Screening glycosynthase libraries with a fluoride chemosensor assay independently of enzyme specificity: identification of a transitional hydrolase to synthase mutant.
Glycosynthases have become efficient tools for the enzymatic synthesis of oligosaccharides, glycoconjugates and polysaccharides. Enzyme-directed evolution approaches are applied to improve the performance of current glycosynthases and engineer specificity for non-natural substrates. However, simple and general screening methods are required since most of the reported assays are specific for each particular enzyme. In the present paper, we report a general screening assay that is independent of enzyme specificity, and implemented in an HTS (high-throughput screening) format for the screening of cell extracts in directed evolution experiments. Fluoride ion is a general by-product released in all glycosynthase reactions with glycosyl fluoride donors. The new assay is based on the use of a specific chemical sensor (a silyl ether of a fluorogenic methylumbelliferone) to transduce fluoride concentration into a fluorescence signal. As a proof-of-concept, it has been applied to a nucleophile saturation mutant library of Bacillus licheniformis 1,3-1,4-β-glucanase. Beyond the expected mutations at the glutamic acid (catalytic) nucleophile, other variants have been shown to acquire glycosynthase activity. Surprisingly, an aspartic acid for glutamic acid replacement renders a highly active glycosynthase, but still retains low hydrolase activity. It appears as an intermediate state between glycosyl hydrolase and glycosynthase.


High levels of interleukin 10 production in vivo are associated with tolerance in SCID patients transplanted with HLA mismatched hematopoietic stem cells.
Transplantation of HLA mismatched hematopoietic stem cells in patients with severe combined immunodeficiency (SCID) can result in a selective engraftment of T cells of donor origin with complete immunologic reconstitution and in vivo tolerance. The latter may occur in the absence of clonal deletion of donor T lymphocytes able to recognize the host HLA antigens. The activity of these host-reactive T cells is suppressed in vivo, since no graft-vs. -host disease is observed in these human chimeras. Here it is shown that the CD4+ host-reactive T cell clones isolated from a SCID patient transplanted with fetal liver stem cells produce unusually high quantities of interleukin 10 (IL-10) and very low amounts of IL-2 after antigen-specific stimulation in vitro. The specific proliferative responses of the host-reactive T cell clones were considerably enhanced in the presence of neutralizing concentrations of an anti-IL-10 monoclonal antibody, suggesting that high levels of endogenous IL-10 suppress the activity of these cells. These in vitro data correlate with observations made in vivo. Semi-quantitative polymerase chain reaction analysis carried out on freshly isolated peripheral blood mononuclear cells (PBMC) of the patient indicated that the levels of IL-10 messenger RNA (mRNA) expression were strongly enhanced, whereas IL-2 mRNA expression was much lower than that in PBMC of healthy donors. In vivo IL-10 mRNA expression was not only high in the T cells, but also in the non-T cell fraction, indicating that host cells also contributed to the high levels of IL-10 in vivo. Patient-derived monocytes were found to be major IL-10 producers. Although no circulating IL-10 could be detected, freshly isolated monocytes of the patient showed a reduced expression of class II HLA antigens. However, their capacity to stimulate T cells of normal donors in primary mixed lymphocyte cultures was within the normal range. Interestingly, similar high in vivo IL-10 mRNA expressions in the T and non-T cell compartment were also observed in three SCID patients transplanted with fetal liver stem cells and in four SCID patients transplanted with T cell-depleted haploidentical bone marrow stem cells. Taken together, these data indicate that high endogenous IL-10 production is a general phenomenon in SCID patients in whom allogenic stem cell transplantation results in immunologic reconstitution and induction of tolerance. Both donor T cells and host accessory cells contribute to these high levels of IL-10, which would suppress the activity of host-reactive T cell in vivo.


Time matters: Resilience of a post-disturbance forest landscape.
Present-day disturbances are transforming European forest landscapes, and their legacies determine the vulnerability and resilience of the emergent forest generation. To understand these legacy effects, we investigated the resilience of the aboveground forest biomass (Babg) to a sequence of disturbances affecting the forest in different recovery phases from the initial large-scale impact. We used the model iLand to simulate windthrows that affected 13-24% of the Babg in a Central European forest landscape. An additional wind event was simulated 20, 40, 60, or 80 years after the initial impact (i.e., sequences of two windthrows were defined). Each windthrow triggered an outbreak of bark beetles that interacted with the recovery processes. We evaluated the resistance of the Babg to and recovery after the impact. Random Forest models were used to identify factors influencing resilience. We found that Babg resistance was the lowest 20 years after the initial impact when the increased proportion of emergent wind-exposed forest edges prevailed the disturbance-dampening effect of reduced biomass levels and increased landscape heterogeneity. This forest had a remarkably high recovery rate and reached the pre-disturbance Babg within 28 years. The forest exhibited a higher resistance and a slower recovery rate in the more advanced recovery phases, reaching the pre-disturbance Babg within 60-80 years. The recovery was enhanced by higher levels of alpha and beta diversity. Under elevated air temperature, the bark beetle outbreak triggered by windthrow delayed the recovery. However, the positive effect of increased temperature on forest productivity caused the recovery rate to be higher under the warming scenario than under the reference climate. We conclude that resilience is not a static property, but its magnitude and drivers vary in time, depending on vegetation feedbacks, interactions between disturbances, and climate. Understanding these mechanisms is an essential step towards the operationalization of resilience-oriented stewardship.


A comparative health care inventory for primary hip arthroplasty between Germany versus the Netherlands. Is there a downside effect to fast-track surgery with regard to patient satisfaction and functional outcome?
Treatment and rehabilitation protocol for hip arthroplasty differs between Germany and the Netherlands. The Dutch system promotes fast-track surgery whereas in Germany conventional care is provided with a longer hospital stay including rehabilitation. Clinical outcome, patient satisfaction and costs in both treatment protocols were compared in a prospective setup. This prospective cohort study included patients allocated for primary THA in 3 German and 1 Dutch hospital in the border region. Patient-reported outcome scores (PROMS) were measured pre- and postoperatively at 6 and 12 months including the Oxford Hip Score, SF12 survey, visual analogue scale for satisfaction and pain. Length of hospitalisation and availability of postoperative rehabilitation were recorded. In addition, a total cost estimation was calculated using health insurers data. A total of 360 consecutive patients were included; 175 THA in Germany compared to 185 THA in the Netherlands. No cross-border healthcare was encountered in both cohorts. Mean length of hospitalisation was 11.3 (range 6-23) days in Germany, compared to 4.4 (range 3-25) days in the Netherlands. In Germany 92% of the patients was discharged with inpatient (72%) or outpatient (20%) rehabilitation, compared to 21% with only inpatient rehabilitation in the Netherlands. No significant differences were measured regarding the PROMS and patient satisfaction between both countries. Due to profound differences in health care financing only a global cost estimation could be made and no major differences were encountered. Germany and the Netherlands both offer highly protocolled care for THA with comparable functional outcome and patient satisfaction with treatment after 12 months. Despite the length of hospitalisation in Germany is significantly longer including a more intensive rehabilitation programme, no significant differences were recorded regarding functional outcome nor patient satisfaction compared to fast-track surgery performed in the Netherlands.


Indole Derivatives as Anti-Tubercular Agents: An Overview on their Synthesis and Biological Activities.
The indole framework is considered as one of the privileged structures in the area of medicinal chemistry and drug discovery because compounds containing this framework have shown to possess a remarkable ability to bind with many receptors or proteins with high affinity. It is therefore not surprising that the indole nucleus is a frequently found moiety in many bioactive agents or drugs. Indole derivatives have also been explored or studied for their anti-tubercular properties for a long time. The growth inhibition of Mycobacterium tuberculosis (MTB) in vitro and in vivo by the gut microbiota metabolite indole propionic acid (IPA) is one of the recent examples. Notably, tuberculosis (TB), an intractable disease and a major cause of death worldwide, has caused an alarming rise in the number of TB cases recently because of two main reasons, i.e., a several-fold rise among HIV-infected patients and increased drug resistance by some bacterial strains. Thus, the identification of new agents or potential drugs against TB is urgently needed. While the specific pharmacological target or mechanism of action (MOA) for antitubercular activities has been reported for many indole derivatives, the MOA is not well defined or known for a number of indole derivatives though they were found to be active against MTB. In the current review article, we have focused on both types of indole derivatives that have shown activities against MTB. The indoles with known MOA are further segregated based on this pharmacological target reported or indicated whereas other indoles are classified based on the type of anti-TB properties shown by them. The literature for the last 20 years as well as related to up to date knowledge and information was searched on Pubmed, Google Scholar, MEDLINE, and various other databases until August 2020. A diverse range of functionalized indole derivatives, such as indole-based alkaloids, simple indoles, fused indoles, amide/peptide derivatives of indole, isatin derivatives, etc., have been reported to possess anti-tubercular activities. The anti-tubercular activities, in silico studies (if reported) and the chemical syntheses (in most of the cases) of representative indole derivatives are presented briefly in the current article. The papers referenced by this review allow a deep analysis of the status of the indole-based anti-tubercular agents explored over the past two decades. This review aims at stimulating renewed interest and effort in the discovery and development of new indole-based agents or potential drugs for the treatment of TB. The emergence of modern methods, especially those based on transition metal-catalyzed reactions, has opened up tremendous opportunities in the area of indole synthesis. The desired goal would be to have utilized these modern methodologies for the identification of potent and promising agents to fight against MTB.


Eigenvalue analysis of peroxisome proliferator-activated receptor gamma agonists.
Eigenvalue analysis (EVA) was conducted on a series of potent agonists of peroxisome proliferator-activated receptor gamma (PPARgamma). Predictive EVA quantitative structure-activity relationship (QSAR) models were established using the SYBYL package, which had conventional r2 and cross-validated coefficient (q2) values up to 0.920 and 0.587 for the AM1 method and 0.863 and 0.586 for the PM3 method, respectively. These models were validated by a test set containing 18 compounds. The capability to predict by these two models for PPARgamma agonists, with the best predictive r2pred value of 0.614 for AM1 and 0.822 for PM3 methods, set a successful example for applying a similar approach in building QSAR models for PPARalpha and -delta that could potentially offer a new opportunity in the design of novel PPAR modulators.


