[Fixation of the tuberculum in multifragment fractures of the proximal humerus treated with hemiarthroplasty].
Hemiarthroplasty is an effective treatment for multifragmentary fractures of the proximal humerus. Purpose of this study was to compare 2 different methods of tuberosity dislocations,which is one of the primary reasons for successful posttraumatic arthroplasty. Between 2006 and 2008, 29 patients with multifragmentary fractures of the proximal humerus underwent hemiarthroplasty of the shoulder. Mean gae was 69y., while mean postoperative follow up was 27 months. We evaluated two different techiques of tuberosity fixation after anatomic reattachment on humeri: (A) intertuberosity suture stabilization around the prothesis, and their suture to the humeri and (B) direct intertuberosity suture and their suture to the humeri Postoperartive Constant score averaged 71 points. Reason for failure of the operative treatment was primary due to the postoperative tuberosity migration It was noted in all patients of group 2 and in 2 patients of group 1. Intertuberosity stabilization using sutures arpund the prothesis and tuberosity fixation to the humeri was proofed to be optimal method, because it decreased strain and mobility of tuberosities, maximize stability of the hemiarthroplasty and facilitate postoperative rehabilitation.


Spaceflight influences gene expression, photoreceptor integrity, and oxidative stress-related damage in the murine retina.
Extended spaceflight has been shown to adversely affect astronaut visual acuity. The purpose of this study was to determine whether spaceflight alters gene expression profiles and induces oxidative damage in the retina. Ten week old adult C57BL/6 male mice were flown aboard the ISS for 35 days and returned to Earth alive. Ground control mice were maintained on Earth under identical environmental conditions. Within 38 (+/-4) hours after splashdown, mice ocular tissues were collected for analysis. RNA sequencing detected 600 differentially expressed genes (DEGs) in murine spaceflight retinas, which were enriched for genes related to visual perception, the phototransduction pathway, and numerous retina and photoreceptor phenotype categories. Twelve DEGs were associated with retinitis pigmentosa, characterized by dystrophy of the photoreceptor layer rods and cones. Differentially expressed transcription factors indicated changes in chromatin structure, offering clues to the observed phenotypic changes. Immunofluorescence assays showed degradation of cone photoreceptors and increased retinal oxidative stress. Total retinal, retinal pigment epithelium, and choroid layer thickness were significantly lower after spaceflight. These results indicate that retinal performance may decrease over extended periods of spaceflight and cause visual impairment.


Excess electrons in ice: a density functional theory study.
We present a density functional theory study of the localization of excess electrons in the bulk and on the surface of crystalline and amorphous water ice. We analyze the initial stages of electron solvation in crystalline and amorphous ice. In the case of crystalline ice we find that excess electrons favor surface states over bulk states, even when the latter are localized at defect sites. In contrast, in amorphous ice excess electrons find it equally favorable to localize in bulk and in surface states which we attribute to the preexisting precursor states in the disordered structure. In all cases excess electrons are found to occupy the vacuum regions of the molecular network. The electron localization in the bulk of amorphous ice is assisted by its distorted hydrogen bonding network as opposed to the crystalline phase. Although qualitative, our results provide a simple interpretation of the large differences observed in the dynamics and localization of excess electrons in crystalline and amorphous ice films on metals.


[Causes of death in patients with rheumatic diseases in Moscow].
To study causes of death of rheumatic patients in Moscow. Autopsy protocols for 1999-2002 were analysed for two pathoanatomic departments of Moscow. Rheumatic diseases were detected in 165 cases (2.0% of overall number of autopsies in these departments). Rheumatic heart disease (RHD) was stated in 99 (60.0%) cases, rheumatic fever relapse (RFR) in 4 (2.4%), rheumatoid arthritis (RA) in 28 (17.0%), systemic lupus erythematosus (SLE) in 8 (4.8%), systemic sclerosis (SS) in 3 (1.8%), ankylosing spondilarthritis (AS) in 2 (1.2%), systemic vasculitis (SV) in 2 (1.2%), osteoarthrosis in 11 (7.3%), gout in 3 (1.8%), polymyositis in 1 (0.6%). RHD patients died of decompensated circulation (DC) (54%), acute heart failure (AHF) (14%), thromboembolism (TE) (6%), other causes (26%). RF patients died of TE (n = 2), DC (n = 1), AHF (n = 1). Out of 28 RA patients, 5 patients died of secondary amyloidosis, 3 of DC, 7 of AHF, 1 of TE, 5 of infectious complications, 7 of other causes. SLE patients died of uremia (n = 2), acute adrenal failure (n = 1), infectious complications (n = 2), AHF (n = 2), brain edema (n = 1). Among rheumatic diseases, rheumatic heart valve disease was most severe as it caused the highest mortality. Cardiovascular pathology caused death in most of the rheumatic patients.


Cellular Chaperones As Therapeutic Targets in ALS to Restore Protein Homeostasis and Improve Cellular Function.
Heat shock proteins (Hsps) are ubiquitously expressed chaperone proteins that enable cells to cope with environmental stresses that cause misfolding and denaturation of proteins. With aging this protein quality control machinery becomes less effective, reducing the ability of cells to cope with damaging environmental stresses and disease-causing mutations. In neurodegenerative disorders such as Amyotrophic Lateral Sclerosis (ALS), such mutations are known to result in protein misfolding, which in turn results in the formation of intracellular aggregates cellular dysfunction and eventual neuronal death. The exact cellular pathology of ALS and other neurodegenerative diseases has been elusive and thus, hindering the development of effective therapies. However, a common scheme has emerged across these "protein misfolding" disorders, in that the mechanism of disease involves one or more aspects of proteostasis; from DNA transcription, RNA translation, to protein folding, transport and degradation via proteosomal and autophagic pathways. Interestingly, members of the Hsp family are involved in each of these steps facilitating normal protein folding, regulating the rate of protein synthesis and degradation. In this short review we summarize the evidence that suggests that ALS is a disease of protein dyshomeostasis in which Hsps may play a key role. Overwhelming evidence now indicates that enabling protein homeostasis to cope with disease-causing mutations might be a successful therapeutic strategy in ALS, as well as other neurodegenerative diseases. Novel small molecule co-inducers of Hsps appear to be able to achieve this aim. Arimoclomol, a hydroxylamine derivative, has shown promising results in cellular and animal models of ALS, as well as other protein misfolding diseases such as Inclusion Body Myositis (IBM). Initial clinical investigations of Arimoclomol have shown promising results. Therefore, it is possible that the long series of unsuccessful clinical trials for ALS may soon be reversed, as optimal targeting of proteostasis in ALS may now be possible, and may deliver clinical benefit to patients.


Prostaglandins and platelets.
Release of arachidonate from cell membrane phospholipids by activation of phospholipase A1 is a key step in the formation of prostaglandins and thromboxanes. In platelets PGH2 and TXA2 are formed from arachidonate and can cause aggregation and the release of granule contents; in vascular tissue, PGI2 is formed instead and, by increasing platelet cAMP, inhibits platelet reactions. There is considerable interest in inhibitors of the enzymes in these pathways as drugs to modify thrombus formation. Results of the clinical trials, however, indicate that drugs which inhibit cyclo-oxygenase may not have a major effect on the thromboembolic complications of arterial disease.


Polymeric display of immunogenic epitopes from herpes simplex virus and transmissible gastroenteritis virus surface proteins on an enteroadherent fimbria.
The strong immunogenicity of bacterial fimbriae results from their polymeric and proteinaceous nature, and the protective role of these immunogens in experimental or commercial vaccines is associated with their capacity to induce antiadhesive antibodies. Fimbria-mediated intestinal colonization by enteropathogens typically leads to similar antibody responses. The possibility of taking advantage of these properties was investigated by determining whether enteroadhesive fimbriae, like the 987P fimbriae of enterotoxigenic Escherichia coli, can serve as carriers for foreign antigens without losing their adhesive characteristics. Random linker insertion mutagenesis of the fasA gene encoding the major 987P subunit identified five different mutants expressing wild-type levels of fimbriation. The linker insertion sites of these mutants were used to introduce three continuous segments of viral surface glycoproteins known to be accessible to antibodies. These segments encode residues 11 to 19 or 272 to 279 of herpes simplex virus type 1 (HSV-1) glycoprotein D [gD(11-19) and gD(272-279), respectively] or residues 379 to 388 of the transmissible gastroenteritis virus (TGEV) spike protein [S(379-388)]. Studies of bacteria expressing fimbriae incorporating mutated FasA subunits alone or together with wild-type FasA subunits (hybrid fimbriae) indicated that foreign epitopes were best exported and displayed on assembled fimbriae when they were inserted near the amino terminus of FasA. Fimbriated bacteria expressing FasA subunits carrying the HSV gD(11-19) or the TGEV S(379-388) epitope inserted between the second and third residues of mature FasA elicited high levels of foreign epitope antibodies in all rabbits immunized parenterally. Antibodies against the HSV epitope were also shown to recognize the epitope in the context of the whole gD protein. Because the 987P adhesive subunit FasG was shown to be present on mutated fimbriae and to mediate bacterial attachment to porcine intestinal receptors, polymeric display of foreign epitopes on 987P offers new opportunities to test the potential beneficial effect of enteroadhesion for mucosal immunization and protection against various enteric pathogens.


Validation of the 8-item Attitudes Towards Gambling Scale (ATGS-8) in a British population survey.
Public opinions concerning gambling are an important factor in shaping public policy. Little empirical attention has been given to assessing gambling attitudes within the general population. The aim of the present study is to validate the 8-item Attitudes Towards Gambling Scale (ATGS-8) in British individuals and to investigate associations of these attitudes with frequency of gambling and gambling problems. Data were derived from 7746 individuals participating in the British Gambling Prevalence Survey 2010, a comprehensive interview-based survey conducted in Great Britain between November 2009 and May 2010. Confirmatory factor analysis and separate regression analyses were applied. The one-dimensional structure of the ATGS-8 was confirmed in the community sample and by gender. Furthermore, more positive attitudes towards gambling were positively related to frequency of gambling and gambling problems. The present study extends the previous evaluations of the scale by providing detailed evidence for the utility and usefulness of the ATGS-8 in a community sample and across gender. The ATGS-8 is a valid instrument to assess public opinion on gambling among the general population.


