A Resident-led Initiative Improves Screening and Treatment for Vitamin D Deficiency in Patients with Hip Fractures.
Acute hip fractures carry a high risk of morbidity and are associated with low vitamin D levels. Improvements in screening and treating low vitamin D levels may lead to lower fall rates and a lower likelihood of additional fragility fractures. However, patients with low vitamin D levels often remain unassessed and untreated, even after they experience these fractures. We wished to determine whether a resident-led initiative can improve (1) screening for and (2) treatment of vitamin D deficiency in patients with acute hip fractures. Our department initiated a housestaff-led, quality improvement project focused on screening and treating vitamin D deficiency in patients with acute hip fractures. Screening encompassed checking serum 25-hydroxyvitamin D level during the acute hospitalization, and treating was defined as starting supplementation before discharge when the serum 25-hydroxyvitamin D level was less than 30 ng/mL. To evaluate the efficacy of this program, an administrative database identified 283 patients treated surgically for an acute hip fracture between July 2010 and June 2014. This period included 2 years before program initiation (Year 1, n = 65 patients; Year 2, n = 61 patients), the initial program year (Year 3, n = 66 patients), and the subsequent program year (Year 4, n = 91 patients). Followup was extended to 6 weeks after treatment with 9.2% (26/282) of patients lost to followup. Eight patients were excluded owing to documented intolerance of vitamin D supplementation. There were no differences regarding patient demographics, fracture type, or treatment rendered across these 4 years. The primary endpoints were the proportion of patients screened and treated for vitamin D deficiency. The secondary endpoint was the continuation of vitamin D supplementation at the patient's 6 week followup, according to the patient's medication list at that visit. This analysis included all patients, assuming those lost to followup had not continued supplementation. ANOVA and chi-square tests were used to evaluate the differences in demographic data and in screening and treating rates. Screening for vitamin D deficiency improved after initiation of the resident-led quality improvement program, with screening performed for 31% of patients in Year 1 (20/65; odds ratio [OR], 0.44; 95% CI, 0.26-0.75), 20% of patients in Year 2 (12/61; OR, 0.24; 95% CI, 0.13-0.46), 46% of patients in Year 3 (30/66; OR, 0.83; 95% CI, 0.51-1.35), and 88% of patients in Year 4 (80/91; OR, 7.27; 95% CI, 3.87-13.7) (p < 0.001). Vitamin D supplementation was initiated for 33% of patients in Year 1 (21/63; OR, 0.5; 95% CI, 0.30-0.84), 28% in Year 2 (17/61; OR, 0.39; 95% CI, 0.22-0.68), 50% in Year 3 (32/64; OR,1.00; 95% CI, 0.61-1.63), and 76% in Year 4 (65/86; OR, 3.10; 95% CI, 1.89-5.06) (p < 0.001). At early postoperative followup, we saw substantial improvement in the proportion of patients who continued receiving vitamin D supplementation: Year 1, 12% (8/64; OR, 0.14; 95% CI, 0.07-0.30); Year 2, 15% (9/61; OR, 0.17; 95% CI, 0.09-0.35); Year 3, 26% (16/64; OR, 0.33; 95% CI, 0.19-0.59); and Year 4, 46% (40/86; OR, 0.87; 95% CI, 0.57-1.33) (p < 0.001). Implementation of a resident-led quality improvement program resulted in higher rates of screening and treating vitamin D deficiency for patients with acute hip fractures. Housestaff-based initiatives may be an effective way to improve care processes that target improvements in bone health.


Interleukin and interleukin receptor gene polymorphisms and susceptibility to melanoma.
Genetic variation in immune-regulating components such as cytokines may lead to interindividual differences in immunosuppression response and susceptibility to melanoma. We evaluated the associations between genetic variants in five interleukins (IL) and IL receptor genes (IL-4, IL-4R, IL-6, IL-6R, and IL-10) and the risk of melanoma. Twenty-five single nucleotide polymorphisms (SNPs) were selected that are functionally relevant or tagging SNPs of each gene. We conducted a nested case-control study of 219 female patients and 219 matched controls within the Nurses' Health Study. We observed that in the IL-6R gene, four SNPs in linkage disequilibrium were associated with an increased risk of melanoma. Three are located in introns (rs6684439, rs4845618, and rs4845622), and one is a nonsynonymous SNP in exon 9 [rs8192284 (Asp358Ala)]. An elevated risk of melanoma was observed in the heterozygous groups of these SNPs with odds ratio of 1.74 [(95% confidence interval, 1.07, 2.81) for rs6684439, 1.72 (1.04, 2.84) for rs4845618, 1.69 (1.03, 2.75) for rs4845622, and 1.68 (1.04, 2.73) for rs8192284]. But these associations were not observed in the homozygous variant group with odds ratios ranging from 0.93 to 1.03. We did not find significant results for the SNPs in the other four genes. These data suggest the involvement of IL-6R in melanoma development. Further studies are needed to confirm these findings.


Vitrified-warmed blastocyst transfer on the 5th or 7th day of progesterone supplementation in an artificial cycle: a randomised controlled trial.
Prospective studies comparing different durations of progesterone supplementation before transfer of vitrified-warmed blastocysts in an artificial cycle are lacking. However, in oocyte donation programmes, the sporadic available evidence demonstrates considerable differences in clinical pregnancy rates according to the duration of progesterone administration. This randomised controlled trial (RCT), included 303 patients undergoing a frozen-thawed embryo transfer (FET) of one or two vitrified-warmed blastocyst(s) in an artificial cycle. Randomisation was performed when the endometrial thickness reached ≥7 mm after oestrogen supplementation. One hundred and fifty two patients in group A received 7 d of vaginal micronised progesterone tablets and 151 patients in group B received 5 d of micronised vaginal progesterone before FET. No differences were seen in clinical pregnancy rate between both groups: 42/152 (27.6%) in group A versus 49/151 (32.5%) in group B. Although no statistically significant difference was observed in clinical pregnancy rates, our study was powered to detect an absolute difference of 16%. In this regard, we cannot exclude that smaller, clinically relevant differences might exist and our study did not have the power to detect this. Patients were also not blinded for the intervention, causing a potential bias.


Identification of novel microRNA signatures linked to human lupus disease activity and pathogenesis: miR-21 regulates aberrant T cell responses through regulation of PDCD4 expression.
MicroRNAs (miRNAs) regulate the expression of genes involved in immune activation. A study was undertaken to characterise the miRNA signature and identify novel genes involved in the regulation of immune responses in systemic lupus erythematosus (SLE). The expression of 365 miRNAs in peripheral blood mononuclear cells of patients with SLE and healthy controls was analysed using TaqMan Low Density Arrays. The results were validated by quantitative real-time PCR and potential target genes were identified using prediction analysis software. The effect of miR-21 on T cell function was assessed by transfection with antago-miR-21 or pre-miR-21. A 27-miRNA signature was identified in patients with SLE; 19 miRNAs correlated with disease activity. Eight miRNAs were deregulated specifically in T cells and four miRNAs in B cells. miR-21 was upregulated and strongly correlated with SLE disease activity (r(2)=0.92). Compared with controls, CD4 T lymphocytes from patients with SLE had higher basal and activation-induced miR-21 expression. Silencing of miR-21 reversed the activated phenotype of T cells from patients with SLE--namely, enhanced proliferation, interleukin 10 production, CD40L expression and their capacity to drive B cell maturation into Ig-secreting CD19+CD38(hi)IgD-(plasma cells. Overexpression of mMiR-21 in normal T cells led to acquisition of an activated phenotype. Investigation of putative gene- targets showed that PDCD4 (a selective protein translation inhibitor) was suppressed by miR-21 and its expression was decreased in active SLE. miRNAs represent potential biomarkers in SLE as their expression reflects underlying pathogenic processes and correlates with disease activity. Upregulated miR-21 affects PDCD4 expression and regulates aberrant T cell responses in human SLE.


Chemical and immunological characterization of a novel amphipathic antigen from biotype B Streptococcus sanguis.
A new type of amphipathic antigen was extracted from whole cells of Streptococcus sanguis ATCC 10557 (biotype B, serotype II) by the phenol/water method. The extract was treated with nuclease P1, and was applied to a column of Sepharose 6B. Each fraction was checked by passive haemagglutination (PHA) and immunodiffusion tests against anti-10557 serum which was obtained by immunizing rabbits with whole cells of strain ATCC 10557. Strong PHA activity was demonstrated in the first hexose-containing peak (peak 1) eluted near the void volume, while the second hexose-containing peak (peak 2) produced a heavy band against anti-10557 serum in an immunodiffusion test. The third peak (peak 3) which partially overlapped with peak 2 reacted with concanavalin A, but not with the antiserum, in agar gel. Peaks 2 and 3 had no PHA activity. Peak 1 contained only 1% phosphorus, indicating that cells of strain ATCC 10557 possess an amphipathic antigen which differs from the lipoteichoic acids that are common in many Gram-positive bacteria. Peak 1 was a fatty acid-substituted heteropolysaccharide composed of glucose, galactose, mannose, glycerol and fatty acids in a molar ratio of approximately 1.0:1.3:2.7:0.3:1.0. PHA activity was inhibited in the presence of polymerized mannose. Peak 2 was composed of glucose, galactose, rhamnose and N-acetylgalactosamine in a molar ratio of approximately 1.0:1.4:0.8:0.8, which was essentially identical to the serotype II carbohydrate antigen reported previously.


Damage detection of structures identified with deterministic-stochastic models using seismic data.
A deterministic-stochastic subspace identification method is adopted and experimentally verified in this study to identify the equivalent single-input-multiple-output system parameters of the discrete-time state equation. The method of damage locating vector (DLV) is then considered for damage detection. A series of shaking table tests using a five-storey steel frame has been conducted. Both single and multiple damage conditions at various locations have been considered. In the system identification analysis, either full or partial observation conditions have been taken into account. It has been shown that the damaged stories can be identified from global responses of the structure to earthquakes if sufficiently observed. In addition to detecting damage(s) with respect to the intact structure, identification of new or extended damages of the as-damaged counterpart has also been studied. This study gives further insights into the scheme in terms of effectiveness, robustness, and limitation for damage localization of frame systems.