Plasticity and dynamical heterogeneity in driven glassy materials.
Many amorphous glassy materials exhibit complex spatio-temporal mechanical response and rheology, characterized by an intermittent stress strain response and a fluctuating velocity profile. Under quasistatic and athermal deformation protocols this heterogeneous plastic flow was shown to be composed of plastic events of various sizes, ranging from local quadrupolar plastic rearrangements to system spanning shear bands. In this paper, through numerical study of a 2D Lennard-Jones amorphous solid, we generalize the study of the heterogeneous dynamics of glassy materials to the finite shear rate (gamma not equal to 0) and temperature case (T not equal to 0). In practice, we choose an effectively athermal limit (T approximately 0) and focus on the influence of shear rate on the rheology of the glass. In line with previous works we find that the model Lennard-Jones glass follows the rheological behavior of a yield stress fluid with a Herschel-Bulkley response of the form, sigma = sigmaY + c1gamma(beta). The global mechanical response obtained through the use of Molecular Dynamics is shown to converge in the limit gamma --> 0 to the quasistatic limit obtained with an energy minimization protocol. The detailed analysis of the plastic deformation at different shear rates shows that the glass follows different flow regimes. At sufficiently low shear rates the mechanical response reaches a shear-rate-independent regime that exhibits all the characteristics of the quasistatic response (finite-size effects, cascades of plastic rearrangements, yield stress, ...). At intermediate shear rates the rheological properties are determined by the externally applied shear rate and the response deviates from the quasistatic limit. Finally at higher shear the system reaches a shear-rate-independent homogeneous regime. The existence of these three regimes is also confirmed by the detailed analysis of the atomic motion. The computation of the four-point correlation function shows that the transition from the shear-rate-dominated to the quasistatic regime is accompanied by the growth of a dynamical cooperativity length scale xi that is shown to diverge with shear rate as xi is proportional to gamma(-nu), with nu approximately 0.2 -0.3. This scaling is compared with the prediction of a simple model that assumes the diffusive propagation of plastic events.


Mini-review: Can non-human leucocyte antigen genes determine susceptibility to severe dengue syndromes?
Dengue viral infections are endemic or epidemic in virtually all tropical countries. Among individuals infected with the dengue virus, severe dengue syndromes (i.e., dengue haemorrhagic fever and dengue shock syndromes) tend to affect only some and this may be due to a combination of host genetic susceptibility and viral factors. In this review article we analyse and discuss the present knowledge of non-human leucocyte antigen host genetic susceptibility to severe dengue syndromes. The relevance of genetic polymorphisms in the pathways of antigen recognition, uptake, processing and presentation, activation of interferon α responses, mast cell and complement activation and T cell activation and dengue disease severity has been reviewed and analysed.


Pulmonary MR imaging with ultra-short TEs: utility for disease severity assessment of connective tissue disease patients.
To evaluate the utility of pulmonary magnetic resonance (MR) imaging with ultra-short echo times (UTEs) at a 3.0 T MR system for pulmonary functional loss and disease severity assessments of connective tissue disease (CTD) patients with interstitial lung disease (ILD). This prospective study was approved by the institutional review board, and written informed consent was obtained from 18 CTD patients (eight men and ten women) and eight normal subjects with suspected chest disease (three men and five women). All subjects underwent thin-section MDCT, pulmonary MR imaging with UTEs, pulmonary function test and serum KL-6. Regional T2 maps were generated from each MR data set, and mean T2 values were determined from ROI measurements. From each thin-section MDCT data set, CT-based disease severity was evaluated with a visual scoring system. Mean T2 values for normal and CTD subjects were statistically compared by using Student's t-test. To assess capability for pulmonary functional loss and disease severity assessments, mean T2 values were statistically correlated with pulmonary functional parameters, serum KL-6 and CT-based disease severity. Mean T2 values for normal and CTD subjects were significantly different (p=0.0019) and showed significant correlations with %VC, %DLCO, serum KL-6 and CT-based disease severity of CTD patients (p<0.05). Pulmonary MR imaging with UTEs is useful for pulmonary functional loss and disease severity assessments of CTD patients with ILD.


Space-based quantum networking in the presence of a nuclear disturbed environment.
Space-based quantum networks provide a means for near-term long-distance transmission of quantum information. This article analyzed the performance of a downlink quantum network between a low-Earth-orbit satellite and an observatory operating in less-than-ideal atmospheric conditions. The effects from fog, haze, and a nuclear disturbed environment on the long-range distribution of quantum states were investigated. A density matrix that estimates the quantum state by capturing the effects from increased signal loss and elevated background noise to estimate the state fidelity of the transmitted quantum state was developed. It was found that the nuclear disturbed environment and other atmospheric effects have a degrading effect on the quantum state. These environments impede the ability to perform quantum communications for the duration of the effects. In the case of the nuclear disturbed environment, the nuclear effects subside quickly, and network performance should return to normal by the next satellite pass.


Adaptations and staff experiences in delivering parenting programmes and other family support services in three community-based organisations in Cape Town, South Africa during the COVID pandemic.
We explore how organisations working on parenting programmes and other types of family support and violence prevention in low-resource settings experienced the pandemic. In August 2020-May 2021, we interviewed (1) staff from three community-based organisations delivering evidence-informed parenting interventions and other psychosocial services for families in Cape Town, South Africa, (2) staff from a parenting programme training organisation and (3) staff from two international organisations supporting psychosocial services in South Africa. Interviews (22) were thematically analysed, with findings in three areas. First, respondents noted changes in the context, including the job losses, food insecurity, and stress experienced by local communities, and reductions in organisational funding. Second, we found that in response to these context changes, the organisations shifted their focus to food provision and COVID prevention. Parenting and psychosocial programmes were adapted - e.g. by changing the physical delivery settings, reducing group sizes, and taking up digital and phone implementation. Participants reported improved perceptions of remote delivery as a feasible approach for working with families - but internet and phone access remained challenging. Third, the pandemic brought new responsibilities for staff, and both the challenges of working from home and the health risks of in-person work.


Differential regulation of MUC5AC/Muc5ac and hCLCA-1/mGob-5 expression in airway epithelium.
This study demonstrates that the two biomarkers, MUC5AC/ Muc5ac and hCLCA1/Gob5, which are frequently associated with surface mucous/goblet cells in asthmatic airways, are differentially regulated. Intratracheal instillation of IL-13 (0.5 mug/mouse lung) elicited 8- and 110-fold induction of Muc5ac and Gob5 messages, respectively, within 24 h in wild-type mouse lung, whereas these inductions were abrogated in Stat6 knockout mice. The induction of MUC5AC/Muc5ac message could not be duplicated in vitro with primary tracheobronchial epithelial (TBE) cells derived from wild-type mice or humans, despite significant inductions still seen for hCLCA1/Gob5. Further studies with JAK inhibitors and STAT6 signaling showed active signaling of the JAK/STAT6 pathway in these primary TBE cultures by IL-13 in the regulation of hCLCA1 expression. Dual immunofluorescent staining with antibodies specific to MUC5AC and hCLCA1 revealed a differential nature of the expression of these two biomarkers by distinct cell types of primary TBE cultures. Finally, MUC5AC expression could be elevated by a bacterial product, peptidoglycan, without any induction of hCLCA1. Thus, these results suggest that the two biomakers of the metaplastic airway mucous cell type are differentially regulated by JAK/STAT6-dependent and -independent pathways.


Congenital posterior angulation of tibia and fibula.
Congenital posterior angulation of the tibia occurs less frequently than congenital anterior bowing. Concomitant deformities of the foot and contractures of the soft tissue usually exist. A review of the literature indicates that nonoperative treatment is usually successful in correcting the deformity. Tibial growth is frequently less than normal and epiphysiodesis of the opposite extermity may be necessary. Three patients are presented who have been followed up for 17 years, two years, and over one year. Improvement is occurring with nonoperative treatment.


Pilot-Scale Test Results of Electrodialysis Bipolar Membrane for Reverse-Osmosis Concentrate Recovery.
In this study, it is aimed to investigate the potential of electrodialysis bipolar membrane (EDBM) systems for the recovery of the concentrate originating from an organized industrial estate (OIE) wastewater treatment system with reverse osmosis (RO). Acids and bases were obtained from a pilot-scale treatment plant as a result of the research. Furthermore, the sustainability and affordability of acids and bases obtained by EDBM systems were investigated. Six cycles were carried out in continuous-flow mode with the EDBM system as batch cycles in the disposal of the concentrate and the production of acids and bases with the EDBM system. For each cycle, the EDBM system was operated for 66, 48, 66, and 80 min, respectively, and the last two cycles were operated for a total of 165 min (70 + 90) with 5 min of waiting. In the EDBM system, a working method was determined such that the cycle flow rate was 180 L/hour, energy to be given to the system was 25 V, and the working pressure was in the range of 0.8-2.5 bar. In the six cycles with the EDBM system, the concentrate, acid and base, conductivity, pH, and pressure increase values were investigated depending on time. Throughout all these studies, the cycles were continued over the products formed in the acid and base chamber. As a result of all the cycles, acid (HCl) production at a level of 1.44% and base (NaOH) production at a level of 2% were obtained.


Prognostic Role of Systemic Inflammatory Indexes in Germ Cell Tumors Treated With High-Dose Chemotherapy.
High-dose chemotherapy (HDCT) has curative potential in relapsed/refractory germ cell tumors (GCT). Due to the complexity of this population and the toxicity of HDCT, we evaluated the association between blood-based systemic inflammatory indexes and the outcome of GCT patients undergoing salvage treatment with HDCT in order to define additional prognostic factors able to orient clinical decision. Baseline characteristics, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and the systemic immune-inflammation index (SII) of 62 patients undergoing HDCT for GCT were retrospectively collected. The aim is to evaluate the correlation between each inflammatory marker (NLR, PLR, and SII) and response to HDCT, overall survival (OS), and progression-free survival (PFS). Using the receiver operating curve to identify the best cutoff values, it was found that patients with GCT with NLR ≥3.3 and SII ≥844,000 had shorter PFS and inferior OS. In the multivariable analysis including inflammatory markers, the International Prognostic Factor Study Group (IPFSG) risk group, age, and previous line of treatment, NLR ≥3.3 and SII ≥844,000 were identified to be independently associated with shorter PFS and OS. Moreover, NLR, PLR, and SII significantly correlate with overall response to HDCT. Associating IPFSG prognostic score to inflammatory markers at baseline of HDCT may improve prognostic information and could help physicians to make more personalized treatment decisions.


