quality of life after prospective accumulation of clinical information including outcome measures for patients with endometrial cancer during the first 5 years of a robotic program, based on the following 3 age categories: women older than 80 years, women between 70 and 80 years, and women younger than 70 years. All consecutive patients with endometrial cancer undergoing robotic surgery (n = 303) were included, with 197 women younger than 70 years, 75 women between 70 and 80 years, and 31 women older than 80 years. There were significantly more patients with advanced stage (stage II to IV in 17%, 34%, and 35%, P = 0.02) and grade 3 disease (26%, 43%, and 58%, P = 0.002) with increasing age. The perioperative data showed similar grade I or II complications (Clavien-Dindo classification) between the groups, but significantly more grade III and IV complications for women older than 80 years compared with women 80 years or younger (10% vs 1%, P = 0.004). The time needed to resume chore activities was significantly shorter for patients 70 years or older than patients younger than 70 years [8.9 (8.7) vs 18.8 (25.5) days, P = 0.048]. Overall, all patients irrespective of age were highly satisfied with the procedure. There was no difference between young and elderly patients for disease-free survival (P = 0.99). Patient's age did not influence minor postoperative morbidity or overall satisfaction after robotic assisted surgery for endometrial cancer. Elderly patients had more major postoperative morbidity but resumed activities quicker than younger patients.


Hip fractures in users of first- vs. second-generation bisphosphonates.
This study compared population hip fracture rates for women with a prior fragility fracture who were treated with first-generation versus second-generation bisphosphonate therapies. The observational study found that, relative to women treated with etidronate, a first-generation bisphosphonate, women treated with the second-generation therapies 'alendronate' or 'risedronate' were equally likely to be admitted to hospital for hip fracture. Our findings must be confirmed in large randomized head-to-head controlled trials. Few studies have examined hip fracture outcomes among users of first- versus second-generation bisphosphonates. We compared hip fracture rates among elderly women with a history of fracture dispensed first- and second-generation bisphosphonates, hypothesizing that hip fracture rates would be higher among users of first- versus second-generation bisphosphonates after adjusting for confounders. Administrative data from Ontario, Canada from 01 April 1998 to 31 March 2002 was used to identify population-based bisphosphonate-naïve cohorts of subjects age 66 years and older initiated on first- (etidronate plus calcium; n = 19,127) or second-generation (alendronate or risedronate; n = 1,460) bisphosphonates. Multivariate Cox proportional hazard models were used for analysis. During over 23,000 person-years of follow-up, we observed 293 hospital admissions for first hip fracture. The unadjusted event rates yielded approximately 12.5 hospital admissions for hip fracture per 1,000 person-years of follow-up in each study group. Relative to the etidronate plus calcium group, females in the alendronate or risedronate group were equally likely to be admitted for hip fracture (adjusted rate ratio [aRR] = 1.0; 95% CI 0.6-1.6). The findings of this study suggest similar rates of hip fracture between the first- and second-generation bisphosphonates when used continuously among elderly females with a prior history of fracture.


[Exploring the lived experiences of middle-aged males diagnosed with chronic kidney disease].
The majority of studies addressing chronic kidney disease (CKD) have been quantitative. The lived experiences of patients with early stage CKD have not been explored in the literature. Due to the lack of symptoms during the early stages of CKD, the progress of this disease is often ignored. This study explores the lived experiences of middle-aged males diagnosed with CKD. Research focuses on the work experiences, interpersonal relationships, and dietary habits of this group. This qualitative study used a narrative research method with purposive sampling. Ten middle-aged male patients were interviewed. We employed the 3 strategies of member checks, peer review, and debriefing in order to enhance the trustworthiness of the data analysis. Four major themes were identified: (1) keeping the same life pace as usual with concerns of deterioration; (2) continuing to support their family and play the role of father; (3) hoping to participate in family and friend relationships despite the illness; and (4) being challenged to maintain a healthy diet. Although the physical status of participants was similar to the general middle-aged adult population, participants were concerned regarding disease deterioration. The results of this study suggest that health professionals: (1) provide appropriate information based on patients' personality, particularly at the first onset or apparent deterioration of the disease; (2) offer specialized healthcare information to patients relevant to their career choices; and (3) understand CKD-patient difficulties in daily life such as eating out habits and challenges faced in maintaining a healthy diet. Results may provide important information to healthcare providers in education planning and implementation and support programs for patients and families.


Neural circuits for jumping in the locust.
The locust jump consists of three distinct phases: Cocking: a rapid flexion of both hindleg tibia and locking of both tibia in full flexion. Co-contraction: simultaneous contractions in hindleg flexor and extensor muscles lasting about 0.5 s resulting in the storage of energy for the jump in elastic elements of the legs and muscles. Triggering: a sudden inhibition of flexor activity to allow the shortening of the contracted extensors and the release of the energy stored during the co-contraction phase. The neural circuitry controlling these three phases is now reasonably well understood. Some of its major features are: (1) pairs of large identifiable interneurons in the thoracic ganglia for evoking the cocking response (C-neurons) and for triggering the jump (M-neurons), (2) a central excitatory pathway from extensor to flexor tibiae motoneurons to ensure simultaneous activation of extensor and flexor motoneurons during the initial part of the co-contraction phase, (3) a positive feedback pathway from cuticular receptors to extensor motoneurons for maintaining extensor activity during the co-contraction phase, (4) proprioceptive feedback to the trigger interneurons for increasing their excitability during the co-contraction phase and thereby allowing a variety of external stimuli to activate the trigger neurons and evoke a jump, (5) presynaptic inhibition of visual pathways to the trigger neurons to ensure that the trigger neurons are not activated by the simultaneous occurrence of visual and auditory stimuli in the absence of proprioceptive input, and (6) a pair of multifunctional visual movement detecting neurons which can initiate cocking or trigger the jump depending on the animal's state.


