Role of warm-water bath in anorectal conditions. The "thermosphincteric reflex".
Why anorectal pain is relieved by sitting in a warm-water bath was investigated in 18 healthy volunteers and 28 patients with painful anorectal disease (18 patients with fissures and 10 with hemorrhoids). Investigations consisted of measuring rectal and interstitial sphincter temperature, rectal and rectal neck pressures, and electromyographic activity of both the external and internal anal sphincters before and after the subjects sat in a warm-water bath at temperatures of 40, 45, and 50 degrees C for 10 min each time. Pain relief was more evident and lasted longer at higher bath temperatures. The rectal and interstitial sphincter temperatures were unchanged before and after bath in both the healthy volunteers and patients. The rectal neck pressure and internal and sphincter electromyographic activity dropped significantly in the bath, but increased gradually to pretest levels 25-70 min after exiting the bath. The higher the bath temperature, the greater the drop in rectal neck pressure and internal sphincter EMG activity, and the longer the time needed to return to pretest levels. Pain relief after Sitz bath seems to be the result of internal anal-sphincter relaxation with a resulting diminution of the rectal neck pressure. The relaxation of the internal sphincter following the warm bath postulates a relationship, but direct action was ruled out. A neural pathway through a "thermosphincteric reflex" seems most likely.


Anaphylaxis during renal transplantation of live donor graft in a child with latex allergy: case report.
Latex allergy is becoming increasingly more frequent, affecting patients and health care professionals. The objective of this report was to present the case of a child with allergy to latex, who developed anaphylaxis during anesthesia for renal transplantation, and emphasize some of the multidisciplinary conducts used to decrease the risk of anaphylactic shock after graft reperfusion. A male child, 5 years and 10 months old, P3 by the ASA classification, with a history of allergy to latex diagnosed after contact with balloons and confirmed by Rast test specific for latex and Prick test, underwent renal transplantation of a live donor graft for end-stage renal disease secondary to urologic malformation. The protocols for patients with Latex Allergy adopted by the Anesthesiology and Nursing Departments of the Hospital das Clínicas da UNICAMP were observed to avoid exposure of the child to latex. They started the day before the surgery by cleaning the operating rooms and substituting of all medical-hospital products by latex-free material. The equipment and materials used during the procedure were latex-free according to a technical report provided by the manufacturers. The surgery was done under general anesthesia and controlled mechanical ventilation. At the end of the surgery, the patient required blood transfusion, which was administered by a pressurizer; he developed cutaneous rash and the blood transfusion was discontinued, hydrocortisone was administered, and the infusion of crystalloids was increased. The child had an immediate and satisfactory response to the treatment. Latex allergy has become a public health problem and the knowledge of specific therapeutic conducts allows immediate treatment and decreases patient risks.


Gender differences in facial imitation and verbally reported emotional contagion from spontaneous to emotionally regulated processing levels.
Previous studies on gender differences in facial imitation and verbally reported emotional contagion have investigated emotional responses to pictures of facial expressions at supraliminal exposure times. The aim of the present study was to investigate how gender differences are related to different exposure times, representing information processing levels from subliminal (spontaneous) to supraliminal (emotionally regulated). Further, the study aimed at exploring correlations between verbally reported emotional contagion and facial responses for men and women. Masked pictures of angry, happy and sad facial expressions were presented to 102 participants (51 men) at exposure times from subliminal (23 ms) to clearly supraliminal (2500 ms). Myoelectric activity (EMG) from the corrugator and the zygomaticus was measured and the participants reported their hedonic tone (verbally reported emotional contagion) after stimulus exposures. The results showed an effect of exposure time on gender differences in facial responses as well as in verbally reported emotional contagion. Women amplified imitative responses towards happy vs. angry faces and verbally reported emotional contagion with prolonged exposure times, whereas men did not. No gender differences were detected at the subliminal or borderliminal exposure times, but at the supraliminal exposure gender differences were found in imitation as well as in verbally reported emotional contagion. Women showed correspondence between their facial responses and their verbally reported emotional contagion to a greater extent than men. The results were interpreted in terms of gender differences in emotion regulation, rather than as differences in biologically prepared emotional reactivity.