Conservation of intrinsic disorder in protein domains and families: II. functions of conserved disorder.
Regions of conserved disorder prediction (CDP) were found in protein domains from all available InterPro member databases, although with varying frequency. These CDP regions were found in proteins from all kingdoms of life, including viruses. However, eukaryotes had 1 order of magnitude more proteins containing long disordered regions than did archaea and bacteria. Sequence conservation in CDP regions varied, but was on average slightly lower than in regions of conserved order. In some cases, disordered regions evolve faster than ordered regions, in others they evolve slower, and in the rest they evolve at roughly the same rate. A variety of functions were found to be associated with domains containing conserved disorder. The most common were DNA/RNA binding, and protein binding. Many ribosomal proteins also were found to contain conserved disordered regions. Other functions identified included membrane translocation and amino acid storage for germination. Due to limitations of current knowledge as well as the methodology used for this work, it was not determined whether these functions were directly associated with the predicted disordered region. However, the functions associated with conserved disorder in this work are in agreement with the functions found in other studies to correlate to disordered regions. We have established that intrinsic disorder may be more common in bacterial and archaeal proteins than previously thought, but this disorder is likely to be used for different purposes than in eukaryotic proteins, as well as occurring in shorter stretches of protein. Regions of predicted disorder were found to be conserved within a large number of protein families and domains. Although many think of such conserved domains as being ordered, in fact a significant number of them contain regions of disorder that are likely to be crucial to their functions.


Identification and characterization of haemofungin, a novel antifungal compound that inhibits the final step of haem biosynthesis.
During recent decades, the number of invasive fungal infections among immunosuppressed patients has increased significantly, whereas the number of effective systemic antifungal drugs remains low and unsatisfactory. The aim of this study was to characterize a novel antifungal compound, CW-8/haemofungin, which we previously identified in a screen for compounds affecting fungal cell wall integrity. The in vitro characteristics of haemofungin were investigated by MIC evaluation against a panel of pathogenic and non-pathogenic fungi, bacteria and mammalian cells in culture. Haemofungin mode-of-action studies were performed by screening an Aspergillus nidulans overexpression genomic library for resistance-conferring plasmids and biochemical validation of the target. In vivo efficacy was tested in the Galleria mellonella and Drosophila melanogaster insect models of infection. We demonstrate that haemofungin causes swelling and lysis of growing fungal cells. It inhibits the growth of pathogenic Aspergillus, Candida, Fusarium and Rhizopus isolates at micromolar concentrations, while only weakly affecting the growth of mammalian cell lines. Genetic and biochemical analyses in A. nidulans and Aspergillus fumigatus indicate that haemofungin primarily inhibits ferrochelatase (HemH), the last enzyme in the haem biosynthetic pathway. Haemofungin was non-toxic and significantly reduced mortality rates of G. mellonella and D. melanogaster infected with A. fumigatus and Rhizopus oryzae, respectively. Further development and in vivo validation of haemofungin is warranted.


Pax6 regulates specification of ventral neurone subtypes in the hindbrain by establishing progenitor domains.
Recent studies have shown that generation of different kinds of neurones is controlled by combinatorial actions of homeodomain (HD) proteins expressed in the neuronal progenitors. Pax6 is a HD protein that has previously been shown to be involved in the differentiation of the hindbrain somatic (SM) motoneurones and V1 interneurones in the hindbrain and/or spinal cord. To investigate in greater depth the role of Pax6 in generation of the ventral neurones, we first examined the expression patterns of HD protein genes and subtype-specific neuronal markers in the hindbrain of the Pax6 homozygous mutant rat. We found that Islet2 (SM neurone marker) and En1 (V1 interneurone marker) were transiently expressed in a small number of cells, indicating that Pax6 is not directly required for specification of these neurones. We also observed that domains of all other HD protein genes (Nkx2.2, Nkx6.1, Irx3, Dbx2 and Dbx1) were shifted and their boundaries became blurred. Thus, Pax6 is required for establishment of the progenitor domains of the ventral neurones. Next, we performed Pax6 overexpression experiments by electroporating rat embryos in whole embryo culture. Pax6 overexpression in the wild type decreased expression of Nkx2.2, but ectopically increased expression of Irx3, Dbx1 and Dbx2. Moreover, electroporation of Pax6 into the Pax6 mutant hindbrain rescued the development of Islet2-positive and En1-positive neurones. To know reasons for perturbed progenitor domain formation in Pax6 mutant, we examined expression patterns of Shh signalling molecules and states of cell death and cell proliferation. Shh was similarly expressed in the floor plate of the mutant hindbrain, while the expressions of Ptc1, Gli1 and Gli2 were altered only in the progenitor domains for the motoneurones. The position and number of TUNEL-positive cells were unchanged in the Pax6 mutant. Although the proportion of cells that were BrdU-positive slightly increased in the mutant, there was no relationship with specific progenitor domains. Taken together, we conclude that Pax6 regulates specification of the ventral neurone subtypes by establishing the correct progenitor domains.


Fast direct Monte Carlo optimization using the inverse kernel approach.
The loss of treatment plan quality after segmentation following fluence optimization is a problem in IMRT. In a previous publication we showed that re-optimization helps to re-establish part of the plan quality. Recently the so-called direct aperture optimization method has been introduced to successfully overcome that difficulty. The aim of the present paper is to present in detail the integration of the inverse kernel method into direct aperture optimization. It can be shown that this integration leads to a system with high performance with regard to time, while Monte Carlo precision is maintained. The integrated simulated annealing optimization algorithm allows easy adaptation to any multi-leaf collimator and it is open to any complex objective function. Investigations of simulated annealing control parameters are performed to improve the performance. The system denoted by direct Monte Carlo optimization (DMCO) is demonstrated on the Carpet phantom and a clinical prostate case as well. Results are compared to inverse kernel optimizations, showing a remarkable time reduction and simultaneously an improvement in plan quality for the Carpet phantom.


"It's a quick way to get what you want": a formative exploration of HIV risk among urban Massachusetts men who have sex with men who attend sex parties.
Community-based studies with men who have sex with men (MSM) suggest that between 8% and 25% of MSM have met recent male sexual partners at private sex parties. Little is known about HIV sexual risk behaviors of MSM who attend sex parties and whether risk reduction interventions can be delivered in this setting. In 2008, 40 MSM who reported attending and/or hosting sex parties in Massachusetts in the past 12 months completed a qualitative interview and quantitative assessment. Participants reported attending a mean number of 10 sex parties in Massachusetts in the past 12 months. A significant percentage (43%) reported also hosting sex parties. Participants had made sexual partner connections across multiple venues, including public cruising areas, bars/clubs, and the Internet. At the most recent sex party attended, the majority had used alcohol (58%) and/or drugs (50%), and one quarter (25%) put themselves at risk of acquiring or transmitting HIV or other sexually transmitted infections (STIs) by having unprotected anal sex with a mean number of three serodiscordant male sex partners. Although many participants perceived that communicating about sexual health in the sex party context would "ruin the mood," the majority (80%) considered some form of HIV prevention at sex parties to be appropriate and necessary, as well as acceptable. Nonintrusive prevention and education activities were especially endorsed (i.e., condoms, lubricants, and coupons for free HIV/STI testing). The majority of participants (75%) expressed some interest in "safer sex" parties. MSM attending sex parties appear to be a subpopulation at high risk for HIV and STI acquisition and transmission. Risk reduction interventions responsive to the needs of MSM who attend sex parties are warranted.


Fabrication and characterization of coated microneedle patches based on PEGDA for transdermal administration of metformin.
Type 2 diabetes is one of the major challenges that the world is facing today. However, metformin (MET) as most type 2 diabetics' first-line oral hypoglycemic drug may cause serious side effects such as gastrointestinal irritation and nausea which reduce the patients' medication compliance. Therefore, the aim of the study was to design a safe and effective self-treatment device for the delivery of MET. Here, a kind of coated microneedle (MN) patches based on poly(ethylene glycol)diacrylate (PEGDA) were prepared by a two-step casting method and photopolymerization process for transdermal administration of MET. The needles wrapped with drug-loaded hyaluronic acid (HA) coating showed promising mechanical properties and drug delivery ability that allowed them to penetrate the skin barrier for rapid drug delivery, and they had no skin irritancy. The in vivo experiment of type 2 diabetic rats showed a satisfying hypoglycemic effect of the coated MN patches. The study shows that the prepared MN patches will be a potential method for the treatment of type 2 diabetes in the future.


Clinical efficacy and safety of combined estradiol valerate and dienogest: a new no-bleed treatment.
A combination of 2 mg estradiol valerate with 2 mg dienogest (E2V/DNG) (Climodien, Schering AG, Berlin, Gemany) is the first continuous combined postmenopausal hormone replacement therapy (HRT) preparation to contain a progestogen with substantial antiandrogenic activity. A study of its clinical efficacy and safety in a comparative study versus a combination of 2 mg estradiol with 1 mg norethisterone acetate (E2/NETA) has shown both preparations to be highly effective in achieving a rapid response in women with postmenopausal symptoms, in terms of hot flushes and the Kupperman index. Biopsy and ultrasound studies have demonstrated that E2V/DNG quickly and effectively achieved endometrial atrophy in the vast majority of subjects, suggesting a protective role in endometrial proliferation. Data on PP-14 (glycodelin) levels may indicate that E2V/DNG is even more effective than E2/NETA in maintaining endometrial atrophy. No-bleed rates with E2V/DNG at 1, 6 and 12 months were at least as favorable as those with other standard HRT products, with evidence that the no-bleed state is attained more quickly with E2V/DNG. The proportions of women with the no-bleed state in a large-scale study (n = 1501) at 1, 6 and 12 months were 71.8%, 76.6% and 86.4%, respectively. Women with irregular bleeding before treatment responded to E2V/DNG in a manner similar to those without bleeding; this concordance was especially marked after five cycles of treatment. In the comparative study, the mean number of days of bleeding over 12 cycles was significantly lower for E2V/DNG than for E2/NETA. Overall, the profiles of adverse events recorded in clinical use were similar in the two preparations, whilst the safety profile of E2V/DNG in the large-scale study was similar to that of other HRT preparations and gave no cause for clinical concern. The 2 mg E2V/2 mg DNG preparation was associated with a favorable lipid profile, whilst a similar combination (2 mg E2V/3 mg DNG) showed no impact on carbohydrate metabolism or hemostasis, compared to placebo. In summary, 2 mg E2V/2 mg DNG is a novel continuous combined HRT preparation that is effective in treating postmenopausal symptoms rapidly, and has a highly favorable bleeding profile. Studies of the safety of 2 mg E2V/2 mg DNG in clinical use have uncovered no factors likely to be disadvantagous in comparison with other HRT products in widespread use.