RNA-Binding Motif Protein 24 (RBM24) Is Involved in Pregenomic RNA Packaging by Mediating Interaction between Hepatitis B Virus Polymerase and the Epsilon Element.
Encapsidation of pregenomic RNA (pgRNA) is a crucial step in hepatitis B virus (HBV) replication. Binding by viral polymerase (Pol) to the epsilon stem-loop (ε) on the 5'-terminal region (TR) of pgRNA is required for pgRNA packaging. However, the detailed mechanism is not well understood. RNA-binding motif protein 24 (RBM24) inhibits core translation by binding to the 5'-TR of pgRNA. Here, we demonstrate that RBM24 is also involved in pgRNA packaging. RBM24 directly binds to the lower bulge of ε via RNA recognition submotifs (RNPs). RBM24 also interacts with Pol in an RNA-independent manner. The alanine-rich domain (ARD) of RBM24 and the reverse transcriptase (RT) domain of Pol are essential for binding between RBM24 and Pol. In addition, overexpression of RBM24 increases Pol-ε interaction, whereas RBM24 knockdown decreases the interaction. RBM24 was able to rescue binding between ε and mutant Pol lacking ε-binding activity, further showing that RBM24 mediates the interaction between Pol and ε by forming a Pol-RBM24-ε complex. Finally, RBM24 significantly promotes the packaging efficiency of pgRNA. In conclusion, RBM24 mediates Pol-ε interaction and formation of a Pol-RBM24-ε complex, which inhibits translation of pgRNA and results in pgRNA packing into capsids/virions for reverse transcription and DNA synthesis.IMPORTANCE Hepatitis B virus (HBV) is a ubiquitous human pathogen, and HBV infection is a major global health burden. Chronic HBV infection is associated with the development of liver diseases, including fulminant hepatitis, hepatic fibrosis, cirrhosis, and hepatocellular carcinoma. A currently approved vaccine can prevent HBV infection, and medications are able to reduce viral loads and prevent liver disease progression. However, current treatments rarely achieve a cure for chronic infection. Thus, it is important to gain insight into the mechanisms of HBV replication. In this study, we found that the host factor RBM24 is involved in pregenomic RNA (pgRNA) packaging and regulates HBV replication. These findings highlight a potential target for antiviral therapeutics of HBV infection.


ApoE and clusterin CSF levels influence associations between APOE genotype and changes in CSF tau, but not CSF Aβ42, levels in non-demented elderly.
Apolipoprotein E (APOE) ε4 genotype is associated with increased cerebral amyloid beta (Aβ) deposition in nondemented elderly and suggested to influence ApoE as well as ApoJ (clusterin [Clu]) and ApoA1 expression. We aimed to assess whether APOE affects early Alzheimer's disease pathophysiology via these apolipoproteins. Cerebrospinal fluid (CSF) ApoE, Clu, ApoA1, and CSF amyloid beta1-42 (Aβ42) and tau levels were assessed in 403 individuals with subjective cognitive decline and mild cognitive impairment using enzyme-linked immunosorbent assay. Whether CSF apolipoprotein levels mediated APOEε4 allele frequency effects on CSF Aβ42 and tau in nondemented elderly was investigated using mediation analysis, with age- and gender-adjusted linear regression analyses. CSF ApoE mediated 48% of the association between APOEε4 and CSF tau, whereas Clu and ApoA1 did not. In addition, CSF Clu partially mediated the relation between CSF ApoE and tau (12%). CSF apolipoproteins did not mediate the inverse relation between APOEε4 and CSF Aβ42, despite a strong association between the latter 2 biomarkers. In summary, our findings suggest that ApoE and Clu are involved in Aβ-independent pathways as part of the cascade leading to Alzheimer pathology.


The associative basis of cue-elicited drug taking in humans.
Drug cues play an important role in motivating human drug taking, lapse and relapse, but the psychological basis of this effect has not been fully specified. To clarify these mechanisms, the study measured the extent to which pictorial and conditioned tobacco cues enhanced smoking topography in an ad libitum smoking session simultaneously with cue effects on subjective craving, pleasure and anxiety. Both cue types increased the number of puffs consumed and craving, but pleasure and anxiety responses were dissociated across cue type. Moreover, cue effects on puff number correlated with effects on craving but not pleasure or anxiety. Finally, whereas overall puff number and craving declined across the two blocks of consumption, consistent with burgeoning satiety, cue enhancement of puff number and craving were both unaffected by satiety. Overall, the data suggest that cue-elicited drug taking in humans is mediated by an expectancy-based associative learning architecture, which paradoxically is autonomous of the current incentive value of the drug.