Emergent Solutions to High-Dimensional Multitask Reinforcement Learning.
Algorithms that learn through environmental interaction and delayed rewards, or reinforcement learning (RL), increasingly face the challenge of scaling to dynamic, high-dimensional, and partially observable environments. Significant attention is being paid to frameworks from deep learning, which scale to high-dimensional data by decomposing the task through multilayered neural networks. While effective, the representation is complex and computationally demanding. In this work, we propose a framework based on genetic programming which adaptively complexifies policies through interaction with the task. We make a direct comparison with several deep reinforcement learning frameworks in the challenging Atari video game environment as well as more traditional reinforcement learning frameworks based on a priori engineered features. Results indicate that the proposed approach matches the quality of deep learning while being a minimum of three orders of magnitude simpler with respect to model complexity. This results in real-time operation of the champion RL agent without recourse to specialized hardware support. Moreover, the approach is capable of evolving solutions to multiple game titles simultaneously with no additional computational cost. In this case, agent behaviours for an individual game as well as single agents capable of playing all games emerge from the same evolutionary run.


Chronic traumatic encephalopathy in sport: a systematic review.
To provide a critical review of chronic traumatic encephalopathy (CTE) by considering the range of clinical presentations, neuropathology and the strength of evidence for CTE as a distinct syndrome. Seven electronic databases were searched using a combination of MeSH terms and key words to identify relevant articles. Specific inclusion and exclusion criteria were used to select studies for review. Data extracted where present included study population, exposure/outcome measures, clinical data, neurological examination findings, cognitive assessment, investigation results and neuropathology results. The data from 158 published case studies were reviewed. Critical differences between the older descriptions of CTE (the 'classic' syndrome) and the recent descriptions (the 'modern' syndrome) exist in the age of onset, natural history, clinical features, pathological findings and diagnostic criteria, which suggests that modern CTE is a different syndrome. The methodology of the current studies does not allow determination of aetiology or risk factors. The clinicopathological differences between the 'classic' CTE syndrome and the 'modern' syndrome suggest that the new syndrome needs a different nomenclature. Further research is required to clearly define the clinical phenotype of the modern CTE syndrome and establish the underlying aetiology. Future research needs to address these issues through large-scale, prospective clinicopathological studies.


Serum visfatin level is associated with complexity of coronary artery disease in patients with stable angina pectoris.
Visfatin is an adipokine that plays a role in the inflammatory process of atherosclerosis. This study aimed to investigate whether adipokine is associated with the extent of stable coronary artery disease (CAD). The study population included 110 patients who underwent elective coronary angiography (CAG) due to stable angina pectoris. The severity of CAD was assessed by the 'Synergy Between Percutaneous Coronary Intervention With TAXUS and Cardiac Surgery (SYNTAX)' score. We evaluated patients in two groups: group 1 with a SYNTAX score <22 (low) and group 2 with a SYNTAX score ⩾22 (intermediate to high). Serum visfatin (8.6 ± 4.2 ng/ml versus 13.4 ± 5.2 ng/ml, p < 0.001) and serum C-reactive protein (CRP) levels [0.46 (0.25-0.77) mg/dl versus 0.71 (0.32-1.10) mg/dl, p < 0.001] were lower in group 1. A positive significant correlation was found between serum visfatin level and SYNTAX score (r = 0.559, p < 0.001). In a multivariate logistic regression analysis, visfatin [odds ratio (OR) 1.22, 95% confidence interval (CI) 1.10-1.36; p < 0.001], CRP (OR 6.22, 95% CI 1.70-22.7; p = 0.006), and diabetes mellitus (OR 3.83, 95% CI 1.10-13.2; p = 0.034) were found to be independent predictors of SYNTAX score. Serum visfatin level was positively correlated with CAD severity in patients with high SYNTAX score. Serum visfatin level can be a useful biomarker for predicting high SYNTAX scores in patients with angina pectoris undergoing CAG.


Aminoacetylation Reaction Catalyzed by Leucyl-tRNA Synthetase Operates via a Self-Assisted Mechanism Using a Conserved Residue and the Aminoacyl Substrate.
Leucyl-tRNA synthetase catalyzes attachment of leucine amino acid to its cognate tRNA. During the second, aminoacetylation, step of the reaction, the leucyl moiety is transferred from leucyl-adenylate to the terminal A76 adenosine of tRNA. In this work, we have investigated the aminoacetylation step catalyzed by leucyl-tRNA synthase, using ab initio quantum chemical/molecular mechanical hybrid potentials in conjunction with reaction-path-location algorithms and molecular dynamics free energy simulations. We have modeled reaction mechanisms arising from both crystallographic studies and computational work. We invoke various groups as potential proton acceptors-namely, the phosphate and leucyl amino groups of leucyl-adenylate, the A76 base of tRNA, and the Asp80 and Glu532 residues of the protein-and consider both metal-assisted and metal-free reactions. Free energy calculations indicate that both the phosphate group of leucyl adenylate and Glu532 are not strong bases. This agrees with the results of the quantum chemical/molecular mechanical reaction path calculations which give high free energy barriers for the studied pathways involving these groups. A self-assisted mechanism with the leucyl amino group and Asp80 as proton acceptors is the most likely. Furthermore, in this mechanism the presence of a metal ion coordinated by the phosphate group and Glu532 strongly activates the reaction.


Survival in cerebro-rhino-orbital zygomycosis and cavernous sinus thrombosis with combined therapy.
An alcoholic man with uncontrolled diabetes mellitus had right conjunctivitis, facial numbness, and periorbital edema progressing to bilateral visual loss, and left ptosis in association with a large necrotic palatal ulcer due to zygomycosis. The infection progressed to bilateral retinal vein engorgement; left-sided ophthalmoplegia, fixed dilated pupil, and absent corneal reflex; and right-sided ophthalmoplegia, ptosis, and facial nerve paralysis. Work-up revealed disease of both ethmoid sinuses and the right maxillary sinus, with bilateral thromboses of the cavernous sinuses. An aggressive combined therapeutic attack (three Caldwell-Luc procedures, exploration of orbit walls, control of diabetes, systemic and local amphotericin therapy) led to survival with a three-year follow-up thus far.


[Bromide-iodide baths and magnetic-laser radiation in the correction of disorders of hemodynamics and of the adaptation processes in metallurgical workers].
Positive changes in cardiovascular function and extracardiac regulation of cardiac rhythm in response to treatment by chloride-sodium bromine-iodine baths combined with magnetolaser radiation of the Zakhar'in-Ged zones suggest the conclusion on adaptogenic effects of the above physiobalneotherapeutic complex. The highest effect of magnetolaser therapy and bromine-iodine baths in metallurgists with adaptive disorders is achieved in early morning hours.


Screen Time and Autism Spectrum Disorder: A Systematic Review and Meta-Analysis.
Contemporary studies raise concerns regarding the implications of excessive screen time on the development of autism spectrum disorder (ASD). However, the existing literature consists of mixed and unquantified findings. To conduct a systematic review and meta-analyis of the association between screen time and ASD. A search was conducted in the PubMed, PsycNET, and ProQuest Dissertation & Theses Global databases for studies published up to May 1, 2023. The search was conducted independently by 2 authors. Included studies comprised empirical, peer-reviewed articles or dissertations published in English with statistics from which relevant effect sizes could be calculated. Discrepancies were resolved by consensus. This study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) reporting guideline. Two authors independently coded all titles and abstracts, reviewed full-text articles against the inclusion and exclusion criteria, and resolved all discrepancies by consensus. Effect sizes were transformed into log odds ratios (ORs) and analyzed using a random-effects meta-analysis and mixed-effects meta-regression. Study quality was assessed using the Grading of Recommendations, Assessment, Development, and Evaluations (GRADE) approach. Publication bias was tested via the Egger z test for funnel plot asymmetry. Data analysis was performed in June 2023. The 2 main variables of interest in this study were screen time and ASD. Screen time was defined as hours of screen use per day or per week, and ASD was defined as an ASD clinical diagnosis (yes or no) or ASD symptoms. The meta-regression considered screen type (ie, general use of screens, television, video games, computers, smartphones, and social media), age group (children vs adults or heterogenous age groups), and type of ASD measure (clinical diagnosis vs ASD symptoms). Of the 4682 records identified, 46 studies with a total of 562 131 participants met the inclusion criteria. The studies were observational (5 were longitudinal and 41 were cross-sectional) and included 66 relevant effect sizes. The meta-analysis resulted in a positive summary effect size (log OR, 0.54 [95% CI, 0.34 to 0.74]). A trim-and-fill correction for a significant publication bias (Egger z = 2.15; P = .03) resulted in a substantially decreased and nonsignificant effect size (log OR, 0.22 [95% CI, -0.004 to 0.44]). The meta-regression results suggested that the positive summary effect size was only significant in studies targeting general screen use (β [SE] = 0.73 [0.34]; t58 = 2.10; P = .03). This effect size was most dominant in studies of children (log OR, 0.98 [95% CI, 0.66 to 1.29]). Interestingly, a negative summary effect size was observed in studies investigating associations between social media and ASD (log OR, -1.24 [95% CI, -1.51 to -0.96]). The findings of this systematic review and meta-analysis suggest that the proclaimed association between screen use and ASD is not sufficiently supported in the existing literature. Although excessive screen use may pose developmental risks, the mixed findings, the small effect sizes (especially when considering the observed publication bias), and the correlational nature of the available research require further scientific investigation. These findings also do not rule out the complementary hypothesis that children with ASD may prioritize screen activities to avoid social challenges.


Misleading first impressions: different for different facial images of the same person.
Studies on first impressions from facial appearance have rapidly proliferated in the past decade. Almost all of these studies have relied on a single face image per target individual, and differences in impressions have been interpreted as originating in stable physiognomic differences between individuals. Here we show that images of the same individual can lead to different impressions, with within-individual image variance comparable to or exceeding between-individuals variance for a variety of social judgments (Experiment 1). We further show that preferences for images shift as a function of the context (e.g., selecting an image for online dating vs. a political campaign; Experiment 2), that preferences are predictably biased by the selection of the images (e.g., an image fitting a political campaign vs. a randomly selected image; Experiment 3), and that these biases are evident after extremely brief (40-ms) presentation of the images (Experiment 4). We discuss the implications of these findings for studies on the accuracy of first impressions.


Lipoxygenase-mediated modification of insect elicitors: generating chemical diversity on the leaf wound surface.
Plants can distinguish mechanical damage from larval folivory through the recognition of specific constituents of larval oral secretions (OS) which are deposited on the surface of leaf wounds during feeding. Fatty acid-amino acid conjugates (FACs) are major constituents of the OS of Lepidopteran larvae and they are strong elicitors of herbivore-induced defense responses in several plant species, including the wild tobacco Nicotiana attenuata. When OS from Manduca sexta larvae is deposited on N. attenuata wounded leaves, the major FAC N-linolenoyl-glutamic acid (18:3-Glu) is modified within seconds by a heat labile process. Some of the major modified forms are oxygenated products derived from 13-lipoxygenase activity and one of these derivatives, 13-oxo-13:2-Glu, is an active elicitor of enhanced JA biosynthesis and differential monoterpene emission in N. attenuata leaves.