Increased frequency of occurrence of bendopnea is associated with poor outcomes in heart failure outpatients.
Relationship between the frequency of occurrence of bendopnea during the daily life of heart failure (HF) outpatients and clinical outcomes has never been evaluated before. Turkish Research Team-Heart Failure (TREAT-HF) is a network between HF centres, which undertakes multicentric observational studies in HF. Herein, the data including stable 573 HF patients with reduced ejection fraction out of seven HF centres were presented. A questionnaire was filled by the patients, with the question 'Do you experience shortness of breath while tying your shoelace?', assessing the presence and frequency of bendopnea. To the question related to bendopnea, 48% of the patients answered 'yes, every time', 31% answered 'yes, sometimes', and 21% answered 'No'. Patients were followed for an average of 24 ± 14 months, and the patients who answered 'yes, every time' and 'yes, sometimes' to the bendopnea question were found having increased risk for both HF-related hospitalisations (HR:3.2, p < .001- HR:2.8, p = .005) and composite outcome consisting of 'HF-related hospitalisations and all-cause death in the multi-variate analysis (HR:3.1, p < .001- HR:3.0, p < .001). Kaplan Meier analysis for HF-related hospitalisation, all-cause death, and the composite of these were provided for these three groups, yielding significant and graded divergence curves with the best prognosis in 'no' group, with the moderate prognosis in 'sometimes' group, and with the worst prognosis in the 'every time' group. For the first time in the literature, our study shows that the increased frequency of bendopnea occurrence in daily life is associated with poor outcomes in HF outpatients.


Dystonin/BPAG1 promotes plus-end-directed transport of herpes simplex virus 1 capsids on microtubules during entry.
During infection by herpes simplex virus 1 (HSV-1), the viral capsid is transported around the cytoplasm along the microtubule (MT) network. Although molecular motors have been implicated in this process, the composition of the molecular machinery required for efficient directional transport is unknown. We previously showed that dystonin (BPAG1) is recruited to HSV-1 capsids by the capsid-bound tegument protein pUL37 to promote efficient cytoplasmic transport of capsids during egress. Dystonin is a cytoskeleton cross-linker which localizes at MT plus ends and has roles in retrograde and anterograde transport in neurons. In this study, we investigated the role of dystonin during the entry stages of HSV-1 infection. Because of the way in which the MT network is organized, capsids are required to change their direction of motion along the MTs as they travel from the point of entry to the nucleus, where replication takes place. Thus, capsids first travel to the centrosome (the principal microtubule organizing center) by minus-end-directed transport and then switch polarity and travel to the nucleus by plus-end-directed transport. We observed that transport of capsids toward the centrosome was slowed, but not blocked, by dystonin depletion. However, transport of capsids away from the centrosome was significantly impaired, causing them to accumulate in the vicinity of the centrosome and reducing the numbers reaching the nucleus. We conclude that, during entry of HSV-1, dystonin has a specific role in plus-ended transport of capsids from the centrosome to the nucleus.


Recurrent corneal erosion after trauma: a statistical study.
Office records of 377 consecutive traumatic corneal abrasions were examined. Twenty-nine cases experienced at least one episode of recurrent corneal erosion (7.69%). There were no recurrences in patients under 20 years of age (P less than .01). The size of the original abrasion could not be correlated with recurrence rate. Injuries with paper, fingernails, or bushes and branches were 4.9 times more likely to recur than injuries from all other materials (P less than .001).


Patient misconceptions concerning lumbar spondylosis diagnosis and treatment.
OBJECT Patient outcome measures are becoming increasingly important in the evaluation of health care quality and physician performance. Of the many novel measures currently being explored, patient satisfaction and other subjective measures of patient experience are among the most heavily weighted. However, these subjective measures are strongly influenced by a number of factors, including patient demographics, level of understanding of the disorder and its treatment, and patient expectations. In the present study, patients referred to a neurosurgery clinic for degenerative spinal disorders were surveyed to determine their understanding of lumbar spondylosis diagnosis and treatment. METHODS A multiple-choice, 6-question survey was distributed to all patients referred to a general neurosurgical spine clinic at a tertiary care center over a period of 11 months as a quality improvement initiative to assist the provider with individualized patient counseling. The survey consisted of questions designed to assess patient understanding of the role of radiological imaging in the diagnosis and treatment of low-back and leg pain, and patient perception of the indications for surgical compared with conservative management. Demographic data were also collected. RESULTS A total of 121 surveys were included in the analysis. More than 50% of the patients indicated that they would undergo spine surgery based on abnormalities found on MRI, even without symptoms; more than 40% of patients indicated the same for plain radiographs. Similarly, a large proportion of patients (33%) believed that back surgery was more effective than physical therapy in the treatment of back pain without leg pain. Nearly one-fifth of the survey group (17%) also believed that back injections were riskier than back surgery. There were no significant differences in survey responses among patients with a previous history of spine surgery compared with those without previous spine surgery. CONCLUSIONS These results show that a surprisingly high percentage of patients have misconceptions regarding the diagnosis and treatment of lumbar spondylosis, and that these misconceptions persist in patients with a history of spine surgery. Specifically, patients overemphasize the value of radiological studies and have mixed perceptions of the relative risk and effectiveness of surgical intervention compared with more conservative management. These misconceptions have the potential to alter patient expectations and decrease satisfaction, which could negatively impact patient outcomes and subjective valuations of physician performance. While these results are preliminary, they highlight a need for improved communication and patient education during surgical consultation for lumbar spondylosis.


Optimizing sparse sequencing of single cells for highly multiplex copy number profiling.
Genome-wide analysis at the level of single cells has recently emerged as a powerful tool to dissect genome heterogeneity in cancer, neurobiology, and development. To be truly transformative, single-cell approaches must affordably accommodate large numbers of single cells. This is feasible in the case of copy number variation (CNV), because CNV determination requires only sparse sequence coverage. We have used a combination of bioinformatic and molecular approaches to optimize single-cell DNA amplification and library preparation for highly multiplexed sequencing, yielding a method that can produce genome-wide CNV profiles of up to a hundred individual cells on a single lane of an Illumina HiSeq instrument. We apply the method to human cancer cell lines and biopsied cancer tissue, thereby illustrating its efficiency, reproducibility, and power to reveal underlying genetic heterogeneity and clonal phylogeny. The capacity of the method to facilitate the rapid profiling of hundreds to thousands of single-cell genomes represents a key step in making single-cell profiling an easily accessible tool for studying cell lineage.


Comparing Randomized Controlled Trials and Real-World Studies in Chronic Obstructive Pulmonary Disease Pharmacotherapy.
Analytic epidemiological studies cover a large spectrum of study methodologies, ranging from noninterventional observational studies (population-based, case-control, or cohort studies) to interventional studies (clinical trials). Herein, we review the different research methodologies or study designs and discuss their advantages and disadvantages in the context of chronic obstructive pulmonary disease (COPD) pharmacotherapy. Although randomized controlled trials (RCTs) are considered the "gold standard" for evaluating the efficacy and safety of an intervention, observational studies conducted in a real-world scenario are useful in providing evidence on the effectiveness of the intervention in clinical practice; understanding both efficacy and effectiveness is important from the clinician's perspective. Pragmatic clinical trials that use real-world data while retaining randomization bridge the gap between explanatory RCTs and noninterventional observational studies. Overall, different study designs have their associated advantages and disadvantages; together, findings from all types of studies bring about progress in clinical research as elucidated through examples from COPD research in this paper.


COVID-19 Reflections and Team-Based Entrustable Professional Activities for Bioevent Preparedness.
This article was migrated. The article was marked as recommended. The coronavirus disease 2019 (COVID-19) outbreak has shaken the entire human race with its unprecedented health and economic outcomes. Researchers and clinicians are trying to get an insight into various aspects of the COVID-19 disease and related aspects, and to some extent, they have succeeded too, but a lot more is yet to be discovered and disclosed. However, it appears that the pandemic is going to stay for a longer duration with varied expressions. In these testing and challenging times when the entire world is battling the massive public health crises, the healthcare workers remain the most vulnerable targets. The current situation unleashes adoption of new structures and processes, but often gloomed with ethical dilemmas. The COVID-19 is not going to be the last major pandemic in the history of human civilization. Henceforth, the million-dollar questions we face now and forever are: How are we planning to prepare the workforce for an health crisis of this cataclysmic proportions? Can we reimagine a curricular framework that could address public health crises arising from epidemics and pandemics? What model could facilitate for advancement in crisis preparedness and management? Do we need an orchestrated team approach in managing and containing any public health crisis of global concern like the COVID-19? What about the entrustable professional activities for a team task? These pertinent questions do not have straight forward answers. It requires a great effort at various levels, but such a change in curricular framework appears to be the need of the hour.


Anxiolytic effects and neuroanatomical targets of estrogen receptor-β (ERβ) activation by a selective ERβ agonist in female mice.
The dichotomous anxiogenic and anxiolytic properties of estrogens have been reported to be mediated by two distinct neural estrogen receptors (ER), ERα and ERβ, respectively. Using a combination of pharmacological and genetic approaches, we confirmed that the anxiolytic actions of estradiol are mediated by ERβ and extended and these observations to demonstrate the neuroanatomical targets involved in ERβ activation in these behavioral responses. We examined the effects of the biologically active S-enantiomer of diarylpropionitrile (S-DPN) on anxiety-related behavioral measures, the corresponding stress hormonal response to hypothalamo-pituitary-adrenal axis reactivity, and potential sites of neuronal activation in mutant female mice carrying a null mutation for ERβ gene (βERKO). S-DPN administration significantly reduced anxiety-like behaviors in the open field, light-dark exploration, and the elevated plus maze (EPM) in ovariectomized wild-type (WT) mice, but not in their βERKO littermates. Stress-induced corticosterone (CORT) and ACTH were also attenuated by S-DPN in the WT mice but not in the βERKO mice. Using c-fos induction after elevated plus maze, as a marker of stress-induced neuronal activation, we identified the anterodorsal medial amygdala and bed nucleus of the stria terminalis as the neuronal targets of S-DPN action. Both areas showed elevated c-fos mRNA expression with S-DPN treatment in the WT but not βERKO females. These studies provide compelling evidence for anxiolytic effects mediated by ERβ, and its neuroanatomical targets, that send or receive projections to/from the paraventricular nucleus, providing potential indirect mode of action for the control of hypothalamo-pituitary-adrenal axis function and behaviors.