Sampling and analytical method development and hand wipe measurements of dermal exposures to polycyclic aromatic hydrocarbons.
This article describes the laboratory assessment of a hand and surface wipe sampling method for polycyclic aromatic hydrocarbons (PAHs). The analytical method employed extraction of the wipe samples into dimethyl sulfoxide (DMSO) and high-performance liquid chromatography (HPLC) flourometric detection of pyrene, a predominant PAH in used gasoline engine oils (UGEO). Recovery of pyrene was evaluated for two different sampling media by first contaminating the hands of a small number of volunteers with UGEO, followed by applying a small amount of corn oil to the palms, and by wiping the skin with a Whatman cellulostic filter paper or a polyester fabric wipe (i.e., Alpha wipes). In summary, using either Whatman or Alpha wipes, the mean recovery of pyrene from the UGEO that was applied to the hands and contained within three consecutive wipes was 69% and 54%, respectively. However, the relative recovery of the first to second wipe was on average 47% and 75% for the two media, respectively. These results indicate that the Alpha wipes were more efficient at recovering pyrene in the first wipe but less efficient overall when all three consecutive samples were included. Even though this sampling was performed in a controlled laboratory environment, the minimum and maximum amount of pyrene recovered in the individual composite samples using either method spanned a range of twofold. Overall, intra-and interpersonal variability, as measured by coefficient of variation, were 22% and 19%, respectively, and were not statistically different by type of media used. This method was used in a pilot field survey to sample the hands of 18 automotive repair technicians and 18 office workers. Detectable amounts of pyrene (>0.2 microg/sample) were found on the hands of 61% and 0% of these two groups, respectively, with the highest measured quantity equal to 1.06 microg. Samples from the upper surfaces of automobile motors were generally low to nondetectable (<0.027 microg/sample), while the median value of 0.047 mkcrlg/50 cm(2)(CV = 160%) and up to 0.640 microg were found on the drip pans.


Human cerebrospinal fluid affects chemoradiotherapy sensitivities in tumor cells from patients with glioblastoma.
Cancers in the central nervous system resist therapies effective in other cancers, possibly due to the unique biochemistry of the human brain microenvironment composed of cerebrospinal fluid (CSF). However, the impact of CSF on cancer cells and therapeutic efficacy is unknown. Here, we examined the effect of human CSF on glioblastoma (GBM) tumors from 25 patients. We found that CSF induces tumor cell plasticity and resistance to standard GBM treatments (temozolomide and irradiation). We identified nuclear protein 1 (NUPR1), a transcription factor hampering ferroptosis, as a mediator of therapeutic resistance in CSF. NUPR1 inhibition with a repurposed antipsychotic, trifluoperazine, enhanced the killing of GBM cells resistant to chemoradiation in CSF. The same chemo-effective doses of trifluoperazine were safe for human neurons and astrocytes derived from pluripotent stem cells. These findings reveal that chemoradiation efficacy decreases in human CSF and suggest that combining trifluoperazine with standard care may improve the survival of patients with GBM.


[A morphological study of primary pulmonary hypertension: an analysis of five autopsies].
Five autopsy cases of typical primary pulmonary hypertension are reported. The histopathological features were as follows: marked medial hypertrophy of muscular pulmonary arteries; muscularization of arterioles; intimal fibrosis and fibroelastosis of muscular pulmonary arteries accompanied by a typical onion skin configuration. The plexigenic pathological change is considered to be the result of response, at the late stage, to pulmonary hypertension rather than due to congenital abnormality or embolism.


Comparative protein modelling by satisfaction of spatial restraints.
We describe a comparative protein modelling method designed to find the most probable structure for a sequence given its alignment with related structures. The three-dimensional (3D) model is obtained by optimally satisfying spatial restraints derived from the alignment and expressed as probability density functions (pdfs) for the features restrained. For example, the probabilities for main-chain conformations of a modelled residue may be restrained by its residue type, main-chain conformation of an equivalent residue in a related protein, and the local similarity between the two sequences. Several such pdfs are obtained from the correlations between structural features in 17 families of homologous proteins which have been aligned on the basis of their 3D structures. The pdfs restrain C alpha-C alpha distances, main-chain N-O distances, main-chain and side-chain dihedral angles. A smoothing procedure is used in the derivation of these relationships to minimize the problem of a sparse database. The 3D model of a protein is obtained by optimization of the molecular pdf such that the model violates the input restraints as little as possible. The molecular pdf is derived as a combination of pdfs restraining individual spatial features of the whole molecule. The optimization procedure is a variable target function method that applies the conjugate gradients algorithm to positions of all non-hydrogen atoms. The method is automated and is illustrated by the modelling of trypsin from two other serine proteinases.