The SH2-containing tyrosine phosphatase corkscrew is required during signaling by sevenless, Ras1 and Raf.
The sevenless gene encodes a receptor tyrosine kinase which is required for the development of the R7 photoreceptor cell in each ommatidium of the Drosophila eye. We have previously used a sensitized genetic screen to identify mutations, designated Enhancers of sevenless (E(sev)), which affect genes that encode components of the sevenless signaling pathway. Here, we report that one of these mutations, E(sev)1Ae0P is a dominantly inhibiting allele of corkscrew, which encodes an SH2 domain-containing protein tyrosine phosphatase (Perkins et al., 1992). We show that corkscrew function is essential for sevenless signaling and that expression of a membrane-targeted form of corkscrew can drive R7 photoreceptor development in the absence of sevenless function. Furthermore, we have used the dominantly inhibiting corkscrew allele to examine the role of corkscrew during signaling by activated forms of Ras1 and Raf. Our analysis indicates that corkscrew function is still required during signaling by activated forms Ras1 and Raf proteins. These results define a function for corkscrew that is either downstream of Ras1 activation or in a parallel pathway that acts with activated Ras1/Raf to specify R7 photoreceptor development.


Screen for ISG15-crossreactive deubiquitinases.
The family of ubiquitin-like molecules (UbLs) comprises several members, each of which has sequence, structural, or functional similarity to ubiquitin. ISG15 is a homolog of ubiquitin in vertebrates and is strongly upregulated following induction by type I interferon. ISG15 can be covalently attached to proteins, analogous to ubiquitination and with actual support of ubiquitin conjugating factors. Specific proteases are able to reverse modification with ubiquitin or UbLs by hydrolyzing the covalent bond between their C-termini and substrate proteins. The tail regions of ubiquitin and ISG15 are identical and we therefore hypothesized that promiscuous deubiquitinating proteases (DUBs) might exist, capable of recognizing both ubiquitin and ISG15. We have cloned and expressed 22 human DUBs, representing the major clades of the USP protease family. Utilizing suicide inhibitors based on ubiquitin and ISG15, we have identified USP2, USP5 (IsoT1), USP13 (IsoT3), and USP14 as ISG15-reactive proteases, in addition to the bona fide ISG15-specific protease USP18 (UBP43). USP14 is a proteasome-associated DUB, and its ISG15 isopeptidase activity increases when complexed with the proteasome. By evolutionary standards, ISG15 is a newcomer among the UbLs and it apparently not only utilizes the conjugating but also the deconjugating machinery of its more established relative ubiquitin. Functional overlap between these two posttranslational modifiers might therefore be more extensive than previously appreciated and explain the rather innocuous phenotype of ISG15 null mice.


Molecular recognition of inositol polyphosphates by intracellular receptors and metabolic enzymes.
The discovery that inositol lipids are fundamentally involved in cell signalling has been one of the most significant recent advances in cell biology. In particular, there is now evidence that certain products of polyphosphoinositide metabolism play second messenger roles in most cells. Inositol 1,4,5-trisphosphate and perhaps inositol 1,3,4,5-tetrakisphosphate bind to specific receptors and regulate Ca2+ release from, and movement between, intracellular stores. The synthesis of novel analogues of these second messengers is now providing clues to the structural requirements at such receptors as well as for molecules with stability towards metabolic enzymes. Stefan Nahorski and Barry Potter discuss these developments with a view to future pharmacological intervention at these sites.


Insomnia: does chiropractic help?
To evaluate the effect of chiropractic care on insomnia. Tripartite pilot study. The expectations of the chiropractic community were canvassed, a retrospective study to recall changes in sleep patterns was undertaken, and a prospective pilot study to monitor sleep patterns after chiropractic care was carried out. Convenience sampling was used. The 221 patients and 15 chiropractors who completed the expectation study tended to believe that patients with sleeping difficulties benefited from chiropractic care. The chiropractors were more guarded in their expectations than participating patients. One third of the 154 patients who completed the semistructured interview reported their sleep pattern was changed immediately after their chiropractic adjustment. All but 1 of these 52 patients reported improvement. Twenty patients with insomnia participated in the prospective study. Although compared with the report in their screening questionnaire, improvement was noted in certain sleep parameters in the 6 days after their adjustment, no temporal trends emerged in the days and/or weeks after the chiropractic consultation. Most patients reported experiencing less or no discomfort during the duration of the study. Although a number of patients do perceive chiropractic care offers temporary respite from their insomnia problem, when changes were more objectively monitored, improvements were erratic and no consistent temporal trends were detectable. Convincing evidence has yet to be produced before routine chiropractic care can be considered adequate intervention for patients with sleeping difficulties. More definitive answers may result from future research being undertaken in sleep laboratories.


