Signal processing in urodynamics: towards high definition urethral pressure profilometry.
Urethral pressure profilometry (UPP) is used in the diagnosis of stress urinary incontinence (SUI) which is a significant medical, social, and economic problem. Low spatial pressure resolution, common occurrence of artifacts, and uncertainties in data location limit the diagnostic value of UPP. To overcome these limitations, high definition urethral pressure profilometry (HD-UPP) combining enhanced UPP hardware and signal processing algorithms has been developed. In this work, we present the different signal processing steps in HD-UPP and show experimental results from female minipigs. We use a special microtip catheter with high angular pressure resolution and an integrated inclination sensor. Signals from the catheter are filtered and time-correlated artifacts removed. A signal reconstruction algorithm processes pressure data into a detailed pressure image on the urethra's inside. Finally, the pressure distribution on the urethra's outside is calculated through deconvolution. A mathematical model of the urethra is contained in a point-spread-function (PSF) which is identified depending on geometric and material properties of the urethra. We additionally investigate the PSF's frequency response to determine the relevant frequency band for pressure information on the urinary sphincter. Experimental pressure data are spatially located and processed into high resolution pressure images. Artifacts are successfully removed from data without blurring other details. The pressure distribution on the urethra's outside is reconstructed and compared to the one on the inside. Finally, the pressure images are mapped onto the urethral geometry calculated from inclination and position data to provide an integrated image of pressure distribution, anatomical shape, and location. With its advanced sensing capabilities, the novel microtip catheter collects an unprecedented amount of urethral pressure data. Through sequential signal processing steps, physicians are provided with detailed information on the pressure distribution in and around the urethra. Therefore, HD-UPP overcomes many current limitations of conventional UPP and offers the opportunity to evaluate urethral structures, especially the sphincter, in context of the correct anatomical location. This could enable the development of focal therapy approaches in the treatment of SUI.


DNA damage-induced replication stress results in PA200-proteasome-mediated degradation of acetylated histones.
Histone acetylation influences protein interactions and chromatin accessibility and plays an important role in the regulation of transcription, replication, and DNA repair. Conversely, DNA damage affects these crucial cellular processes and induces changes in histone acetylation. However, a comprehensive overview of the effects of DNA damage on the histone acetylation landscape is currently lacking. To quantify changes in histone acetylation, we developed an unbiased quantitative mass spectrometry analysis on affinity-purified acetylated histone peptides, generated by differential parallel proteolysis. We identify a large number of histone acetylation sites and observe an overall reduction of acetylated histone residues in response to DNA damage, indicative of a histone-wide loss of acetyl modifications. This decrease is mainly caused by DNA damage-induced replication stress coupled to specific proteasome-dependent loss of acetylated histones. Strikingly, this degradation of acetylated histones is independent of ubiquitylation but requires the PA200-proteasome activator, a complex that specifically targets acetylated histones for degradation. The uncovered replication stress-induced degradation of acetylated histones represents an important chromatin-modifying response to cope with replication stress.


Sequence and developmental expression of amphioxus AmphiNk2-1: insights into the evolutionary origin of the vertebrate thyroid gland and forebrain.
We characterized an amphioxus NK-2 homeobox gene (AmphiNk2-1), a homologue of vertebrate Nkx2-1, which is involved in the development of the central nervous system and thyroid gland. At the early neurula stage of amphioxus, AmphiNk2-1 expression is first detected medially in the neural plate. By the mid-neurula stage, expression is localized ventrally in the nerve cord and also begins in the endoderm. During the late neurula stage, the ventral neural expression becomes transiently segmented posteriorly and is then down-regulated except in the cerebral vesicle at the anterior end of the central nervous system. Within the cerebral vesicle AmphiNk2-1 is expressed in a broad ventral domain, probably comprising both the floor plate and basal plate regions; this pattern is comparable to Nkx2-1 expression in the mouse diencephalon. In the anterior part of the gut, expression becomes intense in the endostyle (the right wall of the pharynx), which is the presumed homologue of the vertebrate thyroid gland. More posteriorly, there is transitory expression in the midgut and hindgut. In sum, the present results help to support homologies (1) between the amphioxus endostyle and the vertebrate thyroid gland and (2) between the amphioxus cerebral vesicle and the vertebrate diencephalic forebrain.


High-resolution ultrasound analysis of subsynovial connective tissue in human cadaver carpal tunnel.
The carpal tunnel contains the median nerve, nine flexor tendons, two synovial bursae, and peritendinous subsynovial connective tissue (SSCT). Fibrosis of the SSCT is the most consistent pathological finding in patients with carpal tunnel syndrome. We investigated the anatomy and gliding characteristics of the flexor digitorum superficialis tendon and its adjacent SSCT with high-resolution ultrasound (15 MHz). Our hypotheses were that tendon and SSCT are distinguishable by ultrasound and that their velocities during tendon excursion are different. Qualitative ultrasound analysis of a flexor tendon and its SSCT was performed on five cadaver wrists and correlated to respective findings after anatomical study of the same cadavers. Quantitative Doppler velocity analysis of eight cadaver wrists was done to assess the sliding movement of the tendon and its SSCT within the carpal tunnel. No significant difference was found between the thickness of SSCT measured by ultrasound and that measured directly after dissection. The SSCT moved slower than its flexor tendon. The SSCT velocities were statistically different from the tendon velocities (t-test, p>0.001). High-resolution ultrasound is a very precise method to display the anatomy of the tendon and SSCT within the carpal tunnel, and their different velocities can be detected with Doppler. Noninvasive assessment of the thickness and velocity of the tenosynovium in carpal tunnel syndrome by high-resolution sonography might be a new diagnostic tool for disorders affecting the SSCT, especially carpal tunnel syndrome.


Removing batch effects in analysis of expression microarray data: an evaluation of six batch adjustment methods.
The expression microarray is a frequently used approach to study gene expression on a genome-wide scale. However, the data produced by the thousands of microarray studies published annually are confounded by "batch effects," the systematic error introduced when samples are processed in multiple batches. Although batch effects can be reduced by careful experimental design, they cannot be eliminated unless the whole study is done in a single batch. A number of programs are now available to adjust microarray data for batch effects prior to analysis. We systematically evaluated six of these programs using multiple measures of precision, accuracy and overall performance. ComBat, an Empirical Bayes method, outperformed the other five programs by most metrics. We also showed that it is essential to standardize expression data at the probe level when testing for correlation of expression profiles, due to a sizeable probe effect in microarray data that can inflate the correlation among replicates and unrelated samples.


Role of macronutrients, vitamins and minerals in the aetiology of squamous-cell carcinoma of the oesophagus.
Between 1992 and 1997 we conducted a case-control study of oesophageal cancer in 3 areas of northern Italy. Cases were 304 patients (29 women), ages 39-77 years (median age 60 years), with a first incident squamous-cell carcinoma (SCC) of the oesophagus. Controls were 743 patients (150 women), ages 35-77 years (median age 60 years), admitted for acute illnesses, unrelated to tobacco and alcohol, to major hospitals of the areas under surveillance. We derived estimates of daily dietary intake of 6 macronutrients, cholesterol, and 20 micronutrients or minerals from a validated food-frequency questionnaire, including 78 food groups and recipes and 15 questions on individual eating patterns. After allowance for age, gender, area of residence, education, body mass index, physical activity, smoking habit, alcohol consumption and energy intake, most micronutrients were inversely associated with oesophageal SCC risk. Highly significant associations emerged for monounsaturated fatty acids [odds ratio (OR) in highest vs. lowest intake quintile = 0.5]; carotene (OR = 0.3); lutein + zeaxanthin (OR = 0.4); vitamin C (OR = 0.4); and niacin (OR = 0.5). Only retinol appeared to be positively related to risk (OR = 1.9). The effect of the above nutrients, expressed as ORs, appeared to be similar in non-smokers and smokers, and non/light drinkers and heavy drinkers.


Gene Expression as a Dosimeter in Irradiated Drosophila melanogaster.
Biological indicators would be of use in radiation dosimetry in situations where an exposed person is not wearing a dosimeter, or when physical dosimeters are insufficient to estimate the risk caused by the radiation exposure. In this work, we investigate the use of gene expression as a dosimeter. Gene expression analysis was done on 15,222 genes of Drosophila melanogaster (fruit flies) at days 2, 10, and 20 postirradiation, with X-ray exposures of 10, 1000, 5000, 10,000, and 20,000 roentgens. Several genes were identified, which could serve as a biodosimeter in an irradiated D. melanogaster model. Many of these genes have human homologues. Six genes showed a linear response (R2 > 0.9) with dose at all time points. One of these genes, inverted repeat-binding protein, is a known DNA repair gene and has a human homologue (XRCC6). The lowest dose, 10 roentgen, is very low for fruit flies. If the lowest dose is excluded, 13 genes showed a linear response with dose at all time points. This includes 5 of 6 genes that were linear with all radiation doses included. Of these 13 genes, 4 have human homologues and 8 have known functions. The expression of this panel of genes, particularly those with human homologues, could potentially be used as the biological indicator of radiation exposure in dosimetry applications.


Blockade of beta-catenin signaling by plant flavonoid apigenin suppresses prostate carcinogenesis in TRAMP mice.
Deregulation of beta-catenin signaling is an important event in the genesis of several human malignancies including prostate cancer. We investigated the effects of apigenin, a naturally occurring plant flavone, on prostate carcinogenesis in TRAMP mice and further elucidated its mechanism of action. Oral intake of apigenin by gavage at doses of 20 and 50 microg/mouse/d, 6 days per week for 20 weeks, significantly decreased tumor volumes of the prostate as well as completely abolished distant-site metastases to lymph nodes, lungs, and liver in TRAMP mice. Apigenin-treated mice had significantly diminished weights of their genitourinary apparatuses and dorsolateral and ventral prostate lobes, compared with the control group, and showed reduced proliferation and increased apoptosis in the dorsolateral prostates, which correlated with elevated plasma apigenin levels. Continuous intake of apigenin up to 50 weeks by TRAMP mice significantly improved their overall survival. P.o. administration of apigenin further resulted in increased levels of E-cadherin and decreased levels of nuclear beta-catenin, c-Myc, and cyclin D1 in the dorsolateral prostates of TRAMP mice. Similar effects were noted in TRAMP mice with established tumors. Treatment of DU145 human prostate cancer cells with 10 and 20 micromol/L apigenin also increased protein levels of E-cadherin by 27% to 74%, inhibited nuclear translocation of beta-catenin and its retention in the cytoplasm, and decreased c-Myc and cyclin D1 levels, an effect similar to the exposure of cells to beta-catenin small interfering RNA. Our results indicate that apigenin effectively suppressed prostate carcinogenesis in TRAMP mice, at least in part, by blocking beta-catenin signaling.