Referral and timing of referral to hospice care in nursing homes: the significant role of staff members.
Given concerns about end-of-life care for many nursing home (NH) residents, this study sought to understand factors influencing hospice referral or nonreferral as well as timing of referral. We conducted semistructured interviews with personnel from seven participating NHs and two hospices. We interviewed NH directors of nursing regarding facility referral practices and conducted interviews with 34 NH nurses, 30 NH aides, and 17 hospice nurses knowledgeable about the factors that led to the hospice status of 32 NH decedents. Selected decedents varied by diagnosis and hospice status (received hospice for >7 days, <or=7 days, or not at all). We audiotaped, transcribed, and coded interviews for themes. NH staff members' recognition of terminal decline, beliefs about hospice, and initiative significantly influenced hospice referral and timing of referral. Staff members' recognition of familiar signs of decline facilitated hospice referral; in contrast, a perception that death was unexpected impeded referral, and a perception of uncertain prognosis delayed referral. Staff members' beliefs that hospice does not add value to NH care or is for crises only impeded referral, and a belief that hospice is only for the "very end" delayed referral. Residents received hospice for longer periods when staff believed that hospice complemented NH care and when staff took the initiative in raising the option of hospice. Enhanced training about recognizing terminal decline, hospice services in NHs, and the role of staff initiative would support NH staff in raising the option of hospice when appropriate.


The superior lateral genicular artery flap for reconstruction of knee and proximal leg defects.
Reconstruction of defects around the knee region requires thin and pliable skin. The superior lateral genicular artery (SLGA) flap provides an excellent alternative to muscle-based flaps. The anatomy and the surgical techniques of the SLGA flap were reviewed and the results of cases using the SLGA flap for coverage of knee and proximal leg defects were analyzed. SLGA flaps were performed in two cases and followed up for at least 6 months. Twelve articles on the use of the SLGA flap were also identified. A review of 39 cases showed that the mean diameter of the perforator supplying the skin of the flap was 1.04 mm, while the mean diameter of the SLGA at its origin was 1.78 mm. The mean length of the pedicle measured from the origin of the popliteal artery was 7.44 cm. The average dimensions of the flap were 14.8×6.6 cm with primary closure of the donor site in 61.5% of cases. Of these cases, 38.5% were due to trauma, 23.1% were post-burn complications, 12.8% were defects after resection of tumors, and 10.3% were for ulcers post-bursectomy. The most common complication was flap tip necrosis. All studies reported favorable outcomes with complete wound healing.


[Study of bovine carbonic anhydrase by 13C-NMR-spectroscopy].
The study of internal mobility in enzymes is of considerable importance for the understanding of their catalytic function, which cannot be adequately described as a property of a rigid protein. [13C]NMR spectroscopy permits simultaneous and selective observation of spectral lines from carbon atoms in many different residues in the enzyme with the chemical shift and relaxation parameters sensitive to structure, conformation and local motion. The changes in internal mobility in bovine carbonic anhydrase B (carbonate hydrolase, EC 22.214.171.124) in the native form and at various stages of denaturation are studied. Measurements of the relaxation parameters (T1, T1 rho) and of the NOE of 13C nuclei in the native protein showed that the extensive beta-sheet together with groups in the active center has a considerable internal librational mobility with tau G about 10(-11) s. This librational mobility is fairly uniform for all the alpha-carbons in the native enzyme. The use of a semiempirical modification of the motional theory proposed by Woessner allows to use simultaneously all the relaxation parameters measured in order to determine reliable values of the various correlation times.


MRSA colonisation and subsequent risk of infection despite effective eradication in orthopaedic elective surgery.
The objective of this study was to determine the effectiveness of screening and successful treatment of methicillin-resistant Staphylococcus aureus (MRSA) colonisation in elective orthopaedic patients on the subsequent risk of developing a surgical site infection (SSI) with MRSA. We screened 5933 elective orthopaedic in-patients for MRSA at pre-operative assessment. Of these, 108 (1.8%) were colonised with MRSA and 90 subsequently underwent surgery. Despite effective eradication therapy, six of these (6.7%) had an SSI within one year of surgery. Among these infections, deep sepsis occurred in four cases (4.4%) and superficial infection in two (2.2%). The responsible organism in four of the six cases was MRSA. Further analysis showed that patients undergoing surgery for joint replacement of the lower limb were at significantly increased risk of an SSI if previously colonised with MRSA. We conclude that previously MRSA-colonised patients undergoing elective surgery are at an increased risk of an SSI compared with other elective patients, and that this risk is significant for those undergoing joint replacement of the lower limb. Furthermore, when an infection occurs, it is likely to be due to MRSA.


Long-Term Treatment of Eosinophilic Esophagitis With Swallowed Topical Corticosteroids: Development and Evaluation of a Therapeutic Concept.
Swallowed topical corticosteroids (STCs) are efficacious in inducing and presumably maintaining remission in patients with active eosinophilic esophagitis (EoE). Hitherto, it has not been evaluated whether long-lasting remission can be achieved, and whether treatment can be stopped once patients have achieved this remission. Since 2007, EoE patients included into a large database at the Swiss EoE Clinics were put on STCs as induction/maintenance therapy. Disease activity was assessed on an annual basis. In patients who achieved long-lasting (≥6 months) clinical, endoscopic, and histological (=deep) remission, treatment was stopped. Data on all patients treated using this therapeutic strategy were analyzed retrospectively. Of the 351 patients, 33 (9.4%) who were treated with STCs achieved deep remission. Median age of remitters at disease onset was 32.6 years (interquartile range (IQR) 19.1-49.3), and diagnostic delay was 5.4 years (IQR 1.2-11.4). Deep remission was achieved after 89.0 weeks (IQR 64.6-173.8). Female gender was the only independent prognostic factor for achieving deep remission (odds ratio (OR) 2.518, 95% confidence interval (CI) 1.203-5.269). Overall, STCs were stopped after 104.7 weeks (IQR 65.5-176.6). No mucosal damage was observed upon histological examination. In 27 of the 33 remitters (81.8%), a clinical relapse occurred after a median of 22.4 weeks (95% CI 5.1-39.7). Six remitters (18.2%) did not experience a clinical relapse during a follow-up of 35.1 weeks (IQR 18.3-44.9). Hence, a total of 1.7% (6/351) patients were able to discontinue STCs in the long term. Long-term EoE treatment with STCs was well tolerated, but only a minority achieved deep remission. Female gender is the only prognostic factor for attainment of such remission. After treatment cessation, the majority experienced a clinical relapse.


Controlled solvent-exchange deposition of phospholipid membranes onto solid surfaces.
Controlled deposition of lipid bilayers plays a key role in creating supported membranes for biosensing devices and biophysical cell studies. The authors adopt a solvent-exchange method in order to deposit a phospholipid bilayer on solid substrates. The basic concept of deposition is to dissolve phospholipids in isopropanol-water mixtures and to increase water content gradually. Shortly before the onset of the micelle-to-vesicle transition, a lipid bilayer nucleates at the solid surface. They investigate the bulk phase behavior and surface coverage using small angle x-ray scattering and attenuated total reflection-Fourier transform infrared spectroscopy. They find a sequence of transitions from inverted-monomeric-micellar and vesicle phases correlating with an increasing amount of lipid on the adsorption layer. Supported lipid bilayers, prepared using this approach, are homogeneous and fluid.


Clinical onset of the Crohn's disease after eradication therapy of Helicobacter pylori infection. Does Helicobacter pylori infection interact with natural history of inflammatory bowel diseases?
There are conflicting reports concerning the prevalence of Helicobacter pylori infection in patients with inflammatory bowel diseases: some studies connected Sulphasalazine therapy and lower incidence of Helicobacter pylori infection, but others showed lower prevalence of Helicobacter pylori infection in inflammatory bowel diseases despite the choice of therapy. A 28-year-old male patient presented in January 1996 with the symptoms of ulcer like dyspepsia. There was no significant abnormality on physical examination, laboratory testing and abdominal ultrasound. Histology examination of the biopsy specimen taken during the upper endoscopy revealed Helicobacter pylori associated active gastritis only in the corporal part of the stomach. After two weeks eradication therapy (Omeprazole, Amoxicillin) he was well. Three months later, at the control endoscopy, granulomatous gastritis of the corporal localization was detected, without Helicobacter pylori present. Antral mucosa appeared normal, both, on endoscopy and histology examination. In July 1996 he started with cramping abdominal pain, mild periodical fever and episodes of watery diarrhea. In laboratory results we found nonspecific signs of inflammation. We repeated upper endoscopy, colonoscopy and enteroclysis--with evidence of segmental stenotic lesions of the upper part of ileum and jejunum. Again, we confirmed granulomatous gastritis and small granuloma in the proximal jejunum. After starting the 5-ASA therapy in combination with Metronidazol, patient was better clinical condition, and laboratory results were normal. We suggested mesalamine maintenance therapy 1 gr. every day, and three years later he is well, in clinical remission of Crohn's disease. The clinical course of the Crohn's disease maybe "sui generis" connected with Helicobacter pylori infection- but the exact mechanisms remain to be discovered.


No bejel among Surinamese, Antillean and Dutch syphilis diagnosed patients in Amsterdam between 2006-2018 evidenced by multi-locus sequence typing of Treponema pallidum isolates.
Treponema pallidum subspecies pallidum (TPA) and subsp. endemicum (TEN) are the causative agents of syphilis and bejel, respectively. TEN shows similar clinical manifestations and is morphologically and serologically indistinguishable from TPA. Recently, bejel was found outside of its assumed endemic areas. Using molecular typing we aimed to discover bejel and characterize circulating TPA types among syphilis cases with Surinamese, Antillean and Dutch ethnicity in Amsterdam. DNA was extracted from 137 ulcer swabs, which tested positive in the in-house diagnostic PCR targeting the polA gene. Samples were collected between 2006 and 2018 from Surinamese, Antillean and Dutch patients attending the Amsterdam STI clinic. Multilocus sequence typing was performed by partial sequence analysis of the tp0136, tp0548 and tp0705 genes. In addition, the 23S rRNA loci were analyzed for A2058G and A2059G macrolide resistance mutations. We found 17 distinct allelic profiles in 103/137 (75%) fully typed samples, which were all TPA and none TEN. Of the strains, 82.5% were SS14-like and 17.5% Nichols-like. The prevalence of Nichols-like strains found in this study is relatively high compared to nearby countries. The most prevalent types were 1.3.1 (42%) and 1.1.1 (19%), in concordance with similar TPA typing studies. The majority of the TPA types found were unique per country. New allelic types (7) and profiles (10) were found. The successfully sequenced 23S rRNA loci from 123/137 (90%) samples showed the presence of 79% A2058G and 2% A2059G mutations. No TEN was found in the samples from different ethnicities residing in Amsterdam, the Netherlands, so no misdiagnoses occurred. Bejel has thus not (yet) spread as a sexually transmitted disease in the Netherlands. The strain diversity found in this study reflects the local male STI clinic population which is a diverse, mixed group.


