Supplementation with (6S)-5-methyltetrahydrofolic acid appears as effective as folic acid in maintaining maternal folate status while reducing unmetabolised folic acid in maternal plasma: a randomised trial of pregnant women in Canada.
Folic acid supplementation is recommended during pregnancy to support healthy fetal development; (6S)-5-methyltetrahydrofolic acid ((6S)-5-MTHF) is available in some commercial prenatal vitamins as an alternative to folic acid, but its effect on blood folate status during pregnancy is unknown. To address this, we randomised sixty pregnant individuals at 8-21 weeks' gestation to 0·6 mg/d folic acid or (6S)-5-MTHF × 16 weeks. Fasting blood specimens were collected at baseline and after 16 weeks (endline). Erythrocyte and serum folate were quantified via microbiological assay (as globally recommended) and plasma unmetabolised folic acid (UMFA) via LC-MS/MS. Differences in biochemical folate markers between groups were explored using multivariable linear/quantile regression, adjusting for baseline concentrations, dietary folate intake and gestational weeks. At endline (n 54), the mean values and standard deviations (or median, inter-quartile range) of erythrocyte folate, serum folate and plasma UMFA (nmol/l) in those supplemented with (6S)-5-MTHF v. folic acid, respectively, were 1826 (sd 471) and 1998 (sd 421); 70 (sd 13) and 78 (sd 17); 0·5 (0·4, 0·8) and 1·3 (0·9, 2·1). In regression analyses, erythrocyte and serum folate did not differ by treatment group; however, concentrations of plasma UMFA in pregnancy were 0·6 nmol/l higher (95 % CI 0·2, 1·1) in those supplementing with folic acid as compared with (6S)-5-MTHF. In conclusion, supplementation with (6S)-5-MTHF may reduce plasma UMFA by ∼50 % as compared with supplementation with folic acid, the biological relevance of which is unclear. As folate is currently available for purchase in both forms, the impact of circulating maternal UMFA on perinatal outcomes needs to be determined.


Conditional senescence in Chlamydomonas reinhardtii (Chlorophyceae).
The mechanisms of microalgal senescence may play an important role in nutrient recycling and enhanced survival. However, the aging physiology of microalgae is an understudied phenomenon. To investigate the patterns of conditional senescence in Chlamydomonas reinhardtii P. A. Dangeard, we used a cell wall-less strain, transformed with a reporter gene to infer changes in photosynthetic gene expression. We examined plastid ultrastructure, photosynthetic function, and photoprotective mechanisms during aging in batch cultures. LHCII transcription levels decreased before the population entered stationary phase, and the characteristic transcriptional light-shift response was lost. A decline in photosynthetic proteins with a concomitant increase in the photoprotective protein, LHCSR, was observed over time. However, nonphotochemical quenching remained stable during growth and stationary phase, and then declined as alternative quenching mechanisms were up-regulated. Photosynthetic efficiency declined, while Fv/Fm remained stable until the death phases. As the culture progressed through stationary phase, disorganization of the chloroplast was observed along with an increase in cytoplasmic oil bodies. We also observed a partial recovery of function and proteins during the final death phase, and attribute this to the release of nutrients into the medium from cell lysis and/or active secretion while cells were senescing. Allowing open gas exchange resulted in high levels of sustained starch production and maintained maximum cell density, prolonging the stationary phase.


A Public Health Response to a Mudslide in Freetown, Sierra Leone, 2017: Lessons Learnt.
On August 14, 2017, a 6-kilometer mudslide occurred in Regent Area, Western Area District of Sierra Leone following a torrential downpour that lasted 3 days. More than 300 houses along River Juba were submerged; 1141 people were reported dead or missing and 5905 displaced. In response to the mudslide, the World Health Organization (WHO) Country Office in Sierra Leone moved swiftly to verify the emergency and constitute an incident management team to coordinate the response. Early contact was made with the Ministry of Health and Sanitation and health sector partners. A Public Health Emergency Operations Center was set up to coordinate the response. Joint assessments, planning, and response among health sector partners ensured effectiveness and efficiency. Oral cholera vaccination was administered to high-risk populations to prevent a cholera outbreak. Surveillance for 4 waterborne diseases was enhanced through daily reporting from 9 health facilities serving the affected population. Performance standards from the WHO Emergency Response Framework were used to monitor the emergency response. An assessment of the country's performance showed that the country's response was well executed. To improve future response, we recommend enhanced district level preparedness, update of disaster response protocols, and pre-disaster mapping of health sector partners.


Elovanoid-N34 modulates TXNRD1 key in protection against oxidative stress-related diseases.
The thioredoxin (TXN) system is an NADPH + H+/FAD redox-triggered effector that sustains homeostasis, bioenergetics, detoxifying drug networks, and cell survival in oxidative stress-related diseases. Elovanoid (ELV)-N34 is an endogenously formed lipid mediator in neural cells from omega-3 fatty acid precursors that modulate neuroinflammation and senescence gene programming when reduction-oxidation (redox) homeostasis is disrupted, enhancing cell survival. Limited proteolysis (LiP) screening of human retinal pigment epithelial (RPE) cells identified TXNRD1 isoforms 2, 3, or 5, the reductase of the TXN system, as an intracellular target of ELV-N34. TXNRD1 silencing confirmed that the ELV-N34 target was isoform 2 or 3. This lipid mediator induces TXNRD1 structure changes that modify the FAD interface domain, leading to its activity modulation. The addition of ELV-N34 decreased membrane and cytosolic TXNRD1 activity, suggesting localizations for the targeted reductase. These results show for the first time that the lipid mediator ELV-N34 directly modulates TXNRD1 activity, underling its protection in several pathologies when uncompensated oxidative stress (UOS) evolves.


Effects of propranolol and yohimbine on periarterial nerve stimulation-induced release of endogenous norepinephrine from the mesenteric vasculature of Wistar Kyoto and spontaneously hypertensive rats.
The overflow of endogenous norepinephrine (NE) from the mesenteric vasculature of the isolated mesentery of Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) was determined in response to periarterial nerve stimulation (PNS) before and after pretreatment with propranolol or yohimbine. Propranolol pretreatment did not significantly alter spontaneous NE overflow, total NE overflow, NE overflow/stimulus or fractional NE overflow in either WKY or SHR mesenteric vascular preparations at any of the PNS frequencies used. Yohimbine pretreatment did not significantly alter spontaneous NE overflow but did significantly increase total NE overflow, NE overflow/stimulus and fractional NE overflow at all PNS frequencies used in both WKY and SHR preparations. The magnitude of the effect of yohimbine on NE overflow/stimulus did not differ between WKY and SHR over the range of PNS frequencies used. The lack of effect of propranolol on NE overflow suggests that corelease of epinephrine is not sufficient to activate beta adrenergic receptor-mediated modulation of noradrenergic neurotransmission in either WKY or SHR mesenteric vascular preparations under the in vitro study conditions used. However, the effects of yohimbine indicate that prejunctional alpha-2 adrenergic receptor-mediated inhibition of noradrenergic neurotransmission is operative in both WKY and SHR mesenteric vascular preparations. These effects of yohimbine also suggest that no significant differences exist between the functional level of prejunctional alpha-2 adrenergic receptor-mediated autoinhibitory modulation of noradrenergic neurotransmission in mesenteric vascular preparations from adult WKY and SHR under the in vitro study conditions used.


The structure of alpha 2-macroglobulin-methylamine after papain digestion as determined by electron microscopy.
alpha 2-Macroglobulin-methylamine (alpha 2M-CH3NH2) was digested with papain at pH 5.0. The major 600 kDa fragment was purified by molecular-exclusion chromatography. In a non-denaturing gel-electrophoresis system, the 600 kDa fragment migrated in a single band at a rate that was comparable with that for the untreated alpha 2M-CH3NH2. The elution volume of the 600 kDa fragment on Superose-6 was slightly increased. In primary cultures of rat hepatocytes, cellular uptake of 125I-alpha 2M-CH3NH2 was not affected by the 600 kDa fragment, confirming the results of other investigators. The 600 kDa fragment was negatively stained with uranyl formate and analysed by transmission electron microscopy. The major structural characteristics of the parent protein (alpha 2M-CH3NH2) remained intact. The most common image included prominent lateral walls and two centrally located regions of stain exclusion termed 'paddle structures'. The distance between the paddle structures was equivalent in alpha 2M-CH3NH2 and the 600 kDa fragment [approximately 13.5 nm (135 A)]. By contrast, the lateral walls in the 600 kDa fragment were decreased in length by approximately 0.37 nm (37 A) (19%). It is proposed that the 600 kDa structure retains the 'hollow cylinder' shape of alpha 2M-CH3NH2. The structure of the cylinder is formed by the lateral walls and four paddle structures (only two are imaged, owing to overlapping). The paddle structures in the 600 kDa fragment are intact and relatively closer to the apices of the molecule, owing to the decrease in lateral wall length. Since the alpha 2M receptor-binding sites are removed by papain digestion, the studies presented here support the location of the receptor-binding sites near the apices of the lateral walls.


Profile of Current Opinion on Arthroscopic Acromioplasty: A Video Survey Study.
To provide a baseline profile of current opinion on use of arthroscopic acromioplasty and evaluate the consistency of surgical decision making on whether or not to perform acromioplasty across different surgeon practices. Twenty-two fellowship-trained orthopaedic shoulder surgeons from the Association of Clinical Elbow and Shoulder Surgeons completed an Internet-based survey on practices associated with acromioplasty, including questions related to 15 arthroscopic videos. Based on video cases, interobserver and intraobserver agreement regarding clinically based decisions related to acromioplasty were assessed. Acromioplasty was uncommonly performed in isolation among this group and was most commonly performed in conjunction with repair of full-thickness rotator cuff tears. Nineteen of 22 (86%) surgeons favored an arthroscopic approach for acromioplasty. Depth of bony resection was determined most commonly based on clinical judgment and experience (68%). The video portion of the survey revealed slight interobserver agreement for classification of acromion morphology (κ = 0.099), need for acromioplasty (κ = 0.020), and adequacy of decompression (κ = 0.1). In contrast, there was fair intraobserver reliability regarding acromion morphology (κ = 0.370) and decision whether to perform acromioplasty in a given case (κ = 0.348) whereas there was moderate intraobserver reliability in the presence of a reparable rotator cuff tear (κ = 0.507) and assessment of the adequacy of decompression (κ = 0.453). Although surgeons had similarities regarding principles of acromioplasty, including indications, surgical approach, and technique, there was lack of consensus when surgeons reviewed the video of clinical cases. Although surgeons may have similar goals in terms of treatment of pathology related to subacromial impingement, individual surgeon thresholds for the need and adequacy of decompression are varied and are not standardized. Level V, expert opinion.