An algorithm for random match probability calculation from peptide sequences.
For the past three decades, forensic genetic investigations have focused on elucidating DNA signatures. While DNA has a number of desirable properties (e.g., presence in most biological materials, an amenable chemistry for analysis and well-developed statistics), DNA also has limitations. DNA may be in low quantity in some tissues, such as hair, and in some tissues it may degrade more readily than its protein counterparts. Recent research efforts have shown the feasibility of performing protein-based human identification in cases in which recovery of DNA is challenged; however, the methods involved in assessing the rarity of a given protein profile have not been addressed adequately. In this paper an algorithm is proposed that describes the computation of a random match probability (RMP) resulting from a genetically variable peptide signature. The approach described herein explicitly models proteomic error and genetic linkage, makes no assumptions as to allelic drop-out, and maps the observed proteomic alleles to their expected protein products from DNA which, in turn, permits standard corrections for population structure and finite database sizes. To assess the feasibility of this approach, RMPs were estimated from peptide profiles of skin samples from 25 individuals of European ancestry. 126 common peptide alleles were used in this approach, yielding a mean RMP of approximately 10-2.


Monitoring spread of Malassezia infections in a neonatal intensive care unit by PCR-mediated genetic typing.
Malassezia furfur and Malassezia pachydermatis were isolated from newborn children and incubators in a neonatal intensive care unit. To assess whether persistence or frequent import of the organisms was the cause of the elevated incidence, genetic typing of the strains was performed by PCR-mediated DNA fingerprinting. By using PCR primers aimed at repeat consensus motifs, six different genotypes could be detected in a collection of six M. furfur reference strains. In the case of 10 M. pachydermatis reference strains, nine different genotypes were detected by three different PCR assays. None of these assays could document genetic differences among the clinical isolates of either M. furfur or M. pachydermatis. On the basis of these results it is concluded that within the neonatal intensive care unit the longitudinal persistence of both an M. furfur and an M. pachydermatis strain has occurred and that Malassezia species can persist on incubator surfaces for prolonged periods of time. It can be concluded that PCR fingerprinting is a Malassezia typing procedure that is to be preferred over the analysis of chromosomal polymorphisms by pulsed-field gel electrophoresis in this genus.


Association of Adipokines and Insulin, Which Have a Role in Obesity, with Colorectal Cancer.
Obesity-related diseases are an important part of public health; and obesity is related with colorectal cancer. Adipocyte hypertrophy and visceral adipose tissue accumulation can cause adipocitis-related diseases and pathogenic adipocyte formation. Adipose tissue has a very important and active role in immune response formation. Cytokines/adipokines, which are secreted from adipose tissue, have an active role in communication between adipocytes and macrophages. Thus, visceral adipocyte is related with low-grade chronic systemic inflammation. Adipocytes have an important role in colorectal cancer pathogenesis because of proinflammatory cytokines, growth factors, and hormones secretion. Most highlighted cytokines are adiponectin, resistin, and ghrelin. Also, insulin resistance, glucose intolerance, increased plasma insulin levels, body mass index, insulin-like growth factor (IGF-1), glucose, and serum free fatty acids levels are considered to be related with colorectal cancer pathogenesis. Thus, in this review, we focus on the role of adipokines and insulin in colorectal cancer.


Role of the pineal gland in diurnal endocrine secretion and rhythm regulation.
The effects of the pineal gland on diurnal endocrine function were studied in male rats. Pineal stimulation by exposure to short daily photoperiods (1L:23D) did not alter plasma levels of testosterone, thyroid-stimulating hormone (TSH) or growth hormone (GH) as compared with those of controls maintained under normal (12L:12D) lighting. Pinealectomy lowered diurnal testosterone levels and increased TSH levels in animals maintained under 1L:23D lighting. Pinealectomy also altered the TSH rhythm and shifted the peak in testosterone levels but did not abolish hormonal rhythms. Active immunization against melatonin and its precursor, N-acetylserotonin (NAS), significantly reduced diurnal androgen levels and elevated TSH levels in animals exposed to short photoperiods, but the 24-hour rhythms persisted. Plasma GH exhibited a diurnal rhythm in animals kept under short photoperiods but GH levels were not affected by any of the above treatments. These data suggest that melatonin and/or NAS may be involved in the maintenance of basal testosterone and TSH levels. The pineal may not be involved in generating hormonal rhythms; however, it may have a role in entraining TSH and testosterone rhythms to environmental lighting.