TGF-β-induced activation of mTOR complex 2 drives epithelial-mesenchymal transition and cell invasion.
In cancer progression, carcinoma cells gain invasive behavior through a loss of epithelial characteristics and acquisition of mesenchymal properties, a process that can lead to epithelial-mesenchymal transition (EMT). TGF-β is a potent inducer of EMT, and increased TGF-β signaling in cancer cells is thought to drive cancer-associated EMT. Here, we examine the physiological requirement for mTOR complex 2 (mTORC2) in cells undergoing EMT. TGF-β rapidly induces mTORC2 kinase activity in cells undergoing EMT, and controls epithelial cell progression through EMT. By regulating EMT-associated cytoskeletal changes and gene expression, mTORC2 is required for cell migration and invasion. Furthermore, inactivation of mTORC2 prevents cancer cell dissemination in vivo. Our results suggest that the mTORC2 pathway is an essential downstream branch of TGF-β signaling, and represents a responsive target to inhibit EMT and prevent cancer cell invasion and metastasis.


Increased hypothalamic content of preproneuropeptide-Y messenger ribonucleic acid in streptozotocin-diabetic rats.
Neuropeptide-Y (NPY) is a 36-amino acid C-terminally amidated peptide found within the hypothalamus that can potently stimulate carbohydrate feeding. Moreover, the hypothalamic content of NPY can be modulated by peripheral hetabolic status. To further evaluate the regulation of NPY synthesis in states of altered metabolic homeostasis, we measured the hypothalamic content of prepro-NPY mRNA in streptozocin (STZ)-diabetic, STZ-diabetic insulin-replaced, and control rats by both nuclease protection and in situ hybridization analyses. Adult male Sprague-Dawley rats received a single injection of STZ (100 mg/kg, ip) or citric acid (control). Beginning 72 h later one group of STZ-treated animals received daily injections of insulin (4 U Ultralente/day). All animals were killed 17-19 days after STZ or control treatment. STZ-treated animals were hyperglycernic and showed growth failure compared to control rats. Glycemic control was restored by insulin replacement, as was partial growth. Nuclease protection analysis revealed an approximately 3- to 4-fold increase in prepro-NPY mRNA levels in the samples from STZ-treated rats vs. control. This increase was returned to control values by insulin replacement. In situ hybridization analysis revealed the STZ-induced increase in hypothalamic prepro-NPY mRNA was detectable in the arcuate nucleus at levels that were in agreement with the nuclease protection results, but that NPY expression in other brain regions appeared to be either unaffected or decreased after STZ treatment. These data suggest that hypothalamic NPY expression is modulated by peripheral metabolic status and provide further explanation for the hyperphagia accompanying STZ-induced diabetes.


Glucose up-regulates expression of the differentiation-associated brush border binding site for enterotoxigenic Escherichia coli colonization factor antigen I in cultured human enterocyte-like cells.
The association of enterotoxigenic Escherichia coli expressing colonization factor antigen I (CFA/I) with the cultured human colon adenocarcinoma cell, a model of the mature enterocyte of the small intestine, is dependent on the binding of CFA/I to a brush border-associated component. Binding of the purified radiolabeled [125I]CFA/I- and 14C-labeled CFA/I-positive bacteria could be displaced by an increasing concentration of unlabeled CFA/I. Moreover, we showed that expression of the specific CFA/I binding developed as a function of cell differentiation in Caco-2 cells, whereas expression of the nonspecific binding did not. Expression of the brush border differentiation-associated component acting as a binding site for CFA/I was up-regulated by glucose. Indeed, the enterocyte-like HT-29 glc- cell subpopulation not expressing the CFA/I binding site when cultured in dialyzed serum and hexose-free medium regained the ability to bind CFA/I when the cells were returned to culture medium containing glucose. Furthermore, expression of the brush border-associated CFA/I binding site in the enterocyte-like Caco-2 cells was repressed when the cells were cultured in hexose-free conditions.


A Quality Framework for Emergency Department Treatment of Opioid Use Disorder.
Emergency clinicians are on the front lines of responding to the opioid epidemic and are leading innovations to reduce opioid overdose deaths through safer prescribing, harm reduction, and improved linkage to outpatient treatment. Currently, there are no nationally recognized quality measures or best practices to guide emergency department quality improvement efforts, implementation science researchers, or policymakers seeking to reduce opioid-associated morbidity and mortality. To address this gap, in May 2017, the National Institute on Drug Abuse's Center for the Clinical Trials Network convened experts in quality measurement from the American College of Emergency Physicians' (ACEP's) Clinical Emergency Data Registry, researchers in emergency and addiction medicine, and representatives from federal agencies, including the National Institute on Drug Abuse and the Centers for Medicare & Medicaid Services. Drawing from discussions at this meeting and with experts in opioid use disorder treatment and quality measure development, we developed a multistakeholder quality improvement framework with specific structural, process, and outcome measures to guide an emergency medicine agenda for opioid use disorder policy, research, and clinical quality improvement.


Human Neurocysticercosis: An Overview.
Human cysticercosis is caused by ingestion of T. solium eggs from taenia carriers. Neurocysticercosis (NCC), defined as the infection of the CNS and the meninges by the larval stage of Taenia solium, is the most common helminthic infection of the CNS worldwide. Parasites may lodge in brain parenchyma, subarachnoid space, ventricular system, or spinal cord, causing pathological changes that account for the pleomorphism of this disease. Seizures/epilepsy are the most common clinical manifestation, but other patients present with headache, focal deficits, intracranial hypertension, or cognitive decline. Accurate diagnosis of NCC is possible after interpretation of clinical data together with findings of neuroimaging studies and results of immunological tests. However, neuroimaging studies are fundamental for diagnosis because immunological test and clinical manifestations only provide circumstantial evidence of NCC. The introduction of cysticidal drugs changed the prognosis of most NCC patients. These drugs have been shown to reduce the burden of infection and to improve the clinical course of the disease in many patients. Efforts should be directed to eradicate the disease through the implementation of control programs against all the steps in the life cycle of T. solium, including carriers of the adult tapeworm, infected pigs, and eggs in the environment.


Functional anthology of intrinsic disorder. 1. Biological processes and functions of proteins with long disordered regions.
Identifying relationships between function, amino acid sequence, and protein structure represents a major challenge. In this study, we propose a bioinformatics approach that identifies functional keywords in the Swiss-Prot database that correlate with intrinsic disorder. A statistical evaluation is employed to rank the significance of these correlations. Protein sequence data redundancy and the relationship between protein length and protein structure were taken into consideration to ensure the quality of the statistical inferences. Over 200,000 proteins from the Swiss-Prot database were analyzed using this approach. The predictions of intrinsic disorder were carried out using PONDR VL3E predictor of long disordered regions that achieves an accuracy of above 86%. Overall, out of the 710 Swiss-Prot functional keywords that were each associated with at least 20 proteins, 238 were found to be strongly positively correlated with predicted long intrinsically disordered regions, whereas 302 were strongly negatively correlated with such regions. The remaining 170 keywords were ambiguous without strong positive or negative correlation with the disorder predictions. These functions cover a large variety of biological activities and imply that disordered regions are characterized by a wide functional repertoire. Our results agree well with literature findings, as we were able to find at least one illustrative example of functional disorder or order shown experimentally for the vast majority of keywords showing the strongest positive or negative correlation with intrinsic disorder. This work opens a series of three papers, which enriches the current view of protein structure-function relationships, especially with regards to functionalities of intrinsically disordered proteins, and provides researchers with a novel tool that could be used to improve the understanding of the relationships between protein structure and function. The first paper of the series describes our statistical approach, outlines the major findings, and provides illustrative examples of biological processes and functions positively and negatively correlated with intrinsic disorder.


Cold-induced brain edema in mice. Involvement of extracellular superoxide dismutase and nitric oxide.
The role of extracellular superoxide in the pathogenesis of vasogenic edema was studied using transgenic mice expressing a 5-fold increase in extracellular superoxide dismutase (EC-SOD) activity in their brains. Increased EC-SOD expression offered significant protection against edema development after cold-induced injury (44% less edema than nontransgenic littermates, p < 0.05). Since iron may contribute to vasogenic edema by catalyzing the production of hydroxyl radical from superoxide and hydrogen peroxide, the effects of the chelator deferoxamine were studied. Deferoxamine reduced edema formation after cold-induced injury (43% less edema than controls, p < 0.05); however, treatment with iron-saturated deferoxamine also reduced edema development in mice (32-48% less edema, p < 0.05). This suggested that the protection offered by deferoxamine was independent of its ability to chelate iron. An iron-independent mechanism by which superoxide can contribute to vasogenic edema is via reaction with nitric oxide to produce the potentially toxic peroxynitrite anion, which is also scavenged by deferoxamine. Mice treated with an inhibitor of nitric oxide synthase were protected against cold-induced edema (37% less edema, p < 0.05). EC-SOD transgenic mice received no additional protection by inhibition of nitric oxide synthesis, supporting this novel alternative mechanism of edema formation.


Pharmacological therapies for patients with human coronavirus infections: a rapid systematic review.
This work aimed to evaluate the effects of drug therapies for coronavirus infections. Rapid systematic review with search in the MEDLINE, EMBASE, Cochrane, BVS, Global Index Medicus, Medrix, bioRxiv, Clinicaltrials.gov and International Clinical Trials Registry Platform databases. Thirty-six studies evaluating alternative drugs against SARS, SARS-CoV-2 and MERS were included. Most of the included studies were conducted in China with an observational design for the treatment of COVID-19. The most studied treatments were with antimalarials and antivirals. In antimalarial, the meta-analysis of two studies with 180 participants did not identify the benefit of hydroxychloroquine concerning the negative viral load via real-time polymerase chain reaction, and the use of antivirals compared to standard care was similar regarding outcomes. The available scientific evidence is preliminary and of low methodological quality, which suggests caution when interpreting its results. Research that evaluates comparative efficacy in randomized, controlled clinical trials, with adequate follow-up time and with the methods properly disclosed and subject to scientific peer review is required. A periodic update of this review is recommended.