CTLA-4 engagement inhibits Th2 but not Th1 cell polarisation.
CTLA-4 deficient mice show severe lymphoproliferative disorders with T helper sub-population skewed toward the Th2 phenotype. In the present work, we investigated the role of CTLA-4 in T helper cell subset differentiation. Naïve CD4+ cells were stimulated with anti-CD3 and anti-CD28 mAbs in the presence of either IL-12 or IL-4 to induce polarisation to Th1 or Th2 cells, respectively. Under these two polarising conditions cells express comparable levels of CTLA-4. CTLA4 was stimulated by plastic-bound mAb. The frequency of IFN-gamma- and IL-4-producing cells were estimated by FACS analysis. In parallel cultures, polarised Th1 and Th2 cells were re-stimulated with anti-CD3 and anti-CD28 mAbs for 48 h and their culture supernatants analysed by ELISA. Results show that CTLA-4 engagement during differentiation inhibits polarisation of naive CD4+ cells to the Th2 but not the Th1 cell subset. At variance, once cells are polarised, CTLA-4 engagement inhibits cytokine production in both effector Th2 and Th1 cells. Altogether these data indicate that CTLA-4 may interfere not only in the signalling involved in acute transcriptional activation of both Th1 and Th2 cells but also in the development of one of the Th cell subsets.


17β-estradiol replacement reverses age-related lung disease in estrogen-deficient C57BL/6J mice.
The role that estrogens play in the aging lung is poorly understood. Remodeling of the aging lung with thickening of the alveolar walls and reduction in the number of peripheral airways is well recognized. The present study was designed to address whether estrogen deficiency would affect age-associated changes in the lungs of female C57BL/6J mice. Lungs isolated from old mice (24 months old, estrogen-deficient) demonstrated decreased lung volume and decreased alveolar surface area. There was no difference in alveolar number in the lungs of old and young mice (6 months old, estrogen-replete). Estrogen replacement restored lung volume, alveolar surface area, and alveolar wall thickness to that of a young mouse. Estrogen receptor-α (ERα) protein expression increased without a change in ERβ protein expression in the lung tissue isolated from old mice. In the lungs of old mice, the number of apoptotic cells was increased as well as the activation of matrix metalloproteinase-2 and ERK. Young mice had the highest serum 17β-estradiol levels that decreased with age. Our data suggest that in the aging female mouse lung, estrogen deficiency and an increase of ERα expression lead to the development of an emphysematous phenotype. Estrogen replacement partially prevents these age-associated changes in the lung architecture by restoration of interalveolar septa. Understanding the role of estrogens in the remodeling of the lung during aging may facilitate interventions and therapies for aging-related lung disease in women.


Novel renal medullary carcinoma cell lines, UOK353 and UOK360, provide preclinical tools to identify new therapeutic treatments.
Renal medullary carcinoma (RMC) is a rare, aggressive disease that predominantly afflicts individuals of African or Mediterranean descent with sickle cell trait. RMC comprises 1% of all renal cell carcinoma diagnoses with a median overall survival of 13 months. Patients are typically young (median age-22) and male (male:female ratio of 2:1) and tumors are characterized by complete loss of expression of the SMARCB1 tumor suppressor protein. Due to the low incidence of RMC and the disease's aggressiveness, treatment decisions are often based on case reports. Thus, it is critical to develop preclinical models of RMC to better understand the pathogenesis of this disease and to identify effective forms of therapy. Two novel cell line models, UOK353 and UOK360, were derived from primary RMCs that both demonstrated the characteristic SMARCB1 loss. Both cell lines overexpressed EZH2 and other members of the polycomb repressive complex and EZH2 inhibition in RMC tumor spheroids resulted in decreased viability. High throughput drug screening of both cell lines revealed several additional candidate compounds, including bortezomib that had both in vitro and in vivo antitumor activity. The activity of bortezomib was shown to be partially dependent on increased oxidative stress as addition of the N-acetyl cysteine antioxidant reduced the effect on cell proliferation. Combining bortezomib and cisplatin further decreased cell viability both in vitro and in vivo that single agent bortezomib treatment. The UOK353 and UOK360 cell lines represent novel preclinical models for the development of effective forms of therapy for RMC patients.


Fostering greater recognition of Caribbean traditional plant knowledge.
The Caribbean is a hotspot of biological and cultural diversity, manifested in traditional plant knowledge of Afrodescendant peoples and other ethnicities. To strengthen the visibility of this knowledge in research, education, and policy making, we propose an eight-step action plan centered on reciprocal relationships with Caribbean plant stewards, especially subsistence farmers.


Psychophysiology and psychoacoustics of music: Perception of complex sound in normal subjects and psychiatric patients.
Perception of complex sound is a process carried out in everyday life situations and contributes in the way one perceives reality. Attempting to explain sound perception and how it affects human beings is complicated. Physics of simple sound can be described as a function of frequency, amplitude and phase. Psychology of sound, also termed psychoacoustics, has its own distinct elements of pitch, intensity and tibre. An interconnection exists between physics and psychology of hearing.Music being a complex sound contributes to communication and conveys information with semantic and emotional elements. These elements indicate the involvement of the central nervous system through processes of integration and interpretation together with peripheral auditory processing.Effects of sound and music in human psychology and physiology are complicated. Psychological influences of listening to different types of music are based on the different characteristics of basic musical sounds. Attempting to explain music perception can be simpler if music is broken down to its basic auditory signals. Perception of auditory signals is analyzed by the science of psychoacoustics. Differences in complex sound perception have been found between normal subjects and psychiatric patients and between different types of psychopathologies.


[Completely implantable infusion pump. A new therapeutic possibility for selected patients with liver tumors].
The action of floxuridine depends greatly on its concentration for treatment of liver metastases of colorectal carcinomas and other tumors. In isolated liver metastases it is best infused regionally in order to achieve a high concentration and to reduce systemic toxicity, particularly as it is almost entirely extracted by the liver. A totally implantable infusion pump has established itself as an effective, safe and reliable device of regional chemotherapy with floxuridine for palliative treatment of selected patients with metastases of colorectal carcinomas and other tumors. Life expectancy of patients with metastases of colorectal carcinomas was prolonged by 12-18 months when compared to a precedent control group without regional chemotherapy. This results require confirmation in a randomised phase III study. Patients with hepatocellular carcinoma can be treated with intraarterial doxorubicine and floxuridine infusions. Preliminary results are encouraging. The implantable system represents a major advance in drug administration, not only for the treatment of malignant disease but also a large number of other diseases. Continuous infusion of drugs using an implantable pump system will probably achieve greater importance in the future in the treatment of cancer.


Evidence by expression analysis of candidate genes for congenital heart defects in the NF1 microdeletion interval.
It was recently reported that congenital heart disease is significantly more frequent in patients with NF1 microdeletion syndrome than in those with classical NF1. The outcome of congenital heart disease in this subset of patients is likely caused by the haploinsufficiency of gene/s in the deletion interval. Following in silico analysis of the deleted region, we found two genes known to be expressed in adult heart, the Joined to JAZF1 (SUZ12) and the Centaurin-alpha 2 (CENTA2) genes, and seven other genes with poorly defined patterns of expression and function. With the aim of defining their expression profiles in human fetal tissues (15th-21st weeks of gestation), expression analysis by RT-PCR and Northern blotting was performed. C17orf40, SUZ12 and CENTA2 were found to be mainly expressed in fetal heart, and following RT-PCR on mouse embryos and embryonic heart and brain at different stages of development, we found that the orthologous genes C17orf40, Suz12 and Centa2 are also expressed in early stages of development, before and during the formation of the four heart chambers. The presence of binding sites for Nkx2-5, a transcription factor expressed early in heart development, in all three mouse orthologous genes was predicted by bioinformatics, thus reinforcing the hypothesis that these genes might be involved in heart development and may be plausible candidates for congenital heart disease.


Practical aspects of decision making in clinical trials: the coronary drug project as a case study. The Coronary Drug Project Research Group.
Arriving at a decision for early termination of a treatment group or of an entire clinical trial, due to either beneficial or adverse results, is a complex process. It may involve, among other things, the necessity to (1) determine whether the observed treatment differences are likely to represent real effects and are not due to chance; (2) weigh the importance of many different response variables, some possible trending in favor of the treatment and some against it; (3) adjust for differences in distributions of baseline characteristics among the treatment groups; (4) discern possible biases (due to the study not being double-blind) in the medical management of patients or in the diagnosis of events; and (5) evaluate treatment effects in subgroups of the study participants. Experiences in the Coronary Drug Project are described in making decisions for early termination of treatment groups. Special methods applied in conjunction with reaching these decisions included: (1) analyses designed to detect possible biases in diagnosing a major endpoint--nonfatal myocardial infarction, e.g.--biases that may have arisen because of unblinding by side effects of the estrogen treatment group; (2) a simulation study to assess the statistical significance of mortality differences between the dextrothyroxine and placebo participants in certain subgroups identified a posteriori; and (3) the development, on the basis of the accumulated data, or new hypotheses relating to treatment differences in specific subgroups and the testing of these hypotheses a few months later on the basis of new events occurring during that interval.


Museum Moving to Inpatients: Le Louvre à l'Hôpital.
Anxiety and depressive symptoms are common in hospitalized patients. Arts and cultural programs were reported to enhance their quality of life. The Le Louvre à l'hôpital study presents a new approach in which the museum moves to the hospital by displaying and discussing artworks with patients interactively. Over one year, four large statues were disposed in the hospital gardens, 30 reprints of large painting were exhibited in the hospital hall, dining rooms, and circulations areas. A total of 83 small-group guided art discussions (90 min) were organized, which 451 patients attended. The 200 small-size reproductions of paintings placed in the patients' rooms were chosen based on their individual preferences. Decreased anxiety after the art sessions was reported by 160 of 201 patients (79.6%). Out of 451 patients, 406 (90%) said the art program had met their expectations, and 372 (82.4%) wished to continue the experience with caregivers (162 paramedics trained for art activity during 66 workshops). In conclusion, moving the museum to the hospital constitutes a valuable way to provide art activities for inpatients in large numbers, which may reduce hospital-related anxiety in many instances.