Donor lymphocyte infusions for patients who relapse after allogeneic stem cell transplantation for chronic myeloid leukaemia.
The understanding of the use of donor lymphocyte infusions (DLI) for graft-versus-leukaemia (GVL) in the treatment of chronic myeloid leukaemia (CML) post haemopoietic stem cell transplant (HSCT) has advanced during the last years. In relapsed leukaemia post-stem cell transplant, DLI can achieve durable remissions in 60-73% patients. Technical improvements in molecular methods of detection of the BCR-ABL transcripts permit the prediction of relapse with increased sensitivity and reproducibility. Use of DLI early at relapse is important since responses to DLI are less likely in the face of bulky or blast-phase disease. Exogenous interleukin-2 may enhance the response to DLI but total cell dose is also relevant to the efficacy of DLI with the effective cell dose (ECD) required being lower in HLA matched unrelated DLI donors compared to siblings. Donor T-lymphocytes target minor histocompatibility (H) antigens and the relative tissue distribution of these may influence the toxicity of DLI, which includes graft-versus-host-disease (GVHD). Modified methods of delivery such as selective deletion of CD8+ cells or escalating cell dosage regimens have reduced the incidence of serious morbidity due to GVHD without compromising the GVL effect mediated by DLI. These approaches have not removed the risk of GVHD entirely and conditional suicide protocols utilising the HSV-tk or fas receptor derived genes are being developed in the clinic. Since significant morbidity and mortality is attributable to the conditioning regimen used prior to HSCT, awareness of the potency of DLI has driven the development of reduced intensity conditioning (RIC) regimens. The purpose of RIC is to enhance tolerisation of the host to the graft while permitting the establishment of donor haemopoiesis. DLI may then be used subsequently to enhance the GVL effect.


[The effect of several surface-active substances on neuromuscular transmission in frogs].
The action of bile salts, saponin and tween-80 on miniature end-plate potentials (MEPP) and electrotonic potentials (ETP) of the muscle fibre membrane was studied in frog Rana ridibunda. Bile salts in a concentration of 10(-4) g/ml increased the frequency and amplitude of MEPP. The input resistance of the muscle fibre membrane decreased under the action of these substances. At higher concentrations (to 10(-3) g/ml) the frequency of the MEPP initially increased, then these potentials were depressed and finally they completely disappeared. Tween-80 did not change noticeably spontaneous synaptic activity, and saponin inhibited it. In Ca-free solutions bile salt were less effective. It is suggested that bile salts change the structure of the presynaptic membrane and at the same time stimulate the release of acetylcholine form the nerve terminals.


Biomarkers in the prevention and treatment of atherosclerosis: need, validation, and future.
Cardiovascular disease (CVD) remains one of the leading causes of morbidity and mortality in the developed world, and there is a clear need to develop novel therapeutic strategies to reduce cardiovascular risk further than is currently possible. Traditionally, the effectiveness of new cardiovascular drugs has been evaluated in clinical trials using cardiovascular outcomes as endpoints. However, such trials require large numbers of patients followed over long periods of time. Clinical trials using surrogate markers for CVD may be shorter in duration and involve fewer participants. Measurement of atherosclerotic progression is an ideal surrogate marker as it is predictive of future cardiovascular events. The "gold standard" for detecting and defining the severity, extent, and rate of atherosclerotic progression has been quantitative coronary angiography. However, this technique has fundamental limitations. More recently, measurement of carotid intima-media thickness using B-mode ultrasound and measurement of atheroma volume using intravascular ultrasound have emerged as more accurate techniques for detecting atherosclerotic progression. Both of these techniques have potential utility as surrogate endpoints in place of cardiovascular outcomes in clinical trials. Their use might facilitate the more rapid development of novel, safe, and effective therapies.


Predictive factors of neck metastases in laryngeal squamous cell carcinoma. Towards an integrated clinico-molecular classification.
Our group has 25 years' experience in the use of molecular predictive markers in head and neck cancer, on a large patient population, enrolled from a single institution, with a long follow-up, and, most of all, homogeneous regarding histology (squamous cell carcinoma) and site (larynx). Among the most frequent malignancies in the US, cancers of the larynx and uterine corpus are the only types not showing an increase in 5-year Survival Rates over the last 30 years. As far as concerns laryngeal squamous cell carcinoma, we can identify several potential reasons for this failure, the most relevant probably lies in the neck. For this reason, a key issue in laryngeal oncology is to assess metastatic potential of squamous cell carcinoma at diagnosis. Nevertheless, the combination of clinical and histological parameters is not sufficiently reliable in the prediction of lymph node metastases. Molecular characterization, by the study of molecular predictive factors, is a clinical approach aimed to define homogeneous subgroups for clinical metastatic behaviour. Defining invasiveness by means of studies on selected molecular markers (among which the most reliable is probably Epidermal Growth Factor Receptor (EGFR)) may be useful in the choice of the most appropriate treatment on both T and on N.


Insulin and glycemic responses in healthy humans to native starches processed in different ways: correlation with in vitro alpha-amylase hydrolysis.
The aim of the study was to elucidate how extracted starches submitted to food processing (or not) can influence plasma insulin and glucose responses in healthy subjects. Native starches from wheat, manihot, smooth peas, or mung beans were tested either raw, as starch gels (boiled and cooled), or cooked and cooled after a preliminary industrial processing: extrusion cooking for wheat, tapioca for manihot, and noodles for mung beans. Eighteen healthy subjects randomly assigned received three different starches under one form of conditioning. All products were submitted to in vitro alpha-amylolysis. Raw manihot starch produced the lowest (p less than 0.05) metabolic responses. Cooking significantly (p less than 0.01) increased plasma responses. However, cooked mung bean noodles gave metabolic responses similar to those of raw products. Close correlations were found between percentages of in vitro starch hydrolysis at 30 min and mean areas under the glycemic curves and the insulinemic curves (r = 0.95, p less than 0.001).


Precipitating/aggravating factors of porphyria cutanea tarda in Spanish patients.
Erythrocyte uroporphyrinogen decarboxylase (UROD) activity was measured to classify 118 Spanish patients with porphyria cutanea tarda (PCT) into three subtypes: sporadic-, familial- and type III-PCT. Seventy-four patients (63%) had eythrocyte UROD activity within the normal range (74% to 126% of the mean activity of 43 healthy controls) and were classified as sporadic-PCT (47%) or as type III-PCT (16%) whenever a family history of PCT was documented. Forty-four patients (37%) had decreased UROD activity and were classified as familial-PCT. The frequency of both familial-PCT and type III-PCT was higher than reported in other countries. The clinical expression of PCT was associated with the coexistence of two or more risk factors in 80% of the sporadic-PCT patients and in 89% of the familial-PCT patients. Hepatitis C virus and alcohol abuse were risk factors frequently found in these patients, being unrelated to age of onset of skin lesions. A heavy alcohol intake was the main risk factor for type III-PCT. Estrogens appeared as a precipitating factor for women with familial-PCT. The H63D mutation in the hemochromatosis type 1 gene was more frequently found than the C282Y mutation. Both mutations appeared to play a role as precipitating factors in sporadic-PCT when associated with hepatitis C virus infection and alcohol abuse.


A robust nonlinear filter for image restoration.
A class of nonlinear regression filters based on robust estimation theory is introduced. The goal of the filtering is to recover a high-quality image from degraded observations. Models for desired image structures and contaminating processes are employed, but deviations from strict assumptions are allowed since the assumptions on signal and noise are typically only approximately true. The robustness of filters is usually addressed only in a distributional sense, i.e., the actual error distribution deviates from the nominal one. In this paper, the robustness is considered in a broad sense since the outliers may also be due to inappropriate signal model, or there may be more than one statistical population present in the processing window, causing biased estimates. Two filtering algorithms minimizing a least trimmed squares criterion are provided. The design of the filters is simple since no scale parameters or context-dependent threshold values are required. Experimental results using both real and simulated data are presented. The filters effectively attenuate both impulsive and nonimpulsive noise while recovering the signal structure and preserving interesting details.


PEGylated interferon beta-1a in the treatment of multiple sclerosis - an update.
Current standard immunomodulatory therapy with interferons (IFNs) for relapsing-remitting multiple sclerosis (MS) exhibits proven, but limited, efficacy and increased side effects due to the need of frequent application of the drug. Therefore, there is a need for more effective and tolerable drugs. Due to their small size, optimization of therapy with IFNs in MS by PEGylation is feasible. PEGylation of an IFN means that at least one molecule of polyethylene glycol (PEG) is covalently added. This modification is a standard procedure to increase the stability, solubility, half-life, and efficacy of a drug, and is applied in several drugs and diseases. Currently, a therapy regimen applying PEG-IFN beta-1a in MS is being developed to achieve an optimized relationship between therapy-related side effects and pharmacokinetic/pharmacodynamic efficacy. Phase I studies demonstrated that subcutaneous PEG-IFN beta-1a at a dose of 125 μg every 2 or 4 weeks might be at least as efficient and safe as the current standard therapy with IFN beta-1a. A global Phase III clinical study is investigating the efficacy of PEG-IFN beta-1a in terms of reduction of the relapse rate in relapsing-remitting MS patients. The latest primary safety and efficacy analysis after 1 year has revealed a favorable risk-benefit profile with no significant difference between dosing regimens. Compared to placebo, the annualized relapse rate was reduced by about one-third and new or newly enlarging T2 brain lesions were reduced by about one-third when dosing every 4 weeks or by two-thirds when dosing every 2 weeks. This presents a significant effect of the dosing interval, favoring administration every 2 weeks. Chronic administration of PEGylated proteins mostly at toxic concentrations causes vacuolation of renal epithelium in animals, which - along with the issue of occurrence of anti-PEG antibodies - has to be addressed by Phase IV studies.