Kappa opiate agonists modulate the hypothalamic-pituitary-adrenocortical axis in the rat.
Systemic injections of opiate agonists were made in male rats to elucidate the involvement of multiple opioid receptors in the stress response. As an index of activity in the hypothalamic-pituitary-adrenocortical axis, plasma corticosterone was measured by radioimmunoassay. Rats were injected with ethylketocyclazocine (EKC), U50488H, MR2034, bremazocine or tifluadom and sacrificed 1 hr later. These kappa agonists produced potent, dose-dependent, stereospecific increases in plasma corticosterone levels at doses far below those needed to elicit analgesia. These effects were reversed by opiate antagonists, naloxone or Win 44441-3, which by themselves caused dose-dependent decreases in plasma corticosterone. Animals made tolerant to the prototype kappa agonist, U50488H, showed an attenuated response to an acute injection of the drug. However, when animals made tolerant to morphine were injected acutely with U50488H, the drug caused a dramatic increase in corticosterone levels. In hypophysectomized rats, U50488H and L-EKC did not increase plasma corticosterone. The agonist/antagonists, butorphanol and cyclazocine, when injected, behaved like kappa agonists and increased plasma corticosterone levels potently. The mu opiates, morphine and etorphine, also had similar effects but were less potent and efficacious than the kappa agonists. The delta agonist D-Ala-D-Leu enkephalin showed similar results, confirming a mu and delta opioid input into the hypothalamic-pituitary-adrenocortical axis. There were concomitant increases in plasma adrenocorticotropin in morphine-, D-Ala-D-Leu enkephalin-, L-EKC- and U50488H-treated rats which were also seen in adrenalectomized rats. D-EKC and D-cyclazocine, which bind to sigma sites, had no effect on corticosterone.(ABSTRACT TRUNCATED AT 250 WORDS)


Bone marrow transplantation improves hepatic fibrosis in Abcb4-/- mice via Th1 response and matrix metalloproteinase activity.
Reports on the effects of bone marrow-derived cells on hepatic fibrosis are contradictory. Impaired fibrosis but increased inflammation has recently been demonstrated 10 weeks after bone marrow transplantation (BM-Tx) in Abcb4-/- mice. It is hypothesised that BM-Tx might have long-term therapeutic potential by altering the immunological and matrix remodelling processes leading to hepatic regeneration. After lethal irradiation of recipient mice, BM cells from GFP+ donor mice (allogeneic Tx) or Abcb4-/- mice (syngeneic Tx) were transplanted via tail vein injection. Readouts were performed 2, 10 and 20 weeks after Tx. Liver integrity was assessed serologically and histologically. Surrogate markers for fibrogenesis, T helper (Th) response, inflammation, graft-versus-host disease and fibrolysis were analysed by quantitative real-time PCR, zymography and immunohistology. 20 weeks after syngeneic and allogeneic BM-Tx, hepatic grading and staging were significantly improved. In contrast, 2 weeks after BM-Tx inflammatory grading, expression of inflammatory cell markers and associated chemokines and their receptors were increased and subsequently declined. In parallel, CD8+/GFP+ donor-derived T cells infiltrated the liver 2 weeks after BM-Tx. The Th1 cyokine interferon γ was increased 2 and 10 weeks after BM-Tx whereas the Th2 associated interleukin 13 was not altered. The gene expression of matrix metalloproteinases MMP-2, MMP-7, MMP-9 and MMP-13 was transiently upregulated and MMP-9 protein remained elevated 20 weeks after BM-Tx with enhanced gelatinase activity located within the fibrotic areas. Neutrophils were identified as major sources of MMP-9. These results show that BM-Tx causes an antifibrotic Th1 response combined with transient inflammatory effects and subsequently upregulated MMP activity. Antifibrotic Th polarisation and prolonged proteolytic activity, especially of MMP-9, might be responsible for long-term amelioration of hepatic fibrosis.


Three-dimensional analysis of CD34 sialomucin distribution on cord blood and bone marrow.
Determining the cellular distribution of key adhesion molecules may aid in understanding haematopoietic progenitor/stem cell (HPSC) homing to bone marrow (BM). CD34, a well-characterized marker for blast-like HPSC, is widely used for the isolation and enumeration of HPSC. Functional studies have yet to identify a ligand for CD34. However, growing evidence suggests that CD34 may aid the regulation of HPSC differentiation and modulate the expression of other HPSC adhesion molecules necessary for homing. This study aimed to determine CD34 distribution on umbilical cord blood (CB) and BM. CD34-selected cells were adhered to positively charged gold slides at room temperature, before indirect fluorescent antibody labelling with fluorescein isothiocyanate. Fluorescent distribution was determined by 1-microm interval, confocal laser scanning microscope Z-sections. Initial analysis showed CD34 distributed within peripheral halos and dense pocket regions. The development of three-dimensional imaging software enabled spatial visualization of CD34 distribution on CB and BM in association with differential interference contrast cell image. This showed that CD34 was distributed within peripheral halos, with magnupodia-associated 'meridian-shaped crescents', extending from points of cell-slide adhesion towards the top of the cell. True CD34 distribution, not previously discernible by confocal laser scanning microscopy alone, suggests a possible role in cell adhesion/homing revealed by three-dimensional imaging.