Response to interferon-beta treatment in multiple sclerosis patients: a genome-wide association study.
Up to 50% of multiple sclerosis (MS) patients do not respond to interferon-beta (IFN-β) treatment and determination of response requires lengthy clinical follow-up of up to 2 years. Response predictive genetic markers would significantly improve disease management. We aimed to identify IFN-β treatment response genetic marker(s) by performing a two-stage genome-wide association study (GWAS). The GWAS was carried out using data from 151 Australian MS patients from the ANZgene/WTCCC2 MS susceptibility GWAS (responder (R)=51, intermediate responders=24 and non-responders (NR)=76). Of the single-nucleotide polymorphisms (SNP) that were validated in an independent group of 479 IFN-β-treated MS patients from Australia, Spain and Italy (R=273 and NR=206), eight showed evidence of association with treatment response. Among the replicated associations, the strongest was observed for FHIT (Fragile Histidine Triad; combined P-value 6.74 × 10-6) and followed by variants in GAPVD1 (GTPase activating protein and VPS9 domains 1; combined P-value 5.83 × 10-5) and near ZNF697 (combined P-value 8.15 × 10-5).


Right size matters! The ideal size of hybrid prosthesis in frozen elephant trunk.
There is abundant of literature emerging to support the use of the frozen elephant trunk prosthesis, but there is still a lack of clear consensus on the sizing of the hybrid prosthesis. There is a general consensus that the stent should not be oversized in acute aortic dissection and chronic aortic dissection. Some surgeons consider that only the true lumen has to be measured while others argue that the entire diameter of the aorta has to be measured, and a few measure the aorta intraoperatively. In regards to thoracic aortic aneurysm, most surgeons oversize the stent-graft by 10% to 20%. A small device may not provide adequate sealing, whereas a larger device may cause new entry points distally. Hence, an appropriate device has to be selected for the optimal outcome.


Secretory factors from human adipose tissue and their functional role.
Obesity is characterized by an expanded adipose tissue mass. Recent data suggest that adipose tissue is a multi-functional organ rather than simply a passive storage site for excess energy. It has been clearly demonstrated that human adipose tissue produces a variety of secretory factors that exert multiple effects at both the local and the systemic level. To date, >100 products, covering a broad range of protein families as well as many fatty acids and prostaglandins, have been reported to be secreted by adipose tissue. The source of these secreted factors is not only mature fat cells but also poorly-identified cells present in the stromal-vascular fraction including macrophages. Secreted factors of particular interest include many cytokines or chemokines, such as TNF-alpha, IL-6, IL-8, as well as plasminogen activator inhibitor-1, angiotensin-II, leptin, and adiponectin. In the obese state the expression and secretion of these factors is disturbed. With the exception of adiponectin, most circulating factors are elevated. From this perspective, obesity can be described as a pro-inflammatory condition. In addition, regional differences in adipose expression of many of these factors have been found. There is now growing evidence that many secretory factors play an important role in the pathophysiology of the metabolic and cardiovascular complications of obesity. The question arising from these observations is how the secretory pattern of adipose tissue can be modified by dietary and pharmacological measures to reduce the health risks of obesity.


The association between otogenic lateral sinus thrombosis and thrombophilia - A long-term follow-up.
Otogenic lateral sinus thrombosis (OLST) is an intracranial, potentially life-threatening complication of acute and chronic otitis media. Since congenital thrombophilic disorders are risk factors for cerebral venous thrombosis, OLST may be related to thrombophilia. The aim of our study was twofold: to evaluate whether patients who suffered from OLST in childhood also have thrombophilia, and whether these patients experienced thromboembolic episodes in future years. Retrospective case series. The medical charts of all children hospitalized for OLST at Soroka University Medical Center of Israel, a tertiary referral hospital, from January 1983 to September 2014 were reviewed. The patients were invited for a follow-up visit and comprehensive medical history was taken along with a physical examination and laboratory work-up for thrombophilia. Seven patients were included in the study. Of these, 3 (43%) had results suggesting thrombophilic disorders manifested by elevated levels of factor IX and decreased levels of protein S activity (n = 1), decreased levels of proteins C and S activity (n = 1), and elevated levels of antibodies to cardiolipin (n = 1). No patients experienced clear thrombophilic events; however, 2 patients (29%) with later proven thrombophilia suffered neurologic sequelae, possibly suggesting thrombophilic events. Pediatric OLST secondary to acute otitis media and mastoiditis may reflect an underlying thrombophilia. Laboratory work-up for thrombophilia should be performed, and anticoagulant treatment may be warranted in managing these patients.


Therapeutic efficacy of L-ornithine-L-aspartate infusions in patients with cirrhosis and hepatic encephalopathy: results of a placebo-controlled, double-blind study.
One hundred twenty-six patients with cirrhosis, hyperammonemia (>50 micromol/L), and chronic (persistent) hepatic encephalopathy (HE), which developed spontaneously without the existence of known precipitating factors, were enrolled in a randomized, double-blind, placebo-controlled clinical trial of intravenously administered L-ornithine-L-aspartate (OA). Patients with subclinical (grade 0, West-Haven criteria) hepatic encephalopathy (SHE), characterized by a prolonged number connection test A (NCT-A) time, and manifest HE (grades I and II, West-Haven criteria) were included in the investigation. The trial was planned as a confirmatory clinical trial OA administered in a dose of 20 g/d, as well as placebo, were dissolved in 250 mL of 5% fructose and infused intravenously for a period of 4 hours during 7 consecutive days with a superimposed protein load at the end of the daily treatment period. Primary variables were postprandial venous ammonia and NCT-A performance time measured following OA or placebo infusions to evaluate the net effect of the treatment on the prevention of the protein-induced hyperammonemia, and on parameters such as NCT-A influenced by hyperammonemia. Mental state gradation, portal systemic encephalopathy index (PSEI), and fasting ammonia levels were estimated as additional efficacy parameters. The data presented are based on the total study sample (intent-to-treat analysis), which included 63 patients in the placebo group and 63 patients in the OA group. Of the 126 patients, 114 met all the criteria for inclusion and completed the trial and treatment as outlined in the protocol (treated-per-protocol analysis). During baseline, the placebo and treatment groups were homogeneous with regard to mental states, NCT-A performance time, fasting venous blood ammonia levels, and Child-Pugh criteria. Although a slight improvement occurred in the placebo group, NCT-A performance times (P < .001) and postprandial venous ammonia concentrations in the OA-treated group showed improvements in comparison with placebo. In addition, venous fasting blood ammonia concentration (P < .01), mental state gradation (P < .001), and PSEI (P < .01), which includes the mental state gradation, NCT-A time, and postprandial venous ammonia in this trial, improved to a much higher degree in the OA group than in the placebo group. In subgroups retrospectively classified according to their initial mental state gradation, OA showed differential but uniformly significant efficacies in patients with manifest HE with respect to ammonia-lowering, improvement in NCT times, and mental state gradation. In patients with initial SHE, OA revealed differences between the medications in the psychometric test used. Adverse events consisting of mild gastrointestinal disturbances were observed in 3 of the OA-treated patients (5%). OA infusion appears to be a safe, effective treatment of chronic (persistent) manifest HE in cirrhotic patients. Additional investigations are required to assess the efficacy of OA in patients with SHE, as well as in patients with more severe grades of HE.


The Maintaining Musculoskeletal Health (MAmMOTH) Study: Protocol for a randomised trial of cognitive behavioural therapy versus usual care for the prevention of chronic widespread pain.
Cognitive behavioural therapy (CBT) has been shown to improve outcomes for patients with fibromyalgia, and its cardinal feature chronic widespread pain (CWP). Prediction models have now been developed which identify groups who are at high-risk of developing CWP. It would be beneficial to be able to prevent the development of CWP in these people because of the high cost of symptoms and because once established they are difficult to manage. We will test the hypothesis that among patients who are identified as at high-risk, a short course of telephone-delivered CBT (tCBT) reduces the onset of CWP. We will further determine the cost-effectiveness of such a preventative intervention. The study will be a two-arm randomised trial testing a course of tCBT against usual care for prevention of CWP. Eligible participants will be identified from a screening questionnaire sent to patients registered at general practices within three Scottish health boards. Those returning questionnaires indicating they have visited their doctor for regional pain in the last 6 months, and who have two of, sleep problems, maladaptive behaviour response to illness, or high number of somatic symptoms, will be invited to participate. After giving consent, participants will be randomly allocated to either tCBT or usual care. We aim to recruit 473 participants to each treatment arm. Participants in the tCBT group will have an initial assessment with a CBT therapist by telephone, then 6 weekly sessions, and booster sessions 3 and 6 months after treatment start. Those in the usual care group will receive no additional intervention. Follow-up questionnaires measuring the same items as the screening survey questionnaire will be sent 3, 12 and 24 months after start of treatment. The main outcome will be CWP at the 12 month questionnaire. This will be the first trial of an intervention aimed at preventing fibromyalgia or CWP. The results of the study will help to inform future treatments for the prevention of chronic pain, and aetiological models of its development. ClinicalTrials.gov ID: NCT02668003URL: Please check that the following URLs are working. If not, please provide alternatives: NCT02668003Alternative is: https://www.clinicaltrials.gov/ct2/show/NCT02668003> . Date registered: 28-Jan-2016.


Protein interaction network revealed by quantitative proteomic analysis links TFIIB to multiple aspects of the transcription cycle.
Although TFIIB is widely regarded as an initiation factor, recent reports have implicated it in multiple aspects of eukaryotic transcription. To investigate the broader role of TFIIB in transcription, we performed quantitative proteomic analysis of yeast TFIIB. We purified two different populations of TFIIB; one from soluble cell lysate, which is not engaged in transcription, and the other from the chromatin fraction which yields the transcriptionally active form of the protein. TFIIB purified from the chromatin exhibits several interactions that explain its non-canonical roles in transcription. RNAPII, TFIIF and TFIIH were the only components of the preinitiation complex with a significant presence in chromatin TFIIB. A notable feature was enrichment of all subunits of CF1 and Rat1 3' end processing-termination complexes in chromatin-TFIIB preparation. Subunits of the CPF termination complex were also detected in both chromatin and soluble derived TFIIB preparations. These results may explain the presence of TFIIB at the 3' end of genes during transcription as well as its role in promoter-termination interaction.