Escin-induced DNA damage promotes escin-induced apoptosis in human colorectal cancer cells via p62 regulation of the ATM/γH2AX pathway.
Escin, a triterpene saponin isolated from horse chestnut seed, has been used to treat encephaledema, tissue swelling and chronic venous insufficiency. Recent studies show that escin induces cell cycle arrest, tumor proliferation inhibition and tumor cell apoptosis. But the relationship between escin-induced DNA damage and cell apoptosis in tumor cells remains unclear. In this study, we investigated whether and how escin-induced DNA damage contributed to escin-induced apoptosis in human colorectal cancer cells. Escin (5-80 μg/mL) dose-dependently inhibited the cell viability and colony formation in HCT116 and HCT8 cells. Escin treatment induced DNA damage, leading to p-ATM and γH2AX upregulation. Meanwhile, escin treatment increased the expression of p62, an adaptor protein, which played a crucial role in controlling cell survival and tumorigenesis, and had a protective effect against escin-induced DNA damage: knockdown of p62 apparently enhanced escin-induced DNA damage, whereas overexpression of p62 reduced escin-induced DNA damage. In addition, escin treatment induced concentration- and time-dependent apoptosis. Similarly, knockdown of p62 significantly increased escin-induced apoptosis in vitro and produced en escin-like antitumor effect in vivo. Overexpression of p62 decreased the rate of apoptosis. Further studies revealed that the functions of p62 in escin-induced DNA damage were associated with escin-induced apoptosis, and p62 knockdown combined with the ATM inhibitor KU55933 augmented escin-induced DNA damage and further increased escin-induced apoptosis. In conclusion, our results demonstrate that p62 regulates ATM/γH2AX pathway-mediated escin-induced DNA damage and apoptosis.


Identification of yeast associated with the planthopper, Perkinsiella saccharicida: potential applications for Fiji leaf gall control.
Yeasts associate with numerous insects, and they can assist the metabolic processes within their hosts. Two distinct yeasts were identified by PCR within the planthopper Perkinsiella saccharicida, the vector of Fiji disease virus to sugarcane. The utility of both microbes for potential paratransgenic approaches to control Fiji leaf gall (FLG) was assessed. Phylogenetic analysis showed one of the microbes is related to yeast-like symbionts from the planthoppers: Laodelphax striatellus, Nilaparvata lugens, and Sogetella furcifera. The second yeast was a member of the Candida genus, a group that has been identified in beetles and recently described in planthoppers. Microscopy revealed the presence of yeast in the fat body of P. saccharicida. The Candida yeast was cultured, and transformation was accomplished by electroporation of Candida albicans codon optimized plasmids, designed to integrate into the genome via homologous recombination. Transgenic lines conferred resistance to the antibiotic nourseothricin and expression of green fluorescent protein was observed in a proportion of the yeast cells. Stably transformed yeast lines could not be isolated as the integrative plasmids presumably replicated within the yeast without integration into the genome. If stable transformation can be achieved, then this yeast may be useful as an agent for a paratransgenic control of FLG.


Review of intern preparedness and education experiences in General Medicine.
Few studies assess the transition from medical student to intern and there is limited understanding of what measures are required to assist intern development. The aim of the study was to assess interns' perception of their preparedness before commencing and on completion of their rotation in General Medicine, and their attitudes towards educational experiences at a tertiary metropolitan teaching hospital. Self-assessed preparedness for the General Medical internship and educational experiences were evaluated using a quantitative 5-point scale (1 = low score and 5 = high score) and qualitatively through interview, on interns based at St Vincent's Hospital (Melbourne). Data were collected at the beginning and at the end of each 10-week rotation (n = 25). Before commencement of the rotation, the interns identified areas where they felt inadequately prepared, particularly resuscitation skills and medico-legal aspects. When resurveyed at the completion of their 10-week rotation, the interns felt they had been better prepared for their role than they initially perceived, both generally and in specific aspects. Nine out of 16 parameters showed a significant increase in preparedness score at week 10 compared to week 1. The educational experiences most valued were peer driven education sessions and informal registrar teaching. Formal consultant teaching and online learning were perceived as being the least useful. Interns at St Vincent's Hospital have been adequately prepared for their role in General Medicine, although many realize this only in retrospect. Deficiencies in educational opportunities for interns have been uncovered that emphasize areas of attention for medical educators.


Acute phase cytokines, TAC1, and toll-like receptor4 mRNA expression and health associated with group size in veal calves.
Chronic stressors are a major health and well-being issue in animals. Immune status of animals under chronic stress is compromised, thus reducing disease resistance and compromising well-being of the animal. The objective of this study was to determine the influence of group size of veal calves on immune status and leukocyte mRNA expression of acute phase cytokines, toll-like receptor 4 (TLR4) and tachykinin 1 (TAC1) over a five-month finishing period. Holstein bull calves (n=168), 44±3 days of age were assigned to one of three treatments; 2, 4, or 8 calves/pen (pen space allowance of 1.82m(2)/calf). Jugular blood samples were collected at the day of grouping and then monthly for 4 months. The differential leukocyte counts were determined and mRNA was extracted from the leukocytes. Reverse transcription-qPCR was used to measure the gene expression of interleukin-1 (IL-1β), IL-1 receptor antagonist (IL-1Ra), tumor necrosis factor (TNF-α), TLR4, and TAC1 in leukocytes. Health was evaluated before grouping and monthly for 4 months. On the 1st month after grouping, veal calves that were housed in groups of 8 have greater expression of IL-1β mRNA than calves housed in groups of 4 or 2 (treatment×month, P=0.04). Also at 1 month, groups of 8 had greater TAC1 expression (P<0.05) than calves housed in groups of 4 or 2. However, the expression of IL-1Ra, TNF-α, and TLR4 were not influenced by group size. In the first month of the trial, calves in groups of 8 coughed more (P<0.05) than calves in groups of 2 and coughed more than calves in groups of 4 and 2 during the 2nd month (treatment×month, P=0.03). Calves housed in groups of 8 tended to have greater neutrophil percentage (P=0.09), neutrophil to lymphocyte ratio (P=0.06), and had lower lymphocyte percentage (P=0.06) than those housed in groups of 4 or 2. In conclusion, the number of veal calves in a group, given the same space during the finishing period did not alter IL-1Ra, TNF-α, and TLR4 mRNA expression. However, housing of calves in groups of 8 was associated with greater expression of IL-1β and TAC1 mRNA in peripheral blood leukocytes, and coughing during the first 2 months after grouping. Therefore, housing of veal calves in larger groups may lead to greater susceptibility to respiratory disease and stress.


Acute Kidney Injury After Subarachnoid Hemorrhage.
Acute kidney injury (AKI) is common in critically ill patients and may contribute to poor outcome. Few data are available on the incidence and impact of AKI in patients suffering from nontraumatic subarachnoid hemorrhage (SAH). We reviewed all patients admitted to our Department of Intensive Care with SAH over a 3-year period. Exclusion criteria were time from SAH symptoms to intensive care unit (ICU) admission >96 hours and ICU stay <48 hours. AKI was defined as sustained oligoanuria (urine output <0.5 mL/kg/h for 24 h) or an increase in plasma creatinine (≥0.3 mg/dL or a 1.5-fold increase from baseline level within 48 h). Neurological status was assessed at day 28 using the Glasgow Outcome Scale (GOS) (from 1=death to 5=good recovery; favorable outcome=GOS 4 to 5). Of 243 patients admitted for SAH during the study period, 202 met the inclusion/exclusion criteria (median age 56 y, 78 male). Twenty-five patients (12%) developed AKI, a median of 8 (4 to 10) days after admission. Independent predictors of AKI were development of clinical vasospasm, and treatment with vancomycin. AKI was more frequent in ICU nonsurvivors than in survivors (11/50 vs. 14/152, P=0.03), and in patients with an unfavorable neurological outcome than in other patients (17/93 vs. 8/109, P=0.03). Nevertheless, in multivariable regression analysis, AKI was not an independent predictor of outcome. AKI occurred in >10% of patients after SAH. These patients had more severe neurological impairment and needed more aggressive ICU therapy; AKI did not significantly influence outcome.


Dynamic Contrast-enhanced MRI Detects Responses to Stroma-directed Therapy in Mouse Models of Pancreatic Ductal Adenocarcinoma.
The dense stroma underlies the drug resistance of pancreatic ductal adenocarcinoma (PDA) and has motivated the development of stroma-directed drugs. Our objective is to test the concept that dynamic contrast-enhanced (DCE) MRI using FDA-approved contrast media, an imaging method sensitive to the tumor microenvironment, can detect early responses to stroma-directed drug. Imaging studies were performed in three mouse models exhibiting high desmoplastic reactions: the autochthonous PDA in genetically engineered mice (KPC), an orthotopic model in syngeneic mice, and a xenograft model of human PDA in athymic mice. An investigational drug, PEGPH20 (pegvorhyaluronidase alfa), which degrades hyaluronan (HA) in the stroma of PDA, was injected alone or in combination with gemcitabine. At 24 hours after a single injection of PEGPH20, Ktrans , a DCE-MRI-derived marker that measures how fast a unit volume of contrast media is transferred from capillaries to interstitial space, increased 56% and 50% from baseline in the orthotopic and xenograft tumors, respectively, compared with a 4% and 6% decrease in vehicle groups (both P < 0.05). Similarly, after three combined treatments, Ktrans in KPC mice increased 54%, whereas it decreased 4% in controls treated with gemcitabine alone (P < 0.05). Consistently, after a single injection of PEGPH20, tumor HA content assessed by IHC was reduced substantially in all three models while drug delivery (measured by paclitaxel accumulation in tumor) was increased by 2.6-fold. These data demonstrated a DCE-MRI marker, Ktrans , can detect early responses to stroma-directed drug and reveal the sustained effect of combination treatment (PEGPH20+ gemcitabine).