Use of Rheumatologic Testing in Patients Who Eventually Receive a Diagnosis of Rheumatoid Arthritis.
Anti-cyclic citrullinated peptide antibody (ACPA) has excellent specificity and prognostic value in patients with early rheumatoid arthritis (RA). The American College of Rheumatology included ACPA in their 2010 classification criteria for RA, but we hypothesize that primary care physicians (PCPs) underuse ACPA, even when clinical suspicion for RA is high. We aimed to describe their use of diagnostic testing in patients who were referred to a rheumatologist and eventually diagnosed as having RA. In this retrospective cohort study, a systematic abstraction tool was used to review the medical records of patients seen between January 1, 2010 and June 15, 2014 in two rheumatology clinics: one private practice and one community health center associated with an academic medical center. For purposes of hypothesis generation, we compared the characteristics of patients with and without testing using unpaired t tests or Fisher exact tests. We identified 173 patients with RA referred from 141 different PCPs: 82.7% were women with a mean ± standard deviation age of 55.5 ± 18.6 years. ACPA and rheumatoid factor were ordered in 28.9% (95% confidence interval 22.6-36.2) and 41.0% (95% confidence interval 33.9-48.6) of patients, respectively. Imaging was underused. Almost half (45.7%, or 37/81) of the patients with documented symptom duration had a delay of at least 1 year before referral; however, ACPA utilization was not associated with the delay to treatment initiation. Most PCPs failed to order diagnostic tests for RA before referring a patient with polyarthritis who eventually received a diagnosis of RA. We also observed delays in diagnosis, with half of the patients waiting >1 year from symptom onset to diagnosis. These findings suggest educational efforts for PCPs should focus on emphasizing earlier diagnostic workups, especially ACPA, in patients suspected to have RA.


Clinical features and prognosis of amyotrophic lateral sclerosis in Africa: the TROPALS study.
We describe and compare the sociodemographic and clinical features, treatments, and prognoses and survival times of patients with amyotrophic lateral sclerosis (ALS) in Africa. We conducted a multicentre, hospital-based cohort study in Africa. Patients with ALS diagnosed in the neurology departments of participating hospitals from 2005 to 2017 were included. Subgroup analysis was performed by subcontinent. Survival analyses were conducted using the Cox proportional hazards model. Nine centres from eight African countries participated. A total of 185 patients with ALS were included: 114 from Northern Africa, 41 from Western Africa and 30 from Southern Africa. A male predominance (male to female ratio 2.9) was evident. The median age at onset was 53.0 years (IQR 44.5-64.0 years). The onset was bulbar in 22.7%. Only 47 patients (26.3%) received riluzole, mainly in Northern and Western Africa. The median survival from the time of diagnosis was 14.0 months (95% CI 10.7 to 17.2 months). The median survival was longer in Northern Africa (19.0 months, 95% CI 10.8 to 27.2 months) than in Western (4.0 months, 95% CI 0.8 to 7.1 months) and Southern (11.0 months, 95% CI 5.6 to 16.4 months) Africa (Breslow test, p<0.0001). Both subcontinental location and riluzole treatment independently affected survival. More African patients with ALS were male and younger and exhibited a lower proportion of bulbar onset compared with patients with ALS from Western nations. Survival was consistent with that in Western registers but far shorter than what would be expected for young patients with ALS. The research improves our understanding of the disease in Africa.


Flagellin treatment prevents increased susceptibility to systemic bacterial infection after injury by inhibiting anti-inflammatory IL-10+ IL-12- neutrophil polarization.
Severe trauma renders patients susceptible to infection. In sepsis, defective bacterial clearance has been linked to specific deviations in the innate immune response. We hypothesized that innate immune modulations observed during sepsis also contribute to increased bacterial susceptibility after severe trauma. A well-established murine model of burn injury, used to replicate infection following trauma, showed that wound inoculation with P. aeruginosa quickly spreads systemically. The systemic IL-10/IL-12 axis was skewed after burn injury with infection as indicated by a significant elevation in serum IL-10 and polarization of neutrophils into an anti-inflammatory ("N2"; IL-10(+) IL-12(-)) phenotype. Infection with an attenuated P. aeruginosa strain (ΔCyaB) was cleared better than the wildtype strain and was associated with an increased pro-inflammatory neutrophil ("N1"; IL-10(-)IL-12(+)) response in burn mice. This suggests that neutrophil polarization influences bacterial clearance after burn injury. Administration of a TLR5 agonist, flagellin, after burn injury restored the neutrophil response towards a N1 phenotype resulting in an increased clearance of wildtype P. aeruginosa after wound inoculation. This study details specific alterations in innate cell populations after burn injury that contribute to increased susceptibility to bacterial infection. In addition, for the first time, it identifies neutrophil polarization as a therapeutic target for the reversal of bacterial susceptibility after injury.


Toddler dietary patterns from the INSIGHT randomized clinical trial comparing responsive parenting versus control: A latent class analysis.
The aim of this study was to examine the effect of a responsive parenting (RP) intervention on toddler diet and explore associations with mothers' infant feeding practices and child weight status. INSIGHT tested an RP intervention designed for the prevention of obesity against a safety control among primiparous mothers and their infants. Mothers reported on feeding practices, as well as toddler diet with a Food Frequency Questionnaire (n = 229). Trained research staff obtained child anthropometrics at age 2 years. Latent class analysis identified three dietary patterns: high fruits and vegetables (HFV, 31%); meat, potatoes, and added sugars (MPAS, 24%); and high juice, low fruits and vegetables (JLFV, 45%). Toddler dietary pattern was not related to study group (RP, control) or child weight status at age 2 years. Mothers who reported more structure-based feeding had toddlers that were more likely to have the healthier, HFV dietary pattern than MPAS and JLFV. Findings for control-based feeding practices were mixed; maternal restriction was associated with the HFV dietary pattern, whereas the use of food as a reward was associated with MPAS and JLFV. Mothers' structure-based feeding practices in infancy, as well as some control-based feeding practices, were associated with later healthier toddler dietary patterns.


A bifurcated continuous field-flow fractionation (BCFFF) chip for high-yield and high-throughput nucleic acid extraction and purification.
We report a bifurcated continuous field-flow fractionation (BCFFF) chip for high-yield and high-throughput (20 min) extraction of nucleic acids from physiological samples. The design uses a membrane ionic transistor to sustain low-ionic strength in a localized region at a junction, such that the resulting high field can selectively isolate high-charge density nucleic acids from the main flow channel and insert them into a standardized buffer in a side channel that bifurcates from the junction. The high local electric field and the bifurcated field-flow design facilitate concentration reduction of both divalent cation (Ca2+) and molecular PCR inhibitors by more than two orders of magnitude, even with high-throughput continuous loading. The unique design with a large (>20 mM mm-1) on-chip ionic-strength gradient allows miniaturization into a high-throughput field-flow fractionation chip that can be integrated with upstream lysing and downstream PCR/sensor modules for various nucleic acid detection/quantification applications. A concentration-independent 85% yield for extraction and an overall post-PCR yield exceeding 60% are demonstrated for a 111 bp dsDNA in 10 μL of human plasma, compared to no amplification with the raw sample. A net yield four times larger than a commercial extraction kit is demonstrated for miR-39 in human plasma.


Detecting 3-D motion field from range image sequences.
An algorithm for computing three-dimensional (3-D) velocity field and motion parameters from a range image sequence is presented. It is based on a new integral 3-D rigid motion invariant feature-the trace of a 3x3 "feature matrix" related to the moment of inertia tensor. This trace can be computed at every point of a moving surface and provides a quantitative measure of the local shape of the surface. Based on the feature's conservation along the trajectory of every moving point, a 3-D Flow Constraint Equation is formulated and solved for the velocity field. The robustness of the feature in presence of noise and discontinuity is analyzed.


Robust multichannel blind deconvolution via fast alternating minimization.
Blind deconvolution, which comprises simultaneous blur and image estimations, is a strongly ill-posed problem. It is by now well known that if multiple images of the same scene are acquired, this multichannel (MC) blind deconvolution problem is better posed and allows blur estimation directly from the degraded images. We improve the MC idea by adding robustness to noise and stability in the case of large blurs or if the blur size is vastly overestimated. We formulate blind deconvolution as an l(1) -regularized optimization problem and seek a solution by alternately optimizing with respect to the image and with respect to blurs. Each optimization step is converted to a constrained problem by variable splitting and then is addressed with an augmented Lagrangian method, which permits simple and fast implementation in the Fourier domain. The rapid convergence of the proposed method is illustrated on synthetically blurred data. Applicability is also demonstrated on the deconvolution of real photos taken by a digital camera.


Binding kinetics of antiricin single domain antibodies and improved detection using a B chain specific binder.
Single domain antibodies are the recombinantly expressed binding fragments derived from heavy chain antibodies found in camels and llamas. These unique binding elements offer many desirable properties such as their small size ( approximately 15 kDa) and thermal stability, which makes them attractive alternatives to conventional monoclonal antibodies. We created a phage display library from llamas immunized with ricin toxoid and selected a number of single domain antibodies. Phage selected on ricin were found to bind to either ricin A chain or the intact molecule; no ricin B chain binders were identified. By panning on B chain, we identified binders and have characterized their binding to the ricin B chain. While they have a poorer affinity than the previously described A chain binders, it was found that they performed dramatically better as capture reagents for the detection of ricin, providing a limit of detection in enzyme linked immunosorbent assay (ELISA) below 100 pg/mL and excellent specificity for ricin versus the highly related RCA 120 (1 to 10 000). We also reevaluated the previously isolated antiricin single domain antibody binding kinetics using surface plasmon resonance and found their K(d)s matched closely to those previously obtained under equilibrium binding conditions measured using the Luminex flow cytometer.


Design and development of an upper extremity motion capture system for a rehabilitation robot.
Human robot interaction is a new and rapidly growing field and its application in the realm of rehabilitation and physical care is a major focus area of research worldwide. This paper discusses the development and implementation of a wireless motion capture system for the human arm which can be used for physical therapy or real-time control of a robotic arm, among many other potential applications. The system is comprised of a mechanical brace with rotary potentiometers inserted at the different joints to capture position data. It also contains surface electrodes which acquire electromyographic signals through the CleveMed BioRadio device. The brace interfaces with a software subsystem which displays real time data signals. The software includes a 3D arm model which imitates the actual movement of a subject's arm under testing. This project began as part of the Rochester Institute of Technology's Undergraduate Multidisciplinary Senior Design curriculum and has been integrated into the overall research objectives of the Biomechatronic Learning Laboratory.