Compromised concentrations of ascorbate in fluid lining the respiratory tract in human subjects after exposure to ozone.
Ozone (O3) imposes an oxidative burden on the lung in two ways. Firstly, directly as a consequence of its oxidising character during exposure, and secondly, indirectly by engendering inflammation. In this study the second pathway was considered by ascertaining the impact of O3 on the redox state of the fluid lining the respiratory tract 6 hours after challenge. Nine subjects were exposed in a double blind crossover control trial to air and 200 ppb O3 for 2 hours with an intermittent exercise and rest protocol. Blood samples were obtained and lung function (forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1)) assessed before, immediately after, and 6 hours after exposure. Bronchoalveolar lavage (BAL) was performed 6 hours after challenge. Inflammation was assessed in BAL fluid (total and differential cell counts, plus myeloperoxidase concentrations), and plasma and BAL fluid redox state were determined by measuring concentrations of antioxidants and markers of oxidative damage. Neutrophil numbers in BAL fluid increased 2.2-fold (p = 0.07) 6 hours after exposure and this was accompanied by increased myeloperoxidase concentrations in BAL fluid (p = 0.08). On the other hand, BAL fluid macrophage and lymphocyte numbers decreased 2.5-fold (p = 0.08) and 3.1-fold (p = 0.08), respectively at this time. Of the antioxidants examined, only ascorbate in BAL fluid was affected by O3, falling in all subjects relative to air values (0.1 (0.0-0.3) v 0.3 (0.2-1.2) mumol/l (p = 0.008)). A marginal decrease in plasma ascorbate was also detected at this time (p < 0.05). Although the decrease in macrophage numbers seemed to be causally related to the increase in neutrophils (R = -0.79), myeloperoxidase concentrations (R = -0.93) and ascorbate concentrations (R = 0.6), no clear associations were apparent between ascorbate changes and neutrophils or myeloperoxidase concentration after O3. Ascorbate in the fluid lining the respiratory tract is depleted as a consequence of O3 exposure at 6 hours after exposure. This was contemporaneous with, although not quantitatively related to the increase in neutrophil numbers and myeloperoxidase concentrations. Decreased macrophage numbers 6 hours after O3 related to the degree of neutrophilic inflammation with populations conserved where ascorbate concentration in the fluid lining the respiratory tract were high after exposure. These results imply that ascorbate has a critical protective role against inflammatory oxidative stress induced by O3.


Gambling Attitudes and Beliefs Predict Attentional Bias in Non-problem Gamblers.
Problem gambling has been identified as a public health concern in Australia, and a considerable proportion of regular gamblers may be at risk of developing gambling related problems. Attentional bias to salient cues has been observed in substance addictions, and to some extent, in problem gamblers. This bias appears to be indicative of an increase in sensitisation to salient cues as a result of continued reforcement of a related behaviour. To test for an attentional bias to gambling-related stimuli in non-problem gamblers, the relationships between gambling frequency, gambling attitudes and beliefs (GABS-23), and attentional bias were investigated. Participants (N = 38) viewed simultaneous pairs of gambling-related and neutral images and performed a dot probe task, during which their eye-movements were recorded. This enabled both direct and indirect measures of attentional bias to be obtained. Gambling frequency and GABS-23 scores predicted both direct and indirect measures of a bias in the maintenance of attention to gambling cues. No bias in attentional engagement was found. These results suggest that regular gamblers who have not yet developed any related problems show signs of sensitisation to gambling cues and may be at risk of progressing further towards problem gambling.


Quality of Life Among Chinese Child Survivors of Heart Failure After Discharge From Pediatric Intensive Care Unit.
This study investigated the current quality of life (QOL) status and related influencing factors in children with heart failure in the pediatric intensive care unit (PICU). One hundred eighty-three children admitted to the PICU with heart failure were consecutively sampled. They were assessed 3 months after discharge and compared with a control group of 160 healthy individuals. The Pediatric Quality of Life Inventory (version 4.0) and Children's Revised Impact of Event Scale were completed, and factors influencing QOL were analyzed. Three months after discharge, the PICU group reported lower average scores on the overall QOL and the physical and emotional functioning scales than the control group. In particular, posttraumatic stress disorder status, age, and caregiver educational level greatly influenced the PICU children's QOL. More interventions and attention are needed to improve the QOL for PICU heart failure patients after discharge.


Randomized clinical trial of micronized flavonoids in the early control of bleeding from acute internal haemorrhoids.
Patients with acute bleeding from internal haemorrhoids often have to be referred by the general practitioner to the surgeon for definitive treatment with invasive outpatient procedures. At the initial consultation, patients frequently seek postponement of immediate surgery to a more convenient time. Effective and rapid non-invasive control of acute bleeding could be of practical use in scheduling surgery to a time convenient to both patient and surgeon. In a 90-day randomized, double-blind study treatment with a micronized purified flavonoid fraction (MPFF) was compared with placebo, in 100 outpatients who presented for treatment of acute internal haemorrhoids of less than 3 days' duration. The primary endpoint was cessation of bleeding on the third day of treatment. Of 50 patients randomized to each group, acute bleeding ceased by the third day in 40 (80 per cent) who received MPFF compared with 19 (38 per cent) who had placebo (P < 0.01). Mean(s.d. ) duration of acute bleeding from onset to cessation of 4.9(1.6) days was 2.1 (95 per cent confidence interval 1.2-2.9) days less than that in patients receiving placebo (P < 0.01). Continued treatment in patients with no bleeding prevented a relapse in 30 of 47 patients, compared with 12 of 30 receiving placebo (P < 0.05). Patients with acute internal haemorrhoids treated with MPFF had rapid cessation of bleeding and a reduced risk of relapse. This could be of value in the more convenient timing of treatment with invasive outpatient procedures.