Facet-Dependent Charge Density of Serpentine: Nanoscopic Implications for Aggregation and Entrainment of Fine Particles.
Deciphering the facet-dependent surface properties of clay minerals holds vital significance in both fundamental research and practical engineering applications. To date, the anisotropic local charge density of serpentine surfaces still remains elusive, and thus, the interaction energies and associated aggregate structures between different crystal planes of serpentine cannot be quantitatively determined. In this work, different crystal planes of serpentine (i.e., SiO basal, MgOH basal, and edge) were selectively exposed, and their surface potentials and charge densities were determined using atomic force microscopy (AFM) force measurements coupled with Derjaguin-Landau-Verwey-Overbeek (DLVO) theory fitting. The SiO and edge planes consistently exhibited a permanently negative surface charge, whereas the point of zero charge (PZC) on the MgOH plane was estimated to be pH 9.0-11.0. Based on the interaction energy calculation between different serpentine planes, the aggregation structures of serpentine were predicted. Combined with scanning electron microscopy observation of freeze-dried samples, SiO-MgOH and MgOH-edge associations were found to dominate the aggregate structures at pH ≤ 9.0, thereby resulting in a stacking or "card-houses" structures. In contrast, all of the plane associations exhibited the repulsive interaction energy at pH 11.0, which led to a completely dispersed system, ultimately causing the most severe fine particle entrainment during froth flotation. Our work provides quantitative clarification of facet-dependent surface properties and aggregate structures of serpentine under different pH conditions, which will help improve the fundamental understanding of colloidal behaviors of clay minerals.


Is peritoneal dialysis the preferred modality in the diabetic patient?
There has been much debate recently whether peritoneal dialysis is an acceptable modality of renal replacement therapy in the diabetic patient. The authors review the recent continuous ambulatory peritoneal dialysis (CAPD) mortality literature and suggest that mortality can be improved by increasing the quantity of peritoneal dialysis delivered. The major disadvantages and advantages of CAPD are reviewed and presented. Peritoneal dialysis remains a viable modality of renal replacement therapy in the appropriate diabetic patient provided an adequate dose of dialysis is maintained.


Targeted broadband ultraviolet b phototherapy produces similar responses to targeted narrowband ultraviolet B phototherapy for vitiligo: a randomized, double-blind study.
Narrowband ultraviolet B (NB-UVB) phototherapy, with a 308-nm xenon chloride excimer laser, and targeted UVB phototherapy have produced encouraging therapeutic results for vitiligo. However, very few studies employing broadband UVB exist. Moreover, there has been no direct comparison study between broadband UVB and NB-UVB for the treatment of vitiligo. The aims of this study were to compare the repigmenting efficacy of targeted broadband UVB phototherapy with that of NB-UVB in an equi-erythemogenic manner. Twenty identical vitiliginous lesions from 10 patients were randomly allocated to receive either targeted broadband UVB or targeted NB-UVB phototherapy. UV fluences were started at 50% of the minimal erythema dose detected within the vitiliginous patches, then increased gradually, in the same manner, to ensure equi-erythemogenic comparison. Treatments were carried out twice weekly for 12 weeks. The results show that grade 1, i.e. 1-25% repigmentation, to grade 2, 26-50% repigmentation, occurred in 6 of 10 subjects. Responses in terms of repigmentation, de-pigmentation, or lack thereof, were similar between lesions receiving broadband and NB-UVB phototherapy. Onset of repigmentation occurred as early as 4 weeks of treatment in most subjects. Treatments were well tolerated, with only minimal erythema and hyperpigmentation. The study was carried out in a small number of patients with skin types III, IV and V. The irradiation device was a targeted UVB device and thus the results may not be applicable to other light sources, such as the excimer laser or total-body irradiation cabinets. In conclusion, targeted broadband UVB produces similar clinical responses to targeted NB-UVB in the treatment of the non-segmental type of vitiligo.


Regulation of gonadotropins by inhibin and activin.
Inhibin, activin, and follistatin were first identified as gonadal hormones that could exert selective effects on follicle-stimulating hormone (FSH) secretion without affecting luteinizing hormone (LH). Although the primary source of inhibin remains the gonad, both activin and follistatin are produced in extragonadal tissues and can exert effects on FSH through an autocrine-paracrine mechanism. These proteins can effect the regulation of the gonadotropins at many levels. First, activin can directly stimulate FSH biosynthesis and release from the gonadotrope cells of the pituitary gland. Second, activin up-regulates gonadotropin-releasing hormone receptor (GnRHR) gene expression, leading to alterations in the synthesis and release of both gonadotropins in response to GnRH. Third, activin can stimulate GnRH release from GnRH neurons in the hypothalamus and thereby affect FSH and LH secretion. Both inhibin and follistatin can negatively regulate these effects by preventing activin binding to the activin receptor at the cell membrane and blocking activation of downstream signal transduction pathways. This review concentrates on the mechanisms through which inhibin, activin, and follistatin regulate the gonadotropins. We discuss the expression of inhibin/activin subunits and receptors throughout the hypothalamus and pituitary and their role in the regulation of FSH and LH. The mechanisms of inhibin and activin signaling are also reported, with particular attention to developments in our understanding of inhibin receptor action and activin-induced transcriptional regulation of the FSHbeta gene promoter. Finally, we present recent findings that other members of the transforming growth factor beta superfamily may also play an important role in transcriptional regulation of the pituitary gonadotropins.


SARS-CoV-2 Nsp5 Demonstrates Two Distinct Mechanisms Targeting RIG-I and MAVS To Evade the Innate Immune Response.
Newly emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global pandemic with astonishing mortality and morbidity. The high replication and transmission of SARS-CoV-2 are remarkably distinct from those of previous closely related coronaviruses, and the underlying molecular mechanisms remain unclear. The innate immune defense is a physical barrier that restricts viral replication. We report here that the SARS-CoV-2 Nsp5 main protease targets RIG-I and mitochondrial antiviral signaling (MAVS) protein via two distinct mechanisms for inhibition. Specifically, Nsp5 cleaves off the 10 most-N-terminal amino acids from RIG-I and deprives it of the ability to activate MAVS, whereas Nsp5 promotes the ubiquitination and proteosome-mediated degradation of MAVS. As such, Nsp5 potently inhibits interferon (IFN) induction by double-stranded RNA (dsRNA) in an enzyme-dependent manner. A synthetic small-molecule inhibitor blunts the Nsp5-mediated destruction of cellular RIG-I and MAVS and processing of SARS-CoV-2 nonstructural proteins, thus restoring the innate immune response and impeding SARS-CoV-2 replication. This work offers new insight into the immune evasion strategy of SARS-CoV-2 and provides a potential antiviral agent to treat CoV disease 2019 (COVID-19) patients. IMPORTANCE The ongoing COVID-19 pandemic is caused by SARS-CoV-2, which is rapidly evolving with better transmissibility. Understanding the molecular basis of the SARS-CoV-2 interaction with host cells is of paramount significance, and development of antiviral agents provides new avenues to prevent and treat COVID-19 diseases. This study describes a molecular characterization of innate immune evasion mediated by the SARS-CoV-2 Nsp5 main protease and subsequent development of a small-molecule inhibitor.


Benefits of a health impact assessment in relation to fuel poverty: assessing Luton's Affordable Warmth Strategy and the need for a national mandatory strategy.
The links between fuel poverty and poor health are well documented, yet there is no statutory requirement on local authorities to develop fuel poverty strategies, which tend to be patchy nationally and differ substantially in quality. Fuel poverty starts from the perspective of income, even though interventions can improve health. The current public health agenda calls for more partnership-based, cost-effective strategies based on sound evidence. Fuel poverty represents a key area where there is currently little local evidence quantifying and qualifying health gain arising from strategic interventions. As a result, this initial study sought to apply the principles of a health impact assessment to Luton's Affordable Warmth Strategy, exploring the potential to identify health impact arising--as a baseline for future research--in the context of the public health agenda. A national strategy would help ensure the promotion of targeted fuel poverty strategies.


[The mechanisms for the decrease in gastric motor-evacuatory function after performing a vagotomy].
In experiment on white rats in was established that after the truncal vagotomy performance in the stress gastric ulcer the lowering of the serotonin and histamine blood content was noted as well as the disorder of ratio of their concentration in gastric wall. Insufficiency of serotonin causes the disorder of gastric motor-evacuation function after the operation. Administration in postoperative period of serotonin adipinate had promoted the raising of serotonin concentration in blood and gastric tissues, histamine level lowering, enhancement of serotonin/histamine ratio.


Disrupting the hydrophobic patches at the antibody variable/constant domain interface: improved in vivo folding and physical characterization of an engineered scFv fragment.
By constructing Fv and single-chain Fv (scFv) fragments of antibodies, the variable domains are taken out of their natural context in the Fab fragment, where they are associated with the constant domains of the light (CL) and heavy chain (CH1). As a consequence, all residues of the former variable/constant domain interface become solvent exposed. In an analysis of 30 non-redundant Fab structures it was found that at the former variable/constant domain interface of the Fv fragment the frequency of exposed hydrophobic residues is much higher than in the rest of the Fv fragment surface. We investigated the importance of these residues for different properties such as folding in vivo and in vitro, thermodynamic stability, solubility of the native protein and antigen affinity. The experimental model system was the scFv fragment of the anti-fluorescein antibody 4-4-20, of which only 2% is native when expressed in the periplasm of Escherichia coli. To improve its in vivo folding, a mutagenesis study of three newly exposed interfacial residues in various combinations was carried out. The replacement of one of the residues (V84D in VH) led to a 25-fold increase of the functional periplasmic expression yield of the scFv fragment of the antibody 4-4-20. With the purified scFv fragment it was shown that the thermodynamic stability and the antigen binding constant are not influenced by these mutations, but the rate of the thermally induced aggregation reaction is decreased. Only a minor effect on the solubility of the native protein was observed, demonstrating that the mutations prevent aggregation during folding and not of the native protein. Since the construction of all scFv fragments leads to the exposure of these residues at the former variable/constant domain interface, this strategy should be generally applicable for improving the in vivo folding of scFv fragments and, by analogy, also the in vivo folding of other engineered protein domains.