Advances in biology and therapy of multiple myeloma.
Even during this past year, further advances have been made in understanding the molecular genetics of the disease, the mechanisms involved in the generation of myeloma-associated bone disease and elucidation of critical signaling pathways as therapeutic targets. New agents (thalidomide, Revimid, Velcade) providing effective salvage therapy for end-stage myeloma, have broadened the therapeutic armamentarium markedly. As evidenced in Section I by Drs. Kuehl and Bergsagel, five recurrent primary translocations resulting from errors in IgH switch recombination during B-cell development in germinal centers involve 11q13 (cyclin D1), 4p16.3 (FGFR3 and MMSET), 6p21 (cyclin D3), 16q23 (c-maf), and 20q11 (mafB), which account for about 40% of all myeloma tumors. Based on gene expression profiling data from two laboratories, the authors propose 5 multiple myeloma (MM) subtypes defined by the expression of translocation oncogenes and cyclins (TC molecular classification of MM) with different prognostic implications. In Section II, Drs. Barillé-Nion and Bataille review new insights into osteoclast activation through the RANK Ligand/OPG and MIP-1 chemokine axes and osteoblast inactivation in the context of recent data on DKK1. The observation that myeloma cells enhance the formation of osteoclasts whose activity or products, in turn, are essential for the survival and growth of myeloma cells forms the basis for a new treatment paradigm aimed at reducing the RANKL/OPG ratio by treatment with RANKL inhibitors and/or MIP inhibitors. In Section III, Dr. Fenton reviews apoptotic pathways as they relate to MM therapy. Defects in the mitochrondrial intrinsic pathway result from imbalances in expression levels of Bcl-2, Bcl-XL and Mcl-1. Mcl-1 is a candidate target gene for rapid induction of apoptosis by flavoperidol. Antisense oglionucleotides (ASO) lead to the rapid induction of caspace activity and apoptosis, which was potentiated by dexamethasone. Similar clinical trials with Bcl-2 ASO molecules alone and in combination with doxorubicin and dexamethasone or thalidomide showed promising results. The extrinsic pathway can be activated upon binding of the ligand TRAIL. OPG, released by osteoblasts and other stromal cells, can act as a decoy receptor for TRAIL, thereby blocking its apoptosis-inducing activity. MM cells inhibit OPG release by stromal cells, thereby promoting osteoclast activation and lytic bone disease (by enhancing RANKL availability) while at the same time exposing themselves to higher levels of ambient TRAIL. Thus, as a recurring theme, the relative levels of pro- versus anti-apoptotic molecules that act in a cell autonomous manner or in the milieu of the bone marrow microenvironment determine the outcome of potentially lethal signals. In Section IV, Dr. Barlogie and colleagues review data on single and tandem autotransplants for newly diagnosed myeloma. CR rates of 60%-70% can be reached with tandem transplants extending median survival to approximately 7 years. Dose adjustments of melphalan in the setting of renal failure and age > 70 may be required to reduce mucositis and other toxicities in such patients, especially in the context of amyloidosis with cardiac involvement. In Total Therapy II the Arkansas group is evaluating the role of added thalidomide in a randomized trial design. While data are still blinded as to the contribution of thalidomide, the overriding adverse importance of cytogenetic abnormalities, previously reported for Total Therapy I, also pertain to this successor trial. In these two-thirds of patients without cytogenetic abnormalities, Total Therapy II effected a doubling of the 4-year EFS estimate from 37% to 75% (P <.0001) and increased the 4-year OS estimate from 63% to 84% (P =.0009). The well-documented graft-vs-MM effect of allotransplants can be more safely examined in the context of non-myeloablative regimens, applied as consolidation after a single autologous transplant with melphalan 200 mg/m(2), have been found to be much better tolerated than standard myeloablative conditioning rege conditioning regimens and yielding promising results even in the high-risk entity of MM with cytogenetic abnormalities. For previously treated patients, the thalidomide congener Revimid and the proteasome inhibitor Velcade both are active in advanced and refractory MM (approximately 30% PR). Gene expression profiling (GEP) has unraveled distinct MM subtypes with different response and survival expectations, can distinguish the presence of or future development of bone disease, and, through serial investigations, can elucidate mechanisms of actions of new agents also in the context of the bone marrow microenvironment. By providing prognostically relevant distinction of MM subgroups, GEP should aid in the development of individualized treatment for MM.


Enzymatic Stability of Myostatin Inhibitory 16-mer Peptides.
Inhibition of myostatin is a promising strategy for treatment of muscle atrophic disorders. A 16-mer myostatin inhibitory linear peptide, MIPE-1686, administered intramuscularly, significantly increases muscle mass and hindlimb grip strength in Duchenne muscular dystrophic model mice. In this paper, we describe our examination of the enzymatic stabilities of this peptide with recombinant human proteases, aminopeptidase N, chymotrypsin C, and trypsin 3. MIPE-1686 was found to be stable in the presence of these enzymes, in contrast to a peptide (1), from which MIPE-1686 was developed. Modification of the peptides at a position distant from the protease cleavage site altered their enzymatic stability. These results suggest the possibility that the stability to proteases of 16-mer myostatin inhibitory peptides is associated with an increase in their known β-sheet formation properties. This study suggests that MIPE-1686 has a potential to serve as a long-lasting agent in vivo.


Treating Treatment-Resistant Patients with Panic Disorder and Agoraphobia Using Psychotherapy: A Randomized Controlled Switching Trial.
Background: Nonresponsiveness to therapy is generally acknowledged, but only a few studies have tested switching to psychotherapy. This study is one of the first to examine the malleability of treatment-resistant patients using acceptance and commitment therapy (ACT). Methods: This was a randomized controlled trial that included 43 patients diagnosed with primary panic disorder and/or agoraphobia (PD/A) with prior unsuccessful state-of-the-art treatment (mean number of previous sessions = 42.2). Patients were treated with an ACT manual administered by novice therapists and followed up for 6 months. They were randomized to immediate treatment (n = 33) or a 4-week waiting list (n = 10) with delayed treatment (n = 8). Treatment consisted of eight sessions, implemented twice weekly over 4 weeks. Primary outcomes were measured with the Panic and Agoraphobia Scale (PAS), the Clinical Global Impression (CGI), and the Mobility Inventory (MI). Results: At post-treatment, patients who received ACT reported significantly more improvements on the PAS and CGI (d = 0.72 and 0.89, respectively) than those who were on the waiting list, while improvement on the MI (d = 0.50) was nearly significant. Secondary outcomes were consistent with ACT theory. Follow-up assessments indicated a stable and continued improvement after treatment. The dropout rate was low (9%). Conclusions: Despite a clinically challenging sample and brief treatment administered by novice therapists, patients who received ACT reported significantly greater changes in functioning and symptomatology than those on the waiting list, with medium-to-large effect sizes that were maintained for at least 6 months. These proof-of-principle data suggest that ACT is a viable treatment option for treatment-resistant PD/A patients. Further work on switching to psychotherapy for nonresponders is clearly needed. © 2015 S. Karger AG, Basel.


Contributions of human liver cytochrome P450 enzymes to the N-oxidation of 4,4'-methylene-bis(2-chloroaniline).
4,4'-Methylene-bis(2-chloroaniline) (MOCA) can produce tumors in rodents and dogs and an increased incidence of bladder tumors has been reported in exposed workers. It is therefore of interest to identify the human cytochrome P450 (P450) enzymes involved in MOCA N-oxidation, the primary reaction involved in the formation of an electrophilic product. Human liver microsomes were fractionated and MOCA N-oxidation activity was monitored through the procedure. The most active enzyme fraction corresponded to P450 3A4, as determined by immunochemical assays and N-terminal amino acid sequence analysis. Yeast recombinant P450 3A4 also had MOCA N-oxidation activity. Purified human liver P450 2A6 showed catalytic activity; however, anti-P450 2A6 inhibited less than 20% of the microsomal activity while anti-P450 3A4 inhibited up to 75%. Levels of marker activities of both P450 3A4 (nifedipine oxidation) and P450 2A6 (coumarin 7-hydroxylation) were measured in a set of human liver microsomes and both were correlated with MOCA N-oxidation rates. Gestodene and troleandomycin inhibited up to half of the microsomal MOCA N-hydroxylation activity but 7,8-benzoflavone showed only slight inhibition. Anti-P450 3A4 inhibited (up to 80% of) the microsomal transformation of MOCA to a product genotoxic as judged by bacterial SOS response. The work indicates that P450 3A4 makes a major contribution to human liver microsomal MOCA N-oxidation, and P450 2A6 has a minor role. P450 1A2, which catalyzes the hydroxylation of many arylamines, does not contribute to a great extent.


p300/CBP-mediated p53 acetylation is commonly induced by p53-activating agents and inhibited by MDM2.
The tumor suppressor p53 is activated in response to many types of cellular and environmental insults via mechanisms involving post-translational modification. Here we demonstrate that, unlike phosphorylation, p53 invariably undergoes acetylation in cells exposed to a variety of stress-inducing agents including hypoxia, anti-metabolites, nuclear export inhibitor and actinomycin D treatment. In vivo, p53 acetylation is mediated by the p300 and CBP acetyltransferases. Overexpression of either p300 or CBP, but not an acetyltransferase-deficient mutant, efficiently induces specific p53 acetylation. In contrast, MDM2, a negative regulator of p53, actively suppresses p300/CBP-mediated p53 acetylation in vivo and in vitro. This inhibitory activity of MDM2 on p53 acetylation is in turn abrogated by tumor suppressor p19(ARF), indicating that regulation of acetylation is a central target of the p53-MDM2-p19(ARF) feedback loop. Functionally, inhibition of deacetylation promotes p53 stability, suggesting that acetylation plays a positive role in the accumulation of p53 protein in stress response. Our results provide evidence that p300/CBP-mediated acetylation may be a universal and critical modification for p53 function.