Plant Nutrient Resource Use Strategies Shape Active Rhizosphere Microbiota Through Root Exudation.
Plant strategies for soil nutrient uptake have the potential to strongly influence plant-microbiota interactions, due to the competition between plants and microorganisms for soil nutrient acquisition and/or conservation. In the present study, we investigate whether these plant strategies could influence rhizosphere microbial activities via root exudation, and contribute to the microbiota diversification of active bacterial communities colonizing the root-adhering soil (RAS) and inhabiting the root tissues. We applied a DNA-based stable isotope probing (DNA-SIP) approach to six grass species distributed along a gradient of plant nutrient resource strategies, from conservative species, characterized by low nitrogen (N) uptake, a long lifespans and low root exudation level, to exploitative species, characterized by high rates of photosynthesis, rapid rates of N uptake and high root exudation level. We analyzed their (i) associated microbiota composition involved in root exudate assimilation and soil organic matter (SOM) degradation by 16S-rRNA-based metabarcoding. (ii) We determine the impact of root exudation level on microbial activities (denitrification and respiration) by gas chromatography. Measurement of microbial activities revealed an increase in denitrification and respiration activities for microbial communities colonizing the RAS of exploitative species. This increase of microbial activities results probably from a higher exudation rate and more diverse metabolites by exploitative plant species. Furthermore, our results demonstrate that plant nutrient resource strategies have a role in shaping active microbiota. We present evidence demonstrating that plant nutrient use strategies shape active microbiota involved in root exudate assimilation and SOM degradation via root exudation.


Effects of some disinfectants on African swine fever virus.
Ten commercially available disinfectants were tested at high pH in 2% sodium hydroxide and low pH in 2% acetic acid as inactivants for African swine fever (ASF) in a protein-rich blood-spleen homogenate. As assayed in leukocyte cultures, sodium hydroxide and acetic acid, sodium meta silicate and Roccal did not inactivate ASF virus in 1 hr at 22 to 25 C. Some viricidal activity as assayed in leukocyte cultures was found with Weladol, Triton X-100 Amphyl, pHisoHex, sodium dodecyl sulfate, LpH, Environ, Environ D, and One-Stroke Environ. Of these, the last four appeared to be most promising. When assayed in pigs, only One-Stroke Environ (1/E) was viricidal. Concentrations of 1.0, 0.75, and 0.5 were effective, but, at 0.25%, virus was not inactivated. The minimal time to inactivate ASF virus by 1% 1/E is 60 min. A room contaminated with ASF virus was made safe for pigs after 1 hr by spraying with 1% 1/E. The most active component of 1/E is o-phenylphenol. Although another component of 1/E, i.e., o-benzyl-p-chlorophenol, also has some activity, the mixture of the active components of 1/E is most effective against ASF virus. One of the soluble antigens associated with ASF virus is destroyed by 1/E.


Insect-mediated transmission of mixed and reassorted cucumovirus genomic RNAs.
Transmissions of virus using the aphid Myzus persicae were performed using plants co-infected with two cucumoviruses, tomato aspermy virus (V-TAV) and cucumber mosaic virus (M-CMV). Five of the aphid-transmitted progeny viruses (3.7%) induced symptoms distinct from those induced by either parental virus. Northern blot hybridization analysis of encapsidated RNAs from these novel progeny demonstrated that all of the RNA profiles were characteristic of pseudo-recombinants, i.e. viruses with reassorted genomic RNAs. The two larger RNAs, 1 and 2, originated from V-TAV, whereas RNA 3 was derived from M-CMV. A more sensitive RNase protection assay analysis of both unencapsidated and encapsidated RNAs revealed the presence of minor populations of V-TAV-derived RNA 3 in all of these novel progeny, and of M-CMV-derived RNA 1 (and presumably RNA 2) in one of the progeny. A bias against the encapsidation of the minor populations of RNAs by the M-CMV coat protein was observed, suggesting that there is specificity or competition with regard to the encapsidation of cucumoviral RNAs in vivo. This study demonstrates that insect vectors can mediate the establishment of pseudorecombinants with mixed populations of RNA 3.


Enteral fluid therapy in 108 horses with large colon impactions and dorsal displacements.
To assess the effect of enteral fluid therapy (EFT) in horses with colic, 78 adult horses with colon impactions and 30 with left dorsal colon displacements received an isotonic electrolyte solution via a nasogastric tube at a rate of 8 to 10 l every two hours until resolution of clinical signs. Clinical progression was monitored closely, and plasma biochemistry was evaluated before, during and after treatment. Volume of fluids, time to resolution, and outcome were also recorded. EFT was well tolerated by 102 of 108 horses and was an effective method to resolve 99 per cent of colon impactions and 83 per cent of displacements. For both groups, the mean (sd) time to resolution was 20.2 (5.2) hours and the volume of fluid administered was 118.6 (34.5) l. No relevant abnormalities were observed in most plasma biochemistry parameters during treatment, except for a mild haemodilution effect in 63 per cent of horses.


Computational approaches to identify sites of phosphorylation.
Due to their oftentimes ambiguous nature, phosphopeptide positional isomers can present challenges in bottom-up mass spectrometry-based workflows as search engine scores alone are often not enough to confidently distinguish them. Additional scoring algorithms can remedy this by providing confidence metrics in addition to these search results, reducing ambiguity. Here we describe challenges to interpreting phosphoproteomics data and review several different approaches to determine sites of phosphorylation for both data-dependent and data-independent acquisition-based workflows. Finally, we discuss open questions regarding neutral losses, gas-phase rearrangement, and false localization rate estimation experienced by both types of acquisition workflows and best practices for managing ambiguity in phosphosite determination. This article is protected by copyright. All rights reserved.