Defining a safe corridor for trans-iliac pin placement in cats.
To identify whether a theoretical predictable safe corridor is available in cats for placement of trans-iliac pins without the use of fluoroscopy. Twenty-one cats with straight orthogonal normal pelvic radiographs were included. Two start points were evaluated: a midpoint and a dorsal point. The midpoint was defined as midway between the dorsal lamina of the sacral vertebral canal and the cranial dorsal iliac spine. The dorsal start point was 2 mm ventral to the cranial dorsal iliac spine. The pin was assumed to be driven at 90 degrees to the lateral face of the ilium, and considered surgeon accuracy was ±4 degrees from the perpendicular. The angular range and the distance between the iliac wings from the ventrodorsal radiograph were used to calculate the possible cross-sectional area and pin exit location if driven from one iliac wing to the other. The corridor was then evaluated for repeatability in six randomly selected cats. Vertebral foramina penetration risk was identified in some cats when using a 1.6 and 2 mm-diameter pin using the mid-iliac wing start point. The dorsal start point decreased the available pin placement area but reduced the risk of entering the hazardous zone for all pin sizes up to 2 mm. A theoretical defined safe corridor is available for trans-iliac pin placement in cats between 2.0 and 5.5 kg. A 1.2-mm pin is the safest if using the mid-iliac wing start point. A more dorsal start point can accommodate up to a 2.0-mm pin if correctly aligned to the sacrum.


Metal Ion-Induced Large Fragment Deactivation: A Different Strategy for Site-Selectivity in a Complex Molecule.
Complex natural product functionalizations generally involve the use of highly engineered reagents, catalysts, or enzymes to react exclusively at a desired site through lowering of a select transition state energy. In this communication, we report a new, complementary strategy in which all transition states representing undesirable sites in a complex ionophore substrate are simultaneously energetically increased through the chelation of a metal ion to the large fragment we wish to neutralize. In the case of an electrophilic, radical based fluorination reaction, charge repulsion (electric field effects), induced steric effects, and electron withdrawal provide the necessary deactivation and proof of principle to afford a highly desirable natural product derivative. We envisage that many other electrophilic or charge based synthetic methods may be amenable to this approach as well.


Technical note: an R program for automating bone cross section reconstruction.
Many recent studies have used long bone cross-sectional geometric properties in various comparative analyses. Methods have been described for reconstructing diaphyseal cross sections from external molds and biplanar radiographs that produce accurate results (within 5% of true values on average). The manual image processing required, however, is both time and labor intensive. A new freely available program developed here for the computational freeware, R, automates much of the process. This study compares cross-sectional properties calculated using the new R program to those from peripheral quantitative CT (pQCT) and the original manual method. We find that the R program works aswell as the original manual image processing for most cross sections eliminates the chance for entry errors at several steps and greatly speeds up data collection.


Analysis of the consequences of car to micromobility user side impact crashes.
The strong rise in modes of travel commonly referred to as micromobility has changed the mobility patterns and lifestyles in cities worldwide, especially after the COVID-19 pandemic. It has led to a significant increase in the number of crashes involving these types of vehicles, especially bicycles and stand-up e-scooters. The risk of crashes is higher at intersections where motor-vehicles perform a turning maneuver crossing a bike lane. The consequences of a passenger car-to-micromobility vehicle side-impact crashes, considering both bicycle and e-scooter, were studied based on the results of the simulation of several scenarios with PC-Crash software. Two injury criteria were applied: Head Injury Criterion (HIC15) and 3 ms chest acceleration criterion. When motor-vehicle speed is lower than 50 km/h, the 3 ms chest acceleration never exceeds the 60 g threshold. However, at 50 km/h, it is close to 50 g in the case of e-scooter rides. At this speed, HIC15 is considerably greater than 1000, both for bicycles and for e-scooters, and the safety margin of 700 is exceeded at 45 km/h for e-scooters. In case of motor vehicle-to-micromobility vehicle side-impact crash, riding a bicycle is safer than riding an e-scooter since the observed HIC15 experienced by the cyclists is lower than that experienced by the e-scooter rider when motor vehicle speed is greater than 30 km/h. To reduce micromobility users injury risk at intersections, motor vehicle speed limit should be equal or lower than 40 km/h. At this impact speed, the activation of hood or bumper airbags could be justified.


Development of Tbet- and CD11c-expressing B cells in a viral infection requires T follicular helper cells outside of germinal centers.
Tbet+CD11c+ B cells arise during type 1 pathogen challenge, aging, and autoimmunity in mice and humans. Here, we examined the developmental requirements of this B cell subset. In acute infection, T follicular helper (Tfh) cells, but not Th1 cells, drove Tbet+CD11c+ B cell generation through proximal delivery of help. Tbet+CD11c+ B cells developed prior to germinal center (GC) formation, exhibiting phenotypic and transcriptional profiles distinct from GC B cells. Fate tracking revealed that most Tbet+CD11c+ B cells developed independently of GC entry and cell-intrinsic Bcl6 expression. Tbet+CD11c+ and GC B cells exhibited minimal repertoire overlap, indicating distinct developmental pathways. As the infection resolved, Tbet+CD11c+ B cells localized to the marginal zone where splenic retention depended on integrins LFA-1 and VLA-4, forming a competitive memory subset that contributed to antibody production and secondary GC seeding upon rechallenge. Therefore, Tbet+CD11c+ B cells comprise a GC-independent memory subset capable of rapid and robust recall responses.


Epidemiology and Outcomes of Lisfranc Injuries Identified at the National Football League Scouting Combine.
Lisfranc injuries are challenging to treat and may have a detrimental effect on athletic performance. (1) Determine the epidemiological characteristics of Lisfranc injuries at the annual National Football League (NFL) Scouting Combine, (2) define player positions at risk for these injuries, and (3) evaluate the impact that these injuries and radiographic findings have on NFL draft position and performance. Cohort study; Level of evidence, 3. All players who sustained a Lisfranc injury prior to Combine evaluation between 2009 and 2015 were evaluated. The epidemiological characteristics, player positions affected, treatment methods, and number of missed collegiate games were recorded. Radiographic outcomes were analyzed via Combine radiograph findings, while NFL performance outcomes were assessed for all Lisfranc injuries (2009-2013) compared with matched controls in the first 2 years of play. A total of 41 of 2162 (1.8%) Combine participants were identified with Lisfranc injuries, of whom 26 of 41 (63.4%) were managed operatively. Players who underwent surgery were more likely to go undrafted compared with players managed nonoperatively (38.5% vs 13.3%, operative vs nonoperative management, respectively; P = .04) and featured a worse NFL draft pick position (155.6 vs 109; P = .03). Lisfranc-injured players when compared with controls were noted to have worse outcomes in terms of NFL draft position (142 vs 111.3, Lisfranc-injured players vs controls, respectively; P = .04), NFL career length 2 years or longer (62.5% vs 69.6%; P = .23), and number of games played (16.9 vs 23.3; P = .001) and started (6.8 vs 10.5; P = .08) within the first 2 years of their NFL career. Radiographs demonstrated that 17 of 41 (41.5%) athletes had residual Lisfranc joint displacement greater than 2 mm compared with the contralateral foot. Lisfranc-injured athletes with greater than 2 mm residual displacement, when compared with matched controls, had worse draft position (156.9 vs 111.2 for Lisfranc-injured players vs controls, respectively; P = .009) and fewer games played (14.4 vs 23.3; P = .001) and started (3.1 vs 10.5; P = .03). Moreover, athletes with greater than 2 mm residual displacement featured worse outcomes across all assessed NFL variables versus athletes with residual displacement of 2 mm or less. Lisfranc injuries identified at the NFL Combine have an adverse effect on an NFL athlete's draft status, draft position, and overall play during initial NFL seasons. In particular, residual displacement of the Lisfranc joint has a detrimental effect on the first 2 seasons of NFL play and may lead to long-lasting negative effects on the athlete's career.


Heart failure guidelines, performance measures, and the practice of medicine: mind the gap.
Guidelines have rather quickly assumed a central role in health care delivery in the U.S. They have become the foundation on which performance measures are built and, therefore, a major player in assessing the quality of care provided by individuals and institutions, the ramifications of which involve reputation, reimbursement, and litigation. We are concerned, however, that in our enthusiasm for collectively endorsing these guidelines, we are marginalizing the importance of physician judgment and inadvertently risking the conversion of guidelines into "cookbooks." We believe that this editorial, while unequivocally acknowledging the fundamental importance of guidelines, simultaneously provides a critically important perspective on the potential for misuse of both guidelines and performance measures. Further, we hope that publication of this commentary will help temper enthusiasm for overzealous conversion of guidelines into performance measures, thereby restoring the vital role of physician judgment and insight into patient management.


Anterior-posterior thoracic force-deflection characteristics measured during cardiopulmonary resuscitation: comparison to post-mortem human subject data.
Comparative data of thoracic compression response between live vs. post mortem human subjects (PMHS) has been reported, but the live subject tests are often at low deflections and include the effects of muscle tensing. Novel technology has been developed that overcomes several of these limitations. Specifically, a load cell and accelerometer has been integrated into a clinical monitor-defibrillator to measure chest compression and applied force during live human cardio-pulmonary resuscitation (CPR). The sensor is interposed between the hands of the person administering CPR and the sternum of the patient. The objective of this study was to compare the thoracic force-deflection measured during adult CPR to that measured during hub-based loading of adult PMHS. CPR represents a unique setting in which to study the mechanics of the chest as the thorax is loaded to a maximum chest deflection similar to that seen in a frontal crash environment and the effects of muscle tensing are minimized. PMHS and CPR data were modeled with a progressive spring in parallel with a viscous damper. Statistical comparison of the model parameters used to describe the compliance of the thorax during large compressions revealed that under the specific loading conditions tested, chest compressions during CPR generated less force at equivalent deflections than the PMHS. Specifically, the spring force at 40 mm deflection was 286 N for the CPR data and 588 N for the PMHS. Differences in area of load application and thoracic structures engaged, however, may partially explain these differences. In addition, the chest response during CPR demonstrated more hysteresis than the PMHS suggesting that the viscoelastic nature of the thorax is more pronounced when tissues are naturally perfused and substantial tissue autolysis has not begun. This study presents fundamental biomechanical data on the mechanical response of the adult thorax that increases our understanding of any potential differences between PMHS and chest response obtained during CPR before substantial tissue autolysis begins. We contend that the thoracic compliance measured during CPR may continue to offer important force-deflection estimates in the future.