The antifungal eugenol perturbs dual aromatic and branched-chain amino acid permeases in the cytoplasmic membrane of yeast.
Eugenol is an aromatic component of clove oil that has therapeutic potential as an antifungal drug, although its mode of action and precise cellular target(s) remain ambiguous. To address this knowledge gap, a chemical-genetic profile analysis of eugenol was done using ∼4700 haploid Saccharomyces cerevisiae gene deletion mutants to reveal 21 deletion mutants with the greatest degree of susceptibility. Cellular roles of deleted genes in the most susceptible mutants indicate that the main targets for eugenol include pathways involved in biosynthesis and transport of aromatic and branched-chain amino acids. Follow-up analyses showed inhibitory effects of eugenol on amino acid permeases in the yeast cytoplasmic membrane. Furthermore, phenotypic suppression analysis revealed that eugenol interferes with two permeases, Tat1p and Gap1p, which are both involved in dual transport of aromatic and branched-chain amino acids through the yeast cytoplasmic membrane. Perturbation of cytoplasmic permeases represents a novel antifungal target and may explain previous observations that exposure to eugenol results in leakage of cell contents. Eugenol exposure may also contribute to amino acid starvation and thus holds promise as an anticancer therapeutic drug. Finally, this study provides further evidence of the usefulness of the yeast Gene Deletion Array approach in uncovering the mode of action of natural health products.


Studies of the role of the Drosophila scs and scs' insulators in defining boundaries of a chromosome puff.
Insulators are DNA elements that establish independent transcriptional domains within eukaryotic genomes. The Drosophila scs and scs' insulators localize near the borders of a structural domain in the polytene chromosomes, known as a puff, produced by transcription of the 87A heat shock protein (hsp) genes. It has been suggested that scs and scs' are boundary elements that delimit this decondensed chromatin domain, reflecting the mechanism by which these sequences act to constrain regulatory interactions. This model was tested using transposons that carried a yellow gene to assess enhancer blocking and an hsp70-lacZ gene to examine the structure of a heat shock puff in the presence and absence of insulators. We found that although scs and scs' blocked enhancer function, these sequences did not prevent the spread of decondensation resulting from hsp70-lacZ transcription. Further analysis of the endogenous 87A locus demonstrated that scs and scs' reside within, not at, the borders of the puff. Taken together, our studies suggest that scs and scs' are not boundary elements that block the propagation of an altered chromatin state associated with puff formation. We propose that these insulators may have a direct role in limiting regulatory interactions in the gene-dense 87A region.


The benefits, costs and feasibility of a low incidence COVID-19 strategy.
In the summer of 2021, European governments removed most NPIs after experiencing prolonged second and third waves of the COVID-19 pandemic. Most countries failed to achieve immunization rates high enough to avoid resurgence of the virus. Public health strategies for autumn and winter 2021 have ranged from countries aiming at low incidence by re-introducing NPIs to accepting high incidence levels. However, such high incidence strategies almost certainly lead to the very consequences that they seek to avoid: restrictions that harm people and economies. At high incidence, the important pandemic containment measure 'test-trace-isolate-support' becomes inefficient. At that point, the spread of SARS-CoV-2 and its numerous harmful consequences can likely only be controlled through restrictions. We argue that all European countries need to pursue a low incidence strategy in a coordinated manner. Such an endeavour can only be successful if it is built on open communication and trust.


Analysis of the Risk Factors for Aerobic Vaginitis: A Case-Control Study.
Aerobic vaginitis (AV) is a newly defined clinical entity which may interfere with women's reproductive health and have negative effects on pregnancy. This study was to identify the risk factors for AV. Participants in this case-control study included healthy women and women with AV. All participants completed a standardized questionnaire covering sociodemographic factors, sexual behaviors, personal hygiene habits and health behaviors. Uni- and multivariate logistic regression analyses were used for statistical evaluation. A total of 290 women of reproductive age were enrolled. In the multivariate analysis, unmarried status (odds ratio [OR] 2.606, 95% confidence interval [CI] 1.257-5.402), use of an intrauterine device (OR 4.989, 95% CI 1.922-12.952), long-term use of antibiotics (OR 11.176, 95% CI 1.363-91.666) and frequent vaginal douching (OR 4.689, 95% CI 1.363-16.135) were independent risk factors for AV. On the contrary, consistent condom use (OR 0.546, 95% CI 0.301-0.991) and college-level education or above (OR 0.255, 95% CI 0.131-0.497) were independent protective factors. Measures that may be considered to prevent AV include enhancing education to improve women's knowledge related to reproductive health, especially unmarried women, encouraging them to consistently use condoms as a contraceptive method, to avoid long-term use of antibiotics and to stop frequent vaginal douching. © 2015 S. Karger AG, Basel.