Determinants of acute kidney injury after cardiac surgery: a systematic review.
Acute kidney injury following cardiac surgery (CS-AKI) represents a severe postoperative complication, negatively impacting short-term and long-term mortality. Due to the lack of a specific treatment, effective prevention remains the most powerful tool to overcome the CS-AKI burden. Improving the preventive strategies is possible by establishing appropriate preoperative risk profiles. Various clinical models were proposed as a means to assist physicians in stratifying the risk of CS-AKI. However, these models are used for predicting severe forms of CS-AKI, while their predictive power for mild forms is insufficient. Our paper represents the first systematic approach to review all proposed preoperative risk factors and their predictive power. Our strategy is the starting point for selecting and comparing the predictive elements to be integrated into future risk models. Heart failure, chronic hyperglycemia, anemia, obesity, preoperative exposure to nephrotoxic drugs or contrast media, inflammation, proteinuria, and pre-existing kidney disease were systematically reviewed and were found to be associated with an increased risk of postoperative CS-AKI. As no externally validated and universally accepted risk models currently exist, the clinical judgment and a good knowledge of the preoperative risk factors in the light of new evidence may help personalize preoperative risk profiles as the cornerstone of prevention measures.


Bistability in a simple fluid network due to viscosity contrast.
We study the existence of multiple equilibrium states in a simple fluid network using Newtonian fluids and laminar flow. We demonstrate theoretically the presence of hysteresis and bistability, and we confirm these predictions in an experiment using two miscible fluids of different viscosity-sucrose solution and water. Possible applications include blood flow, microfluidics, and other network flows governed by similar principles.


Triage abilities of nurse practitioner vs pediatrician.
The ability of ediatric nurse practitioners (PNPs) to perform triage functions for ill children in a busy pediatric outpatient service was compared to that of a group of pediatricians. One hundred thirteen children were seen separately on arrival by a PNP and by a pediatrician. The PNPs' evaluations agreed with the pediatricians' 84% of the time. Only two significant differences were shown: PNPs were more diligent in recording patient problems, while pediatricians were better able to anticipate the need for roentgenograms and laboratory studies. The results indicate that valuable physician time may be conserved by using the PNP to perform triage in large pediatric outpatient services.


Clinical Features and Outcomes of 23 Patients with Wiskott-Aldrich Syndrome: A Single-Center Experience
Wiskott-Aldrich syndrome (WAS) is an X-linked primary immune deficiency characterized by microthrombocytopenia, eczema, and recurrent infections. We aimed to evaluate the clinical features and outcomes of a WAS cohort. We retrospectively evaluated the clinical courses, immunological features, treatments, and outcomes in a total of 23 WAS patients together with data related to 11 transplanted cases among them between 1982 and 2019. Before admission, 11 patients (48%) were misdiagnosed with immune thrombocytopenia. WAS scores were mostly 4 or 5. Eleven patients were transplanted and they had an overall survival rate of 100% during a median follow-up period of 8.5 years (range: 8 months to 20 years). Five patients who were not transplanted died at a median of 7 years (range: 2-26 years). Nontransplanted patients had high morbidity due to organ damage, mostly caused by autoimmunity, bleeding, and infections. Two novel mutations were also defined. All male babies with microthrombocytopenia should be evaluated for WAS. Hematopoietic stem cell transplantation should be performed at the earliest age with the best possible donors.


Crystal structure and local order in Co(6)Al(11-x)Si(6+x).
The ternary compound Co(6)Al(11-x)Si(6+x) (epsilon phase) was prepared from the elements by arc melting and subsequent heat treatment, and then characterized by single-crystal X-ray diffraction (XRD), electron-probe microanalysis (EPMA), differential thermal analysis (DTA) and transmission electron microscopy (TEM). This new structure type consists of planar layers with the composition [Co(6)Al(10)Si(4)], which are penetrated by perpendicular (Si-Si-Al) chains. While the layers are well described by an orthorhombic model (space group Pnma, Pearson symbol oP46), the chains exhibit doubled periodicity, thus yielding a superstructure. Two alternative ordering models (space group Cmc2(1), oC184, and space group P2(1)/c, mP92) are presented and discussed based on XRD and TEM results. The (Si-Si-Al) chains are located in pentagonal antiprismatic ;channels' which reveal the similarity of the Co(6)Al(11-x)Si(6+x) structure to Al-rich transition-metal compounds such as Co(4)Al(13), Co(2)Al(5), Fe(4)Al(13), V(7)Al(45), V(4)Al(23) and VAl(10), which also exhibit this type of pentagonal ;channels' in their crystal structures. The phase shows only a very small homogeneity range.


Asthma diagnosis: into the fourth dimension.
Asthma is the most common chronic respiratory disease in the UK; however, the misdiagnosis rate is substantial. The lack of consistency in national guidelines and the paucity of data on the performance of diagnostic algorithms compound the challenges in asthma diagnosis. Asthma is a highly rhythmic disease, characterised by diurnal variability in clinical symptoms and pathogenesis. Asthma also varies day to day, seasonally and from year to year. As much as it is a hallmark for asthma, this variability also poses significant challenges to asthma diagnosis. Almost all established asthma diagnostic tools demonstrate diurnal variation, yet few are performed with standardised timing of measurements. The dichotomous interpretation of diagnostic outcomes using fixed cut-off values may further limit the accuracy of the tests, particularly when diurnal variability straddles cut-off values within a day, and careful interpretation beyond the 'positive' and 'negative' outcome is needed. The day-to-day and more long-term variations are less predictable and it is unclear whether performing asthma diagnostic tests during asymptomatic periods may influence diagnostic sensitivities. With the evolution of asthma diagnostic tools, home monitoring and digital apps, novel strategies are needed to bridge these gaps in knowledge, and circadian variability should be considered during the standardisation process. This review summarises the biological mechanisms of circadian rhythms in asthma and highlights novel data on the significance of time (the fourth dimension) in asthma diagnosis.


Functional signatures of oral dysbiosis during periodontitis progression revealed by microbial metatranscriptome analysis.
Periodontitis is a polymicrobial biofilm-induced inflammatory disease that affects 743 million people worldwide. The current model to explain periodontitis progression proposes that changes in the relative abundance of members of the oral microbiome lead to dysbiosis in the host-microbiome crosstalk and then to inflammation and bone loss. Using combined metagenome/metatranscriptome analysis of the subgingival microbiome in progressing and non-progressing sites, we have characterized the distinct molecular signatures of periodontitis progression. Metatranscriptome analysis was conducted on samples from subgingival biofilms from progressing and stable sites from periodontitis patients. Community-wide expression profiles were obtained using Next Generation Sequencing (Illumina). Sequences were aligned using 'bowtie2' against a constructed oral microbiome database. Differential expression analysis was performed using the non-parametric algorithm implemented on the R package 'NOISeqBio'. We summarized global functional activities of the oral microbial community by set enrichment analysis based on the Gene Ontology (GO) orthology. Gene ontology enrichment analysis showed an over-representation in the baseline of active sites of terms related to cell motility, lipid A and peptidoglycan biosynthesis, and transport of iron, potassium, and amino acids. Periodontal pathogens (Tannerella forsythia and Porphyromonas gingivalis) upregulated different TonB-dependent receptors, peptidases, proteases, aerotolerance genes, iron transport genes, hemolysins, and CRISPR-associated genes. Surprisingly, organisms that have not been usually associated with the disease (Streptococcus oralis, Streptococcus mutans, Streptococcus intermedius, Streptococcus mitis, Veillonella parvula, and Pseudomonas fluorenscens) were highly active transcribing putative virulence factors. We detected patterns of activities associated with progression of clinical traits. Among those we found that the profiles of expression of cobalamin biosynthesis, proteolysis, and potassium transport were associated with the evolution towards disease. We identified metabolic changes in the microbial community associated with the initial stages of dysbiosis. Regardless of the overall composition of the community, certain metabolic signatures are consistent with disease progression. Our results suggest that the whole community, and not just a handful of oral pathogens, is responsible for an increase in virulence that leads to progression. NCT01489839, 6 December 2011.


Hemorrhage in newborn lambs: effects on arterial blood pressure, ACTH, cortisol, and vasopressin.
Arterial blood pressure, central venous pressure, and plasma concentrations of ACTH, cortisol, and vasopressin (AVP) were monitored in chronically prepared, unanesthetized newborn and weanling lambs at rest and during and after hemorrhage of 15% of estimated blood volume at 1.5%/min. Differences in the endocrine and blood pressure responses to hypovolemia were noted in the two groups of animals. Hemorrhage did not change arterial mean pressure, reduced central venous pressure, and caused a delayed increase in the plasma concentrations of ACTH, cortisol, and AVP in the newborn lambs. In weanling lambs, hemorrhage reduced arterial mean pressure and central venous pressure and promptly increased plasma ACTH and cortisol levels while plasma AVP concentrations again showed a delayed increase. The data indicate that certain hormonal mechanisms for the defense of blood volume are present and operational within 3 days of birth and that age-related differences in the responses to hemorrhage exist in the lamb.


Coupling of mitochondria to store-operated Ca(2+)-signaling sustains constitutive activation of protein kinase B/Akt and augments survival of malignant melanoma cells.
Mitochondria are emerging as a major hub for cellular Ca(2+)-signaling, though their contribution to Ca(2+)-driven growth- and survival-promoting events in cancer is poorly understood. Here employing flow cytometry to monitor mitochondrial and cytosolic Ca(2+), we assessed trans-mitochondrial Ca(2+)-transport and store-operated Ca(2+)-influx (store-operated channels (SOC)) in malignant vs. non-malignant B16BL6 melanoma clones. Remarkably, mitochondrial Ca(2+)-fluxes measured in whole cells or in isolated mitochondria were accelerated in the malignant clones compared to their non-malignant counterpart clones. This coincided with enhanced SOC-mediated Ca(2+)-influx and high levels of constitutively active protein kinase B/Akt (PKB). Interruption of trans-mitochondrial Ca(2+)-transport in the malignant cells with an antagonist of the mitochondrial Na(+)/Ca(2+) exchanger, CGP-37157, abolsihed SOC-mediated Ca(2+)-influx, inactivated PKB, retarded cell growth and increased vulnerability to apoptosis. Similarly, direct SOC blockade by silencing Stim1 inhibited PKB, indicating that the crosstalk between SOC and mitochondria is essential to preserve PKB in constitutively active state. Finally, the retraction of mitochondria from sub-plasmalemmal micro-domains triggered by Fis1 over-expression inhibited SOC-coupled trans-mitochondrial Ca(2+)-flux, Ca(2+)-entry via SOC and PKB activity. Taken together, our data show that in the malignant melanoma cells, the functional and spatial relationship of up-regulated mitochondrial Ca(2+)-transport to the SOC sustains the robust Ca(2+)-responses and down-stream signaling critical for apoptosis-resistance and proliferation.