[Dientamoeba fragilis infection in patients with gastrointestinal system complaints].
In this study, we aimed to investigate the incidence of Dientamoeba fragilis with different diagnostic methods in patients with gastrointestinal symptoms and determine the sensitivity and specificity of existing diagnostic methods. Fecal samples collected from 101 patients with gastrointestinal complaints (especially upper abdominal pain, abdominal and pelvic pain, nausea and vomiting, gastroenteritis and colitis, unexplained fever and diarrhea) and 20 control cases from various clinics were included in the study. Samples were first examined with native-Lugol (N-L) method and cultured in Robinson medium. All 121 stool and culture samples were stained with iron hematoxylin stain (IHS) and trichrome stain (TS) methods and examined by PCR and QPCR for D.fragilis. Among 121 stool samples 13 (10.7%), 2 (1.7%), 7 (5.7%) 13 (10.7%), and 7 (5.8%), 4 (3.3%), 2 (1.7%), 3 (2.5%) of cultured samples were determined positive with IHS, TS, PCR, QPCR respectively. Fifteen of the 121 stool samples were determined as diarrheal. All diarrheal stool samples were negative with IHS and TS. One of the diarrheal stools and 6 (4.9%) of the non-diarrheal stools were positive by PCR. All of the diarrheal stools were negative. Thirteen of the non-diarrheal stool samples (10.7%) were positive by QPCR. When the QPCR method was considered as gold standard, sensitivity and specificity values were determined as 46% and 93% in IHS, 0% and 99% in TS, 54% and 100% by PCR and sensitivity and specificity values were 67% and 96% in IHS, 33% and 98% in TS, 67% and 100% by PCR among cultured stool samples. As a result, it was determined that there was a statistically significant difference between the samples of the patients and the control groups and the sensitivity and specificity of the conventional and molecular methods (IHS, TS, PCR and QPCR) determined in this study supported the results of other compared studies. It has been determined that staining methods used for the diagnosis of D.fragilis gave false positivite or negativite results. In addition, the QPCR method is more advantageous in terms of time saving for the diagnosis and initiation of the treatment and in cases where QPCR is not available, IHS and conventional PCR methods should be used together. In our opinion, this study will contribute to the results of epidemiological and scientific studies on D.fragilis in Turkey.


Risk of Contraceptive Lapse and New Sexual Partnership Among Female University Students Traveling Internationally.
International travel is common and may influence women's sexual and reproductive health through myriad pathways. Existing studies focus on travelers' risk of acquiring STDs, but not their pregnancy prevention behaviors. Further, it is unclear whether sexual and reproductive health outcomes observed for female travelers are associated with travel itself, because few studies compare travelers with nontravelers. An online survey was conducted in 2016-2017 among females attending a U.S. university who reported a history of sex with men; the sample comprised 340 students who had traveled internationally in the past three months (travelers) and 170 who intended to travel in the next three months (pretravelers). Multivariable modified Poisson regression models assessed associations between travel status and two outcomes during travelers' recent trip or a period of similar duration for pretravelers: contraceptive lapse (incorrect or inconsistent use of any method) and new sexual partnership. Travelers and pretravelers reported similar levels of contraceptive lapse (29% and 32%, respectively) and new sexual partnerships (17% and 12%). Multivariable analysis confirmed that travelers were no more likely than pretravelers to have contraceptive lapses, but indicated that they were more likely to report a new male partner (relative risk, 1.7). Most participants (80%) had a regular source of sexual and reproductive health care; 42% of travelers had seen a health care provider in preparation for their trip. If the findings are corroborated by additional research, they may suggest a role for health care providers in students' pretravel period.


Disruption of the ascending arousal system and cortical attention networks in post-stroke delirium and spatial neglect.
Delirium is an acute attention and cognitive dysfunction, adversely affecting functional outcomes and mortality. As many as half of hospitalized right brain stroke survivors may develop delirium. Further, about 50% of right stroke patients experience spatial neglect, impairing safety and recovery. In this review we explore the brain mechanisms, which may explain the high incidence of delirium and spatial neglect after right-brain stroke. We suggest that brain networks for spatial attention and arousal, composed of ascending projections from the midbrain nuclei and integrating dorsal and ventral cortical and limbic components, may underlie impairments in delirium and spatial neglect. We propose that lateralized deficits in spatial neglect may arise because cortical and limbic components of these functional networks are disproportionally impaired by right-brain strokes, and that spatial neglect may lower the threshold for developing delirium. An improved understanding of the brain basis of delirium and spatial neglect could provide a critical biomarker for initiating preventive care in stroke patients at high risk of hospital morbidity and loss of independence.


[Immunogenicity and the protective effectiveness of the subunit trivalent influenza vaccine Grippovac].
In the process of the immunological approbation of several experimental batches of the triyalent subunit influenza vaccine Grippovac, the pronounced immunogenicity of the antigens of all strains contained in the vaccine was established; most of the vaccinees were found to retain sufficiently high antibody titers for a year, and the essential total increase of antibody titers was found to occur after a single booster immunization. The serological checking of the diagnosed cases of influenza among immunized and nonimmunized children, carried out in two boarding schools during the influenza epidemic in the winter of 1984-1985 provided the proofs of the high effectiveness of Grippovac in preventing viral influenza, types A and B: the decrease of influenza morbidity among the immunized children reached 79,5-67,2-66,5% and the total morbidity rate in influenza in these two boarding scholls dropped, on the average, 3,0-3,5 times.


"SHOUT" to improve the quality of care delivered to patients with acute kidney injury at Great Western Hospital.
Acute kidney injury (AKI) affects up to 20% of all patients admitted to hospital, and is associated with a higher risk of adverse clinical outcomes, increased healthcare costs, as well as long term risks of chronic kidney disease and end stage renal failure. The aim of this project was to improve the quality of care for patients with AKI admitted to the acute medical unit (AMU) at the Great Western Hospital (GWH). We assessed awareness and self reported confidence among physicians in our Trust, in addition to basic aspects of care relevant to AKI on our AMU. A multifaceted quality improvement strategy was developed, which included measures to improve awareness such as a Trust wide AKI awareness day, and reconfiguring the admission proforma on our AMU in order to enhance risk assessment, staging, and early response to AKI. Ancillary measures such as the dissemination of flashcards for lanyards containing core information were also used. Follow up assessments showed that foundation year one (FY1) doctors' self reported confidence in managing AKI increased from 2.8 to 4.2, as measured on a five point Likert scale (P=0.0003). AKI risk assessment increased from 13% to 57% (P=0.07) following a change in the admission proforma. Documentation of the diagnosis of AKI increased from 66% to 95% (P=0.038) among flagged patients. Documentation of urine dip results increased from 33% to 73% (P=0.01), in addition to a rise in appropriate referral for specialist input, although this was not statistically significant. Our results suggest that using the twin approaches of improving awareness, and small changes to systemic factors such as modification of the admission proforma, can lead to significant enhancements in the quality of care of patients with AKI.


Role of S-layer proteins in the biosorption capacity of lead by Lactobacillus kefir.
The role of S-layer proteins (SLP) on the Pb(2+) sequestrant capacity by Lactobacillus kefir CIDCA 8348 and JCM 5818 was investigated. Cultures in the stationary phase were treated with proteinase K. A dot blot assay was carried out to assess the removal of SLP. Strains with and without SLP were exposed to 0-0.5 mM Pb(NO3)2. The maximum binding capacity (q max ) and the affinity coefficient (b) were calculated using the Langmuir equation. The structural effect of Pb(2+) on microorganisms with and without SLP was determined using Raman spectroscopy. The bacterial interaction with Pb(2+) led to a broadening in the phosphate bands (1,300-1,200 cm(-1) region) and strong alterations on amide and carboxylate-related bands (νCOO(-) as and νCOO(-) s). Microorganisms without SLP removed higher percentages of Pb(2+) and had higher q max than those bearing SLP. Isolated SLP had much lower q max and also removed lower percentages of Pb(2+) than the corresponding whole microorganisms. The hydrofobicity of both strains dramatically dropped when removing SLP. When bearing SLP, strains do not expose a large amount of charged groups on their surfaces, thus making less efficient the Pb(2+) removal. On the contrary, the extremely low hydrofobicity of microorganisms without SLP (and consequently, their higher capacity to remove Pb(2+)) can be explained on the basis of a greater exposure of charged chemical groups for the interaction with Pb(2+). The viability of bacteria without SLP was not significantly lower than that of bacteria bearing SLP. However, microorganisms without SLP were more prone to the detrimental effect of Pb(2+), thus suggesting that SLP acts as a protective rather than as a sequestrant layer.


Can a pediatric trauma center improve the response to a mass casualty incident?
Recent events including the 2001 terrorist attacks on New York; Hurricane Katrina; the 2010 Haitian and Chilean earthquakes; and the 2011 earthquake, tsunami, and nuclear disaster in Japan have reminded disaster planners and responders of the tremendous scale of mass casualty disasters and their resulting human devastation. Although adult disaster medicine is a well-developed field with roots in wartime medicine, we are increasingly recognizing that children may comprise up to 50% of disaster victims, and response mechanisms are often designed without adequate preparation for the number of pediatric victims that can result. In this short educational review, we explore the differences between the pediatric and adult disaster and trauma populations, the requirements for designation of a site as a pediatric trauma center (PTC), and the magnitude of the problem of pediatric disaster patients as described in the literature, specifically as it pertains to the availability and use of designated PTCs as opposed to trauma centers in general. We also review our own experience in planning and simulating pediatric mass casualty events and suggest strategies for preparedness when there is no PTC available. We aim to demonstrate from this brief survey that the availability of a designated PTC in the setting of a mass casualty disaster event is likely to significantly improve the outcome for the pediatric demographic of the affected population. We conclude that the relative scarcity of disaster data specific to children limits epidemiologic study of the pediatric disaster population and offer suggestions for strategies for future study of our hypothesis. Systematic review, level III.


Biodiversity and ecosystem function: making sense of numerous species interactions in multi-species communities.
Understanding the biodiversity and ecosystem function relationship can be challenging in species-rich ecosystems. Traditionally, species richness has been relied on heavily to explain changes in ecosystem function across diversity gradients. Diversity-Interactions models can test how ecosystem function is affected by species identity, species interactions, and evenness, in addition to richness. However, in a species-rich system, there may be too many species interactions to allow estimation of each coefficient, and if all interaction coefficients are estimable, they may be devoid of any sensible biological meaning. Parsimonious descriptions using constraints among interaction coefficients have been developed but important variability may still remain unexplained. Here, we extend Diversity-Interactions models to describe the effects of diversity on ecosystem function using a combination of fixed coefficients and random effects. Our approach provides improved standard errors for testing fixed coefficients and incorporates lack-of-fit tests for diversity effects. We illustrate our methods using data from a grassland and a microbial experiment. Our framework considerably reduces the complexities associated with understanding how species interactions contribute to ecosystem function in species-rich ecosystems.