Social cognitive development during adolescence.
Social relationships are particularly important during adolescence. In recent years, histological and MRI studies have shown that the brain is subject to considerable structural development during adolescence. Brain regions that are implicated in social cognition, including parts of prefrontal, parietal and superior temporal cortex, undergo the most pronounced and prolonged change. However, the development of social cognition during adolescence and its neural underpinnings remains poorly understood. Here, we begin by outlining how the brain changes between childhood and adulthood. We then describe findings that have emerged from behavioural and neuroimaging studies of the recognition of facial expression during adolescence. Finally, we present new data that demonstrate development of emotional perspective taking during adolescence. In this study, 112 participants, aged 8-36 years, performed a computerised task that involved taking an emotional perspective either from the participant's own point of view or from that of another person. The results showed that average difference in reaction time (RT) to answer questions in the first person perspective (1PP) and third person perspective (3PP) significantly decreased with age. The RT difference of adults tended to cluster close to the zero line (3PP = 1PP), while a greater proportion of pre-adolescents had higher difference values in both the positive (3PP > 1PP) and negative direction (1PP > 3PP) of the scale. The data suggest that the efficiency, and possibly strategy, of perspective taking develop in parallel with brain maturation and psychosocial development during adolescence.


From clinical microbiology to infection pathogenesis: how daring to be different works for Staphylococcus lugdunensis.
Staphylococcus lugdunensis has gained recognition as an atypically virulent pathogen with a unique microbiological and clinical profile. S. lugdunensis is coagulase negative due to the lack of production of secreted coagulase, but a membrane-bound form of the enzyme present in some isolates can result in misidentification of the organism as Staphylococcus aureus in the clinical microbiology laboratory. S. lugdunensis is a skin commensal and an infrequent pathogen compared to S. aureus and S. epidermidis, but clinically, infections caused by this organism resemble those caused by S. aureus rather than those caused by other coagulase-negative staphylococci. S. lugdunensis can cause acute and highly destructive cases of native valve endocarditis that often require surgical treatment in addition to antimicrobial therapy. Other types of S. lugdunensis infections include abscess and wound infection, urinary tract infection, and infection of intravascular catheters and other implanted medical devices. S. lugdunensis is generally susceptible to antimicrobial agents and shares CLSI antimicrobial susceptibility breakpoints with S. aureus. Virulence factors contributing to this organism's heightened pathogenicity remain largely unknown. Those characterized to date suggest that the organism has the ability to bind to and interact with host cells and to form biofilms on host tissues or prosthetic surfaces.


Antigenicity of trophoblast and possible antigen-masking effects during pregnancy.
The interaction between cultured human trophoblast cells and materanl lymphocytes was used as an in vitro model to investigate trophoblast antigenicity. Cytotoxic effects in the trophoblast monolayer were apparent after 72 hr incubation and depended on the presence of non-lymphoid cell types in addition to lymphocytes. Lysis of trophoblast was preceded by blast cell formation and apparently involved close contact between the maternal cells and the trophoblast cells. The nature of the cytotoxic reaction suggested the presence of histocompatibility factors on trypsinized trophoblast cells. This manifestation could be due to removal of fibrinoid or enhancing antibody from the cultured cells. The ability of trypsinized trophoblast cells to synthesize mucoprotein was investigated by the Hale colloidal iron test and found to be unimpaired. The effect of maternal serum on the recognition of human trophoblast antigens by maternal lymphocytes was used as an in vitro model to investigate the occurrence of enhancing antibody in maternal serum. The cytotoxic effects of maternal lymphocytes on trophoblast were completely prevented by the presence of maternal serum, this protective effect being reduced significantly by removal of IgG from the maternal serum. A slight protective effect of allogeneic pregnancy serum was also observed. It is suggested that these findings support a role for immunological enhancement in maintaining the foetal allograft.