Possibilities of the exposure reduction of hands during the preparation and application of radiopharmaceuticals.
The current routinely used methods of estimating the skin equivalent dose relies on the finger dosimetry which usually largely underestimates the real maximum exposure and thus appropriate correction factors have to be used. The group under the investigation consisted of 10 workers preparing and 5 workers administering radiopharmaceuticals labelled with 18F. The monitoring was carried out using 12 pairs of thermoluminiscent dosimeters (TLDs) placed on each hand of the worker. A total of 46 measurements were completed. The maximum exposure of the skin of hands, defined in terms of the quantity of the personal dose equivalent Hp(0.07), was related to the unit activity of radiopharmaceutical with which the worker came into the contact during the measurement. The exposure of the hands of workers handling 18F-labelled radiopharmaceuticals showed significant inhomogeneity. Out of 15 workers, in 53 % of cases, the maximum skin exposure was observed on the tip of their index finger. It was estimated that in about 60 % of the cases (during the preparation and administration of radiopharmaceuticals), the exposure may exceed the 3/10 of the annual dose limit. Moreover, in 40 % of all cases, the exposure may even be higher than this dose limit. The established relevant correction factors reached the values up to 8 (as for preparations) and 13 (as for administrations). The study resulted in the establishment of the appropriate correction factors and in the recommendations of procedures aimed at the further reduction of the exposure of extremities (Tab. 3, Fig. 2, Ref. 17).


The urban house mouse (Mus domesticus) as a reservoir of infection for the human parasite Toxoplasma gondii: an unrecognised public health issue?
Toxoplasma gondii is a protozoan parasite capable of infecting almost all warm-blooded animals. The cat is the definitive host and becomes infected by consuming contaminated meat or infected prey. Humans can act as intermediate hosts and in healthy individuals the infection is mild and self-limiting. In pregnant women it can cause spontaneous abortions and foetal abnormalities and is capable of inducing serious illness in immuno-compromised patients. In infested dwellings, mice could act as intermediate hosts and play a role in the persistence/propagation of the disease. A total of 200 mice were trapped alive in 27 infested properties in Manchester, UK, and screened for Toxoplasma infection; 59% tested positive. Evidence of vertical transmission from infected dams to foetus was found, possibly maintaining the infection in urban areas. These findings have important implications when considering approaches to rodent control.


Risk of Death among HIV Co-Infected Multidrug Resistant Tuberculosis Patients, Compared To Mortality in the General Population of South Africa.
Even though highly effective drugs are available in South Africa, multidrug resistant tuberculosis (MDR-TB) patients with HIV infection have higher mortality compared to HIV-uninfected MDR-TB patients. This trend has been observed in similar countries with high HIV prevalence. This study sought to determine excess mortality attributable to HIV among MDR-TB patients in South Africa using relative survival methods. Data available were from a cohort of 2079 MDR-TB patients enrolled in a Standardized Programmatic Management of MDR-TB from 2000 to 2004 in South Africa. A Poisson-based model adjusted for age, gender, year of diagnosis, TB history, and resistance to ethambutol, anti-TB injectable drugs and fluoroquinolones antibiotics was constructed to assess the excess mortality among HIV co-infected MDR-TB patients. Excess hazard ratios (EHRs) were used to describe the effect of the predictors on net mortality, controlling for the general mortality in the South African population. Death was recorded on 1619 patients, of whom 367 (22.7%) had died within 2 years. Out of the 1413 patients that tested for HIV infection, 554 (39.2%) tested positive. Excess mortality was higher in HIV infected, compared to HIV uninfected, MDR-TB patients (adjusted excess hazard ratio, 5.6 [95% CI, 3.2-9.7]); in patients whose TB isolates' resistance to ethambutol and kanamycin was unknown (3.7 [2.1-6.2] and 4.87 [1.9-13.3], respectively) vs. known. There were no differences in excess mortality between age and gender of the patient, year of diagnosis and TB history. Adjusting for some important predictors, MDR-TB patients with HIV infection experienced higher excess mortality compared to HIV-uninfected MDR-TB patients, after accounting for the general mortality in South Africa. An appropriate, though complex method has produced predictor effect estimates similar to those obtained from classical methods. Thus, the use of relative survival methods should be encouraged in the analysis of causespecific mortality, when ascertainment of cause of death is inaccurate or unknown.


Renal function and risk stratification of diabetic and nondiabetic patients undergoing evaluation for coronary artery disease.
The aim of this study was to evaluate the impact of renal function by estimated glomerular filtration rate (eGFR) on risk stratification of diabetic and nondiabetic patients undergoing myocardial perfusion imaging (MPI) by single-photon emission computed tomography for suspected ischemia. Coronary artery disease is the leading cause of death among diabetic persons; however, diabetic persons are a very heterogeneous group in terms of cardiovascular risk, necessitating further risk stratification. Patients (n = 1,747, age 65 +/- 10 years, 37% diabetic) undergoing MPI were followed for cardiac death (CD) for a mean of 2.15 +/- 0.8 years. Chronic kidney disease (CKD) was defined by an eGFR <60 ml/min. In the presence of a normal scan, annual CD rate was 0.9% for those with no diabetes mellitus (DM) and no CKD, 0.5% in the DM alone group, 2.35% in CKD alone, and 2.9% in those with both DM and CKD (p < 0.001). Patients with DM+CKD had a 2.7-fold risk of CD compared with no DM no CKD (p = 0.001) after controlling for age, ejection fraction, history of coronary artery disease, and other risk factors. The risk of CD increased as a function of the presence and severity of perfusion defects, regardless of CKD or DM status. Presence of CKD conferred a several-fold higher risk of CD for the various strata of perfusion defects. Log-rank test for difference in probability of CD was nonsignificant for comparison between patients with no DM no CKD and those with DM alone (p = 0.73) but was significant for comparison between patients with no DM no CKD and patients with CKD alone (p < 0.001) or DM+CKD (p < 0.001). MPI and eGFR provide valuable risk stratification for diabetic and nondiabetic patients. Diabetic patients without CKD seem to have similar short-term cardiac outcomes compared with nondiabetic patients. Underlying CKD seems to identify a high-risk subgroup of diabetic patients.


Attraction of Phlebotomus papatasi to common fruit in the field.
Sand flies have been reported feeding on various plant organs including stems, leaves, and flowers but the attraction of sand flies to sugar-rich fruits has received little attention. In this study, we tested 24 commercially available fruits for their attractiveness to sand flies, and found that the top three attractive fruits were nectarine (Prunus persica var. nectarina), cactus fruit, (Opuntia ficus-indica), and guava (Psidium guajava). These fruits were fed upon equally by both males and females. There were slight differences in the order of preference to the less-attractive fruits by males and females, but these were not statistically significant. The knowledge of fruit preference may help to improve existing methods that use plant phytochemicals to attract and kill biting flies.


Reduction of (per)chlorate by a novel organism isolated from paper mill waste.
As part of a study on the microbiology of chlorate reduction, several new dissimilatory chlorate-reducing bacteria were isolated from a broad diversity of environments. One of these, strain CKB, was selected for a more complete characterization. Strain CKB was enriched and isolated from paper mill waste with acetate as the sole electron donor and chlorate as the sole electron acceptor. Strain CKB is a completely oxidizing, non-fermentative, Gram-negative, facultative anaerobe. Cells of strain CKB are 0.5 x 2 microm and are highly motile, with a single polar flagellum. In addition to acetate, strain CKB can use propionate, butyrate, lactate, succinate, fumarate, malate or yeast extract as electron donors, with chlorate as the sole electron acceptor. Strain CKB can also couple chlorate reduction to the oxidation of ferrous iron, sulphide, or the reduced form of the humic substances analogue 2,6-anthrahydroquinone disulphonate. Fe(II) is oxidized to insoluble amorphous Fe(II) oxide, whereas sulphide is oxidized to elemental sulphur. Growth is not associated with this metabolism, even when small quantities of acetate are added as a potential carbon source. In addition to chlorate, strain CKB can also couple acetate oxidation to the reduction of oxygen or perchlorate. Chlorate is completely reduced to chloride. Strain CKB has an optimum temperature of 35 degrees C, a pH optimum of 7.5 and a salinity optimum of 1% NaCl. Strain CKB can grow in chlorate and perchlorate concentrations of 80 or 20 mM respectively. Under anaerobic conditions, strain CKB can dismutate chlorite into chloride and O2, and is only the second organism shown to be capable of this metabolism. Oxidized minus reduced spectra of whole-cell suspensions of strain CKB showed absorbance maxima at 423, 523 and 552nm, which are indicative of the presence of c-type cytochrome(s). Analysis of the complete sequence of the 16S rDNA indicates that strain CKB is a member of the beta subclass of the Proteobacteria. The phototroph Rhodocyclus tenuis is the closest known relative. When tested, strain CKB could not grow by phototrophy and did not contain bacteriochlorophyll. Phenotypically and phylogenetically, strain CKB differs from all other described bacteria and represents the type strain of a new genus and species.


Application of 13C flux analysis to identify high-productivity CHO metabolic phenotypes.
Industrial bioprocesses place high demands on the energy metabolism of host cells to meet biosynthetic requirements for maximal protein expression. Identifying metabolic phenotypes that promote high expression is therefore a major goal of the biotech industry. We conducted a series of 13C flux analysis studies to examine the metabolic response to IgG expression during early stationary phase of CHO cell cultures grown in 3L fed-batch bioreactors. We examined eight clones expressing four different IgGs and compared with three non-expressing host-cell controls. Some clones were genetically manipulated to be apoptosis-resistant by expressing Bcl-2Δ, which correlated with increased IgG production and elevated glucose metabolism. The metabolic phenotypes of the non-expressing, IgG-expressing, and Bcl-2Δ/IgG-expressing clones were fully segregated by hierarchical clustering analysis. Lactate consumption and citric acid cycle fluxes were most strongly associated with specific IgG productivity. These studies indicate that enhanced oxidative metabolism is a characteristic of high-producing CHO cell lines.


Metoclopramide and pimozide in Parkinson's disease and levodopa-induced dyskinesias.
Metoclopramide is an antiemetic drug which occasionally produced acute dystonic reactions. Although known to interfere with central dopamine mechanisms, it is frequently used in Parkinson's disease to prevent levodopa-induced nausea and vomiting. In this study metoclopramide did not increase Parkinsonism or reduce levodopa-induced involuntary movements in patients with Parkinson's disease. Pimozide, by contrast, increased Parkinsonism and reduced involuntary movements. The capacity of metoclopramide to produce acute dyskinesias while being apparently free of Parkinsonism effects is pharmacologically unique and differentiates this drug from the phenothiazines and butyrophenones.