Tongue Coating and the Salivary Microbial Communities Vary in Children with Halitosis.
Halitosis is a common symptom mainly caused by microbial activities in the oral cavity. Here, we used 16S rRNA gene pyrosequencing and metagenomic sequencing to examine oral microbial compositions and their functional variations in children with halitosis. We found that the tongue coating of subjects with halitosis had greater bacterial richness than those of healthy subjects. The relative abundance and prevalence of Leptotrichia wadei and Peptostreptococcus stomatis were higher in tongue coating samples from children with halitosis than those from children without halitosis; Prevotella shahii had higher relative abundance and prevalence in saliva samples from children with halitosis. We present the first comprehensive evaluation of the co-occurrence networks of saliva and tongue coating communities under healthy and halitosis conditions, and investigated patterns of significant differences between these communities. Moreover, we observed that bacterial genes associated with responses to infectious diseases and terpenoid and polyketide metabolism were enriched in subjects with halitosis, but not in healthy subjects. Hydrogen sulphide (H2S)-related metabolic pathways suggested that there was higher microbial production and less usage of H2S in subjects with halitosis. Thus, the mechanism of halitosis was implied for the first time via metagenomic sequencing.


Cross-Adaptation: Heat and Cold Adaptation to Improve Physiological and Cellular Responses to Hypoxia.
To prepare for extremes of heat, cold or low partial pressures of oxygen (O2), humans can undertake a period of acclimation or acclimatization to induce environment-specific adaptations, e.g. heat acclimation (HA), cold acclimation (CA), or altitude training. While these strategies are effective, they are not always feasible due to logistical impracticalities. Cross-adaptation is a term used to describe the phenomenon whereby alternative environmental interventions, e.g. HA or CA, may be a beneficial alternative to altitude interventions, providing physiological stress and inducing adaptations observable at altitude. HA can attenuate physiological strain at rest and during moderate-intensity exercise at altitude via adaptations allied to improved O2 delivery to metabolically active tissue, likely following increases in plasma volume and reductions in body temperature. CA appears to improve physiological responses to altitude by attenuating the autonomic response to altitude. While no cross-acclimation-derived exercise performance/capacity data have been measured following CA, post-HA improvements in performance underpinned by aerobic metabolism, and therefore dependent on O2 delivery at altitude, are likely. At a cellular level, heat shock protein responses to altitude are attenuated by prior HA, suggesting that an attenuation of the cellular stress response and therefore a reduced disruption to homeostasis at altitude has occurred. This process is known as cross-tolerance. The effects of CA on markers of cross-tolerance is an area requiring further investigation. Because much of the evidence relating to cross-adaptation to altitude has examined the benefits at moderate to high altitudes, future research examining responses at lower altitudes should be conducted, given that these environments are more frequently visited by athletes and workers. Mechanistic work to identify the specific physiological and cellular pathways responsible for cross-adaptation between heat and altitude, and between cold and altitude, is warranted, as is exploration of benefits across different populations and physical activity profiles.


[Factors determining the instability of the conformation of human mutant hemoglobins. A method for evaluating the instability].
A method that allows to estimate the influence of single amino acid substitutions on the stability of proteins with known three-dimensional structure was developed. The role of 20 different physico-chemical properties of amino acids and some characteristics of the three-dimensional structure in the destabilization of protein structure caused by mutations was analyzed. Analysis of 54 proteins led us to reveal the characteristics of the three-dimensional structure. The parameters of the method were calculated on the basis of experimental data for 330 mutant human hemoglobins. The prediction accuracy of the method is on average 81%. The method may be used for the analysis of the total mutational spectra of the proteins and also for the estimation of effects of the single amino acid substitution on the protein structure in gene-engineering experiments.


[Current treatment of venous thrombembolism].
Deep vein thrombosis and pulmonary embolism, considered to be different manifestations of the same disease - venous thromboembolism, have few differences regarding the anticoagulant treatment. However, there are some issues which will be discussed. The therapy objectives in patients with venous thromboembolism include: prevention of death by pulmonary embolism, relieving symptoms in the affected leg, preventing morbidity and prevention of recurrent thromboembolism or postthrombotic syndrome, or minimize symptoms of post-thrombotic syndrome. For most patients, treatment goals are achieved using appropriate anticoagulant therapy, reducing the risk of recurrence in the first three months after diagnosis from over 25% to under 4%. Using of compression socks, providing a gradient of 30-40 mmHg at the ankle for 2 years after the diagnosis, reduce the risk of postthrombotic syndrome. Thrombolysis, applied either systemic or directly by catheter, is indicated in selected cases to prevent onset of postthrombotic syndrome or remove quickly the symptoms due to high venous obstruction. Thrombolytic therapy should be continued with anticoagulant therapy to prevent recurrence of venous thromboembolism. The use of an inferior vena cava filter is indicated for prevention of death by pulmonary embolism in patients who have contraindications to anticoagulant therapy, or anticoagulant treatment that was properly administered remains inefficient. Surgical treatment is recommended in case of chronic pulmonary hypertension, due to thromboembolic disease.


Age of the host and other factors affecting the production with urethane of pulmonary adenomas in mice.
Young, rapidly growing mice are greatly more responsive to the adenoma-inducing influence of urethane than are those just arriving at maturity. This is manifest both in the proportion of animals developing the tumors and in their number per individual. An amount of urethane per gram body weight which suffices to induce adenomas in only an occasional 8-week-old animal will cause them to appear in quantity in more than half the 3-week-old mice injected. There is an almost absolute inverse correlation between the rate of growth of the pulmonary tissue between the ages of 2 and 10 weeks and the response to urethane in terms of adenomas. Hence the conclusion seems justified that the natural proliferative activity of the alveolar cells during youth plays a major part in the formation of the tumors. After the 6th week the age differences become relatively slight, yet there is reason to think that they continue in some degree as life goes on. Urethane has no effect to promote multiplication of the cells it has rendered neoplastic, its whole role being to initiate neoplastic change. The abnormalities induced by urethane in the nucleus of normal and neoplastic cells, as observed by previous workers, have suggested that the substance brings about the adenomatous state by acting upon the nucleus. But colchicine, also a karyolytic poison causing pronounced nuclear changes, does not alter in the least the yield of adenomas to urethane when administered concurrently. Nor does fasting influence the yield, though it markedly reduces mitotic activity. The meaning of these facts is discussed.


Indexing electron backscatter diffraction patterns with a refined template matching approach.
Electron backscatter diffraction (EBSD) is a well-established method of characterisation for crystalline materials. Using this technique, we can rapidly acquire and index diffraction patterns to provide phase and orientation information about the crystals on the material surface. The conventional analysis method uses signal processing based on a Hough/Radon transform to index each diffraction pattern. This method is limited to the analysis of simple geometric features and ignores subtle characteristics of diffraction patterns, such as variations in relative band intensities. A second method, developed to address the shortcomings of the Hough/Radon transform, is based on template matching of a test experimental pattern with a large library of potential patterns. In the present work, the template matching approach has been refined with a new cross correlation function that allows for a smaller library and enables a dramatic speed up in pattern indexing. Refinement of the indexed orientation is performed with a follow-up step to allow for small alterations to the best match from the library search. The refined template matching approach is shown to be comparable in accuracy, precision and sensitivity to the Hough based method, even exceeding it in some cases, via the use of simulations and experimental data collected from a silicon single crystal and a deformed α-iron sample. The speed up and pattern refinement approaches should increase the widespread utility of pattern matching approaches.


Effects of respiratory muscle training (RMT) in children with infantile-onset Pompe disease and respiratory muscle weakness.
Respiratory muscle weakness is a primary therapeutic challenge for patients with infantile Pompe disease. We previously described the clinical implementation of a respiratory muscle training (RMT) regimen in two adults with late-onset Pompe disease; both demonstrated marked increases in inspiratory and expiratory muscle strength in response to RMT. However, the use of RMT in pediatric survivors of infantile Pompe disease has not been previously reported. We report the effects of an intensive RMT program on maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP) using A-B-A (baseline-treatment-posttest) single subject experimental design in two pediatric survivors of infantile Pompe disease. Both subjects had persistent respiratory muscle weakness despite long-term treatment with alglucosidase alfa. Subject 1 demonstrated negligible to modest increases in MIP/MEP (6% increase in MIP, d=0.25; 19% increase in MEP, d=0.87), while Subject 2 demonstrated very large increases in MIP/MEP (45% increase in MIP, d=2.38; 81% increase in MEP, d=4.31). Following three-month RMT withdrawal, both subjects maintained these strength increases and demonstrated maximal MIP and MEP values at follow-up. Intensive RMT may be a beneficial treatment for respiratory muscle weakness in pediatric survivors of infantile Pompe disease.


Equilibration time improves the sperm variables of wild ruminant ejaculated and epididymal sperm cryopreserved by ultra-rapid freezing.
This work examines the effect of equilibration time with extender on ultra-rapidly frozen-thawed wild ruminant epididymal (origin: Iberian ibex) and ejaculated (origin: mouflon) sperm variables. Sperm samples were prepared either without prior equilibration, or equilibrated for 30 min before freezing. Higher quality (p < 0.05) frozen-thawed spermatozoa were obtained when equilibration was allowed, for ejaculated sperm in terms of sperm motility, acrosome apical ridge integrity, sperm viability, and percentage of normal cells, and for epididymal sperm in terms of linearity and straightness of sperm movement. The sperm head area, head perimeter, head length and head width were smaller (p < 0.01) in the equilibrated than non-equilibrated frozen-thawed epididymal sperm; no such dimensional changes were recorded for ejaculated sperm. In conclusion, equilibration prior to ultra-rapid freezing improves the cryoresistance of sperm cells, although viable sperm cells can be obtained without equilibration. The epididymal sperm showed greater cryoresistance, supporting the idea that it is more resistant to freeze-thawing than ejaculated sperm.