Efficacy and safety of high versus standard dose ibuprofen for patent ductus arteriosus treatment in preterm infants: A systematic review and meta-analysis.
Several small randomized controlled trials (RCTs) and observational studies have compared high (15-20/7.5-10/7.5-10 mg/kg/dose) versus standard dose (10/5/5 mg/kg/dose) ibuprofen for patent ductus arteriosus (PDA) closure, with limited evidence on efficacy and safety. To systematically review and meta-analyze studies of high versus standard dose ibuprofen for the closure of PDA in preterm infants. Databases were searched for RCTs and observational studies assessing high compared to standard dose of ibuprofen for PDA closure for preterm infants until August 2021. The primary outcome was failure of PDA closure after the first course of ibuprofen. The secondary outcomes were the failure of PDA closure after a second course of ibuprofen, rates of PDA ligation, all-cause mortality prior to hospital discharge, bronchopulmonary dysplasia, necrotizing enterocolitis, bleeding disorders, oliguria, and serum creatinine after treatment. There were 6 studies with 369 patients (3 RCT, N = 190; 3 observational studies, N = 179). Compared to standard dose, high dose ibuprofen did not significantly decrease the failure rate of PDA closure in preterm infants after the first course (Relative risk (RR) 0.74, 95% confidence interval (CI) 0.53 -1.03, 6 studies, N = 369). High dose ibuprofen significantly decreased the rates of PDA ligation compared to standard dose (RR 0.33, 95% CI 0.16 -0.70, 5 studies, N = 309). Based on low-grade evidence, high dose ibuprofen may more effectively reduce rates of PDA ligation compared to standard dose with no increase in adverse effects, neonatal morbidities and mortality.


Dynamic masquerade with morphing three-dimensional skin in cuttlefish.
Masquerade is a defence tactic in which a prey resembles an inedible or inanimate object thus causing predators to misclassify it. Most masquerade colour patterns are static although some species adopt postures or behaviours to enhance the effect. Dynamic masquerade in which the colour pattern can be changed is rare. Here we report a two-step sensory process that enables an additional novel capability known only in cuttlefish and octopus: morphing three-dimensional physical skin texture that further enhances the optical illusions created by coloured skin patterns. Our experimental design incorporated sequential sensory processes: addition of a three-dimensional rock to the testing arena, which attracted the cuttlefish to settle next to it; then visual processing by the cuttlefish of physical textures on the rock to guide expression of the skin papillae, which can range from fully relaxed (smooth skin) to fully expressed (bumpy skin). When a uniformly white smooth rock was presented, cuttlefish moved to the rock and deployed a uniform body pattern with mostly smooth skin. When a rock with small-scale fragments of contrasting shells was presented, the cuttlefish deployed mottled body patterns with strong papillae expression. These robust and reversible responses indicate a sophisticated visual sensorimotor system for dynamic masquerade.


Comparative genome structure, secondary metabolite, and effector coding capacity across Cochliobolus pathogens.
The genomes of five Cochliobolus heterostrophus strains, two Cochliobolus sativus strains, three additional Cochliobolus species (Cochliobolus victoriae, Cochliobolus carbonum, Cochliobolus miyabeanus), and closely related Setosphaeria turcica were sequenced at the Joint Genome Institute (JGI). The datasets were used to identify SNPs between strains and species, unique genomic regions, core secondary metabolism genes, and small secreted protein (SSP) candidate effector encoding genes with a view towards pinpointing structural elements and gene content associated with specificity of these closely related fungi to different cereal hosts. Whole-genome alignment shows that three to five percent of each genome differs between strains of the same species, while a quarter of each genome differs between species. On average, SNP counts among field isolates of the same C. heterostrophus species are more than 25× higher than those between inbred lines and 50× lower than SNPs between Cochliobolus species. The suites of nonribosomal peptide synthetase (NRPS), polyketide synthase (PKS), and SSP-encoding genes are astoundingly diverse among species but remarkably conserved among isolates of the same species, whether inbred or field strains, except for defining examples that map to unique genomic regions. Functional analysis of several strain-unique PKSs and NRPSs reveal a strong correlation with a role in virulence.


Galangin (GG) combined with cisplatin (DDP) to suppress human lung cancer by inhibition of STAT3-regulated NF-κB and Bcl-2/Bax signaling pathways.
Lung cancer represents a significant problem for public health worldwide. Galangin (GG), a natural active compound 3, 5, 7-trihydroxyflavone, is a type of bioflavonoid, which is isolated from the Alpinia galangal root and suggested to induce apoptosis in various cancers. We investigated the ability of Galangin (GG) to attenuate the drug resistance of human lung cancer cells, resistant to treatment of cisplatin (DDP). DDP is a pyrimidine analog, widely used in cancer treatment. Galangin and DDP co-treatment resulted in a dose-dependent suppression of the cell proliferation. Decreasing of p-STAT3 was included in p65 suppression by GG with DDP in combination. Additionally, the presence of GG potentiated the effects of DDP on apoptosis induction through suppressing Bcl-2 in DDP-resistant lung cancer cells. The pro-apoptotic proteins of Bax and Bid were up-regulated, accompanied with Caspases cleavage, leading to apoptosis. Moreover, in mice xenograft models, the combined therapy inhibited tumor growth compared to the GG or DDP treatment alone. Our data indicated a novel therapeutic strategy to potentiate DDP-induced anti-tumor effect in lung cancer cells with DDP resistance by GG through inactivating p-STAT3/p65 and Bcl-2 pathways.