[Prevalence of metabolic syndrome estimated with the new World Health Organization recommendations. The HERMEX study].
The unification of criteria for the diagnosis of metabolic syndrome, together with the subsequent World Health Organization (WHO) proposal to eliminate diabetes and cardiovascular diseases from the diagnostic criteria, will change estimates of the known prevalence of this syndrome. The aim of this study was to determine the prevalence of metabolic syndrome in a health area of Badajoz (Spain) using the latest consensus criteria and eliminating diabetes and cardiovascular disease. We performed a cross-sectional population-wide study of randomly selected individuals aged between 25 and 79 years old in a health area of Badajoz. In all patients, data on their history of cardiovascular risk factors were gathered, waist circumference and blood pressure were measured and a fasting blood sample was collected. The prevalence of metabolic syndrome, following recent criteria, was compared by age and gender. We recruited 2,833 individuals (46.5% men). The mean age was 51.2 years The prevalence of metabolic syndrome was 33.6% and was significantly higher in men (36.7% vs 30.9%; p < 0.001). The prevalence of metabolic syndrome fell significantly after exclusion of patients with diabetes or cardiovascular disease (20.8%; p < 0.001). The difference in prevalence between the distinct criteria was significant for the whole population and by sex (p < 0.000). A significant difference in prevalence between genders was observed from the age of 45-54 years in men and 55-64 years in women The prevalence of metabolic syndrome in a health area of Badajoz is among the highest reported in population-based studies in Spain. Although estimates of the prevalence are decreased by the new international recommendations, a considerable proportion of the young population requires preventive measures.


Nucleation and Growth of Nanocrystals Investigated by In Situ Transmission Electron Microscopy.
Nanocrystals play a key role in the modern energy, catalysis, semiconductor, and biology industries due to their unique structures and performances. However, controllable fabrication of ideal nanocrystals with the desired structures and properties is still challenging, which needs a deep understanding of their nucleation and growth process. Here, the research on nucleation and growth of nanocrystals studied by in situ transmission electron microscopy (TEM) is reviewed, mainly focusing on the atomic migration dynamics, interface evolution, and structure transformation. In addition, the challenges in the study of nanocrystal growth by TEM are discussed and the perspective on the future development of advanced in situ TEM techniques is provided. It is hoped that the review can give a deep insight into the nanocrystal nucleation and growth process, and further contribute to the rational design and precise fabrication of high-performance functional nanocrystals.


Fictive temperature, structural relaxation, and reality of residual entropy.
By determining the fictive temperature, Tf, in two ways from the same Cp data, we investigate whether the residual entropy, Sres, of a glass could be an artifact of using the Cp d ln(T) integral in the glass-liquid temperature range. Although the integral gives only the upper and lower limits of the real entropy change, it is still useful and is distinguished as Delta(sigma). We determine Tf(sigma) from Delta(sigma) and the usual TfH from the Cp dT integral for two metal alloy glasses, a basalt composition glass and a spray-quenched propylene glycol glass from the available data, and find that Tf9sigma is about the same as TfH within errors. To substantiate it, we report a differential scanning calorimetry study performed during cooling of the Mg65Cu25Tb10 and Pd40Ni10Cu30P20 melts and on heating their glassy states at the same rates. In addition, we simulate Cp-T plots from a known model for nonexponential, nonlinear relaxation and analyze the data. The quantity Delta(sigma) on cooling the liquid and heating the glass differs negligibly; that is, net change in a temperature cycle between glass and its melt is close to zero, a characteristic of a nearly reversible change. We conclude that spontaneous enthalpy release has little effect on the entropy change determined from the Cp d ln(T) integral and, contrary to recent suggestions, Sres is real.


The primary physician: generalist or specialist?
Dr W W Yellowlees makes out a strong case against the creation of general practitioner 'specialoids'. He emphasizes the limitations of the specialists' organ-orientation, without the generalized knowledge of the patient as a person and of the influence of his environment on his illness.Dr Paul Sadler takes another point of view and describes the successful division of his housing-estate practice into four specialist departments: pxediatrics, geriatrics, obstetrics and general medicine. He further details the organization and advantages of his own department, pediatrics.


Photochemical pathways in nucleobases measured with an X-ray FEL.
The conversion of light energy into other molecular energetic degrees of freedom is often dominated by ultrafast, non-adiabatic processes. Femtosecond spectroscopy with optical pulses has helped in shaping our understanding of crucial processes in molecular energy-conversion. The advent of new, ultrashort and bright X-ray free electron laser sources opens the possibility to use X-ray-typical element and site sensitivity for ultrafast molecular research. We present two types of spectroscopy, ultrafast Auger and ultrafast X-ray absorption spectroscopy, and discuss their sensitivity to molecular processes. While Auger spectroscopy is able to monitor bond distance changes in the vicinity of an X-ray created core hole, near-edge absorption spectroscopy can deliver high-fidelity information on non-adiabatic transitions involving lone-pair orbitals. We demonstrate these features on the example of the UV-excited nucleobase thymine, investigated at the oxygen K-edge. We find a C-O bond elongation in the Auger data in addition to ππ*/ nπ* non-adiabatic transition in X-ray near-edge absorption. We compare the results from both methods and draw a conclusive scenario of non-adiabatic molecular relaxation after UV excitation. This article is part of the theme issue 'Measurement of ultrafast electronic and structural dynamics with X-rays'.


Ferredoxin-1 mRNA is destabilized by changes in photosynthetic electron transport.
In transgenic tobacco, pea Ferredoxin-1 (Fed-1) mRNA accumulates rapidly in response to photosynthesis even when the transgene is driven by a constitutive promoter. To investigate the role of photosynthesis on Fed-1 mRNA stability, we used the tetracycline repressible Top10 promoter system to specifically shut off transcription of the Fed-1 transgene. The Fed-1 mRNA has a half-life of approximately 2.4 hr in the light and a half-life of only 1.2 hr in the dark or in the presence of the photosynthetic electron transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). These data indicate that cessation of photosynthesis, either by darkness or DCMU results in a destabilization of the Fed-1 mRNA. Furthermore, the Fed-1 mRNA half-life is reduced immediately upon transfer to darkness, suggesting that Fed-1 mRNA destabilization is a primary response to photosynthesis rather than a secondary response to long-term dark adaptation. Finally, the two different methods for efficient tetracycline delivery reported here generally should be useful for half-life measurements of other mRNAs in whole plants.


Direct nitrate reductase assay versus microscopic observation drug susceptibility test for rapid detection of MDR-TB in Uganda.
The most common method for detection of drug resistant (DR) TB in resource-limited settings (RLSs) is indirect susceptibility testing on Lowenstein-Jensen medium (LJ) which is very time consuming with results available only after 2-3 months. Effective therapy of DR TB is therefore markedly delayed and patients can transmit resistant strains. Rapid and accurate tests suitable for RLSs in the diagnosis of DR TB are thus highly needed. In this study we compared two direct techniques--Nitrate Reductase Assay (NRA) and Microscopic Observation Drug Susceptibility (MODS) for rapid detection of MDR-TB in a high burden RLS. The sensitivity, specificity, and proportion of interpretable results were studied. Smear positive sputum was collected from 245 consecutive re-treatment TB patients attending a TB clinic in Kampala, Uganda. Samples were processed at the national reference laboratory and tested for susceptibility to rifampicin and isoniazid with direct NRA, direct MODS and the indirect LJ proportion method as reference. A total of 229 specimens were confirmed as M. tuberculosis, of these interpretable results were obtained in 217 (95%) with either the NRA or MODS. Sensitivity, specificity and kappa agreement for MDR-TB diagnosis was 97%, 98% and 0.93 with the NRA; and 87%, 95% and 0.78 with the MODS, respectively. The median time to results was 10, 7 and 64 days with NRA, MODS and the reference technique, respectively. The cost of laboratory supplies per sample was low, around 5 USD, for the rapid tests. The direct NRA and MODS offered rapid detection of resistance almost eight weeks earlier than with the reference method. In the study settings, the direct NRA was highly sensitive and specific. We consider it to have a strong potential for timely detection of MDR-TB in RLS.


[Ultrasound-guided blockade of axillary plexus brachialis for hand surgery].
Blockade of the axillary plexus brachialis is an integral part of the anesthetic-specific procedures in the field of hand surgery. Less side-effects with high success rates are demanded of the surgeons and required by the patients. Normally, blockade of the axillary plexus brachialis is performed by using nerve stimulation. An alternative technique described here as a retrospective study uses ultrasound for detection of the nerves. At the level of axilla, anatomical structures (axillary vein and artery, triceps/coracobrachialis muscles, ulnar/radial/median nerves) were visualized by ultrasound (linear scanner, 10-MHz transducer). At this position, a puncture needle was inserted into the nerves and local anesthetics were injected (50 ml prilocaine 1 % or 40 prilocaine 1 % and 20 ropivacaine 0.5 %). Parameters like onset time of blockade (min), complete block (yes/no) and possible complications were documented. In 30 months 221 ultrasound-guided blocks of axillary plexus brachialis were performed by eight anesthetists with different levels of training. In 96.8 % of the documented cases blockade was complete and did not require any supplementation. Only in seven cases was a conversion of the anesthetic technique performed. Adverse effects like toxic-allergic reactions or nerve damage caused by the punctures were not documented in any case. The average time to onset of the blockade was 2.2 minutes. Ultrasound-guided blockade of the axillary plexus brachialis represents an alternative to procedures using nerve stimulation, even in patients with anatomic abnormalities. The described method proved to be effective, especially in the field of hand surgery.