Moments of transfer of stimulus control in practical assembly tasks by mentally retarded adults.
Twenty vocational rehabilitation clients, divided into two aptitude groups, were trained to assemble three different 10-part apparatuses (lawn mower engine, electric drill, and bicycle brake) using three progressive prompt delay intervals (1, 3, and 5 seconds). The experimenter modeled selection and placement of each part on Trial 1 (0-second delay). On Trial 2, the modeling prompt was delayed 1, 3, or 5 seconds. On subsequent trials, a correct response increased the delay interval while an incorrect response shortened the delay interval. The 1-second delay condition produced fewest errors, most trials with no errors, and earliest acquisition. The patterns of the "moments of transfer" (shift from prompt dependent to independent responding) were similar for the high- and low-aptitude groups.


Episiotomy in modern obstetrics--necessity versus malpractice.
Episiotomy is now one of the most common procedures performed in obstetrics. At the beginning of its existence, it was performed very carefully and used in exceptional circumstances. In the second half of the twentieth century, its use became so widespread that it was almost regarded as a standard procedure in labor rooms. Authors intend to provide answers to the question as to whether it was an appropriate move in this discussion. Undoubtedly, there are reasons for which an incision is an appropriate decision, sometimes necessary, but in recent years its usefulness and relevance, in particular its routine, too widespread use are starting to be increasingly questioned and subjected to doubt, both by various women's organizations, individuals interested in the issue, stakeholders, as well as professionals. Poland is still one of the few European countries where routine episiotomy is so far regarded as an important and recognized part of patient management during almost every childbirth taking place in a hospital setting. This topic currently causes broad discussion in the media, the press and among the public. Hence, the aim of this work is to discuss key issues on episiotomy, the arguments for and against episiotomy based on literature review and available studies and reports. It is also going to present the opinion of different authors and the existing differences in their views on the above issue.


Potential molecular pathways of angiotensin receptor blockers in the brain toward cognitive improvement in dementia.
The alarming rise of cognitive impairment and memory decline and limited effective solutions present a worldwide concern for dementia patients. The multivariant role of the renin-angiotensin system (RAS) in the brain offers strong evidence of a role for angiotensin receptor blockers (ARBs) in the management of memory impairment by modifying glutamate excitotoxicity, downregulating inflammatory cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)α, inhibiting kynurenine aminotransferase (KAT)-II, nucleotide-binding domain, leucine-rich-containing family and pyrin-domain-containing-3 (NLRP3) inflammasomes, boosting cholinergic activity, activating peroxisome proliferator-activated receptor (PPAR)-γ, countering cyclooxygenase (COX) and mitigating the hypoxic condition. The present work focuses on the intricate molecular mechanisms involved in brain-RAS, highlighting the role of ARBs, connecting links between evidence-based unexplored pathways and investigating probable biomarkers involved in dementia through supported preclinical and clinical literature.


Cellular measurements on early and advanced squamous cell carcinoma of the esophagus in Northern China.
Cellular measurements and microstructural changes are reported in 68 proven esophageal squamous cell carcinomas including 29 cases (1,100 cells measured) of early cancer (in situ or early invasive) and 39 cases (1,170 cells measured) of advanced cancer. The cytoplasmic diameters were 20.72 +/- 0.20 micrometers in the early cancers and 22.98 +/- 0.28 micrometers in the advanced cancers (P less than 0.001). The nuclear diameters were 13.52 +/- 0.09 micrometers (early) and 13.78 +/- 0.1 micrometers (advanced) (P less than 0.05). An increase in the amount of cytoplasm was the predominant variant in cellular size with progression of the disease. Irregularities of the nuclear membrane were not significantly different between cells of the two groups. The number of nuclei and nucleoli was increased in advanced cancer. In 31.72% of the early cancer cells, the nucleoli were indistinct. The largest nucleoli were 6.75 micrometers in diameter. Overall, the most common chromatin pattern was the reticulated; however, the percentage of clumped, coarse granular and combined (coarse granular and clumped) chromatin patterns increased as the disease progressed (P less than 0.001).


Induction of apoptosis in leukemic cells by homovanillic acid derivative, capsaicin, through oxidative stress: implication of phosphorylation of p53 at Ser-15 residue by reactive oxygen species.
Capsaicin (N-vanillyl-8-methyl-1-nonenamide) is a homovanillic acid derivative found in pungent fruits. Several investigators have reported the ability of capsaicin to inhibit events associated with the promotion of cancer. However, the effects of capsaicin on human leukemic cells have never been investigated. We investigated the effects of capsaicin on leukemic cells in vitro and in vivo and further examined the molecular mechanisms of capsaicin-induced apoptosis in myeloid leukemic cells. Capsaicin suppressed the growth of leukemic cells, but not normal bone marrow mononuclear cells, via induction of G(0)-G(1) phase cell cycle arrest and apoptosis. Capsaicin-induced apoptosis was in association with the elevation of intracellular reactive oxygen species production. Interestingly, capsaicin-sensitive leukemic cells were possessed of wild-type p53, resulting in the phosphorylation of p53 at the Ser-15 residue by the treatment of capsaicin. Abrogation of p53 expression by the antisense oligonucleotides significantly attenuated capsaicin-induced cell cycle arrest and apoptosis. Pretreatment with the antioxidant N-acetyl-L-cystein and catalase, but not superoxide dismutase, completely inhibited capsaicin-induced apoptosis by inhibiting phosphorylation of Ser-15 residue of p53. Moreover, capsaicin effectively inhibited tumor growth and induced apoptosis in vivo using NOD/SCID mice with no toxic effects. We conclude that capsaicin has potential as a novel therapeutic agent for the treatment of leukemia.


Elucidation of Mechanisms and Selectivities of Metal-Catalyzed Reactions using Quantum Chemical Methodology.
Quantum chemical techniques today are indispensable for the detailed mechanistic understanding of catalytic reactions. The development of modern density functional theory approaches combined with the enormous growth in computer power have made it possible to treat quite large systems at a reasonable level of accuracy. Accordingly, quantum chemistry has been applied extensively to a wide variety of catalytic systems. A huge number of problems have been solved successfully, and vast amounts of chemical insights have been gained. In this Account, we summarize some of our recent work in this field. A number of examples concerned with transition metal-catalyzed reactions are selected, with emphasis on reactions with various kinds of selectivities. The discussed cases are (1) copper-catalyzed C-H bond amidation of indoles, (2) iridium-catalyzed C(sp(3))-H borylation of chlorosilanes, (3) vanadium-catalyzed Meyer-Schuster rearrangement and its combination with aldol- and Mannich-type additions, (4) palladium-catalyzed propargylic substitution with phosphorus nucleophiles, (5) rhodium-catalyzed 1:2 coupling of aldehydes and allenes, and finally (6) copper-catalyzed coupling of nitrones and alkynes to produce β-lactams (Kinugasa reaction). First, the methodology adopted in these studies is presented briefly. The electronic structure method in the great majority of these kinds of mechanistic investigations has for the last two decades been based on density functional theory. In the cases discussed here, mainly the B3LYP functional has been employed in conjunction with Grimme's empirical dispersion correction, which has been shown to improve the calculated energies significantly. The effect of the surrounding solvent is described by implicit solvation techniques, and the thermochemical corrections are included using the rigid-rotor harmonic oscillator approximation. The reviewed examples are chosen to illustrate the usefulness and versatility of the adopted methodology in solving complex problems and proposing new detailed reaction mechanisms that rationalize the experimental findings. For each of the considered reactions, a consistent mechanism is presented, the experimentally observed selectivities are reproduced, and their sources are identified. Reproducing selectivities requires high accuracy in computing relative transition state energies. As demonstrated by the results summarized in this Account, this accuracy is possible with the use of the presented methodology, benefiting of course from a large extent of cancellation of systematic errors. It is argued that as the employed models become larger, the number of rotamers and isomers that have to be considered for every stationary point increases and a careful assessment of their energies is therefore necessary in order to ensure that the lowest energy conformation is located. This issue constitutes a bottleneck of the investigation in some cases and is particularly important when analyzing selectivities, since small energy differences need to be reproduced.


Functional interaction between the cytoplasmic leucine-zipper domain of HIV-1 gp41 and p115-RhoGEF.
The long cytoplasmic tail of the human immunodeficiency virus (HIV)-1 transmembrane protein gp41 (gp41C) is implicated in the replication and cytopathicity of HIV-1 [1]. Little is known about the specific functions of gp41C, however. HIV-1 or simian immunodeficiency virus (SIV) mutants with defective gp41C have cell-type- or species-dependent phenotypes [2] [3] [4] [5] [6]. Thus, host factors are implicated in mediating the functions of gp41C. We report here that gp41C interacted with the carboxy-terminal regulatory domain of p115-RhoGEF [7], a specific guanine nucleotide exchange factor (GEF) and activator of the RhoA GTPase, which regulates actin stress fiber formation, activation of serum response factor (SRF) and cell proliferation [8] [9]. We demonstrate that gp41C inhibited p115-mediated actin stress fiber formation and activation of SRF. An amphipathic helix region with a leucine-zipper motif in gp41C is involved in its interaction with p115. Mutations in gp41C leading to loss of interaction with p115 impaired HIV-1 replication in human T cells. These findings suggest that an important function of gp41C is to modulate the activity of p115-RhoGEF and they thus reveal a new potential anti-HIV-1 target.


Hung out to dry: choice of priority ecoregions for conserving threatened neotropical anurans depends on life-history traits.
In the Neotropics, nearly 35% of amphibian species are threatened by habitat loss, habitat fragmentation, and habitat split; anuran species with different developmental modes respond to habitat disturbance in different ways. This entails broad-scale strategies for conserving biodiversity and advocates for the identification of high conservation-value regions that are significant in a global or continental context and that could underpin more detailed conservation assessments towards such areas. We identified key ecoregion sets for anuran conservation using an algorithm that favors complementarity (beta-diversity) among ecoregions. Using the WWF's Wildfinder database, which encompasses 700 threatened anuran species in 119 Neotropical ecoregions, we separated species into those with aquatic larvae (AL) or terrestrial development (TD), as this life-history trait affects their response to habitat disturbance. The conservation target of 100% of species representation was attained with a set of 66 ecoregions. Among these, 30 were classified as priority both for species with AL and TD, 26 were priority exclusively for species with AL, and 10 for species with TD only. Priority ecoregions for both developmental modes are concentrated in the Andes and in Mesoamerica. Ecoregions important for conserving species with AL are widely distributed across the Neotropics. When anuran life histories were ignored, species with AL were always underrepresented in priority sets. The inclusion of anuran developmental modes in prioritization analyses resulted in more comprehensive coverage of priority ecoregions-especially those essential for species that require an aquatic habitat for their reproduction-when compared to usual analyses that do not consider this life-history trait. This is the first appraisal of the most important regions for conservation of threatened Neotropical anurans. It is also a first endeavor including anuran life-history traits in priority area-selection for conservation, with a clear gain in comprehensiveness of the selection process.