"Willed action": a functional MRI study of the human prefrontal cortex during a sensorimotor task.
Functional MRI (fMRI) was used to examine human brain activity within the dorsolateral prefrontal cortex during a sensorimotor task that had been proposed to require selection between several responses, a cognitive concept termed "willed action" in a positron emission tomography (PET) study by Frith et al. [Frith, C. D., Friston, K., Liddle, P. F. & Frackowiak, R. S. J. (1991) Proc. R. Soc. London Ser. B 244, 241-246]. We repeated their sensorimotor task, in which the subject chooses to move either of two fingers after a stimulus, by fMRI experiments in a 2.1-T imaging spectrometer. Echo-planar images were acquired from four coronal slices in the prefrontal cortex from nine healthy subjects. Slices were 5 mm thick, centers separated by 7 mm, with nominal in-plane spatial resolution of 9.6 x 5.0 mm2 for mean data. Our mean results are in agreement with the PET results in that we saw similar bilateral activations. The present results are compared with our previously published fMRI study of a verbal fluency task, which had also been proposed by Frith et al. to elicit a "willed action" response. We find a clear separation of activation foci in the left dorsolateral prefrontal cortex for the sensorimotor (Brodmann area 46) and verbal fluency (Brodmann area 45) tasks. Hence, assigning a particular activated region to "willed action" is not supported by the fMRI data when examined closely because identical regions are not activated with different modalities. Similar modality linked activations can be observed in the original PET study but the greater resolution of the fMRI data makes the modality linkages more definite.


Revisiting salt and water retention: new diuretics, aquaretics, and natriuretics.
Diuretics continue to be a mainstay in patients with CHF. Conventional diuretic therapy is associated, however, with potentially deleterious neurohumoral activation and renal impairment. It is not known to what extent these neurohumoral effects are offset by concurrent therapy with ACE-I, beta-blockers, and other agents. In the past, there was no alternative to conventional diuretic therapy, so their potential for adverse outcome in the long term could not be assessed. Enhancement of the natriuretic peptide system could provide us with a better strategy to treat sodium and water retention. In a unique way, the natriuretic peptides combine several of the beneficial actions of the other diuretics, but without the associated cost. Natriuretic peptides, like conventional diuretics, are natriuretic and diuretic. There are important differences, however. First, unlike conventional diuretics, NPs do not activate RAAS. Activation of this system is associated with progression of CHF. Second, NPs inhibit the sympathetic nervous system, the activation of which is associated with heart failure progression, myocyte necrosis and apoptosis, and arrhythmias. Third, unlike conventional diuretics that lead to a decrease in GFR by reflex mechanisms. NPs maintain or even improve GFR. We now appreciate that some "old" drugs may be beneficial to CHF patients in a new way, as is the case with spironolactone. The survival benefit of this aldosterone antagonist is clear: its usefulness, however, may be more a result of both its antifibrotic actions in addition to its tradional role as a potassium-sparing and natriuretic agent. It is hoped that the SARAs will provide the same survival benefit, but with fewer of the sex-steroid side effects. In addition, AVP-receptor antagonists may become useful tools in the treatment of patients with hyponatremia. Likewise, the A1 AR antagonists may find a role in the CHF armamentarium by providing good diuresis and natriuresis while at the same time maintaining GFR through inhibition of TGF. Many questions remain unanswered, and studies are needed to demonstrate that the positive results seen in basic research translate into improved morbidity and mortality.


Self-interaction correction schemes for non-collinear spin-density-functional theory.
We extend some of the well-established self-interaction correction (SIC) schemes of density-functional theory-the Perdew-Zunger SIC and the average-density SIC-to the case of systems with noncollinear magnetism. Our proposed SIC schemes are tested on a set of molecules and metallic clusters in combination with the widely used local spin-density approximation. As expected from the collinear SIC, we show that the averaged-density SIC works well for improving ionization energies but fails to improve more subtle quantities like the dipole moments of polar molecules. We investigate the exchange-correlation magnetic field produced by our extension of the Perdew-Zunger SIC, showing that it is not aligned with the local total magnetization, thus producing an exchange-correlation torque.


Piecewise linearity of approximate density functionals revisited: implications for frontier orbital energies.
In the exact Kohn-Sham density-functional theory, the total energy versus the number of electrons is a series of linear segments between integer points. However, commonly used approximate density functionals produce total energies that do not exhibit this piecewise-linear behavior. As a result, the ionization potential theorem, equating the highest occupied eigenvalue with the ionization potential, is grossly disobeyed. Here, we show that, contrary to conventional wisdom, most of the required piecewise linearity of an arbitrary approximate density functional can be restored by careful consideration of the ensemble generalization of density-functional theory. Furthermore, the resulting formulation introduces the desired derivative discontinuity to any approximate exchange-correlation functional, even one that is explicitly density dependent. This opens the door to calculations of the ionization potential and electron affinity, even without explicit electron removal or addition. All these advances are achieved while neither introducing empiricism nor changing the underlying functional form. The power of the approach is demonstrated on benchmark systems using the local density approximation as an illustrative example.


The Strong, Silent Type(ology): Examining Intersections of Alexithymia and the Drive for Muscularity.
This study used latent profile analysis to explore the interrelations of drive for muscularity (DM) and alexithymia among 505 men. Four unique classes of men emerged: (1) low endorsement of DM and alexithymia, (2) moderate endorsement of alexithymia and DM, (3) moderate endorsement of alexithymia and high endorsement of attitudinal and behavioral DM, and (4) high endorsement of DM attitudes and alexithymia, and low endorsement of DM behaviors. We explored relations between class membership and avoidant attachment, social pressure to be muscular, help-seeking attitudes, and body appreciation, using Class 1 as the reference group. Alexithymia and attitudinal DM, in the absence of behavioral DM, appear to be a particularly unhealthful constellation of alexithymia and DM.


Acute lymphoblastic leukemia in adults: recent progress, future directions.
The data reviewed indicate substantial recent progress in the treatment of ALL in adults. Most patients, greater than 70% to 80%, will achieve a remission following induction chemotherapy with vincristine, prednisone, and an anthracycline. Additional drugs do not convincingly increase the remission rate; their effect on remission duration is unknown. Although continuation therapy has not been critically evaluated in randomized trials, the longest remissions have been reported in individuals receiving consolidation/intensification, particularly in the immediate postremission period. Presently, a disease-free survival is 35% to 45% at greater than 5 years when this type of consolidation/intensification therapy is used. The precise value of conventional maintenance chemotherapy in adult ALL is unknown. CNS prophylaxis is necessary; the standard of treatment remains cranial radiation and intrathecal methotrexate. Alternative approaches are currently being studied such as systemic treatment with high doses of methotrexate or cytarabine. Recently, prognostic factors have been better defined. In recent studies with intensive treatment, time to response, age, WBC, immunologic subtype and cytogenetic data can be used to divide patients into a low-risk group with a projected disease-free survival of 60% or more and a high-risk group with a disease-free survival of 20% to 25%. The definition of risk groups in adult ALL may be useful in determining future further optimal therapeutic approaches.


[Results of surgical in undifferentiated small cell cancer of the lung].
The results of surgical treatment of 36 patients suffering from undifferentiated small-cell carcinoma of lung are discussed. During the first two years, 28 patients (78%) died as a result of tumor progression, while 8 patients (22%) survived for 3--20 years after operation. Retrospective study established that the majority of the patients had a small -- size peripheral tumor without metastases in regional lymph nodes. Histological examination revealed a considerable development of stroma having a structure of mature fibrous tissue, few mitoses and no necroses. The findings show that in some cases of early stages of undifferentiated small-cell carcinoma of the lung, surgery may assure a long survival of tumor patients.


Arabidopsis DMR6 encodes a putative 2OG-Fe(II) oxygenase that is defense-associated but required for susceptibility to downy mildew.
The Arabidopsis mutant downy mildew resistant 6 (dmr6) carries a recessive mutation that results in the loss of susceptibility to Hyaloperonospora parasitica. Here we describe the map-based cloning of DMR6 (At5g24530), which was found to encode a 2-oxoglutarate (2OG)-Fe(II) oxygenase of unknown function. DMR6 transcription is locally induced during infections with both compatible and incompatible H. parasitica isolates. High DMR6 transcript levels were also observed in constitutive defense mutants and after treatment with salicylic acid analog BTH, suggesting that DMR6 has a role during plant defense. Expression analysis of dmr6 mutants, using DNA microarrays and quantitative PCR, showed the enhanced expression of a subset of defense-associated genes, including DMR6 itself, suggesting dmr6-mediated resistance results from the activation of plant defense responses. Alternatively, resistance could be caused by the accumulation of a toxic DMR6 substrate, or by the absence of a DMR6 metabolic product that is required for H. parasitica infection.


Self-association of an insect β-1,3-glucan recognition protein upon binding laminarin stimulates prophenoloxidase activation as an innate immune response.
Insect β-glucan recognition protein (βGRP), a pathogen recognition receptor for innate immune responses, detects β-1,3-glucan on fungal surfaces via its N-terminal carbohydrate-binding domain (N-βGRP) and triggers serine protease cascades for the activation of prophenoloxidase (pro-PO) or Toll pathways. Using biophysical and biochemical methods, we characterized the interaction of the N-terminal domain from Manduca sexta βGRP2 (N-βGRP2) with laminarin, a soluble form of β-1,3-glucan. We found that carbohydrate binding by N-βGRP2 induces the formation of two types of protein-carbohydrate complexes, depending on the molar ratio of carbohydrate to protein ([C]/[P]). Precipitation, analytical ultracentrifugation, and chemical cross-linking experiments have shown that an insoluble aggregate forms when the molar ratio of carbohydrate to protein is low ([C]/[P] ∼ 1). In contrast, a soluble complex, containing at least five N-βGRP2 molecules forms at a higher molar ratio of carbohydrate/protein ([C]/[P] >5). A hypothesis that this complex is assembled partly due to protein-protein interactions was supported by chemical cross-linking experiments combined with LC-MS/MS spectrometry analysis, which permitted identification of a specific intermolecular cross-link site between N-βGRP molecules in the soluble complex. The pro-PO activation in naive plasma was strongly stimulated by addition of the insoluble aggregates of N-βGRP2. The soluble complex with laminarin formed in the plasma also stimulated pro-PO activation, but at a lower level. Taken together, these results provide experimental evidence for novel mechanisms in which associations of βGRP with microbial polysaccharide promotes assembly of βGRP oligomers, which may form a platform needed to trigger the pro-PO pathway activation cascade.