Decreased estimated blood loss in lateral trans-psoas versus anterior approach to lumbar interbody fusion for degenerative spondylolisthesis.
The goal of the current study was to compare the perioperative and post-operative outcomes of eXtreme lateral trans-psoas approach (XLIF) versus anterior lumbar interbody fusion (ALIF) for single level degenerative spondylolisthesis. The ideal approach for degenerative spondylolisthesis remains controversial. Consecutive patients undergoing single level XLIF (n=21) or ALIF (n=54) for L4-5 degenerative spondylolisthesis between 2008-2012 from a single academic center were retrospectively reviewed. Groups were compared for peri-operative data (estimated blood loss, operative time, adjunct procedures or additional implants), radiographic measurements (L1-S1 cobb angle, disc height, fusion grade, subsidence), 30-day complications (infection, DVT/PE, weakness/paresthesia, etc.), and patient reported outcomes (leg and back Numerical Rating Scale, and Oswestry Disability Index). Estimated blood loss was significantly lower for XLIF [median 100; interquartile range (IQR), 50-100 mL] than for ALIF (median 250; IQR, 150-400 mL; P<0.001), including after adjusting for significantly higher rates of posterior decompression in the ALIF group. There were no significant differences in rates of complications within 30 days, radiographic outcomes, or in re-operation rates. Both groups experienced significant pain relief post-operatively. The lateral trans-psoas approach is associated with diminished blood loss compared to the anterior approach in the treatment of degenerative spondylolisthesis. We were unable to detect differences in radiographic outcomes, complication rates, or patient reported outcomes. Continued efforts to directly compare approaches for specific indications will minimize complications and improve outcomes. Further studies will continue to define indications for lateral versus anterior approach to lumbar spine for degenerative spondylolisthesis.


Janus Kinase 3, a Novel Regulator for Smooth Muscle Proliferation and Vascular Remodeling.
Vascular remodeling because of smooth muscle cell (SMC) proliferation is a common process occurring in several vascular diseases, such as atherosclerosis, aortic aneurysm, post-transplant vasculopathy, restenosis after angioplasty, etc. The molecular mechanism underlying SMC proliferation, however, is not completely understood. The objective of this study is to determine the role and mechanism of Janus kinase 3 (JAK3) in vascular remodeling and SMC proliferation. Platelet-derived growth factor-BB, an SMC mitogen, induces JAK3 expression and phosphorylation while stimulating SMC proliferation. Janex-1, a specific inhibitor of JAK3, or knockdown of JAK3 by short hairpin RNA, inhibits the SMC proliferation. Conversely, ectopic expression of JAK3 promotes SMC proliferation. Mechanistically, JAK3 promotes the phosphorylation of signal transducer and activator of transcription 3 and c-Jun N-terminal kinase in SMC, 2 signaling pathways known to be critical for SMC proliferation and vascular remodeling. Blockade of these 2 signaling pathways by their inhibitors impeded the JAK3-mediated SMC proliferation. In vivo, knockdown of JAK3 attenuates injury-induced neointima formation with attenuated neointimal SMC proliferation. Knockdown of JAK3 also induces neointimal SMC apoptosis in rat carotid artery balloon injury model. Our results demonstrate that JAK3 mediates SMC proliferation and survival during injury-induced vascular remodeling, which provides a potential therapeutic target for preventing neointimal hyperplasia in proliferative vascular diseases.


A new window of opportunity in echocardiography.
Improvements in echocardiographic technology have made technically difficult studies a rare entity. However, physical barriers such as bandages, inability for patients to turn because of intubation, arterial lines, and organ and life support machines make echocardiographic imaging challenging. We performed echocardiographic imaging from left and right posterior thoracic approach using acoustic properties of pleural fluid to assist in obtaining good imaging windows in patients who had pleural effusion (PE). In this study we describe one author's (T. Z. N.) experience with the mid to lower posterior thoracic window in 18 patients who had PE and in whom conventional transthoracic windows either provided suboptimal images or incomplete clinical information. The posterior approach allowed excellent differentiation of pericardial effusion versus PE, detection of pericardial disease and pericardial infiltration, and excellent endocardial border definition of left and right ventricle in those with poor anterior transthoracic windows. Native and prosthetic aortic valve gradients could be assessed adequately as a result of perfectly parallel Doppler alignment beam to left ventricular outflow tract and aortic valve. In addition, right posterior thoracic window provided views comparable with subcostal view and allowed visualization of inferior vena cava, right atrium, and liver. In patients with PE, imaging from low to midposterior thorax can provide additional diagnostic echocardiographic images and should be used in patients in whom conventional images are technically difficult or require additional information.


Exploiting Isospin Symmetry to Study the Role of Isomers in Stellar Environments.
Proton capture on the excited isomeric state of ^{26}Al strongly influences the abundance of ^{26}Mg ejected in explosive astronomical events and, as such, plays a critical role in determining the initial content of radiogenic ^{26}Al in presolar grains. This reaction also affects the temperature range for thermal equilibrium between the ground and isomeric levels. We present a novel technique, which exploits the isospin symmetry of the nuclear force, to address the long-standing challenge of determining proton-capture rates on excited nuclear levels. Such a technique has in-built tests that strongly support its veracity and, for the first time, we have experimentally constrained the strengths of resonances that dominate the astrophysical ^{26m}Al(p,γ)^{27}Si reaction. These constraints demonstrate that the rate is at least a factor ∼8 lower than previously expected, indicating an increase in the stellar production of ^{26}Mg and a possible need to reinvestigate sensitivity studies involving the thermal equilibration of ^{26}Al.


Is Dementia More Fatal Than Previously Estimated? A Population-based Prospective Cohort Study.
Dementia increases the risk of mortality (ROM) in the elderly and estimates of hazard ratio (HR) of dementia for mortality have ranged from 1.7 to 6.3. However, previous studies may have underestimated ROM of dementia due to length bias, which occurs when failing to include the persons with rapidly progressive diseases, who died before they could be included in the study. This population-based prospective cohort study conducted on 6,752 randomly sampled Koreans, aged 60 years or older (the Korean Longitudinal Study on Cognitive Aging and Dementia). Cognitive disorders were evaluated at baseline and 2-year follow-up using the Korean version of the Consortium to Establish a Registry for Alzheimer's Disease Assessment Packet (CERAD-K), and prevalent and incident cases of dementia were identified. The participants' deaths were confirmed through the National Mortality Database of Statistics Korea. We compared the ROM between prevalent and incident dementia, and estimated HR of dementia for mortality using Cox proportional hazards model. Of the 5,097 responders to the 2-year follow-up assessment, 150 participants had dementia from the baseline (prevalent dementia), and 95 participants developed dementia during the 2-year follow-up period (incident dementia). The ROM of participants with incident dementia was about 3 times higher than the ROM of those with prevalent dementia (HR = 3.04, 95% confidence interval [CI] = 1.34-6.91). Compared to cognitively normal participants at both the baseline and 2-year follow-up assessments, the ROM of those with incident dementia approximately 8 times higher (HR = 8.37, 95 % CI = 4.23-16.54). In conclusion, the ROM of dementia using prevalent cases was underestimated due to length bias, and dementia may be much more fatal than previously estimated. In clinical settings, the ROM of dementia warrants the attention of physicians, particularly in recently incident dementia cases.


Role of an ethyl radical and the problem of HF(v) bimodal vibrational distribution in the F(2P) + C2H6→ HF(v) + C2H5 reaction.
A theoretical study of the dynamics of the F(2P) + C2H6 hydrogen abstraction reaction was presented using quasi-classical trajectories propagated on an ab initio fitted global potential energy surface, PES-2018. The results were compared with experimental information at a collision energy of 3.2 kcal mol-1. We found a small fraction of available energy, 13%, deposited as C2H5 internal energy, the largest fraction, 67%, being deposited as HF(v) vibration, where the HF(v,j) rotational distribution is colder when the vibrational level increases. These results reproduce the experimental evidence. In addition, the reaction cross section scarcely changes with energy, and the scattering distribution shifts from isotropic to forward when the HF(v) vibrational state increases. These last two findings await experimental confirmation. Finally, two important issues related to the title reaction were analysed: the role of an ethyl radical, and the theory/experiment controversy about the HF(v) bimodal vibrational distribution. We found that in spite of its low internal energy, the ethyl product does not act as a spectator of the reaction, and that the controversy can be explained by the net result of two opposite effects: strong couplings between vibrational modes, which are the rule and complicate dynamics analysis in polyatomic systems, and an artificial energy transfer from the ethyl radical to the HF molecule due to the classical nature of the quasi-classical trajectory calculations. As is seen, the ethyl radical actively participates in both effects.


Free-breathing Accelerated Cardiac MRI Using Deep Learning: Validation in Children and Young Adults.
Background Obtaining ventricular volumetry and mass is key to most cardiac MRI but challenged by long multibreath-hold acquisitions. Purpose To assess the image quality and performance of a highly accelerated, free-breathing, two-dimensional cine cardiac MRI sequence incorporating deep learning (DL) reconstruction compared with reference standard balanced steady-state free precession (bSSFP). Materials and Methods A DL algorithm was developed to reconstruct custom 12-fold accelerated bSSFP cardiac MRI cine images from coil sensitivity maps using 15 iterations of separable three-dimensional convolutions and data consistency steps. The model was trained, validated, and internally tested in 10, two, and 10 adult human volunteers, respectively, based on vendor partner-supplied fully sampled bSSFP acquisitions. For prospective external clinical validation, consecutive children and young adults undergoing cardiac MRI from September through December 2019 at a single children's hospital underwent both conventional and highly accelerated short-axis bSSFP cine acquisitions in one MRI examination. Two radiologists scored overall and volumetric three-dimensional mesh image quality of all short-axis stacks on a five-point Likert scale and manually segmented endocardial and epicardial contours. Scan times and image quality were compared using the Wilcoxon rank sum test. Measurement agreement was assessed with intraclass correlation coefficient and Bland-Altman analysis. Results Fifty participants (mean age, 16 years ± 4 [standard deviation]; range, 5-30 years; 29 men) were evaluated. The mean prescribed acquisition times of accelerated scans (non-breath-held) and bSSFP (excluding breath-hold time) were 0.9 minute ± 0.3 versus 3.0 minutes ± 1.9 (P < .001). Overall and three-dimensional mesh image quality scores were, respectively, 3.8 ± 0.6 versus 4.3 ± 0.6 (P < .001) and 4.0 ± 1.0 versus 4.4 ± 0.8 (P < .001). Raters had strong agreement between all bSSFP and DL measurements, with intraclass correlation coefficients of 0.76 to 0.97, near-zero mean differences, and narrow limits of agreement. Conclusion With slightly lower image quality yet much faster speed, deep learning reconstruction may allow substantially shorter acquisition times of cardiac MRI compared with conventional balanced steady-state free precession MRI performed for ventricular volumetry. © RSNA, 2021 Online supplemental material is available for this article.