CD44v(3,8-10) is involved in cytoskeleton-mediated tumor cell migration and matrix metalloproteinase (MMP-9) association in metastatic breast cancer cells.
In the present study, we have employed a unique breast cancer cell line (Met-1, which was derived from a high metastatic potential tumor in transgenic mice expressing polyomavirus middle T oncogene) to study the role of CD44 variant isoform(s) in the regulation of metastatic breast tumor cell behavior. The results of reverse transcriptase-polymerase chain reaction, Southern blot, nucleotide sequencing, immunoprecipitation, and immunoblot analyses indicated that these cells express a major CD44 isoform (molecular weight approximately 260 kDa) containing a v3,8-10 exon insertion (designated as CD44v3,8-10). In addition, we have determined that CD44v3,8-10 binds specifically to the cytoskeletal proteins such as ankyrin. Biochemical analyses, using competition binding assays and a synthetic peptide identical to NGGNGTVEDRKPSEL (a sequence located between aa480 and aa494 of CD44v3,8-10) indicate that this 15-amino acid peptide binds specifically to the cytoskeletal protein ankyrin (but not to fodrin or spectrin). This peptide competes effectively for ankyrin binding to CD44v3,8-10. Therefore, we believe that the sequence 480NGGNGTVEDRKPSE494L, located at the cytoplasmic domain of CD44v3,8-10, is required for the ankyrin binding. We have also detected that CD44v3,8-10-containing Met-1 cells are capable of forming membrane spikes or "invadopodia" structures and undergo active migration processes. Treatments of Met-1 cells with certain agents including anti-CD44v3 antibody, cytochalasin D (a microfilament inhibitor), and W-7 (a calmodulin antagonist), but not colchicine (a microtubule disrupting agent) effectively inhibit "invadopodia" formation and subsequent tumor cell migration. Further analyses using zymography assays and double immunofluorescence staining indicated that CD44v3,8-10 is closely associated with the active form of matrix metalloproteinase, MMP-9, in a complex within "invadopodia" structures. These findings suggest that CD44v3,8-10 plays an important role in linking ankyrin to the membrane-associated actomyosin contractile system required for "invadopodia" formation (coupled with matrix degradation activities) and tumor cell migration during breast cancer progression.


[Follow-up of patients that use lithium].
Lithium has been among the most important pharmacological treatments of psychiatric disease for more than 50 years. The main indication is treatment and prophylaxis of bipolar disease. Lithium is also used in the treatment of schizophrenia conditions with affective symptoms. The therapeutic effect of lithium is well documented, but it can also cause serious adverse effects. The drug has a narrow therapeutic range and intoxications can occur at normal or low doses. This article reviews guidelines for lithium treatment, including side effects, interactions and intoxications.


On-chip testing device for electrochemotherapeutic effects on human breast cells.
A microfabricated cell-based testing device for electrochemotherapy (ECT) has been developed by miniaturizing the widely used clinical electroporator with a two-needle array into two-dimensional planar electrodes while keeping the similarity of the electric field strength distribution. In this device, all the biological processes from cell culture to electroporation and final cell-based assays were carried out on a chip using a conventional 2D cell culture method, and the multiple electrochemotherapeutic assays could be realized by exploiting the six electroporation sites in a single device. With the proposed platform, the electroporation rate was evaluated with propidium iodide and cell proliferation after 48 h of electrochemotherapy with bleomycin was determined with T47D human breast ductal carcinoma cell line in various electric field strengths and drug concentrations. This microsystem has several advantages over conventional cuvette type electroporation assay, such as multiple assays on a chip, on-chip based operation from cell culture to final assay, and having similar electric field distribution as that of the clinical electroporator. As the clinical trials of electrochemotherapy are being carried out, this new platform is expected to have valuable applications in basic in vitro ECT studies, drug discovery, and development of clinical ECT equipment.


The Development of Biocatalysis as a Tool for Drug Discovery.
Enzymes are versatile biocatalysts capable of performing selective reactions. The advantages of enzymes in comparison to classical chemistry including chemical catalysts are the generally milder process conditions and avoidance of harmful reactants. Their high selectivity and specificity are especially beneficial for the enzymatic synthesis of new products with potential applications in drug research. Therefore, in the past decades, the utilization of isolated enzymes or whole-cell biocatalysts has spread through a growing number of biotechnological industries. The applications comprise the production of chiral building blocks for the pharmaceutical and fine chemical industry, the enzymatic synthesis of drug metabolites for testing of toxicity, function, biological activity, degradation and the production of biocatalytically modified natural products, which all play a role in drug discovery. Especially Oreste Ghisalba's contributions, which paved the way for the industrial use of enzymes, will be considered in this review.


Fluorometric Detection of Low-Abundance EGFR Exon 19 Deletion Mutation Using Tandem Gene Amplification.
Epidermal growth factor receptor (EGFR) mutations are not only genetic markers for diagnosis but also biomarkers of clinical-response against tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). Among the EGFR mutations, the in-frame deletion mutation in EGFR exon 19 kinase domain (EGFR exon 19-del) is the most frequent mutation, accounting for about 45% of EGFR mutations in NSCLCs. Development of sensitive method for detecting the EGFR mutation is highly required to make a better screening for drug-response in the treatment of NSCLC patients. Here, we developed a fluorometric tandem gene amplification assay for sensitive detection of low-abundance EGFR exon 19-del mutant genomic DNA. The method consists of pre-amplification with PCR, thermal cycling of ligation by Taq ligase, and subsequent rolling circle amplification (RCA). PCR-amplified DNA from genomic DNA samples was used as splint DNA to conjugate both ends of linear padlock DNA, generating circular padlock DNA template for RCA. Long stretches of ssDNA harboring multiple copies of G-quadruplex structure was generated in RCA and detected by thioflavin T (ThT) fluorescence, which is specifically intercalated into the G-quadruplex, emitting strong fluorescence. Sensitivity of tandem gene amplification assay for detection of the EGFR exon 19-del from gDNA was as low as 3.6 pg, and mutant gDNA present in the pooled normal plasma was readily detected as low as 1% fraction. Hence, fluorometric detection of low-abundance EGFR exon 19 deletion mutation using tandem gene amplification may be applicable to clinical diagnosis of NSCLC patients with appropriate TKI treatment.


Radiographic anatomical relationship between spinous process and pedicle in thoracolumbar and lumbar spine.
Pedicle screws are widely used in spinal surgeries, but it remains technically demanding to place. There are numerous studies on the anatomy of pedicle; however, there is very little insight on the relationship between the pedicle and the spinous process, which is an important part of the spinal posterior column.The aim of the study was to investigate the radiographic anatomical relationship between spinous processes and pedicles in the thoracolumbar and lumbar spine, in order to reveal a novel entrance point for pedicle screw insertion.Sixty candidates were enrolled in this study; cases were excluded with degenerative disorders and other disorders as osteoporosis, deformity, and tumor. Radiographs and computer tomography scans between T10 and L5 were obtained on each case. The distance was measured that between the superior margin of spinous process root and the superior border, the inferior border and the axis of pedicle. In laboratory, 5 fresh cadavers were used to imitate the pedicle screw insertion.The basic reference point was supposed as the intersection between the horizontal line of superior margin of spinous process root and the central vertical line of the superior facet. For T10 to T12, the pedicle axis was 5 mm beyond the reference point. For L1 to L4, the pedicle axis was at the reference point. At L5, the pedicle axis was 5 mm beneath the reference point. In laboratory, 80 screws were all inserted into pedicles successfully according to the newly referred entrance point.The study reveals the radiographic anatomical relationship between the pedicle and the spinous process. The pedicle axis is around the horizontal line of the superior margin of spinous process root. It provides a new anatomic mark of pedicle screw entrance point.


The natural history of classic galactosemia: lessons from the GalNet registry.
Classic galactosemia is a rare inborn error of carbohydrate metabolism, caused by a severe deficiency of the enzyme galactose-1-phosphate uridylyltransferase (GALT). A galactose-restricted diet has proven to be very effective to treat the neonatal life-threatening manifestations and has been the cornerstone of treatment for this severe disease. However, burdensome complications occur despite a lifelong diet. For rare diseases, a patient disease specific registry is fundamental to monitor the lifespan pathology and to evaluate the safety and efficacy of potential therapies. In 2014, the international Galactosemias Network (GalNet) developed a web-based patient registry for this disease, the GalNet Registry. The aim was to delineate the natural history of classic galactosemia based on a large dataset of patients. Observational data derived from 15 countries and 32 centers including 509 patients were acquired between December 2014 and July 2018. Most affected patients experienced neonatal manifestations (79.8%) and despite following a diet developed brain impairments (85.0%), primary ovarian insufficiency (79.7%) and a diminished bone mineral density (26.5%). Newborn screening, age at onset of dietary treatment, strictness of the galactose-restricted diet, p.Gln188Arg mutation and GALT enzyme activity influenced the clinical picture. Detection by newborn screening and commencement of diet in the first week of life were associated with a more favorable outcome. A homozygous p.Gln188Arg mutation, GALT enzyme activity of ≤ 1% and strict galactose restriction were associated with a less favorable outcome. This study describes the natural history of classic galactosemia based on the hitherto largest data set.


Acne vulgaris, probiotics and the gut-brain-skin axis: from anecdote to translational medicine.
Acne vulgaris has long been postulated to feature a gastrointestinal mechanism, dating back 80 years to dermatologists John H. Stokes and Donald M. Pillsbury. They hypothesised that emotional states (e.g. depression and anxiety) could alter normal intestinal microbiota, increase intestinal permeability, and contribute to systemic inflammation. They were also among the first to propose the use of probiotic Lactobacillus acidophilus cultures. In recent years, aspects of this gut-brain-skin theory have been further validated via modern scientific investigations. It is evident that gut microbes and oral probiotics could be linked to the skin, and particularly acne severity, by their ability to influence systemic inflammation, oxidative stress, glycaemic control, tissue lipid content, and even mood. This intricate relationship between gut microbiota and the skin may also be influenced by diet, a current area of intense scrutiny by those who study acne. Here we provide a historical background to the gut-brain-skin theory in acne, followed by a summary of contemporary investigations and clinical implications.