A clinical feature of hyperlipidemia in patients with central diabetes insipidus.
In this study, we analyzed plasma lipid and lipoprotein levels before and after treatment with 1-desamino-8-D-arginine vasopressin (DDAVP) in subjects with partial and complete central diabetes insipidus (DI) in order to determine how a shortage and supplement of this hormone affect plasma lipid metabolism. The subjects consisted of 6 patients with partial and 6 with complete central DI. After treatment with DDAVP through nasal cavity, plasma total cholesterol (TC) level did not decrease either in complete or partial form. Plasma triglyceride (TG) levels decreased from 306+/-175 mg/dl to 198+/-91 (35% decrease, p=0.027) in complete form, while TG did not change significantly in partial form. A detailed investigation of plasma lipoprotein metabolism during treatment with DDAVP was carried out in 3 of the 6 subjects with complete form of DI. Lipoprotein lipase activity and mass in post-heparin plasma from those three subjects tended to increase after treatment with DDAVP, along with the complete disappearance of an unusual lipoprotein between low density lipoprotein (LDL) and very low density lipoprotein (VLDL) as analyzed by polyacrylamide gel electrophoresis. These results suggest that the DDAVP treatment has a favorable effect on lipid and lipoprotein metabolism, especially triglyceride-rich lipoproteins, either directly or through modifying factors contributing to lipid metabolism.


[Temporal and spatial fluctuations in the count of soil bacteria considered by a luminescence method].
Spatial and temporal (within a day or a month and daily) variations in the number of bacteria in soddy-podzolic soil were determined by the direct count of cells under the luminescent microscope. No reliable change in the number of bacteria in soil was detected in the course of a day whereas the effect of the spatial factor was statistically significant. Daily increases and decreases in the number of bacteria were registered only in the summer. Seasonal changes in the number of bacterial population were considerable (4--5 times).


[Therapy of severe migraine attacks: practical tips].
Severe migraine attacks are treated with triptans. The forms of administration of the active substances depend on the drug profile. A standard therapy does not exist. The rate of side effects is low. Triptans are contraindicated for patients with coronary, cerebral and peripheral vascular diseases. Under no circumstances are they to be combined with ergot alkaloids. Triptans are all the more effective, the earlier they are taken. For determining the optimal time point for taking the medication, the so-called triptan threshold is helpful. Triptans are not suitable for treating status migrainosus. Sumatriptan can be administered electively and in compliance with the contraindications for the emergency treatment.


Expression of Syntaxin 2 in Bovine Sperm.
Syntaxins are membrane integrated Q-SNARE proteins known to participate in exocytosis. Vesicle docking involves the binding of two plasma membrane proteins, syntaxin and SNAP-25, to the vesicle membrane protein VAMP to form a stable trimeric core complex; synaptophysin is thought to regulate the formation of this complex. Although the members of Q-SNARE proteins are characterized in somatic cells, it is not known whether related proteins function in the sperm acrosome reaction. The objective of the present study is to identify and localize syntaxin in bovine epididymal spermatozoa and to determine the fate of syntaxin 2 during the acrosome reaction. Western blots of caput and cauda sperm lysates and plasma membrane fractions, stained with anti-syntaxin 2, revealed the presence of a 31kDa band in both sperm lysates and plasma membrane fractions, respectively. Indirect immunofluorescence localized syntaxin 2 to the anterior but not the equatorial regions of the acrosomal segment. Several biochemical analyses demonstrated that syntaxin 2 is an integral component of bovine cauda sperm plasma membranes. Our immunoblot data reveals that syntaxin 2 of bovine cauda sperm is released after lysophosphatidylcholine (LPC)-induced acrosomal exocytosis. It is assumed that syntaxin 2 may be involved in triggering the acrosome reaction through a ligand-receptor mediated signal transduction pathway.


Structural basis of substrate specificity in malate dehydrogenases: crystal structure of a ternary complex of porcine cytoplasmic malate dehydrogenase, alpha-ketomalonate and tetrahydoNAD.
The structural basis for the extreme discrimination achieved by malate dehydrogenases between a variety of closely related substrates encountered within the cell has been difficult to assess because of the lack of an appropriate catalytically competent structure of the enzyme. Here, we have determined the crystal structure of a ternary complex of porcine cytoplasmic malate dehydrogenase with the alternative substrate alpha-ketomalonate and the coenzyme analogue 1,4,5,6-tetrahydronicotinamide. Both subunits of the dimeric porcine heart, and from the prokaryotes Escherichia coli and Thermus flavus. However, large changes are noted around the active site, where a mobile loop now closes to bring key residues into contact with the substrate. This observation substantiates a postulated mechanism in which the enzyme achieves high levels of substrate discrimination through charge balancing in the active site. As the activated cofactor/substrate complex has a net negative charge, a positive counter-charge is provided by a conserved arginine in the active site loop. The enzyme must, however, also discriminate against smaller substrates, such as pyruvate. The structure shows in the closed (loop down) catalytically competent complex two arginine residues (91 and 97) are driven into close proximity. Without the complimentary, negative charge of the substrate side-chain of oxaloacetate or alpha-ketomalonate, charge repulsion would resist formation production of this catalytically productive conformation, hence minimising the effectiveness of pyruvate as a substrate. By this mechanism, malate dehydrogenase uses charge balancing to achieve fivefold orders of magnitude in discrimination between potential substrates.