Calcineurin and synaptophysin in the human spinal cord of normal individuals and patients with familial dysautonomia.
This report concerns the immunohistochemical demonstration of two neuronal Ca2(+)-binding proteins, calcineurin and synaptophysin, in the spinal cord of normal controls and from patients with familial dysautonomia. In controls, calcineurin immunoreactivity was highly concentrated in small nerve cells and fibers of the substantia gelatinosa. Synaptophysin immunoreactivity was normally distributed throughout the spinal cord gray matter, being highly concentrated in the substantia gelatinosa, the dorsal nucleus of Clarke and the anterior horn. In patients with familial dysautonomia, no apparent changes in calcineurin immunoreactivity were found in the substantia gelatinosa. By contrast, there was a significant depletion of synaptophysin-positive axon terminals in the substantia gelatinosa and in the dorsal nucleus of Clarke of patients with familial dysautonomia.


Single-cell approaches identify the molecular network driving malignant hematopoietic stem cell self-renewal.
Recent advances in single-cell technologies have permitted the investigation of heterogeneous cell populations at previously unattainable resolution. Here we apply such approaches to resolve the molecular mechanisms driving disease in mouse hematopoietic stem cells (HSCs), using JAK2V617F mutant myeloproliferative neoplasms (MPNs) as a model. Single-cell gene expression and functional assays identified a subset of JAK2V617F mutant HSCs that display defective self-renewal. This defect is rescued at the single HSC level by crossing JAK2V617F mice with mice lacking TET2, the most commonly comutated gene in patients with MPN. Single-cell gene expression profiling of JAK2V617F-mutant HSCs revealed a loss of specific regulator genes, some of which were restored to normal levels in single TET2/JAK2 mutant HSCs. Of these, Bmi1 and, to a lesser extent, Pbx1 and Meis1 overexpression in JAK2-mutant HSCs could drive a disease phenotype and retain durable stem cell self-renewal in functional assays. Together, these single-cell approaches refine the molecules involved in clonal expansion of MPNs and have broad implications for deconstructing the molecular network of normal and malignant stem cells.


Strong 3D Printing by TPMS Injection.
3D printed objects are rapidly becoming prevalent in science, technology and daily life. An important question is how to obtain strong and durable 3D models using standard printing techniques. This question is often translated to computing smartly designed interior structures that provide strong support and yield resistant 3D models. In this paper we suggest a combination between 3D printing and material injection to achieve strong 3D printed objects. We utilize triply periodic minimal surfaces (TPMS) to define novel interior support structures. TPMS are closed form and can be computed in a simple and straightforward manner. Since TPMS are smooth and connected, we utilize them to define channels that adequately distribute injected materials in the shape interior. To account for weak regions, TPMS channels are locally optimized according to the shape stress field. After the object is printed, we simply inject the TPMS channels with materials that solidify and yield a strong inner structure that supports the shape. Our method allows injecting a wide range of materials in an object interior in a fast and easy manner. Results demonstrate the efficiency of strong printing by combining 3D printing and injection together.


Marginal integrity of turkom-cera compared to other all-ceramic materials: effect of finish line.
The aim of this study was to evaluate the marginal adaptation of Turkom-Cera all-ceramic crowns compared to In-Ceram and Procera AllCeram systems. The influence of finish line design (chamfer or shoulder) on the marginal adaptation of Turkom-Cera all-ceramic crowns was also investigated. Thirty human premolars were prepared with chamfer margins and assigned to either the Turkom-Cera, In-Ceram, or Procera system group. In addition, 10 premolars were prepared with rounded shoulder finish lines and assigned to an additional Turkom-Cera group. Ceramic copings (0.6-mm thick) were fabricated for each group following the manufacturers' instructions. The copings were seated on abutments using a special holding device that facilitated uniform loading, and marginal adaptation was assessed using a stereomicroscope. Data were analyzed using analysis of variance, the Tukey HSD post hoc test, and an independent samples t test. There was a statistically significant difference regarding marginal adaptation among the three all-ceramic systems (P < .05). There were no significant differences in the mean marginal discrepancies of Turkom-Cera crowns among chamfer and shoulder finish line groups (P > .05). Within the limitations of this study, the marginal discrepancies were all within the clinically acceptable standard. Int J Prosthodont 2011;24:379-381.


Alcohol intake and systemic markers of inflammation--shape of the association according to sex and body mass index.
To assess the association of alcohol intake with high-sensitivity C-reactive protein (hs-CRP), uric acid and leukocyte count in blood, and whether sex and body mass index (BMI) modify these associations. Individuals aged > or =18 years were randomly selected from the population of Porto, Portugal (70% of participation). A total of 840 women and 490 men with reliable information on inflammatory markers and alcohol intake, obtained from a validated food frequency questionnaire, were studied. Associations and their respective trends were estimated from generalized linear models, with adjustment for potential confounders. Analyses were stratified by sex and BMI. In women, adjusted hs-CRP levels (mg/l) were 2.69 in non-drinkers, 2.25 in drinkers of >0-15 g alcohol/day, 2.32 in drinkers of >15-30 g alcohol/day and 3.18 in drinkers of >30 g alcohol/day (P-value for the quadratic trend <0.001). In men, the association between alcohol intake and hs-CRP was positive and linear (P-value for the linear trend = 0.014). Alcohol intake was also positively and linearly associated with uric acid in each sex. Body weight modified these associations, which remained statistically significant only in normal-weight (BMI <25 kg/m(2)) women and overweight (BMI > or =25 kg/m(2)) men for hs-CRP, and in normal-weight individuals for uric acid. No significant association between alcohol intake and leukocyte count was found. The association of alcohol intake with hs-CRP was J-shaped in women but positive and linear-shaped in men. Alcohol intake was directly associated with uric acid in men and women. BMI modifies the effect of alcohol on hs-CRP and uric acid levels in each sex.


Ictal asystole during long-term video-EEG; semiology, localization, and intervention.
Ictal arrhythmias are disturbances of cardiac conduction that occur during clinical or electrographic seizures. Ictal asystole (IA) is rare, and its incidence can range from 0.3-0.4% in patients with epilepsy who were monitored by video-EEG (van der Lende et al., 2015). We report on ten patients (six males and four females) with an age ranging from 31 to 70 years old) who were monitored in our video-EEG (VEEG) unit over the last eight years. These patients were selected based on the history of documented ictal asystole during inpatient VEEG monitoring). In our series the mean latency from the seizure onset to the onset of ictal asystole was 22 seconds and the mean duration of the IA was 15.8 seconds. During the asystolic phase the seizures may clinically continue or syncopal signs may supervene. In our case series all the patients had either left or right temporal lobe epilepsy, six of which were lesional. We found two patterns of ictal semiology in our series. The first group of patients included five patients who experienced a rapid onset of IA in their seizure and the second group where the latency of ictal asystole was relatively late. All our cohort had a permanent pacemaker following the diagnosis, six of these patients have been event free since placement.


The Relationship Between Reductions in WHO Risk Drinking Levels During Treatment and Subsequent Healthcare Costs for the ACTIVE Workgroup.
Abstinence has historically been considered the target outcome for alcohol use disorder (AUD) treatment, yet recent work has found drinking reductions after AUD treatment, as measured by World Health Organization (WHO) risk drinking levels, are associated with meaningful improvements in functioning, physical health, and quality of life. This study extends previous analyses of AUD treatment outcomes by estimating the association between changes in WHO risk drinking levels (very high, high, medium, and low, based on average daily alcohol consumption) and healthcare costs. Secondary data analysis of the COMBINE study, a multisite randomized clinical trial of acamprosate, naltrexone and behavioral interventions for AUD. Generalized gamma regression models were used to estimate relationships between WHOrisk drinking level reductions over the course of treatment and healthcare costs in the year after treatment (N = 964) and up to 3 years following treatment (N = 651). SustainedWHOrisk drinking reductions of 2 or more levels throughout treatment were associated with 52.0% lower healthcare costs ( P < 0.001) in the year following treatment, and 44.0% lower costs ( P < 0.0025) over 3 years. A reduction of exactly 1 level was associated with 34.8% lower costs over 3 years, which was not significant ( P = 0.05). Cost reductions were driven by lower inpatient behavioral health and emergency department utilization. Reduction in WHO risk drinking levels of at least 2 levels was associated with lower healthcare costs over 1 and 3 years. Our results add to literature showing drinking reductions are associated with improvement in health.


Non-union of isolated displaced triquetral body fracture--a case report.
Triquetral fractures are the second most common carpal fractures. Triquetral body fractures are the less common type of triquetral fractures but they can be missed on plain radiographs and a non-union can be associated with considerable morbidity and reduction in functional activities. We report a unique case of displaced isolated triquetral body fracture that was initially missed on plain radiographs and resulted in non-union. The fracture non-union was diagnosed on magnetic resonance imaging and was treated successfully with open reduction and internal fixation using compression screws, without bone grafting, and early mobilisation of the wrist allowing a return to premorbid activity levels. To our knowledge, this has not been previously described in the literature. We advocate early clinical suspicion of triquetral fractures in patients with persistent ulnar-sided wrist pain following trauma, and early evaluation using computed tomography or magnetic resonance imaging.


The primary transcription unit of the human alpha 2 globin gene defined by quantitative RT/PCR.
We have set up an experimental system to map the primary transcription unit of the human alpha 2 globin gene. The duplicated human alpha globin genes (alpha 2-alpha 1) were linked to the alpha globin locus Positive Regulatory Element (PRE) and stably transfected into murine erythroleukaemia cells. We then developed a quantitative reverse transcriptase, polymerase chain reaction assay to map alpha 2 primary transcripts using primer pairs derived from different parts of the alpha 2 globin gene and its 3' flanking region. This approach has revealed the presence of steady state nuclear RNA past the poly(A) site of the alpha 2 globin gene at approximately 40% of the level of unspliced intron transcript. Furthermore, these 3' flanking transcripts diminish 500 bp into the 3' flanking region, identifying this part of the alpha 2 globin gene as the principal region of termination of transcription.