Field and laboratory tests of new insecticides against the apple maggot, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae).
Laboratory bioassays and field tests were conducted to compare the effectiveness of the new insecticides, imidacloprid, indoxacarb, pyriproxyfen, spinosad, thiacloprid, and thiamethoxam, against apple maggot. The activity ranking of the compounds in reducing oviposition in laboratory bioassays was: imidacloprid, 95% reduction at 11 ppm > thiamethoxam, 91% and thiacloprid, 89% reduction at 100 ppm > spinosad, 98% reduction at 316 ppm > indoxacarb, 80% reduction at 1000 ppm > pyriproxyfen, 0% reduction at 38 ppm. In laboratory bioassays, the only insecticides that were toxic to flies at concentrations equal to or below the recommended field rates were imidacloprid, (50% of flies at 11 ppm), spinosad (90% of flies at rates > 10 ppm), and thiamethoxam (approximately 50% of flies at 32 ppm). In field trials, thiacloprid was the only material that consistently controlled apple maggot fruit infestation that was comparable to standard treatments of organophosphate insecticides. Spinosad applied at weekly intervals, and indoxacarb applied as biweekly sprays provided adequate control of apple maggot damage when infestation levels in the field were low, but were not effective in preventing damage in small plots when apple maggot pressure was high.


An assessment of DHS maternal mortality indicators.
This study presents an assessment of the quality of data relating to maternal mortality collected in 14 Demographic and Health Surveys (DHS) for 13 countries that included a complete sibling history. Four aspects of data quality are considered: completeness of the data for reported events, evidence of omission in the reporting of events, plausibility of the pattern of sibling deaths, and sampling errors of the maternal mortality estimates. Although the data relating to reported events are complete for most variables, comparisons of sibling-history-based estimates of adult mortality for both males and females with other independent estimates suggest that sibling estimates are more likely to be underestimates than overestimates. The downward bias is probably greater for female mortality than for male mortality. The sampling errors associated with maternal mortality ratios are substantially larger than those associated with other frequently used DHS indicators. This lack of precision precludes the use of these data for trend analysis and has led to the recommendation that this DHS module not be used more than once every ten years in the same country.


Mouse senile amyloid deposition is suppressed by adenovirus-mediated overexpression of amyloid-resistant apolipoprotein A-II.
Apolipoprotein A-II (apoA-II), the second most abundant apolipoprotein of serum high density lipoprotein, deposits as an amyloid fibril (AApoAII) in old mice. Mouse strains with a high incidence of senile amyloidosis have the type C apoA-II gene (Apoa2(c)), whereas the strains with a low incidence of amyloidosis have the type B apoA-II gene (Apoa2(b)). In this study, to investigate whether the type B apoA-II protein inhibits the extension of amyloid fibrils, we constructed an adenovirus vector bearing the Apoa2(b) cDNA (Adex1CATApoa2(b)), which is expressed under the control of a hepatocyte-specific promoter. The mice were infected with Adex1CATApoa2(b) before induction of amyloidosis by the injection of AApoAII amyloid fibril seeds. Compared with the mice infected with the control virus, amyloid deposition was suppressed significantly in the mice infected with Adex1CATApoa2(b). Fluorometry using thioflavine T also revealed that AApoAII fibril extension was inhibited by the addition of type B apoA-II in vitro. Thus, we propose that Apoa2(b) contributes as an active inhibitor of amyloid fibril extension and overexpression of amyloid-resistant gene variant may be an attractive therapeutic target in amyloidosis.


Computational Models of the Intestinal Environment. 3. The Impact of Cholesterol Content and pH on Mixed Micelle Colloids.
In this study, we use molecular dynamics (MD) and experimental techniques (nephelometry and dynamic light scattering) to investigate the influence of cholesterol content and pH on the colloidal structures that form in the gastrointestinal (GI) tract upon lipid digestion. We demonstrate that the ionization state of the molecular species is a primary driver for the self-assembly of aggregates formed by model bile and therefore should be considered when performing in silico modeling of colloidal drug delivery systems. Additionally, the incorporation of physiological concentrations of cholesterol within the model systems does not affect size, number, shape, or dynamics of the aggregates to a significant degree. The MD data shows a reduction in aggregate size with increasing pH, a preference for glycodeoxycholate (GDX) to occupy the aggregate surface, and that the mixed micellar aggregates are oblate spheroids (disc-like). The results obtained assist in understanding the process by which pH and cholesterol influence self-assembly of mixed micelles within the GI tract. The MD approach provides a platform for investigation of interactions of drugs and formulation excipients with the endogenous contents of the GI tract.


Insulin attenuates vasopressin-induced calcium transients and a voltage-dependent calcium response in rat vascular smooth muscle cells.
Insulin attenuates the contractile responses of vascular smooth muscle (VSM) to various agonists. Insulinopenic and insulin-resistant rats lack this normal attenuation of vascular contractile responses. To study this attenuating mechanism, the effects of insulin on calcium (Ca2+) responses of cultured VSM cells (a7r5) to arginine vasopressin (AVP) and membrane potential were investigated. Insulin (1 and 100 mU/ml) shifted AVP dose-response curves to the right, reducing relative potency of AVP by 16-fold and 220-fold, respectively. Responses to AVP were significantly attenuated within 30 min of insulin application. The AVP-elicited rise in [Ca2+]i was partially dependent upon extracellular Ca2+. AVP-elicited inward current was reduced by 90 min of insulin treatment (100 mU/ml), from a peak current of -103 +/- 27 pA (normal) to -37 +/- 15 pA (insulin treated). Peak voltage-dependent Ca(2+)-dependent inward current was unaffected by insulin; however, the current-voltage curve was shifted 16 +/-3 mV to the right by insulin. Thus, insulin may reduce VSM contractile responses by attenuating agonist-mediated rises in [Ca2+]i mediated, in part, by reductions in Ca2+ influx through both receptor- and voltage-operated channels.


Sensitivity of genetically engineered organisms to selective media.
Eighteen strains of Escherichia coli used in genetic studies were tested for their ability to grow on several selective media. Highest recoveries were obtained with m-T7 agar. The SOS system, particularly the recA gene, may play some role in the sensitivity of E. coli to selective agents. These results may be important in the selection of media used to detect genetically engineered organisms released into the environment.


The anti-angiogenic effect of sinomenine.
Sinomenine is an alkaloid extracted from the Chinese medicinal plant, Sinomenium acutum, which has been utilized to treat rheumatoid arthritis (RA) in China for over 2000 years. Sinomenine has been shown to mediate a wide range of pharmacological actions which includes anti-inflammatory and anti-rheumatic effects. RA has been classified as a chronic immune-mediated disease that exhibits overlapping manifestation of inflammatory, abnormal cellular and hormonal immune responses with synovial hyperplasia. Since, angiogenesis is recognized to play a critical role in the development of RA and anti-angiogenic therapy has been proposed as a new therapeutic strategy for treatment of RA, we would like to see if sinomenine possesses anti-angiogenic property. In this study, sinomenine inhibited bFGF-induced proliferation of human umbilical vein endothelial cells (HUVEC) and arrested its cell cycle in G1 phase. Sinomenine disrupted tube formation of HUVEC on Matrigel and suppressed the chemotaxis of HUVEC. In addition, sinomenine reduced neovascularization in Matrigel plug assay as well as microvascular outgrowth in rat aorta ring sprouting assay. These results suggest that sinomenine inhibited bFGF-induced angiogenesis in vitro and in vivo. As the leukocytes-endothelial adhesive interactions also play an important role in inflammation, we found that sinomenine reduced the transmigration of granulocytic differentiated HL60 cells across IL-1beta activated HUVEC monolayer. Therefore, the inhibition of leukocytes migration across blood vessel walls and the anti-angiogenic effect of sinomenine may contribute towards its therapeutic mechanisms in alleviating the pathogenesis of RA.


Risks and benefits of vegan and vegetarian diets in children.
In parallel with increased public awareness of the health and environmental benefits of consuming a plant-based diet, the numbers of people who identify as vegan has increased sharply. The question of whether vegetarian and vegan diets are appropriate for children is a longstanding and unresolved controversy. The more restrictive the diet and the younger the child, the greater the risk of nutritional deficiency. Nutrients of potential concern are protein quantity and quality, iron, zinc, selenium, calcium, riboflavin, vitamins A, D, B12 and essential fatty acids. Although intakes and status of some nutrients (e.g. vitamin D and iron) are low in many children, vegan children are particularly susceptible due to inadequate supply and/or excess dietary fibre as well as other components that limit bioavailability. Although position papers from North America state that well-planned vegetarian and vegan diets, supplemented appropriately, are suitable for all life stages, European statements include strong recommendations to parents that vegan diets should not be adopted by children without medical and dietetic supervision. Case histories of malnutrition and serious harm persist, including irreversible neurological damage due to vitamin B12 deficiency among un-supplemented children. The evidence available to evaluate the nutritional appropriateness of vegetarian diets for children is inadequate and dated. Although nutritionally adequate vegetarian diets are more easily achieved, successful provision of a complete vegan diet for a young child requires substantial commitment, expert guidance, planning, resources and supplementation.


CircUBAP2-mediated competing endogenous RNA network modulates tumorigenesis in pancreatic adenocarcinoma.
We investigated the role of the competing endogenous RNA (ceRNA) network in the development and progression of pancreatic adenocarcinoma (PAAD). We analyzed the expression profiles of PAAD and normal pancreatic tissues from multiple GEO databases and identified 457 differentially expressed circular RNAs (DEcircRNAs), 19 microRNAs (DEmiRNAs) and 1993 mRNAs (DEmRNAs). We constructed a ceRNA network consisting of 4 DEcircRNAs, 3 DEmiRNAs and 149 DEmRNAs that regulates the NF-kappa B, PI3K-Akt, and Wnt signaling pathways. We then identified and validated five hub genes, CXCR4, HIF1A, ZEB1, SDC1 and TWIST1, which are overexpressed in PAAD tissues. The expression of CXCR4, HIF1A, ZEB1, and SDC1 in PAAD was regulated by circ-UBAP2 and hsa-miR-494. The expression of CXCR4 and ZEB1 correlated with the levels of M2 macrophages, T-regulatory cells (Tregs) and exhausted T cells in the PAAD tissues. The expression of CXCR4 and ZEB1 positively correlated with the expression of CTLA-4 and PD-1. This suggests that CXCR4 and ZEB1 proteins inhibit antigen presentation and promote immune escape mechanisms in PAAD cells. In summary, our data suggest that the circUBAP2-mediated ceRNA network modulates PAAD by regulating the infiltration and function of immune cells.