Androgens induce sebaceous differentiation in sebocyte cells expressing a stable functional androgen receptor.
Androgens act through non-genomic and androgen receptor (AR)-dependent genomic mechanisms. AR is expressed in the sebaceous gland and the importance of androgens in the sebaceous function is well established. However, the in vitro models used to date have failed to evidence a clear genomic effect (e.g., modification of gene expression profile) of androgens on human sebocyte cells. In order to study the impact of active androgens in sebocytes, we constructed a stable human sebocyte cell line derived from SEBO662 [17] constitutively expressing a fully functional AR. In these SEBO662 AR+ cells, dihydrotestosterone (DHT) induced AR nuclear translocation and the strong modulation of a set of transcripts (RASD1, GREB1...) known to be androgen-sensitive in other androgenic cells and tissues. Moreover, we observed that DHT precociously down-regulated markers for immature follicular cells (KRT15, TNC) and for hair lineage (KRT75, FST) and up-regulated the expression of genes potentially related to sebocyte differentiation (MUC1/EMA, AQP3, FADS2). These effects were fully confirmed at the protein level. In addition, DHT-stimulated SEBO662 AR+, cultured in a low-calcium defined keratinocyte medium without serum or any complement, neosynthesize lipids, including sebum lipids, and store increased amounts of triglycerides in lipid droplets. DHT also induces morphological changes, increases cell size, and treatments over 7 days lead to a time-dependent increase in the population of apoptotic DNA-fragmented cells. Taken together, these results show for the first time that active androgens alone can engage immature sebocytes in a clear lipogenic differentiation process (Graphical abstract). These effects depend on the expression of a functional AR in these cells. This model should be of interest for revisiting the mechanisms of the sebaceous function in vitro and for the design of relevant pharmacological models for drug or compound testing.


Comparison of dimensional accuracy of conventionally and digitally manufactured intracoronal restorations.
Advances have been made in digital dentistry for the fabrication of dental prostheses, but evidence regarding the efficacy of digital techniques for the fabrication of intracoronal restorations is lacking. The purpose of this in vitro study was to compare the dimensional accuracy of intracoronal restorations fabricated with digital and conventional techniques. A sound mandibular molar tooth received a standard onlay preparation, and onlays were fabricated with 1 of 3 fabrication techniques. In group CC, the onlays were made after conventional impression and conventional fabrication of a resin pattern. In group CP, the onlays were made after conventional impression and 3-dimensional (3D) printing of the pattern. In group IP, the onlays were made after intraoral scanning, and 3D printing produced the resin pattern. Ten specimens in each group (N=30) were evaluated. Glass-ceramic restorations were fabricated using the press technique. The replica technique was used to assess the marginal fit. Each replica was assessed at 8 points. One-way ANOVA was used to compare the marginal discrepancy among the 3 groups. The Tukey honest significant differences test was applied for pairwise comparisons of the groups (α=.05). No significant differences were noted in the marginal discrepancy at the gingival margin among the 3 groups (P=.342), but significant differences were noted among the 3 groups in the pulpal (P=.025) and lingual (P=.031) areas. Comparison of the absolute discrepancy among the 3 groups revealed that only groups CC and CP were significantly different (P=.020) from each other. Within the limitations of this in vitro study, the conventional method yielded more accuracy than the 3D printing method, and no differences were found between the methods which used the 3D printer (groups CP and IP).


European consensus on grading bone marrow fibrosis and assessment of cellularity.
Quantification of characteristic bone marrow biopsy features includes basic parameters such as cellularity and fiber content. These are important to assess the dynamics of disease processes with a significant impact on risk stratification, survival patterns and, especially, therapy-related changes. A panel of experienced European pathologists and a foreign expert evaluated, at a multi-headed microscope, a large number of representative slides of trephine biopsies from patients with myelofibrosis in an attempt to reach a consensus on how to grade cellularity and fibrosis. This included a critical evaluation of previously described scoring systems. During the microscopic analysis and subsequent discussion and voting, the importance of age-dependent decrease in cellularity was recognized. Grading of myelofibrosis was simplified by using four easily reproducible categories including differentiation between reticulin and collagen. A consensus was reached that the density of fibers must be assessed in relation to the hematopoietic tissue. This feature is especially important in order to avoid a false impression of a reduced fiber content in fatty and/or edematous bone marrow samples after treatment. The consensus for measuring myelofibrosis by clear and reproducible guidelines achieved by our group should allow for precise grading during the disease process and after therapy.