Carbon-Restricted Anoxic Zone as an Overlooked Anammox Hotspot in Municipal Wastewater Treatment Plants.
The anoxic zone serves as the core functional unit in municipal wastewater treatment plants (MWWTPs). Unfortunately, in most cases, the downstream range of the anoxic zone is severely lacking in available organic carbon and thus contributes little to the removal of nutrients. This undesirable range is termed the "carbon-restricted anoxic zone", representing an insurmountable drawback for traditional MWWTPs. This study uncovers a previously overlooked role for the carbon-restricted anoxic zone: a hotspot for anaerobic ammonium oxidation (anammox). In a continuous-flow pilot-scale plant treating municipal wastewater (55 m3/d), virgin biocarriers were introduced into the carbon-restricted anoxic zone (downstream 25% of the anoxic zone with BOD5 of 5.9 ± 2.3 mg/L). During the 517-day monitoring, anammox bacteria highly self-enriched within the biofilms, with absolute and relative abundance reaching up to (9.4 ± 0.1) × 109 copies/g-VSS and 6.17% (Candidatus Brocadia), respectively. 15N isotopic tracing confirmed that anammox overwhelmingly dominated nitrogen metabolism, responsible for 92.5% of nitrogen removal. Following this upgrade, the contribution ratio of the carbon-restricted anoxic zone to total nitrogen removal increased from 9.2 ± 4.1% to 19.2 ± 4.2% (P < 0.001), while its N2O emission flux decreased by 84.5% (P < 0.001). These findings challenge stereotypes about the carbon-restricted anoxic zone and highlight the multiple environmental implications of this newfound anammox hotspot.


Matching ICD-11 personality status to clinical management in a community team-The Boston (UK) Personality Project: Study protocol.
Epidemiological studies show 30% to 50% of all patients in community mental health teams have personality disorders. These are normally comorbid with other psychiatric disorders, often as Galenic syndromes, and are seldom identified. In the Boston (UK) Personality Project all patients under a community health service in Boston in Lincolnshire will be asked to agree to have their personality status assessed using scales recording the new ICD-11 classification, together with clinical ratings, social function and satisfaction. A control group of 100 patients from an adjacent service of similar demographics (Spalding) will also have similar ratings but no personality assessments. Changes in clinical status, social function and service satisfaction will be made after 6 and 12 months in both groups. The patients in the Boston group will be offered matched interventions using a stepped care approach for both the severity of disorder and its domain structure. These interventions will include shorter versions of existing psychological treatments, environmental therapies including nidotherapy, adaptive and acceptance models, drug reduction and social prescribing. Full costs of psychiatric care will be measured in both groups. The main hypothesis is that greater awareness of personality function will lead to better clinical outcomes and satisfaction.


Clostridium frigidicarnis sp. nov., a psychrotolerant bacterium associated with 'blown pack' spoilage of vacuum-packed meats.
Two strains of a psychrotolerant Clostridium, isolated from vacuum-packed, temperature-abused beef, were characterized using a multiphasic approach. The strains were Gram-positive motile rods producing elliptical subterminal spores during early stationary growth phase. The strains were psychrotolerant. At pH 7.0, they grew between 3.8 and 40.5 degrees C; their optimum growth temperature was 30.0-38.5 degrees C. At 30 degrees C, the pH range for growth was between 4.7 and 9.5; the optimum pH for growth was 6.4-7.2. The organisms were proteolytic and saccharolytic, lecithinase-positive and hydrolysed gelatin. The fermentation products formed in peptone/yeast extract/glucose/starch broth were acetate, ethanol, butyrate, isovalerate, butanol, isobutyrate, oxalacetate, lactate, hydrogen and carbon dioxide. The DNA G + C compositions of the two meat strains were 27.3 and 28.4 mol%. Phylogenetic analyses indicated that the strains belong to Cluster I of the genus Clostridium (sensu Collins et al., 1994). The new strains differed from phylogenetically related clostridia in terms of cellular fatty acid composition, soluble protein profiles and phenotypic properties. On the basis of phenotypic and genotypic characterization data, the strains were assigned to a new species for which the name Clostridium frigidicarnis is proposed; strain SPL77AT (= DSM 12271T) is the type strain.


DNABERT-based explainable lncRNA identification in plant genome assemblies.
Long non-coding ribonucleic acids (lncRNAs) have been shown to play an important role in plant gene regulation, involving both epigenetic and transcript regulation. LncRNAs are transcripts longer than 200 nucleotides that are not translated into functional proteins but can be translated into small peptides. Machine learning models have predominantly used transcriptome data with manually defined features to detect lncRNAs, however, they often underrepresent the abundance of lncRNAs and can be biased in their detection. Here we present a study using Natural Language Processing (NLP) models to identify plant lncRNAs from genomic sequences rather than transcriptomic data. The NLP models were trained to predict lncRNAs for seven model and crop species (Zea mays, Arabidopsis thaliana, Brassica napus, Brassica oleracea, Brassica rapa, Glycine max and Oryza sativa) using publicly available genomic references. We demonstrated that lncRNAs can be accurately predicted from genomic sequences with the highest accuracy of 83.4% for Z. mays and the lowest accuracy of 57.9% for B. rapa, revealing that genome assembly quality might affect the accuracy of lncRNA identification. Furthermore, we demonstrated the potential of using NLP models for cross-species prediction with an average of 63.1% accuracy using target species not previously seen by the model. As more species are incorporated into the training datasets, we expect the accuracy to increase, becoming a more reliable tool for uncovering novel lncRNAs. Finally, we show that the models can be interpreted using explainable artificial intelligence to identify motifs important to lncRNA prediction and that these motifs frequently flanked the lncRNA sequence.


Use of Folk Therapy in Taiwan: A Nationwide Cross-Sectional Survey of Prevalence and Associated Factors.
Background. This study investigates the prevalence of and factors associated with users of folk therapy in Taiwan. Methods. Using data from the 2005 National Health Interview Survey and the National Health Insurance Research Database, we identified 16,750 adults aged 20 years and older. Sociodemographic factors, lifestyle, medical utilization, and health behaviors were compared between people using and not using folk therapy. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) of factors associated with folk therapy were analyzed. Results. The one-month prevalence of folk therapy use was 6.8%, which was significantly associated with ages of 30-59 years (OR = 1.98, 95% CI = 1.49-2.63), women (OR = 1.63, 95% CI = 1.40-1.90), nonindigenous population (OR = 1.90, 95% CI = 1.14-3.17), having two or more unhealthy lifestyle habits (OR = 1.51, 95% CI = 1.26-1.81), high density of traditional Chinese medicine (TCM) physicians (OR = 1.40, 95% CI = 1.20-1.62), and being ill without receiving medical care in past six months (OR = 2.11, 95% CI = 1.76-2.53). Medical care utilization of TCM and Western medicine were also associated factors for folk therapy. Conclusions. The use of folk therapy is correlated with sociodemographics, lifestyle and health behaviors.


Thyroid, gastric, and adrenal autoimmunities associated with insulin-dependent diabetes mellitus.
Thyroid microsomal, gastric parietal, and adrenocortical autoantibodies were sought in 1456 Caucasian and 240 black patients with insulin-dependent diabetes mellitus (IDDM), in 1467 of the Caucasian patients' immediate family members, and in 1519 normal Caucasian control subjects. Positive clinical significances and predictive values of these autoantibodies for associated gland dysfunctions were found. In all groups, thyroid and gastric autoantibodies were more common in female subjects. In the control group, thyroid and gastric autoantibodies were more frequent with advancing age, affecting more than one-third of Caucasian women after age 60 yr. This age-augmented increase occurred very prematurely in patients with IDDM and their relatives. Among the patients with IDDM and thyroid and gastric autoantibodies, these autoantibodies appeared by the time of onset of IDDM in the large majority of cases; however, they were more common among patients with later ages of onset of IDDM. Occurrence of thyroid and gastric autoantibodies in 404 siblings of patients with IDDM was not affected by their degree of HLA-haplotype sharing with their diabetic sibling, suggesting that the inherited predisposition to thyroid and gastric autoimmunity is not HLA-related. In light of this, the increased frequencies of these antibodies in patients with IDDM and their relatives suggest that "thyroid and gastric autoimmunity genes" may also predispose to IDDM.


Faecal Particle Size in Free-Ranging Proboscis Monkeys, Nasalis larvatus: Variation between Seasons.
Reducing the size of food particles is crucial for herbivores. Seasonal dietary changes are known to influence animals' chewing efficiency. Proboscis monkeys (Nasalis larvatus) are foregut fermenters, with a high chewing efficiency allowing them to achieve very fine faecal particles. In this study, we investigated how proboscis monkeys' chewing efficiency varies between wet and dry seasons, hypothesising differences possibly related to diet change. Faecal particle size analysis is an established approach to estimate chewing efficiency in mammalian herbivores. We analysed 113 proboscis monkey faecal samples collected in the Lower Kinabatangan Wildlife Sanctuary, between 2015 and 2017. By following standard sieve analysis protocols, we measured a mean particle size MPS0.025-8 of 0.45 ± 0.14 mm, and confirmed a previous result that proboscis monkeys have a very low faecal MPS. This study highlights a seasonal influence on proboscis monkeys' chewing efficiency, with smaller MPS (better chewing efficiency) during the wet season. During that time of the year, individuals may potentially change their diet, as all faecal samples contained intact seeds. Whether the seasonal MPS difference in proboscis monkeys is smaller than in other colobines due to their "rumination" strategy remains to be investigated.