Chronic lateral ankle instability surgical repairs: the long term prospective.
The present study sought to assess the clinical and radiological results and long-term joint impact of different techniques of lateral ankle ligament reconstruction. A multicenter retrospective review was performed on 310 lateral ankle ligament reconstructions, with a mean 13-year-follow-up (minimum FU: 5 years). Male subjects (53%) and sports trauma (78%) predominated. Mean duration of instability was 92 months; mean age at surgery was 28 years. Twenty-eight percent of cases showed subtalar joint involvement. Four classes of surgical technique were distinguished: C1, direct capsular ligamentous complex reattachment; C2, augmented repair; C3, ligamentoplasty using part of the peroneus brevis tendon and C4, ligamentoplasty using the whole peroneus brevis tendon. Clinical and functional assessment used Karlsson and Good-Jones-Livingstone scores; radiologic assessment combined centered AP and lateral views, hindfoot weight-bearing Méary views and dynamic views (manual technique, Telos or self-imposed varus). The majority of results (92%) were satisfactory. The mean Karlsson score of 90 [19-100] (i.e., 87% good and very good results) correlated with the subjective assessment, and did not evolve over time. Postoperative complications (20%), particularly when neurologic, were associated with poorer results. Control X-ray confirmed the very minor progression in degenerative changes, with improved stability; there was, however, no correlation between functional result and residual laxity on X-ray. Unstable and painful ankles showed poorer clinical results and more secondary osteoarthritis. Analysis by class of technique found poorer results in C4-type plasties and poorer control of laxity on X-ray in C1-type tension restoration. The present results confirm the interest of lateral ankle ligamentoplasty in the management of instability and protection against secondary osteoarthritis, and of precise lesion assessment (CT-scan/MRI) to adapt surgery to the ligamentary and associated lesions. Level IV. Retrospective therapeutic study.


Influence of a 60 Hz, 3 microT, electromagnetic field on the somatic maturation of wistar rat offspring fed a regional basic diet during pregnancy.
The aim of the present study was to observe how the exposition of pregnant rats to an electromagnetic field (EMF), with frequency of 60 Hz, and a magnetic field of 3 microT for 2 hours per day and/or using the so-called Regional Basic Diet (RBD), influenced the somatic maturation in their offspring. Four groups were formed: Group A (casein), B (casein and EMF), C (RBD) and D (RBD and EMF). The diet manipulation occurred during pregnancy. The somatic maturation indexes--assessed daily between 12:00 AM and 2:00 PM--were: Eye Opening (EO), Auricle Opening (AO), Auditory Canal Opening (ACO), Low Incisor Eruption (LIE), and Upper Incisor Eruption (UIE). The association between EMF and deficient diet caused a delay in all Somatic Maturation Indexes (SMI) and the RBD caused delay only in the AO. Furthermore, the EMF caused delay in AO, ACO, LIE. In relation to the body weight, the EMF associated with the deficient diet caused change in the twenty-first day of life. The RBD, during pregnancy, caused lower body weight in the offspring in the first and third day of life. The body weight of the offspring whose mothers were fed casein and exposed to the EMF during pregnancy was lower in the third and sixth day of life. In conclusion, the EMF associated with under-nutrition caused delay in all SMI. In relation to the body weight, the EMF associated with under-nutrition caused a decrease in the body weight at the sixth day of life.


An active role for basement membrane assembly and modification in tissue sculpting.
Basement membranes are a dense, sheet-like form of extracellular matrix (ECM) that underlie epithelia and endothelia, and surround muscle, fat and Schwann cells. Basement membranes separate tissues and protect them from mechanical stress. Although traditionally thought of as a static support structure, a growing body of evidence suggests that dynamic basement membrane deposition and modification instructs coordinated cellular behaviors and acts mechanically to sculpt tissues. In this Commentary, we highlight recent studies that support the idea that far from being a passive matrix, basement membranes play formative roles in shaping tissues.


Types of multidimensional vulnerability and well-being among the retired in the U.S.
An extensive study investigated the risk factors for low well-being in post-retirement. Most previous studies have taken a unidimensional perspective, focusing on single factors such as financial status, physical health, and mental health. Drawing on the vulnerability framework, we first identify and describe the empirical subgroups of vulnerability among retirees in the United States across four major domains of later life: material, physical, social, and mental vulnerability. Then, we investigate the association between vulnerability profiles and well-being. The sample included 3,158 retirees aged 65+ who participated in the Health and Retirement Study (HRS). Latent class analysis was utilized to identify the heterogeneous subgroups of vulnerability, and then a series of OLS regression analyses was conducted to examine the relationship between patterns of vulnerability and well-being. Five vulnerability patterns were identified: material vulnerable (12%), health & social vulnerable (14%), material, health & social vulnerable (6%), least vulnerable (34%), and social vulnerable (35%). The health & social vulnerable group had the strongest negative influence on well-being among all subgroups. As the largest subgroup, the social vulnerable group's negative influence on well-being stood out, with a stronger effect than that of material privation experienced by those in the material vulnerable group. By empirically identifying subgroups of differential vulnerability patterns among retirees, this study showed that post-retirement vulnerability reflects complex interactions among multiple disadvantages. Findings of this study enhance understanding of the disparities in well-being within the retired population, pointing to the possibility of targeted policy and program development.


Structural MRI Analysis of Chronic Pain Patients Following Interdisciplinary Treatment Shows Changes in Brain Volume and Opiate-Dependent Reorganization of the Amygdala and Hippocampus.
The present study examined pre- to post-treatment changes in volumes for brain structures known to be associated with pain processing (thalamus, caudate, putamen, pallidum, hippocampus, amygdala, and accumbens) following an interdisciplinary pain management program. Twenty-one patients participating in a four-week interdisciplinary pain management program completed the study. The program consisted of individual and group therapies with the following disciplines: physical therapy, occupational therapy, pain psychology, biofeedback/relaxation training, nursing lectures, and medical management. All patients underwent functional magnetic resonance imaging of the brain before the start and at completion of the program. They also completed standard outcome measures assessing pain, symptoms of central sensitization, disability, mood, coping, pain acceptance, and impressions of change. Our results showed a significant increase in total brain volume, as well as increased volumes in the thalamus, hippocampus, and amygdala. As expected, we also found significant improvements in our standard outcome measures. The majority of patients rated themselves as much or very much improved. The increase in volume in the hippocampus was significantly associated with patient perceptions of change. However, the correlations were in the unexpected direction, such that greater increases in hippocampal volume were associated with perceptions of less improvement. Further exploratory analyses comparing patients by their opioid use status (use vs no use) showed differential program effects on volume increases in the hippocampus and amygdala. These findings show that a four-week interdisciplinary pain management program resulted in changes in the brain, which adds objective findings further demonstrating program efficacy.


Influence of prior transient ischaemic attack on stroke prognosis.
To evaluate potential neuroprotection afforded by prior transient ischaemic attack (TIA) on functional and survival outcomes after ischaemic stroke. All cases of first-ever ischaemic strokes, diagnosed between 1985 and 2008, were identified from the Dijon Stroke Registry. Patients were analysed in three groups according to the time interval between prior TIA and stroke (<4 weeks, ≥ 4 weeks, no TIA) or the duration of TIA (≤ 30 min, >30 min, no TIA). Outcomes were severe functional handicap (unable to walk, bedridden or death) at hospital discharge or at outpatient consultation, and 1-month and 1-year any-cause mortality. Stratified analyses were performed by stroke subtypes (non-lacunar, lacunar). Generalised linear mixed models and Cox proportional hazard models with a sandwich covariance matrix accounting for the treatment centre as a random effect were used for multivariate analyses. Among the 3015 patients with first-ever ischaemic stroke, 389 had had a prestroke TIA <4 weeks and 97 a prestroke TIA ≥ 4 weeks. Patients with TIAs had better ambulatory status (adjusted OR 0.61, 95% CI 0.45 to 0.81; p = 0.008) and better survival at 1 month (adjusted HR 0.76, 95% CI 0.65 to 0.89; p = 0.0006) and at 1 year (adjusted HR 0.72, 95% CI 0.67 to 0.76; p<0.0001) than those with no TIAs. Prestroke TIA <4 weeks and TIA duration ≤ 30 min also significantly improved the outcomes in overall, non-lacunar and lacunar strokes. Recent prestroke TIA was associated with better functional outcome and lower 1-month and 1-year mortality after stroke, suggesting a neuroprotective effect.


Assessment of a fully 3D Monte Carlo reconstruction method for preclinical PET with iodine-124.
Iodine-124 is a radionuclide well suited to the labeling of intact monoclonal antibodies. Yet, accurate quantification in preclinical imaging with I-124 is challenging due to the large positron range and a complex decay scheme including high-energy gammas. The aim of this work was to assess the quantitative performance of a fully 3D Monte Carlo (MC) reconstruction for preclinical I-124 PET. The high-resolution small animal PET Inveon (Siemens) was simulated using GATE 6.1. Three system matrices (SM) of different complexity were calculated in addition to a Siddon-based ray tracing approach for comparison purpose. Each system matrix accounted for a more or less complete description of the physics processes both in the scanned object and in the PET scanner. One homogeneous water phantom and three heterogeneous phantoms including water, lungs and bones were simulated, where hot and cold regions were used to assess activity recovery as well as the trade-off between contrast recovery and noise in different regions. The benefit of accounting for scatter, attenuation, positron range and spurious coincidences occurring in the object when calculating the system matrix used to reconstruct I-124 PET images was highlighted. We found that the use of an MC SM including a thorough modelling of the detector response and physical effects in a uniform water-equivalent phantom was efficient to get reasonable quantitative accuracy in homogeneous and heterogeneous phantoms. Modelling the phantom heterogeneities in the SM did not necessarily yield the most accurate estimate of the activity distribution, due to the high variance affecting many SM elements in the most sophisticated SM.