Lgr4 protein deficiency induces ataxia-like phenotype in mice and impairs long term depression at cerebellar parallel fiber-Purkinje cell synapses.
Cerebellar dysfunction causes ataxia characterized by loss of balance and coordination. Until now, the molecular and neuronal mechanisms of several types of inherited cerebellar ataxia have not been completely clarified. Here, we report that leucine-rich G protein-coupled receptor 4 (Lgr4/Gpr48) is highly expressed in Purkinje cells (PCs) in the cerebellum. Deficiency of Lgr4 leads to an ataxia-like phenotype in mice. Histologically, no obvious morphological changes were observed in the cerebellum of Lgr4 mutant mice. However, the number of PCs was slightly but significantly reduced in Lgr4(-/-) mice. In addition, in vitro electrophysiological analysis showed an impaired long term depression (LTD) at parallel fiber-PC (PF-PC) synapses in Lgr4(-/-) mice. Consistently, immunostaining experiments showed that the level of phosphorylated cAMP-responsive element-binding protein (Creb) was significantly decreased in Lgr4(-/-) PCs. Furthermore, treatment with forskolin, an adenylyl cyclase agonist, rescued phospho-Creb in PCs and reversed the impairment in PF-PC LTD in Lgr4(-/-) cerebellar slices, indicating that Lgr4 is an upstream regulator of Creb signaling, which is underlying PF-PC LTD. Together, our findings demonstrate for first time an important role for Lgr4 in motor coordination and cerebellar synaptic plasticity and provide a potential therapeutic target for certain types of inherited cerebellar ataxia.


Relationship satisfaction and dating experience: factors affecting future time orientation in relationships with the opposite sex.
This study was conducted to examine the effects of relationship satisfaction and dating experience on future time orientation in relationships with the opposite sex. Data from a sample of 226 undergraduate students from Middle East Technical University indicated that individuals who were highly satisfied in their relationships with the opposite sex tended to seek temporary relationships more and tended to be less future focused in their relationships than those who were less satisfied. Results also indicated that present dating experience did not show any significant effect on future time orientation; however, previous dating experience did show an effect in the interaction with relationship satisfaction and different factors of future time orientation. Implications of these results are discussed.


Synthesis and antiinflammatory activity of some imidazolinylpyrazole derivatives.
Some new imidazolinyl-pyrazoles (4) were synthesized by condensation of 2-imidazolylhydrazine hydroiodid (1a) with beta-ketoesters to afford the intermediate 3 that, as free base, ring closes only to 4. The attempt to obtain the imidazo-triazepinone 5, starting from 2-imidazolinyl-2-methylhydrazine, was accomplished, in low yield, only in reacting with ethyl acetoacetate. The pharmacological evaluation of antiinflammatory and analgesic activities of pyrazoles 4a-f revealed an appreciable antiinflammatory activity.


The phases and stages of labour.
During recent decades the understanding and practice of obstetric care have evolved at a rapid pace. Consequently definitions that have promoted the principles of practice have had to be greatly modified or discarded. My thesis is that the time has come for the concepts of phases and stages of labour to be abandoned on the grounds that they now retard, rather than advance, progress in the care of the labouring woman.


Reconstructing seasonal range expansion of the tropical butterfly, Heliconius charithonia, into Texas using historical records.
While butterfly responses to climate change are well studied, detailed analyses of the seasonal dynamics of range expansion are few. Therefore, the seasonal range expansion of the butterfly Heliconius charithonia L. (Lepidoptera: Nymphalidae) was analyzed using a database of sightings and collection records dating from 1884 to 1992 from Texas. First and last sightings for each year were noted, and residency time calculated, for each collection locality. To test whether sighting dates were a consequence of distance from source (defined as the southernmost location of permanent residence), the distance between source and other locations was calculated. Additionally, consistent directional change over time of arrival dates was tested in a well-sampled area (San Antonio). Also, correlations between temperature, rainfall, and butterfly distribution were tested to determine whether butterfly sightings were influenced by climate. Both arrival date and residency interval were influenced by distance from source: butterflies arrived later and residency time was shorter at more distant locations. Butterfly occurrence was correlated with temperature but not rainfall. Residency time was also correlated with temperature but not rainfall. Since temperature follows a north-south gradient this may explain the inverse relationship between residency and distance from entry point. No long-term directional change in arrival dates was found in San Antonio. The biological meaning of these findings is discussed suggesting that naturalist notes can be a useful tool in reconstructing spatial dynamics.


Physical exercise augmented cognitive behaviour therapy for older adults with generalised anxiety disorder (PEXACOG): a feasibility study for a randomized controlled trial.
Generalised anxiety disorder (GAD) is a frequent and severe disorder among older adults. For older adults with GAD the effect of the recommended treatment, cognitive behaviour therapy (CBT), is reduced. Physical exercise (PE) may enhance the effect of CBT by improving cognitive function and increasing levels of brain-derived neurotrophic factor (BDNF), a predictor of the effect of CBT in patients with anxiety. The aim of the study was to assess the feasibility of a randomized controlled trial (RCT) investigating treatment effect of the combination of CBT and PE for GAD in a sample of older adults, including procedures for assessment and treatment. Four participants aged 62-70 years (M = 65.5, SD = 3.2) with a primary diagnosis of GAD were included. Participants received 15 weeks of PE in combination with 10 weeks of CBT. Participants completed self-report measures, and clinical, biological, physiological and neuropsychological tests at pre-, interim- and post-treatment. Procedures, protocols, and results are presented. One participant dropped out during treatment. For the three participants completing, the total adherence to PE and CBT was 80% and 100%, respectively. An independent assessor concluded that the completers no longer fulfilled the criteria for GAD after treatment. Changes in self-report measures suggest symptom reduction related to anxiety and worry. The sample is considered representative for the target population. The results indicate that combining CBT and PE for older adults with GAD is feasible, and that the procedures and tests are suitable and manageable for the current sample. ClinicalTrials.gov, NCT02690441. Registered on 24 February 2016, https://clinicaltrials.gov/ct2/show/NCT02690441 .


A heat shock transcription factor with reduced activity suppresses a yeast HSP70 mutant.
Strains carrying deletions in both the SSA1 and SSA2 HSP70 genes of Saccharomyces cerevisiae exhibit pleiotropic phenotypes, including the inability to grow at 37 degrees C or higher, reduced growth rate at permissive temperatures, increased HSP gene expression, and constitutive thermotolerance. A screen for extragenic suppressors of the ssa1 ssa2 slow-growth phenotype identified a spontaneous dominant suppressor mutation, EXA3-1 (R.J. Nelson, M. Heschl, and E.A. Craig, Genetics 131:277-285, 1992). Here we report that EXA3-1 is an allele of HSF1, which encodes the heat shock transcription factor (HSF). Strains containing the EXA3-1 allele in a wild-type background exhibit a 10- to 15-fold reduction in HSF activity during steady-state growth conditions as well as a delay in the accumulation of the SSA4, HSP26, and HSP104 mRNAs after a heat shock. EXA3-1-mediated suppression is the result of a single amino acid substitution of a highly conserved residue in the HSF DNA-binding domain which drastically reduces the ability of HSF to bind to heat shock elements as evaluated by band shift analysis. Together, these results indicate that the poor growth of ssa1 ssa2 strains is the result, at least in part, of the overproduction of a deleterious heat shock protein(s). This conclusion is supported by the fact that the levels of at least some heat shock proteins are reduced in ssa1 ssa2 cells containing the EXA3-1 allele. Surprisingly, strains containing the EXA3-1 allele in a wild-type HSP70 background grow early as well as the wild-type strain over a wide temperature range, displaying only a slight reduction in growth rate at 37 degrees Celsius, indicating that cells contain significantly more HSF activity than is require for growth under steady-state conditions.


Neurogenic bowel dysfunction in patients with spinal cord injury, myelomeningocele, multiple sclerosis and Parkinson's disease.
Exciting new features have been described concerning neurogenic bowel dysfunction, including interactions between the central nervous system, the enteric nervous system, axonal injury, neuronal loss, neurotransmission of noxious and non-noxious stimuli, and the fields of gastroenterology and neurology. Patients with spinal cord injury, myelomeningocele, multiple sclerosis and Parkinson's disease present with serious upper and lower bowel dysfunctions characterized by constipation, incontinence, gastrointestinal motor dysfunction and altered visceral sensitivity. Spinal cord injury is associated with severe autonomic dysfunction, and bowel dysfunction is a major physical and psychological burden for these patients. An adult myelomeningocele patient commonly has multiple problems reflecting the multisystemic nature of the disease. Multiple sclerosis is a neurodegenerative disorder in which axonal injury, neuronal loss, and atrophy of the central nervous system can lead to permanent neurological damage and clinical disability. Parkinson's disease is a multisystem disorder involving dopaminergic, noradrenergic, serotoninergic and cholinergic systems, characterized by motor and non-motor symptoms. Parkinson's disease affects several neuronal structures outside the substantia nigra, among which is the enteric nervous system. Recent reports have shown that the lesions in the enteric nervous system occur in very early stages of the disease, even before the involvement of the central nervous system. This has led to the postulation that the enteric nervous system could be critical in the pathophysiology of Parkinson's disease, as it could represent the point of entry for a putative environmental factor to initiate the pathological process. This review covers the data related to the etiology, epidemiology, clinical expression, pathophysiology, genetic aspects, gastrointestinal motor dysfunction, visceral sensitivity, management, prevention and prognosis of neurogenic bowel dysfunction patients with these neurological diseases. Embryological, morphological and experimental studies on animal models and humans are also taken into account.