Unusual mode of firearm injury from the recoiled rear end of a gun barrel.
Atypical gunshot wounds are caused by a diverse set of parameters relating to weapons and ammunition. We report a previously-unreported and atypical mode of gunshot wound produced by a detached rear end of the barrel of a gun following accidental gun fire, and discuss the difficulties in the management. A 36-year-old man presented to the emergency department with an alleged history of injury on the forehead with the rear end of a gun barrel following accidental gunfire while cleaning the nozzle. Since the time of injury, the patient was in an altered sensorium and had weakness on the right side of the body. There was minimal but continuous bleeding from the wound, with extrusion of brain matter. Skull radiograph showed that the rear end of the barrel had entered the left frontal bone, with associated depressed fracture of the frontal bone. The patient underwent a bicoronal, bifrontal craniotomy with a T-shaped extension towards the barrel to facilitate the reflection of the scalp flap and to avoid any movement of the barrel as it might further injure the brain. Necrotic brain, dura and bone pieces were removed. The patient was doing well at follow-up except for mild residual motor deficits. This case illustrates that while working with limited facilities, particularly in underdeveloped countries, a careful clinical assessment, interpretation of available images and a judicious operative approach can help to save the patient.


Reduced flavins promote oxidative DNA damage in non-respiring Escherichia coli by delivering electrons to intracellular free iron.
When cells are exposed to external H(2)O(2), the H(2)O(2) rapidly diffuses inside and oxidizes ferrous iron, thereby forming hydroxyl radicals that damage DNA. Thus the process of oxidative DNA damage requires only H(2)O(2), free iron, and an as-yet unidentified electron donor that reduces ferric iron to the ferrous state. Previous work showed that H(2)O(2) kills Escherichia coli especially rapidly when respiration is inhibited either by cyanide or by genetic defects in respiratory enzymes. In this study we established that these respiratory blocks accelerate the rate of DNA damage. The respiratory blocks did not substantially affect the amounts of intracellular free iron or H(2)O(2), indicating that that they accelerated damage because they increased the availability of the electron donor. The goal of this work was to identify that donor. As expected, the respiratory inhibitors caused a large increase in the amount of intracellular NADH. However, NADH itself was a poor reductant of free iron in vitro. This suggests that in non-respiring cells electrons are transferred from NADH to another carrier that directly reduces the iron. Genetic manipulations of the amounts of intracellular glutathione, NADPH, alpha-ketoacids, ferredoxin, and thioredoxin indicated that none of these was the direct electron donor. However, cells were protected from cyanide-stimulated DNA damage if they lacked flavin reductase, an enzyme that transfers electrons from NADH to free FAD. The K(m) value of this enzyme for NADH is much higher than the usual intracellular NADH concentration, which explains why its flux increased when NADH levels rose during respiratory inhibition. Flavins that were reduced by purified flavin reductase rapidly transferred electrons to free iron and drove a DNA-damaging Fenton system in vitro. Thus the rate of oxidative DNA damage can be limited by the rate at which electron donors reduce free iron, and reduced flavins become the predominant donors in E. coli when respiration is blocked. It remains unclear whether flavins or other reductants drive Fenton chemistry in respiring cells.


Protein induced by vitamin K absence or antagonist II: Experience to date and future directions.
Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer with high mortality. The realization of precision medicine in HCC relies upon efficient biomarkers. Protein induced by vitamin K absence or antagonist II (PIVKA-II) is an immature prothrombin with insufficient coagulation activity, overexpressing in HCC cells. Previous evidence confirmed the role of PIVKA-II in screening and diagnosing HCC. However, the increased PIVKA-II was observed not only in HCC, but also in non-HCC individuals such as vitamin K deficiency. The joint detection of PIVKA-II and other biomarkers could significantly improve diagnostic accuracy in HCC. Furthermore, PIVKA-II serves as a valuable prognostic predictor, transplantation eligibility, resectability, tumor recurrence, therapeutic efficacy, and malignant tumor behaviors. Additionally, PIVKA-II represents a potential target for agent development to establish new therapeutic strategies. Besides HCC, PIVKA-II also serves as a biomarker of vitamin K status. In this review, we assess the role of PIVKA-II in diagnosis, prediction, and treatment. Over the past decades, substantial progress has been achieved in the application of PIVKA-II. Exploration and innovation are required for further advances in the field of PIVKA-II investigation.


Potential of UV-B radiation in drought stress resilience: A multidimensional approach to plant adaptation and future implications.
The escalating impact of climate change and ultraviolet (UV) radiation is subjecting plants to unique combinations of UV-B and drought stress. These combined stressors could have additive, synergistic, or antagonistic effects, but the precise nature of these impacts remains uncertain, hampering our ability to predict plant adaptations approach towards stressors. Our analysis of various studies shows that UV-B or drought conditions detrimentally influence plant growth and health metrics by the enhanced generation of reactive oxygen species causing damage to lipids, proteins, carbohydrates and DNA. Further reducing biomass accumulation, plant height, photosynthetic efficiency, leaf area, and water transpiration, while enhancing stress-related symptoms. In response to UV-B radiation and drought stress, plants exhibit a notable up-regulation of specific acclimation-associated metabolites, including proline, flavonoids, anthocyanins, unsaturated fatty acids, and antioxidants. These metabolites play a pivotal role in conferring protection against environmental stresses. Their biosynthesis and functional roles are potentially modulated by signalling molecules such as hydrogen peroxide, abscisic acid, jasmonic acid, salicylic acid, and ethylene, all of which have associated genetic markers that further elucidate their involvement in stress response pathways. In comparison to single stress, the combination of UV-B and drought induces the plant defence responses and growth retardation which are less-than-additive. This sub-additive response, consistent across different study environments, suggests the possibility of a cross-resistance mechanism. Our outlines imply that the adverse effects of increased drought and UV-B could potentially be mitigated by cross-talk between UV-B and drought regimes utilizing a multidimensional approach. This crucial insight could contribute significantly to refining our understanding of stress tolerance in the face of ongoing global climate change.


Immunocytochemical and autoradiographic localization of GABAergic neurons in the goldfish retina.
The putative neurotransmitter gamma-aminobutyric acid (GABA) was localized in goldfish retina by using an antiserum directed against GABA itself. The same types of cells were stained with this antibody as were labelled with an antiserum directed against the synthesizing enzyme for GABA, glutamic acid decarboxylase. Stained neurites of these cells were located throughout the inner plexiform layer (IPL) but staining was more intense in the proximal IPL. The GABA-immunoreactive staining could be reduced or completely abolished by preabsorbing the primary antibody with GABA. Uptake of [3H]-GABA or the GABA agonist [3H]-muscimol was localized in GABA-stained retinas using light microscope autoradiography. These experiments demonstrated that all types of GABA-immunoreactive amacrine cells had high-affinity uptake mechanisms for both [3H]-GABA and -muscimol. Thirty percent of proximal inner nuclear layer (INL) and some cells in the ganglion cell layer (GCL) were labelled by all three GABAergic markers. Most GABA-immunoreactive amacrine cells were lightly labelled due to [3H]-GABA uptake but a few amacrines (Ab) were heavily labelled. These findings demonstrate that the autoradiographic localization of [3H]-GABA or [3H]-muscimol uptake and the immunocytochemical localization of GAD or GABA are appropriate methods for localizing GABAergic neurons in the retina. Few GABA-immunoreactive amacrine cells accumulated the putative amino acid transmitter [3H]-glycine, verifying that the goldfish retina contains distinct subpopulations of glycinergic and GABAergic amacrine cells.


The large-scale cultivation of diploid cell strains in microcarrier culture. Improvement of microcarriers.
The use of diploid cell strains for virus vaccine production is mostly retarded by the inefficient culture techniques used for the propagation of these cells. The microcarrier culture system might be a good solution for this problem. DEAE-gels appear to be the most satisfactory microcarrier. The best results are obtained by using DEAE-gels with a rather low anionic exchange capacity or at higher capacity after coating with polymers.


Ultrafine Mo2C nanoparticles embedded in an MOF derived N and P co-doped carbon matrix for an efficient electrocatalytic oxygen reduction reaction in zinc-air batteries.
Exploring high-activity electrocatalysts for an oxygen reduction reaction (ORR) is of great significance for a variety of renewable energy conversion and storage technologies. Here, ultrafine Mo2C nanoparticles assembled in N and P-co-doped carbon (Mo2C@NPC) was developed from ZIF-8 encapsulated molybdenum-based polyoxometalates (PMo12) as a highly efficient ORR electrocatalyst and shows excellent performance for zinc-air batteries. The well distribution of the PMo12 in ZIF-8 results in the formation of ultrafine Mo2C nanocrystallites encapsulated in a porous carbon matrix after pyrolysis. Significantly, from experimental and theoretical investigations, the highly porous structure, highly dispersed ultrafine Mo2C and the N and P co-doping in the Mo2C@NPC lead to the remarkable ORR activity with an onset potential of ∼1.01 V, a half-wave potential of ∼0.90 V and a Tafel slope of 51.7 mV dec-1 at 1600 rpm in 0.1 M KOH. In addition, the Mo2C@NPC as an ORR catalyst in zinc-air batteries achieved a high power density of 266 mW cm-2 and a high specific capacity of 780.9 mA h g-1, exceeding that driven by commercial Pt/C. Our results revealed that the porous architecture and ultrafine Mo2C nanocrystallites of the electrocatalysts could facilitate mass transport and increase the accessibility of active sites, thus optimizing their performances in an ORR. The present study provides some guidelines for the design and synthesis of efficient nanostructured electrocatalysts.


Impact of Sublobar Resection on Pulmonary Function: Long-Term Results from American College of Surgeons Oncology Group Z4032 (Alliance).
Sublobar resection (SR) in high-risk operable patients may result in a long-term decrease in pulmonary function. We previously reported 3-month pulmonary function outcomes from a randomized phase III study of SR alone compared with SR with brachytherapy in patients with non-small cell lung cancer. We now report long-term pulmonary function after SR. Pulmonary function was measured at baseline and at 3, 12, and 24 months. A decline of 10% or more from baseline in the percentage predicted forced expiratory volume of 1 percentage or in the diffusion capacity of the lung for carbon monoxide was considered clinically meaningful. The effect of study arm, tumor location, size, approach (video-assisted thoracoscopic surgery vs thoracotomy), and SR type (wedge vs segmentectomy) on pulmonary function was assessed using a Wilcoxon rank sum test. A generalized estimating equation model was used to assess the effect of each factor on longitudinal data, including all four time points. Complete pulmonary function data at all time points was available in 69 patients. No significant differences were observed in pulmonary function between SR and SR with brachytherapy, thus the study arms were combined for all analyses. A decline of 10% or more (p = 0.02) in the percentage predicted forced expiratory volume in 1 second was demonstrated for lower-lobe resections at 3 months but was not at 12 or 24 months. A decline of 10% or more (p = 0.05) in the percentage predicted diffusion capacity of the lung for carbon monoxide was seen for thoracotomy at 3 months but was not at 12 or 24 months. Clinically meaningful declines in pulmonary function occurred after lower lobe resection and after thoracotomy at 3 months but subsequently recovered. This study suggests that SR does not result in sustained decreased pulmonary function in high-risk operable patients.