Sesquiterpene lactones isolated from indigenous Middle Eastern plants inhibit tumor promoter-induced transformation of JB6 cells.
Sesquiterpene lactones (SL) are plant secondary metabolites that are known for their anti-fungal, anti-bacterial, anti-inflammatory, and anti-tumor properties. Considering that several SL-derived drugs are currently in cancer clinical trials, we have tested two SL molecules, 3-β-methoxy-iso-seco-tanapartholide (β-tan) isolated from Achillea falcata and salograviolide A (Sal A) isolated from Centaurea ainetensis, for their anti-tumor properties. We used the mouse epidermal JB6P + cells as a model for tumor promotion and cellular transformation. Key players that are involved in cellular transformation and tumorigenesis are the AP-1 and NF-κB transcription factors; therefore, we assessed how β-tan and Sal A modulate their signaling pathways in JB6P + cells. The effects of β-tan and Sal A on the growth of normal and neoplastic keratinocytes and on the tumor promotion-responsive JB6P + cells were determined using the MTT assay. Anchorage-independent cell growth transformation assays were used to evaluate the anti-tumor promoting properties of these SL molecules in JB6P + cells and dual luciferase reporter assays and western blot analysis were used to investigate their effects on tumor promoter-induced AP-1 and NF-κB activities and protein levels of key AP-1 and NF-кB target genes. β-tan and Sal A selectively inhibited tumor promoter-induced cell growth and transformation of JB6P + cells at concentrations that do not affect JB6P + and primary keratinocytes basal cell growth. In addition, both molecules reduced basal and tumor promoter-induced NF-κB transcriptional activities, differentially regulated basal and tumor promoter-induced AP-1 transcriptional activities, and modulated key players of the AP-1 and NF-κB signaling pathways. These results highlight the anti-tumor promoting properties of β-tan and Sal A. These SL molecules isolated from two plant species native to the Middle East may provide opportunities for complementary medicine practices.


TMEM14C is required for erythroid mitochondrial heme metabolism.
The transport and intracellular trafficking of heme biosynthesis intermediates are crucial for hemoglobin production, which is a critical process in developing red cells. Here, we profiled gene expression in terminally differentiating murine fetal liver-derived erythroid cells to identify regulators of heme metabolism. We determined that TMEM14C, an inner mitochondrial membrane protein that is enriched in vertebrate hematopoietic tissues, is essential for erythropoiesis and heme synthesis in vivo and in cultured erythroid cells. In mice, TMEM14C deficiency resulted in porphyrin accumulation in the fetal liver, erythroid maturation arrest, and embryonic lethality due to profound anemia. Protoporphyrin IX synthesis in TMEM14C-deficient erythroid cells was blocked, leading to an accumulation of porphyrin precursors. The heme synthesis defect in TMEM14C-deficient cells was ameliorated with a protoporphyrin IX analog, indicating that TMEM14C primarily functions in the terminal steps of the heme synthesis pathway. Together, our data demonstrate that TMEM14C facilitates the import of protoporphyrinogen IX into the mitochondrial matrix for heme synthesis and subsequent hemoglobin production. Furthermore, the identification of TMEM14C as a protoporphyrinogen IX importer provides a genetic tool for further exploring erythropoiesis and congenital anemias.


[Asthma and rhinoconjunctivitis caused by castor bean dust].
A 57 years old trader of agricultural goods with chronic asthma had attended our medical department for diagnosis and therapy. Since more than 35 years the patient had always experienced work-related asthma episode and rhinoconjunctivities when he worked in workplace where castor-bean oil cake as fertilizer was sold. With the intensive contact with castor-bean containing goods beginning about eight years ago, his asthma attack had become chronic. The allergological diagnosis confirmed that he had high-degree type I-allergy to castor-bean allergens.


Finding the right thermal limit: a framework to reconcile ecological, physiological and methodological aspects of CTmax in ectotherms.
Upper thermal limits (CTmax) are frequently used to parameterize the fundamental niche of ectothermic animals and to infer biogeographical distribution limits under current and future climate scenarios. However, there is considerable debate associated with the methodological, ecological and physiological definitions of CTmax. The recent (re)introduction of the thermal death time (TDT) model has reconciled some of these issues and now offers a solid mathematical foundation to model CTmax by considering both intensity and duration of thermal stress. Nevertheless, the physiological origin and boundaries of this temperature-duration model remain unexplored. Supported by empirical data, we here outline a reconciling framework that integrates the TDT model, which operates at stressful temperatures, with the classic thermal performance curve (TPC) that typically describes biological functions at permissive temperatures. Further, we discuss how the TDT model is founded on a balance between disruptive and regenerative biological processes that ultimately defines a critical boundary temperature (Tc) separating the TDT and TPC models. Collectively, this framework allows inclusion of both repair and accumulation of heat stress, and therefore also offers a consistent conceptual approach to understand the impact of high temperature under fluctuating thermal conditions. Further, this reconciling framework allows improved experimental designs to understand the physiological underpinnings and ecological consequences of ectotherm heat tolerance.


Th17 cells, HIV and the gut mucosal barrier.
We will present recent studies on a subset of CD4 T helper cells, Th17 cells, that appears to be critical for regulating gut mucosal immune responses against extracellular microbial pathogens and may serve as a link between innate and adaptive immune responses. Implications of the loss of Th17 CD4 T cells in HIV infection will be discussed in relation to the chronic immune activation and HIV pathogenesis. Severe depletion of CD4 T cells occurs in the gut mucosa during primary HIV and simian immunodeficiency virus infections. A pronounced loss of mucosal Th17 CD4 T cells in the simian immunodeficiency virus-infected rhesus macaque model of AIDS is linked to impaired immune responses in the gut mucosa to an enteric pathogen, Salmonella typhimurium, leading to the lack of local control of the pathogen and its translocation. Recovery of the gut mucosal immune system during highly active antiretroviral therapy is slow and incomplete compared with the peripheral blood compartment. Recent studies suggest that the replenishment of Th17 CD4 T cells in the gut mucosa during highly active antiretroviral therapy, or during nonpathogenic simian immunodeficiency virus infections in the nonhuman primate models, correlates with better restoration and function of the gut mucosal immune system. A better understanding of the role of Th17 CD4 cells in the generation of mucosal immune responses to enteric pathogens and maintenance of the intestinal epithelial integrity in HIV-infected patients will help in the development of novel strategies to modulate and enhance mucosal immune system and its function.


The capability of trapping gaseous microemboli of two pediatric arterial filters with pulsatile and nonpulsatile flow in a simulated infant CPB model.
The study objective was to test the capability of Medtronic Affinity and Terumo Capiox pediatric arterial filters to trap gaseous microemboli in a simulated infant cardiopulmonary bypass (CPB) model. The filters were used in parallel pattern. The circuit was primed with lactated ringer's solution (700 ml) and postfilter pressure was maintained at 100 mm Hg using a Hoffman clamp. Trials were conducted at flow rates ranging from 500 to 1,250 ml/min. After introducing 20 ml air into the venous line via an 18-G needle, 2-minute segments of data were recorded. This entire process was repeated 6 times for each unique combination of arterial filter, flow rate and perfusion mode, yielding a total of 96 experiments. More than 80% of gaseous microemboli were trapped by the two pediatric arterial filters. With increased flow rates and pulsatile mode, more gaseous microemboli passed through the arterial filters. There were no differences in terms of the percentage of gaseous microemboli trapped and pressure drops between Medtronic Affinity and Terumo Capiox pediatric arterial filters. Results demonstrated that Medtronic Affinity and Terumo Capiox pediatric arterial filters could trap the majority of gaseous microemboli in this particular setting of an open arterial filter purge line in a simulated infant CPB circuit with pulsatile and nonpulsatile flow.


Relatedness between numerically small Dutch Red dairy cattle populations and possibilities for multibreed genomic prediction.
Red dairy breeds are a valuable cultural and historical asset, and often a source of unique genetic diversity. However, they have difficulties competing with other, more productive, dairy breeds. Improving competitiveness of Red dairy breeds, by accelerating their genetic improvement using genomic selection, may be a promising strategy to secure their long-term future. For many Red dairy breeds, establishing a sufficiently large breed-specific reference population for genomic prediction is often not possible, but may be overcome by adding individuals from another breed. Relatedness between breeds strongly decides the benefit of adding another breed to the reference population. To prioritize among available breeds, the effective number of chromosome segments (Me) can be used as an indicator of relatedness between individuals from different breeds. The Me is also an important parameter in determining the accuracy of genomic prediction. The Me can be estimated both within a population and between 2 populations or breeds, as the reciprocal of the variance of genomic relationships. We investigated relatedness between 6 Dutch Red cattle breeds, Groningen White Headed (GWH), Dutch Friesian (DF), Meuse-Rhine-Yssel (MRY), Dutch Belted (DB), Deep Red (DR), and Improved Red (IR), focusing primarily on the Me, to predict which of those breeds may benefit from including reference animals of the other breeds. All of these breeds, except MRY, are under high risk of extinction. Our results indicated high variability of Me, especially between Me ranging from ∼3,500 to ∼17,400, indicating different levels of relatedness between the breeds. Two clusters are especially important, one formed by MRY, DR, and IR, and the other comprising DF and DB. Although relatedness between breeds within each of these 2 clusters is high, across-breed genomic prediction is still limited by the current number of genotyped individuals, which for many breeds is low. However, adding MRY individuals would increase the reference population of DR substantially. We estimated that between 11 and 133 individuals from other breeds are needed to achieve accuracy of genomic prediction equivalent to using one additional individual from the same breed. Given the variation in size of the breeds in this study, the benefit of a multibreed reference population is expected to be lower for larger breeds than for the smaller ones.


Intestinal pseudo-obstruction.
Pseudo-obstruction syndromes are increasingly recognized in clinical practice. They result from impairment of intrinsic neuromuscular or extrinsic control of gut motility. Typically, pseudo-obstruction syndromes result in features suggestive of mechanical obstruction and bowel dilatation in the absence of any demonstrable obstruction or mucosal disease. The syndrome may affect any region of the gut. Less severe variants without bowel dilatation are diagnosed by measurement of gastrointestinal transit and pressure profiles. The aims of treatment are restoration of nutrition and hydration, symptom relief, normalization of intestinal propulsion with prokinetics, and suppression of bacterial overgrowth. Surgery plays a limited role, adjunctive to medical treatment, facilitating enteral nutrition and decompression by means of jejunostomy. Infrequently, resection of localized disease or intestinal transplantation are indicated. The roles of intestinal pacemakers (interstitial cells of Cajal) and genetic mutations in the etiology of pseudo-obstruction, as well as the cost-benefit ratio of transplantation for pseudo-obstruction, will be clarified in the future.