Introducing HL7 FHIR Genomics Operations: a developer-friendly approach to genomics-EHR integration.
Enabling clinicians to formulate individualized clinical management strategies from the sea of molecular data remains a fundamentally important but daunting task. Here, we describe efforts towards a new paradigm in genomics-electronic health record (HER) integration, using a standardized suite of FHIR Genomics Operations that encapsulates the complexity of molecular data so that precision medicine solution developers can focus on building applications. FHIR Genomics Operations essentially "wrap" a genomics data repository, presenting a uniform interface to applications. More importantly, operations encapsulate the complexity of data within a repository and normalize redundant data representations-particularly relevant in genomics, where a tremendous amount of raw data exists in often-complex non-FHIR formats. Fifteen FHIR Genomics Operations have been developed, designed to support a wide range of clinical scenarios, such as variant discovery; clinical trial matching; hereditary condition and pharmacogenomic screening; and variant reanalysis. Operations are being matured through the HL7 balloting process, connectathons, pilots, and the HL7 FHIR Accelerator program. Next-generation sequencing can identify thousands to millions of variants, whose clinical significance can change over time as our knowledge evolves. To manage such a large volume of dynamic and complex data, new models of genomics-EHR integration are needed. Qualitative observations to date suggest that freeing application developers from the need to understand the nuances of genomic data, and instead base applications on standardized APIs can not only accelerate integration but also dramatically expand the applications of Omic data in driving precision care at scale for all.


Lignin degradation in wood-feeding insects.
The aromatic polymer lignin protects plants from most forms of microbial attack. Despite the fact that a significant fraction of all lignocellulose degraded passes through arthropod guts, the fate of lignin in these systems is not known. Using tetramethylammonium hydroxide thermochemolysis, we show lignin degradation by two insect species, the Asian longhorned beetle (Anoplophora glabripennis) and the Pacific dampwood termite (Zootermopsis angusticollis). In both the beetle and termite, significant levels of propyl side-chain oxidation (depolymerization) and demethylation of ring methoxyl groups is detected; for the termite, ring hydroxylation is also observed. In addition, culture-independent fungal gut community analysis of A. glabripennis identified a single species of fungus in the Fusarium solani/Nectria haematococca species complex. This is a soft-rot fungus that may be contributing to wood degradation. These results transform our understanding of lignin degradation by wood-feeding insects.


Effect of Inspiratory Muscle Training with Royal Jelly Supplement on Iron Metabolism in Cigarette Addicts.
The strength or weakness of the respiratory muscles compared with other skeletal muscles owing to their more specialized structures is considered as an indicator or cause of a disease. This study aimed to investigate the effects of inspiratory muscle training with royal jelly (RJ) supplement on iron metabolism in cigarette addicts. A total of 40 male volunteers participated in the study who were divided into 4 groups before the study as follows: smoker control group (n=10), inspiratory muscle training (IMT) group (n=10), RJ group (n=10), and RJ+IMT (n=10) group. Blood samples were taken from all the participants for analysis of iron, iron binding, and total iron binding capacity. Maximal inspiratory pressure (MIP) and maximal expiratory pressure (MEP) measurements for the groups to perform training were carried out with 40% of their MIP values. The training sessions were carried out at the same time every day for 4 weeks (5 days per week). The Statistical Package for Social Sciences version 22.0 program was used for statistical analyses. The results of the analysis found that the iron, iron binding, and total iron binding capacities were in favor of post-tests in the RJ supplement IMT, and RJ+IMT groups compared with those in the control group (p<0.05). There was a statistically significant difference between the RJ supplement, IMT, and RJ+IMT groups compared with the control group (p<0.05). Therefore, it could be concluded that the IMT and RJ supplements positively affected these parameters by altering the iron metabolism of the cigarette addicts.


Recent Patterns in Shared Decision Making for Prostate-Specific Antigen Testing in the United States.
Previous studies report infrequent use of shared decision making for prostate-specific antigen (PSA) testing. It is unknown whether this pattern has changed recently considering increased emphasis on shared decision making in prostate cancer screening recommendations. Thus, the objective of this study is to examine recent changes in shared decision making. We conducted a retrospective cross-sectional study among men aged 50 years and older in the United States using 2010 and 2015 National Health Interview Survey (NHIS) data (n = 9,598). Changes in receipt of shared decision making were expressed as adjusted prevalence ratios (aPR) and 95% confidence intervals (CI). Analyses were stratified on PSA testing (recent [in the past year] or no testing). Elements of shared decision making assessed included the patient being informed about the advantages only, advantages and disadvantages, and full shared decision making (advantages, disadvantages, and uncertainties). Among men with recent PSA testing, 58.5% and 62.6% reported having received ≥1 element of shared decision making in 2010 and 2015, respectively (P = .054, aPR = 1.04; 95% CI, 0.98-1.11). Between 2010 and 2015, being told only about the advantages of PSA testing significantly declined (aPR = 0.82; 95% CI, 0.71-0.96) and full shared decision making prevalence significantly increased (aPR = 1.51; 95% CI, 1.28-1.79) in recently tested men. Among men without prior PSA testing, 10% reported ≥1 element of shared decision making, which did not change with time. Between 2010 and 2015, there was no increase in shared decision making among men with recent PSA testing though there was a shift away from only being told about the advantages of PSA testing towards full shared decision making. Many men receiving PSA testing did not receive shared decision making.


[Acute asthmatic crisis in children].
Acute asthma attack in children is an attack responsible for life-threatening acute respiratory distress with partial or no response to bronchodilator drugs. The severity of the episode needs to be quickly evaluated. This presupposes a perfect knowledge of the clinical signs of severity. Treatment is urgent and first based on the administration of high doses of inhaled short-acting beta 2-agonists. In the more obstructed children, anti-cholinergic drugs can be added to nebulized beta 2-agonists. Because of their delayed effect, systemic steroids require an early prescription. Symptomatic treatments are: urgent hospitalization, oxygen if needed, proper hydratation. Continuous nebulization or intravenous perfusion of beta 2-agonists are prescribed with cardiac monitoring when no objective improvement is noted. Admission into the pediatric intensive care unit when bronchial obstruction continues will permit the association of bronchodilator drugs and the proposal of mechanical ventilation if needed. When the episode is resolved, a prophylactic treatment using inhaled corticosteroids must be prescribed. Clinical and spirometric follow-up has to be organized, and the patient and his/her family have to be educated.


Complex carbohydrates occurring in the digestive apparatus of Umbrina cirrosa (L.) fry.
The glycosoaminoglycans in the digestive apparatus of immature fish have important biological functions and are involved in morphofunctional differentiation. The aim of this study was to investigate the glycoconjugate histochemistry in the different parts of the digestive apparatus (oesophagus, stomach, intestine) of Umbrina cirrosa (L.) fry using classical histochemical reactions (periodic acid-Schiff, Alcian blue pH 2.5, Alcian blue/periodic acid-Schiff, high iron diamine, low iron diamine) in conjunction with glycolytic digestions that degrade different classes of glycosoaminoglycans. No differences were observed in the reactivity to conventional histochemical staining of the oesophagus, stomach or intestine among 27-, 34- or 44-day-old fry. In the oesophagus, the mucopolysaccharides contained chondroitin sulphates B and A and/or C, heparan sulphate and chondroitin. In the stomach, only neutral glycoconjugates were revealed, whereas in the intestine there were only chondroitin sulphates. Some differences in the type and content of glycoconjugates were found in Umbrina cirrosa (L.) fry compared to those of adult subjects, probably related to different dietary habits and to changes in the environmental conditions.


Selection for longer limbs in mice increases bone stiffness and brittleness, but does not alter bending strength.
The ability of a bone to withstand loads depends on its structural and material properties. These tend to differ among species with different modes of locomotion, reflecting their unique loading patterns. The evolution of derived limb morphologies, such as the long limbs associated with jumping, may compromise overall bone strength. We evaluated bone mechanical properties in the Longshanks mouse, which was selectively bred for increased tibia length relative to body mass. We combined analyses of 3D shape and cross-sectional geometry of the tibia, with mechanical testing and bone composition assays, to compare bone strength, elastic properties and mineral composition in Longshanks mice and randomly bred controls. Our data show that, despite being more slender, cortical geometry and predicted bending strength of the Longshanks tibia were similar to controls. In whole bone bending tests, measures of bone bending strength were similar across groups; however, Longshanks tibiae were significantly more rigid, more brittle, and required less than half the energy to fracture. Tissue-level elastic properties were also altered in Longshanks mice, but the bones did not differ from the control in water content, ash content or density. These results indicate that while Longshanks bones are as strong as control tibiae, selection for increased tibia length has altered its elastic properties, possibly through changes in organic bony matrix composition. We conclude that selection for certain limb morphologies, and/or selection for rapid skeletal growth, can lead to tissue-level changes that can increase the risk of skeletal fracture, which in turn may favor the correlated evolution of compensatory mechanisms to mitigate increased fracture risk, such as delayed skeletal maturity.


Subunit composition of Rhodothermus marinus respiratory complex I.
The basic structural characterization of complex I is still not trivial due to its complexity, not only in the number of its protein constituents but especially because of the different properties of the several subunits. Bacterial complex I generally contains 14 subunits: 7 hydrophilic proteins located in the peripheral arm and 7 hydrophobic proteins present in the membrane arm. It is the identification of the hydrophobic proteins that makes the characterization of complex I, and of membrane proteins in general, very difficult. In this article, we report the identification of the subunits of complex I from Rhodothermus marinus. The original approach, presented here, combined several protein and peptides separation strategies (different reversed phase materials, high-performance liquid chromatography, and gel electrophoresis) with different identification methods (electrospray ionization-tandem mass spectrometry, matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry, and Edman degradation analysis) and represents a step forward in the characterization of membrane proteins that studies are still technically highly challenging. The combination of the different methodologies allowed the identification of complex I canonical subunits and also a possible novel subunit, namely a pterin-4α-carbinolamine dehydratase (PCD). This was the first time that a PCD was suggested to be part of complex I, and its possible regulatory role is discussed.


{5,5'-Dihydroxy-2,2'-[(2-hydroxypropane-1,3-diyl)bis(nitrilomethanylyli-dene)]diphenolato}nickel(II) dihydrate.
In the title complex, [Ni(C17H16N2O5)]·2H2O, the Ni(II) ion is four-coordinated by two azomethine N and two phenolato O atoms of the tetradentate Schiff base ligand in a slightly distorted square-planar geometry. In the six-membered ring containing the metal, the azomethine N atoms and the three C atoms of the connecting 1,3-di-amino-propane-2-ol, all atoms except the metal are disordered over two sets of sites with an occupacy ratio of 0.566 (3):0.434 (3). The central C atom of the major component is significantly out of the mean plane of the remaing atoms while the conformation of this ring in the minor component is noticeably different. In the crystal, O-H⋯O hydrogen bonds involving the lattice water mol-ecules and the hy-droxy groups are observed.