The clinical utility of CA 19-9 in pancreatic adenocarcinoma: diagnostic and prognostic updates.
CA 19-9 and CEA are the most commonly used biomarkers for diagnosis and management of patients with pancreatic cancer. Since the original compendium by Steinberg in 1990, numerous studies have reported the use of CA 19-9 and, to a lesser extent, CEA in the diagnosis of pancreatic cancer. Here we update an evaluation of the accuracy of CA 19-9 and CEA, and, unlike previous reviews, focus on discrimination between malignant and benign disease instead of normal controls. In 57 studies involving 3,285 pancreatic carcinoma cases, the combined sensitivity of CA 19-9 was 78.2% and in 37 studies involving 1,882 cases with benign pancreatic disease the specificity of CA 19-9 was 82.8%. From the combined analysis of studies reporting CEA, the sensitivity was 44.2% (1,324 cases) and the specificity was 84.8% (656 cases). These measurements more appropriately reflect the expected biomarker accuracy in the differential diagnosis of patients with periampullary diseases. We also present a summary of the use of CA 19-9 as a prognostic tool and evaluate CA 19-9 diagnostic and prognostic utility in a 10-year, single institution experience.


Addressing Unmet Basic Needs to Improve Colposcopy Adherence Among Women With Abnormal Cervical Cancer Screening.
The aims of the study were to identify unmet basic needs (BNs) among women referred to colposcopy, to assess patient acceptability/satisfaction with assistance from a navigator to address unmet BNs, and to estimate adherence to colposcopy. Women were recruited between September 2017 and January 2019 from 2 academic colposcopy centers, one serving a rural and one an urban area. Basic needs were assessed by phone before colposcopy appointments and considered unmet if unlikely to resolve in 1 month. Colposcopy adherence prestudy and poststudy implementation was abstracted over 4-6 months from administrative records. After a lead-in phase of 25 patients at each site, a BN navigator was offered to new participants with 1 or more unmet BNs. Primary outcome was adherence to initial appointment. Among 100 women, 59% had 1 or more unmet BNs, with similar prevalence between urban and rural sites. Adherence to initial colposcopy was 83% overall, 72% at the rural clinic, and 94% at the urban clinic (p = .006). These adherence rates were improved from 4 months before study launch (30/59 [51%] rural clinic and 68/137 [50%] urban clinic). Although acceptability of BN navigation was greater than 96% and women felt that it helped them get to their colposcopy visit, having a navigator was not associated with adherence. Women reporting no unmet BNs had the lowest adherence compared with women with 1 or more unmet BNs, regardless of navigator assistance (p = .03). Disadvantaged women who need colposcopy have unmet BNs and value navigator assistance for initial appointments. However, when appointment scheduling includes telephone reminders and inquiring about BNs, a navigator may not add value.


Mainstream tobacco smoke causes paternal germ-line DNA mutation.
Despite the presence of known mutagens and carcinogens in cigarette smoke, there is currently no evidence to show that smoking, or exposure to cigarette smoke, can result in heritable genetic mutation. We show that male mice exposed to mainstream tobacco smoke (MTS) exhibit a significant increase in germ-line mutation frequency in spermatogonial stem cells. We exposed mature male mice to MTS for 6 or 12 weeks and investigated mutations arising in exposed spermatogonial stem cells at the expanded simple tandem repeat locus Ms6-hm. A generalized score test showed a significant treatment effect (P = 0.0214). Ms6-hm mutation frequency was 1.4 and 1.7 times higher in mice exposed to MTS for 6 and 12 weeks, respectively, compared with sham controls. The data suggest that mutations accumulate in the spermatogonial stem cells with extended exposures. Mutation spectra were identical between exposed and sham individuals, supporting the hypothesis that tandem repeat mutations arise through indirect mechanisms of mutation. Mutations in sperm that are passed on to offspring cause permanent, irreversible changes in genetic composition and can persist in future generations. Our research suggests that the consequences of smoking extend beyond the smoker to their nonsmoking descendents.


Impact of poverty on serum phosphate concentrations in the Third National Health and Nutrition Examination Survey.
Increased serum phosphate is associated with adverse health outcomes. High intake of inexpensive processed and fast foods is common in impoverished communities, and is linked with excessive dietary phosphorus intake and elevated serum phosphate concentrations in chronic kidney disease patients. We examined the impact of socioeconomic status on dietary phosphorus intake and serum phosphate concentrations in the general population. Cross-sectional study. A total of 14,261 adult participants in the Third National Health and Nutrition Examination Survey. PREDICTORS AND OUTCOMES: Poverty to income ratio (PIR; family income indexed to the federal poverty level) was the primary index of socioeconomic status. Serum phosphate was the primary outcome variable. Although estimated phosphorus intake decreased with decreasing quartiles of PIR (P < .001), serum phosphate was inversely associated with PIR (P = .003). The relationship between lower PIR and higher serum phosphate remained significant after adjustment for demographic, laboratory, and dietary intake characteristics (P = .02). Compared with participants in the highest PIR quartile (income >300% of the federal poverty level), participants in the lowest quartile (income < the federal poverty level) had more than twice the odds of hyperphosphatemia (≥ 4.4 mg/dL) in unadjusted and multivariable-adjusted logistic regression analyses (OR, 2.2; 95% CI, 1.5 to 3.2). Although lower income was associated with decreased estimated phosphorus intake, increasing poverty was independently linked with increased serum phosphate and higher likelihood of hyperphosphatemia. These findings may indicate that conventional dietary instruments underestimate phosphorus intake, especially among impoverished individuals. Further studies are needed to explore these possibilities.


[Anatomopathologic-embryologic correlation in atrioventricular connection absence].
In order to offer a pathogenetic explanation for the absence of atrioventricular connexion, a correlation was made between the pathologic anatomy of this cardiac malformation and the embryonic processes which take part in the septation of the atrioventricular canal and the development of atrioventricular connections. The correspondence was made between the development of the canal's atrioventricular cushions, the septation of the common atrioventricular canal, the right and left atrioventricular canals and the morphogenesis of the mitral valve, all these processes were correlated with the anatomic elements derived from them. This led to infer that the malposition of the atrioventricular cushions divide the common atrioventricular canal unequally, giving rise to a narrow canal becoming atretic and a big canal where the mitral valve is evolved. The extreme lateralization of the atrioventricular septum to the right side would led to the absence of the right atrioventricular connection and the same process but to the left side, would give rise to the left absence of the atrioventricular connection. This ectopic septation process is supported by similar ones which can occur in other segments of the heart such as in tetralogy of Fallot and the transposition of the great arteries. This hypothesis explains sufficiently the pathologic anatomy of this type of congenital heart disease.


Resolution of metabolic columns by a double-label 2-DG technique: interdigitation and coincidence in visual cortical areas of the same monkey.
A double-label modification of the 2-deoxyglucose (DG) technique uses both 3H-2-DG and 14C-2-DG and allows for metabolic activity engaged by 2 distinct experimental conditions to be dissociated throughout the brain of a single subject. In the present study, we used this double-label method to examine the relationship between metabolic columns subserving the 2 eyes in cortical visual areas V1 and V2. The left and right eye's ocular dominance columns were separately activated in the same monkey to demonstrate that the double-label 2-DG method can resolve metabolic differences at the level of the cortical column. In 2 monkeys, 14C-2-DG was injected first and one eye was occluded while the other eye was visually stimulated for 30 min. Then 3H-2-DG was injected, and the occluder was switched so that the alternate eye was stimulated with the same pattern for 30 min. Autoradiographs depicting the 2 labels separately were obtained by exploiting the differential sensitivity of X-ray film and Ultrofilm to 3H and 14C emissions and by applying a radioactivity-subtraction algorithm to pairs of digitized images of the same section (Friedman et al., 1987). Columnar regions of increased activity were evident throughout V1, excepting the representations of the optic disk and monocular crescent. Superimposition of the 3H and 14C images from the same sections demonstrated that columns of increased 3H label were interdigitated with columns of increased 14C label in V1. In contrast, bands of increased 3H-2-DG uptake in extrastriate area V2 were largely coincident with the bands of increased 14C-2-DG uptake. These results illustrate the value of the double-label 2-DG technique for studying fluctuations of metabolic activity under different experimental conditions in the same subject. In the present example, the demonstration that ocular dominance columns are interdigitated in V1, whereas metabolically active bands are coincident in V2, would not have been fully appreciated by comparing 2-DG labeling across separate animals.


Mediators in thermal injury.
The combined goal of each of the mediators discussed here is homeostasis in a stressful situation. The associated immunosuppression and immunohyperactivity are unfortunate sequelae of massive upregulation of mediators normally produced in miniscule quantities. Perhaps they were once necessary to regulate survival of severely injured patients. However, Stoner's question about whether these mediators are beneficial for modern medical practices is very relevant. Our significant challenge is now to counteract the body's own efforts to heal itself. Understanding the interactions initiates this task; discovering therapeutic control mechanisms must be the next step.


Switching of oral anticoagulants in patients with nonvalvular atrial fibrillation: A narrative review.
Approval of direct-acting oral anticoagulants (DOACs) for stroke prevention in atrial fibrillation (AF) was an important milestone, providing a wider range of treatment options and creating the possibility for drug switching after initiation. In addition to improved utilisation of oral anticoagulants (OACs) for stroke prevention, reports of switching among OACs are growing in the literature; switching may influence clinical outcomes, healthcare costs and patient satisfaction. This review aimed to summarise the current literature on the pattern of OAC switching in patients with AF, including reasons for switching and clinical consequences following switching. A literature search was conducted in PubMed, Scopus and Embase on 27 June 2020. We included 39 articles published after 2013, following the introduction of apixaban. The review found that switching among OACs was common in clinical practice, significantly varying with the type of OAC. Studies reporting the reason for switching and clinical outcomes were comparatively limited. The decision to switch was often related to safety issues (usually bleeding), poor anticoagulation control and ease of use. Patient characteristics, clinical conditions and drug interactions were found to be associated with switching from OACs. Findings regarding bleeding outcomes following switching were inconsistent, possibly confounded by the rationale for switching and the switching protocol. Noting the limited number of studies included and their relatively short follow-up periods, switching did not have a significant impact on the risk of stroke and other thrombotic outcomes. Further prospective studies are needed to understand better potential rationales for switching and the clinical outcomes.