Integrin clustering enables anandamide-induced Ca2+ signaling in endothelial cells via GPR55 by protection against CB1-receptor-triggered repression.
Although the endocannabinoid anandamide is frequently described to act predominantly in the cardiovascular system, the molecular mechanisms of its signaling remained unclear. In human endothelial cells, two receptors for anandamide were found, which were characterized as cannabinoid 1 receptor (CB1R; CNR1) and G-protein-coupled receptor 55 (GPR55). Both receptors trigger distinct signaling pathways. It crucially depends on the activation status of integrins which signaling cascade becomes promoted upon anandamide stimulation. Under conditions of inactive integrins, anandamide initiates CB1R-derived signaling, including Gi-protein-mediated activation of spleen tyrosine kinase (Syk), resulting in NFkappaB translocation. Furthermore, Syk inhibits phosphoinositide 3-kinase (PI3K) that represents a key protein in the transduction of GPR55-originated signaling. However, once integrins are clustered, CB1R splits from integrins and, thus, Syk cannot further inhibit GPR55-triggered signaling resulting in intracellular Ca2+ mobilization from the endoplasmic reticulum (ER) via a PI3K-Bmx-phospholipase C (PLC) pathway and activation of nuclear factor of activated T-cells. Altogether, these data demonstrate that the physiological effects of anandamide on endothelial cells depend on the status of integrin clustering.


Nature or Nurture? Role of the Bone Marrow Microenvironment in the Genesis and Maintenance of Myelodysplastic Syndromes.
Myelodysplastic syndrome (MDS) are clonal haematopoietic stem cell (HSC) disorders driven by a complex combination(s) of changes within the genome that result in heterogeneity in both clinical phenotype and disease outcomes. MDS is among the most common of the haematological cancers and its incidence markedly increases with age. Currently available treatments have limited success, with <5% of patients undergoing allogeneic HSC transplantation, a procedure that offers the only possible cure. Critical contributions of the bone marrow microenvironment to the MDS have recently been investigated. Although the better understanding of the underlying biology, particularly genetics of haematopoietic stem cells, has led to better disease and risk classification; however, the role that the bone marrow microenvironment plays in the development of MDS remains largely unclear. This review provides a comprehensive overview of the latest developments in understanding the aetiology of MDS, particularly focussing on understanding how HSCs and the surrounding immune/non-immune bone marrow niche interacts together.


E2A proteins promote development of lymphoid-primed multipotent progenitors.
The first lymphoid-restricted progeny of hematopoietic stem cells (HSCs) are lymphoid-primed multipotent progenitors (LMPPs), which have little erythromyeloid potential but retain lymphoid, granulocyte, and macrophage differentiation capacity. Despite recent advances in the identification of LMPPs, the transcription factors essential for their generation remain to be identified. Here, we demonstrated that the E2A transcription factors were required for proper development of LMPPs. Within HSCs and LMPPs, E2A proteins primed expression of a subset of lymphoid-associated genes and prevented expression of genes that are not normally prevalent in these cells, including HSC-associated and nonlymphoid genes. E2A proteins also restricted proliferation of HSCs, MPPs, and LMPPs and antagonized differentiation of LMPPs toward the myeloid fate. Our results reveal that E2A proteins play a critical role in supporting lymphoid specification from HSCs and that the reduced generation of LMPPs underlies the severe lymphocyte deficiencies observed in E2A-deficient mice.


Effects of ions on ligand binding to pyruvate kinase: mapping the binding site with infrared spectroscopy.
The effects of mono- and divalent ions (Li(+), K(+), Na(+), Cs(+), Mg(2+), Ca(2+), Mn(2+), Zn(2+)) on the binding of phosphoenolpyruvate (PEP) to rabbit muscle pyruvate kinase (PK) were studied by attenuated total reflection infrared spectroscopy in combination with a dialysis accessory. The experiments assessed the structural change of the protein as well as the binding mode of PEP. They indicated that a signal at 1638 cm(-1) assigned to a β sheet was perturbed differently with Na(+) as compared to the other monovalent ions. Otherwise, we obtained similar conformational changes in the presence of different monovalent cations, and therefore, it seems unlikely that the ion effects on activity are due to an ion effect on the structure of the PEP:PK complex. With different divalent cations, a particularly large conformational change was observed with Mn(2+) and attributed to a more closed conformation of the complex. The absorption of bound PEP was also detected. The antisymmetric stretching vibration of the carboxylate group of bound PEP indicates a more homogeneous binding mode for Mn(2+) compared to the other divalent ions. The symmetric stretching vibration depends on both monovalent and divalent ions, indicating that the dihedral angle O-C(1)-C(2)-O is affected by the ions in the catalytic site. Little change in the bond strengths of PEP is observed, indicating that the PEP:PK complex does not adopt a reactive conformation.


[The medico-social characteristic of persons with multiple injuries].
The analysis of a medico-social characteristic of victims with multiple injuries in Omsk revealed the nature of multiple and associated injuries distribution among socially-adapted and socially-disadapted groups of population. The top place was taken by multiple injuries received as a result of road accidents. The sections of population most open to multiple injuries were determined. The age peculiarity of such injuries, the accompanying diseases influencing the nature of injury received and the future prognosis in victims were identified.


Strong Superexchange in a d^{9-δ} Nickelate Revealed by Resonant Inelastic X-Ray Scattering.
The discovery of superconductivity in a d^{9-δ} nickelate has inspired disparate theoretical perspectives regarding the essential physics of this class of materials. A key issue is the magnitude of the magnetic superexchange, which relates to whether cuprate-like high-temperature nickelate superconductivity could be realized. We address this question using Ni L-edge and O K-edge spectroscopy of the reduced d^{9-1/3} trilayer nickelates R_{4}Ni_{3}O_{8} (where R=La, Pr) and associated theoretical modeling. A magnon energy scale of ∼80 meV resulting from a nearest-neighbor magnetic exchange of J=69(4) meV is observed, proving that d^{9-δ} nickelates can host a large superexchange. This value, along with that of the Ni-O hybridization estimated from our O K-edge data, implies that trilayer nickelates represent an intermediate case between the infinite-layer nickelates and the cuprates. Layered nickelates thus provide a route to testing the relevance of superexchange to nickelate superconductivity.


The effect of cycloheximide on dictyosome activity in Tradescantia pollen tubes determined using cytochalasin D.
Dictyosome activity in Tradescantia pollen tubes has been determined using a recently developed method based on the assumption that the rate of vesicle accumulation around the dictyosomes, after treatment with cytochalasin D, is equivalent to the actual rate of vesicle production. In tubes germinated in the presence of 1.0 micrograms/ml cycloheximide, reduced dictyosome activity could be detected as early as 10 min after sowing, although tube extension was not halted until later. After 30 min vesicle production had completely ceased. These observations are discussed in relation to previous reports on the effect of cycloheximide on pollen tube growth, and in relation to the synthesis and transfer of membrane proteins to secretory vesicles and the plasma membrane. It is concluded that the ability of pollen to germinate and produce short tubes in the presence of cycloheximide, does not necessarily indicate that protein synthesis is not a requirement for early pollen tube growth, as protein shortages would not be expected to become apparent over time periods less than the dictyosome turnover time and the secretory vesicle residence time.


Response of the small intestine to the application of a hypertonic solution.
The morphologic and functional alterations caused by a commonly used hypertonic radiographic dye (Hypaque(R)-50%) were compared with changes observed during the absorption of 150 mM saline in closed segments of the ileum of 2- to 3-kg rabbits. Hypertonic dye caused a rapid decrease in height and width of the villi, a decrease in height of the epithelial cells and closure of the intercellular space. Concomitantly, the tissue fluid content of the bowel wall and the volume of venous outflow from the segment of ileum decreased, presumably in response to the osmotic gradient between ileal lumen and blood. The fluid added to the luminal contents was hypotonic and contained sodium, potassium, chloride and bicarbonate. In contrast, the ileum exposed to 150 mM saline had prominent intercellular spaces between adjacent epithelial cells and absorbed the solution at isotonic conditions. These studies indicate that production of diarrheal fluid by this hypertonic solution is different from that reported for enteric pathogens.


Factors influencing frequency and appropriateness of utilization of the emergency room by the elderly.
The authors analyzed predictors of the frequency and appropriateness of utilization of an emergency room by a random sample of 75 noninstitutionalized persons aged 65 years and older in a large city in Canada, using the Andersen and Newman model of medical care utilization as a framework. The total amount of variance explained in the frequency of utilization (47%) exceeded that explained by other studies of utilization of various health services by the elderly. Attitude toward health care, prior experience with the hospital system, and number of sources of health care were important predictors of the frequency of presentation to the emergency room. Almost one half (46.7%) were appropriate users; younger age and prior experience were important predictors of appropriateness of use. The results suggest that the elderly are more appropriate users of the emergency room than younger persons; educational interventions by family physicians could contribute to further improvement.


Interpreting conformational effects in protein nano-ESI-MS spectra.
Nano-electrospray-ionization mass spectrometry (nano-ESI-MS) is employed here to describe equilibrium protein conformational transitions and to analyze the influence of instrumental settings, pH, and solvent surface tension on the charge-state distributions (CSD). A first set of experiments shows that high flow rates of N(2) as curtain gas can induce unfolding of cytochrome c (cyt c) and myoglobin (Mb), under conditions in which the stability of the native protein structure has already been reduced by acidification. However, it is possible to identify conditions under which the instrumental settings are not limiting factors for the conformational stability of the protein inside ESI droplets. Under such conditions, equilibrium unfolding transitions described by ESI-MS are comparable with those obtained by other established biophysical methods. Experiments with the very stable proteins ubiquitin (Ubq) and lysozyme (Lyz) enable testing of the influence of extreme pH changes on the ESI process, uncoupled from acid-induced unfolding. When HCl is used for acidification, Ubq and Lyz mass spectra do not change between pH~7 and pH 2.2, indicating that the CSD is highly characteristic of a given protein conformation and not directly affected by even large pH changes. Use of formic or acetic acid for acidification of Ubq solutions results in major spectral changes that can be interpreted in terms of protein unfolding as a result of the increased hydrophobicity of the solvent. On the other hand, Lyz, cyt c, and Mb enable direct comparison of protein CSD (corresponding to either the folded or the unfolded protein) in HCl or acetic acid solutions at low pH. The values of surface tension for these solutions differ significantly. Confirming indications already present in the literature, we observe very similar CSD under these solvent conditions for several proteins in either compact or disordered conformations. The same is true for comparison between water and water-acetic acid for folded cyt c and Lyz. Thus, protein CSD from water-acetic solutions do not seem to be limited by the low surface tension of acetic acid as previously suggested. This result could reflect a general lack of dependence of protein CSD on the surface tension of the solvent. However, it is also possible that the effect of acetic acid on the precursor ESI droplets is smaller than generally assumed.