Reactive species mediated injury of human lung epithelial cells after hypoxia-reoxygenation.
This study tested the hypothesis that hypoxia exposure predisposed lung epithelial cells to reactive oxygen species-(ROS) mediated cellular injury. Human lung carcinoma cells (ATCC line H441) having epithelial characteristics (including lamellar bodies, surfactant protein [SP]-A, and SP-B) were cultured in air (air/5% CO(2)) or hypoxia (< 1% O(2)/5% CO(2)) for 0 to 24 hours before imposition of oxidant stress. Cellular manganese superoxide dismutase (MnSOD) activity (units/mg protein) decreased significantly after 24 hours of hypoxia. In normoxic culture after hypoxia, the cells produced increased ROS, detected as dichlorofluorescein (DCF) fluorescence and H(2)O(2) accumulation in medium. Antioxidants N-acetylcysteine (N-Ac) and ebselen inhibited increased DCF fluorescence after hypoxia. To test their ability to tolerate oxidant stress, some cells were incubated with antimycin A (100 microM) and trifluorocarbonylcyanide phenylhydrazone (10 microM) (anti A + FCCP), a mitochondrial complex III inhibitor and respiratory chain uncoupler, which together increase mitochondrial superoxide (O(2)(-)) and H(2)O(2) production. Lung epithelial cells preexposed to hypoxia released more lactate dehydrogenase (LDH) than normoxic controls in response to increased O(2)(-) production. Increased LDH release from hypoxia-preexposed cells treated with anti A + FCCP was inhibited by 1 mM N-Ac. Rotenone and myxothiazole increased DCF oxidation more in hypoxic than in normoxic cells, suggesting that mitochondrial electron transport complex I may have been altered by hypoxia preexposure.


Natural language processing: state of the art and prospects for significant progress, a workshop sponsored by the National Library of Medicine.
Natural language processing (NLP) is crucial for advancing healthcare because it is needed to transform relevant information locked in text into structured data that can be used by computer processes aimed at improving patient care and advancing medicine. In light of the importance of NLP to health, the National Library of Medicine (NLM) recently sponsored a workshop to review the state of the art in NLP focusing on text in English, both in biomedicine and in the general language domain. Specific goals of the NLM-sponsored workshop were to identify the current state of the art, grand challenges and specific roadblocks, and to identify effective use and best practices. This paper reports on the main outcomes of the workshop, including an overview of the state of the art, strategies for advancing the field, and obstacles that need to be addressed, resulting in recommendations for a research agenda intended to advance the field.


PAPP-A and the IGF system in atherosclerosis: what's up, what's down?
Pregnancy-associated plasma protein-A (PAPP-A) is a metalloproteinase with a well-established role in releasing bioactive insulin-like growth factor-1 (IGF-1) from IGF-binding protein-2, -4, and -5 by proteolytic processing of these. The IGF system has repeatedly been suggested to be involved in the pathology of atherosclerosis, and both PAPP-A and IGF-1 are proposed biomarkers and therapeutic targets for this disease. Several experimental approaches based on atherosclerosis mouse models have been undertaken to obtain causative and mechanistic insight to the role of these molecules in atherogenesis. However, reports seem conflicting. The literature suggests that PAPP-A is detrimental, while IGF-1 is beneficial. This raises important questions that need to be addressed. Here we summarize the various studies and discuss potential underlying explanations for this seemingly inconsistency with the objective of better understanding complexities and limitations when manipulating the IGF system in mouse models of atherosclerosis. A debate clarifying what's up and what's down is highly warranted going forward with the ultimate goal of improving atherosclerosis therapy by targeting the IGF system.


Mechanism of the formation and proteolytic release of H2O2-induced dityrosine and tyrosine oxidation products in hemoglobin and red blood cells.
Oxyhemoglobin exposed to a continuous flux of H(2)O(2) underwent oxidative modifications, including limited release of fluorescent fragmentation products. The main fragments formed were identified as oxidation products of tyrosine, including dopamine, dopamine quinone, and dihydroxyindol. Further release of these oxidation products plus dityrosine was only seen after proteolytic degradation of the oxidatively modified hemoprotein. A possible mechanism is proposed to explain the formation of these oxidation products that includes cyclization, decarboxylation, and further oxidation of the intermediates. Release of dityrosine is proposed as a useful technique for evaluating selective proteolysis after an oxidative stress, because dityrosine is metabolically stable, and it is only released after enzymatic hydrolysis of the oxidatively modified protein. The measurement can be accomplished by high performance liquid chromatography with fluorescence detection or by high efficiency thin layer chromatography. Comparable results, in terms of dityrosine release, were obtained using red blood cells of different sources after exposing them to a flux of H(2)O(2). Furthermore, dityrosine has been reported to occur in a wide variety of oxidatively modified proteins. These observations suggest that dityrosine formation and release can be used as a highly specific marker for protein oxidation and selective proteolysis.


Current perspective on assessing site of digestion in ruminants.
The site of nutrient digestion in the gastrointestinal tract of ruminants affects nutrient availability and the nature of digestive end-products supplied to the host animal. Methods to study site of digestion in vivo provide a tool to obtain data that enhance the ability to interpret or predict animal responses to some feeding practices. Examples are discussed for which site of digestion data provided insights that might not have been evident from other approaches. The use of site of digestion techniques may provide interpretation regarding digestion of N by ruminants different from those derived from measurement of total tract N digestion. Site of digestion measurements have been particularly important in studying effects of heat processing of protein sources and in understanding the nature of digestion of N in high-quality, fresh forages. Moreover, site of digestion techniques have been instrumental in interpreting the influences of supplemental fat sources on ruminal digestion and ruminal biohydrogenation and small intestinal digestion of long-chain fatty acids.


Germplasm characterization and SDS-PAGE analysis of caper (Capparis spinosa L.) from different provenances.
Capparis spinosa L. is a typical desert plant that is resistant to high temperatures and drought, and at the same time is rich in medicinal and food values. The objective of this study is to explore the variations in nutrient composition, morphological characteristics, and SDS-PAGE patterns of caper seeds from different provenances, aiming to provide insights for the selection of superior seed provenances. In this experiment, there were significant differences in the morphological characteristics and major nutritional components of caper seeds from different provenances. Seeds from the YKL (Karayagaqi Township, Yining County) and YKG (G218, KashiTown, Yining County) regions were larger in size compared to seeds from other regions. Among the four measured nutritional components, crude fat had the highest content, especially in the YKL and YKG region. The results of correlation analysis showed that crude fat was negatively correlated with soluble sugar and soluble protein but significantly positively correlated with starch content. As longitude increased from east to west, the morphological characteristics gradually increased. Based on the principal component analysis of all the parameters of the seeds, the eight provenances could be classified into three groups. HM (Hami), TGS (S202, Gaochang District, Turpan), HYW (Wubao Town, Yizhou District, Hami), TQQ (Qiquanhu Town, Turpan), and TLF (Turpan) were a group with higher soluble protein, soluble sugar, and water content. YKL and YKG were in one group, which had larger seed grains with high crude fat and starch content. AKS (Aksu) was in a separate group. The protein fractions from seeds of eight regions were extracted using Osborne fractionation method, it was found that glutelin content was the highest, while albumin content was the lowest. After these proteins were analyzed by SDS-PAGE, the electrophoretic patterns showed that the protein molecular weights were relatively small, and there were differences in protein bands among different provenances. According to the PCA results, the eight seed provenances could be divided into three groups. There were both geographically distant ones clustered into one group, and those close to each other were also divided into one group. There were differences in seed morphology, nutrient content and SDS-PAGE profiles among the different seed sources. This difference might be caused by a combination of geographic and climatic factors. In addition, YKL and YKG were roughly selected as good seed provenances, which provided a theoretical basis for the development of C. spinosa L. germplasm resources.


Clinical trial master file migration: A preordained step for a centralized electronic trial master file.
The need to speed up clinical trial processes in a cost-effective manner, increased importance of data integrity, and ensuring timely compliance to regulatory requirement updates regarding the Trial Master File (TMF), has made the pharmaceutical industry delineate the requirement to maintain a centralized TMF with quality control. With the exponential increase in the number of sponsors using centralized electronic TMF (eTMF), the shift of trend positively impacts the need for data migration requirements in the TMF space. With an objective to serve the readers handling migration projects, this review article discusses the data migration requirements in clinical operations and eTMF in clinical trials, possible techniques to consider avoiding anticipated roadblocks, and a few other key points. The article also focuses on steps to be taken post migration to ensure meeting the quality of the migrated data in terms of regulatory compliance.


Is dangerousness an issue for physicians in emergency commitment?
The laws of many of the states require a physician to determine that a mentally ill individual presents a danger to others before the disturbed person can be civilly committed for involuntary treatment. The author contends that the prediction of dangerousness is not and should not be within the competence of medicine, but that physicians are competent to judge whether or not the severity of mental illness impairs a patient's competence to make an informed decision regarding treatment. The basic issue in emergency commitment is the patient's welfare, not his potential dangerousness. Dr. Alan A. Stone comments on Dr. Peszke's presentation.


Frequent allelic loss of 21q11.1 approximately q21.1 region in advanced stage oral squamous cell carcinoma.
A fine mapping of loss of heterozygosity (LOH) was performed in oral squamous cell carcinoma (OSCC), using 12 markers on 21q11.1 approximately q21.1. We studied 43 resected primary invasive tumors and their paired normal tissues, concurrent dysplasia or carcinoma in situ in separate areas from 8 of the specimens, and 6 local recurrent carcinomas. LOH status was compared between lesions of different phases of progression within the same patient. A high frequency of LOH was observed for D21S1410, D21S120, and D21S1433 (60% each) in the primary lesions, constituting two interstitial deleted regions encompassing eight known genes. Cases showing LOH of D21S120 were significantly associated with advanced clinical stages (III and IV; P=0.02). Consistent allelic loss was observed in 64.2% of the informative cases between the precursor lesions and their corresponding invasive tumors, and in 59.5% of those between the primary lesions and their recurrent counterparts. Fewer than half of the different lesions within a given patient showed discordant allelic loss for tested markers. Our results suggest that 21q11.1 approximately q21.1 harbors tumor suppressor genes in OSCC. Genetic divergence may develop during tumor clone evolution.