Hypothalamic alterations in Huntington's disease patients: comparison with genetic rodent models.
Unintended weight loss, sleep and circadian disturbances and autonomic dysfunction are prevalent features of Huntington's disease (HD), an autosomal dominantly inherited neurodegenerative disorder caused by an expanded CAG repeat sequence in the HTT gene. These features form a substantial contribution to disease burden in HD patients and appear to be accompanied by a number of neuroendocrine and metabolic changes, pointing towards hypothalamic pathology as a likely underlying mechanism. Neuronal inclusion bodies of mutant huntingtin, which are hallmarks of the disease, occur throughout the hypothalamus, and indicate local mutant huntingtin expression that could interfere with hypothalamic neuropeptide production. Also, several genetic rodent models of HD show features that could be related to hypothalamic pathology, such as weight loss and circadian rhythm disturbances. In these rodents, several hypothalamic neuropeptide populations are affected. In the present review, we summarise the changes in genetic rodent models of HD for individual hypothalamic nuclei, compare these observations to the hypothalamic changes that occur in HD patients, and make an inventory of the work that still needs to be done. Surprisingly, there is only limited overlap in the hypothalamic changes reported in HD patients and genetic rodent models. At present, the only similarity between the hypothalamic alterations in HD patients and genetic rodent models is a decrease in the number of orexin-expressing neurones in the lateral hypothalamus. Possible reasons for these discrepancies, as well as potential consequences for the development of novel therapeutic strategies, are discussed.


Will You Protect Me or Make the Situation Worse?: Teachers' Responses to School Violence Against Students With Disabilities.
School violence is a global public health epidemic, with students with disabilities at a significantly greater risk than their non-disabled peers. Students with disabilities are more vulnerable to school violence from peers, teachers, and school staff due to stereotypes and prejudice. Teachers are pivotal in preventing violence and intervening, but literature on the role that teachers play in responding to disability-based violence is limited. Guided by the social-ecological framework of bullying, this qualitative study explored educators' responses to school violence against students with disabilities in Zambia. Data generation included document review, interviews, and focus groups with 33 teachers and 12 parents, and child-friendly methods with 90 students with disabilities. Findings illuminated that students with disabilities are less safe in schools. Teachers are not responding to violence seen or heard about due to stigmatizing beliefs and cultural norms surrounding disability and violence, with students with disabilities blamed for the violence and the response being their burden. Students with disabilities felt protected by special education teachers; however, disability-based stigma did not end with the student. By association, special education teachers were experiencing stigma from other teachers and were discouraged to respond. This stigma undermined the support special education teachers could provide to decrease school violence. Findings provide direction so teachers can respond to school violence in prosocial ways that create an environment where students with disabilities feel safe.


Novel process of fermenting black soybean [Glycine max (L.) Merrill] yogurt with dramatically reduced flatulence-causing oligosaccharides but enriched soy phytoalexins.
Black soybeans [Glycine max (L.) Merrill] were germinated under fungal stress with food grade R. oligosporus for 3 days and were homogenized and fermented with lactic acid bacteria (LAB) to produce soy yogurt. Fungal stress led to the generation of oxylipins [oxooctadecadienoic acids (KODES) isomers and their respective glyceryl esters] and glyceollins--a type of phytoalexins unique to soybeans. In soy yogurt, the concentrations of total KODES and total glyceollins were 0.678 mg/g (dry matter) and 0.953 mg/g, respectively. The concentrations of other isoflavones (mainly genistein and daidzein and their derivatives) in soy yogurt remained largely unchanged after the processes compared with the control soy yogurt. Germination of black soybean under fungal stress for 3 days was sufficient to reduce stachyose and raffinose (which cause flatulence) by 92 and 80%, respectively. With a pH value of 4.42, a lactic acid content of 0.262%, and a maximum viable cell count of 2.1 x 10 (8) CFU/mL in the final soy yogurt, soy milk from germinated soybeans under fungal stress was concluded to be a suitable medium for yogurt-making. The resulting soy yogurt had significantly altered micronutrient profiles with significantly reduced oligosaccharides and enriched glyceollins.


Early and prolonged exposure to reward delay: effects on impulsive choice and alcohol self-administration in male rats.
Naturally occurring impulsive choice has been found to positively predict alcohol consumption in rats. However, the extent to which experimental manipulation of impulsive choice may modify alcohol consumption remains unclear. In the present study, we sought to: (a) train low levels of impulsive choice in rats using early, prolonged exposure to reward delay, and (b) determine the effects of this manipulation on subsequent alcohol consumption. During a prolonged training regimen, three groups of male, adolescent Long-Evans rats (21-22 days old at intake) responded on a single lever for food rewards delivered after either a progressively increasing delay, a fixed delay, or no delay. Posttests of impulsive choice were conducted, as was an evaluation of alcohol consumption using a limited-access, two-bottle test. Following delay-exposure training, both groups of delay-exposed rats made significantly fewer impulsive choices than did rats in the no-delay group. In addition, fixed-delay rats consumed significantly more alcohol during daily, 30-min sessions than no-delay rats. Possible mechanisms of these effects are discussed, as is the significance of these findings to nonhuman models of addiction.


Role of Intratympanic Dexamethasone for Intractable Posterior Canal Benign Paroxysmal Positional Vertigo.
Benign paroxysmal positional vertigo is a frequent diagnosed disorder, most of the patients are successfully treated with reposition maneuvers. In between 3-12.5% of these patients remain symptomatic. Recent studies support the use of intratympanic corticosteroid for intractable vertigo with promising results. Patients diagnosed with benign paroxysmal positional vertigo between June 2017 and December 2019 in a tertiary university hospital and in two private hospitals were included in the study and analyzed prospectively. They were treated and followed with repositioning maneuvers and intratympanic dexamethasone injections if the criteria was met. 4 out 72 patients included in the study developed criteria for intractable vertigo after at least 6 repositioning maneuvers. The posterior semicircular canal was affected in all cases, 3 out of 4 patients experienced symptom resolution, after two, four and five intratympanic dexamethasone injections respectively. The use of intratympanic steroids to treat patients with refractory benign paroxysmal positional vertigo showed encouraging results. We believe a multicenter randomized clinical trial should be performed to assess the efficacy of intratympanic steroids in the treatment of this pathology.


Seizure types, epilepsy syndromes, etiology, and diagnosis.
The clinical manifestation of epileptic seizures may vary widely from patient to patient, depending on the region of the brain involved. Over the centuries, many seizure classification systems have been used, and the current most widely used classification system is that of the International League Against Epilepsy (ILAE). The ILAE system divides seizures into those of partial onset and those of generalized onset, depending on whether the initial clinical manifestations indicate that one cortical region or both hemispheres are involved at the onset of the seizure. Partial seizures are then divided into simple partial seizures, in which a fully conscious state is retained, or complex partial seizures, in which consciousness is impaired. A more recent classification system based purely on symptom features and signs has been proposed, and this system may provide advantages for localization, and especially for surgical evaluation. Epilepsy is a condition characterized by recurrent unprovoked seizures. Epilepsy may be idiopathic, cryptogenic, or symptomatic. Idiopathic epilepsies are generally genetic, and while many such syndromes have been described, advances in molecular genetics will undoubtedly reveal many more syndromes in the near future. Cryptogenic epilepsies are those in which an underlying cause is suspected, but the etiology remains undetected. Epilepsies for which there is an underlying structural cause or major metabolic derangement are considered symptomatic. Common causes and diagnostic evaluation are described in this article.


[Characteristics of the responses of auditory cortex neurons in the cat to tonal stimulation during nembutal anesthesia and after recovery from it].
Responses of auditory (AI) cortex neurons to tonal (1-25 kHz) stimulation were studied in cats under and after nembutal anaesthesia. Patterns of responses recorded during first hours and 10-30 hours after nembutal injection differed essentially. Most (89%) of neurons of anaesthetized cat had no background activity and produced stereotypical on-responses to tonal stimulation with the best frequency. Off-effects were usually absent. Variability of responses increased substantially after cessation of anesthesia. On-, on-off- and off-responses were obvious in 76% of neurons, approximately 21% of neurons tonically increased or decreased spike activity during the stimulus action. Duration of the stimulus action was reflected in response characteristics of the overwhelming majority of AI cortical neurons.


Biochemical and immunological characterization of multiple glycoforms of mouse mast cell protease 1: comparison with an isolated murine serosal mast cell protease (MMCP-4).
Five highly soluble, chymotrypsin-like, neutral serine proteases, with molecular masses in the range 30-33 kDa, were isolated from Trichinella spiralis-infected mouse small intestine. These enzymes were closely related antigenically on Western blotting and by Ouchterlony double diffusion using a polyclonal, cross-absorbed, sheep antibody raised against mouse mast cell protease-1 (MMCP-1) and on the basis of N-terminal amino acid sequence analysis, were identified as variant forms of MMCP-1. Substrate and inhibitor analysis confirmed that the five variants (MMCP-1 A-E) had similar characteristics, although highly significant (P = 0.025 to P < 0.0001) variations in Km and kcat, were detected. Against human alpha 1-proteinase inhibitor the Ki for MMCP-1C (45 pM) was significantly (P < 0.0001) greater than those for the other proteases (0.76-2.2 pM). The differences in electrophoretic mobility are probably a result of variable glycosylation, since removal of N-linked carbohydrate produced a polypeptide of approx. 28 kDa in each case which was, like the native enzyme, immunoreactive on Western blotting. A much less soluble 28 kDa enzyme was isolated from serosal mast cells and identified as MMCP-4 by N-terminal amino acid sequencing. Like MMCP-1 it has chymotrypsin-like substrate specificities with activity at neutral pH. However, it was antigenically distinct from MMCP-1 and, using sheep anti-MMCP-1, was not detected on Western blotting or by Ouchterlony double diffusion, e.l.i.s.a. or immunohistochemistry. This last technique established that the MMCP-1 variants were uniquely present in enteric mast cells, thereby providing a highly selective means of distinguishing the mucosal and connective tissue mast cell subsets in the mouse.