Clinical and Radiologic Responses to Cladribine for the Treatment of Erdheim-Chester Disease.
While cladribine is best known for the treatment of hairy cell leukemia and other lymphoid cancers, it also has activity against myeloid neoplasms, such as Erdheim-Chester disease (ECD). To assess the efficacy of cladribine (2-chloro-2'-deoxyadenosine) in the treatment of ECD. This study was a single-institution retrospective medical record review from January 1, 1998, to April 6, 2016, at a tertiary academic medical center. In all eligible cases, the diagnosis of ECD was made using clinical criteria in conjunction with histopathologic findings. Cladribine therapy in first-line treatment or later. Two response criteria were used, clinical and radiological. For clinical response, the following criteria were used: complete response (complete resolution of symptoms attributed to ECD), partial response (partial resolution of symptoms attributed to ECD), stable disease (no change in symptoms attributed to ECD), and progressive disease (worsening of symptoms attributed to ECD). For radiological response, the following categories were used: complete response (complete resolution of proven or suspected lesion due to ECD), partial response (partial resolution of proven or suspected lesion due to ECD), stable disease (no significant change in proven or suspected lesion due to ECD for ≥3 months), and progressive disease (progression or worsening of proven or suspected lesion due to ECD). A total of 63 adult patients with confirmed ECD were identified. Their median age at diagnosis of ECD was 54 years (age range, 18-80 years), and 67% (42 of 63) were male. Cladribine was the most commonly used chemotherapeutic agent and was administered in 21 of 63 patients (33%). Their median age at the time of cladribine therapy was 62 years (age range, 40-78 years). Cladribine was used as the first-line treatment in 9 patients and as later-line treatment in the remaining 12 patients. The median number of cycles of cladribine administered was 2.5 (range, 1-6). The overall clinical response rate was 52% (9 of 17) (6% [1 of 17] complete response and 46% [8 of 17] partial response), with 18% (3 of 17) stable disease and 30% (5 of 17) progressive disease. Among patients who responded to cladribine therapy, the median duration of clinical response was 9 months (range, 6-129 months), with ongoing response in 2 patients. The overall radiological response rate was 54% (8 of 15) (all partial response), with 26% (4 of 15) stable disease and 20% (2 of 15) progressive disease. Treatment-related adverse effects included 2 infectious complications (pneumonia and central line infection, both requiring hospitalization) and 2 hematologic adverse effects (grade 4 neutropenia and thrombocytopenia, and grade 3 neutropenia, both requiring therapy discontinuation). Cladribine has moderate clinical efficacy in the treatment of ECD and can be considered a treatment option in cases without the BRAF V600E mutation. It is generally well tolerated and may result in a durable response.


Maxillary, mandibular, and chin advancement: treatment planning based on airway anatomy in obstructive sleep apnea.
Surgical correction of obstructive sleep apnea (OSA) syndrome involves understanding a number of parameters, of which the 3-dimensional airway anatomy is important. Visualization of the upper airway based on cone beam computed tomography scans and automated computer analysis is an aid in understanding normal and abnormal airway conditions and their response to surgery. The goal of surgical treatment of OSA syndrome is to enlarge the velo-oropharyngeal airway by anterior/lateral displacement of the soft tissues and musculature by maxillary, mandibular, and possibly, genioglossus advancement. Knowledge of the specific airway obstruction and characteristics based on 3-dimensional studies permits a directed surgical treatment plan that can successfully address the area or areas of airway obstruction. The end occlusal result can be improved when orthodontic treatment is combined with the surgical plan. The individual with OSA, though, is more complicated than the usual orthognathic patient, and both the medical condition and treatment length need to be judiciously managed when OSA and associated conditions are present. The perioperative management of the patient with OSA is more complex and the margin for error is reduced, and this needs to be taken into consideration and the care altered as indicated.


Oral anticoagulants for primary prevention, treatment and secondary prevention of venous thromboembolic disease, and for prevention of stroke in atrial fibrillation: systematic review, network meta-analysis and cost-effectiveness analysis.
Warfarin is effective for stroke prevention in atrial fibrillation (AF), but anticoagulation is underused in clinical care. The risk of venous thromboembolic disease during hospitalisation can be reduced by low-molecular-weight heparin (LMWH): warfarin is the most frequently prescribed anticoagulant for treatment and secondary prevention of venous thromboembolism (VTE). Warfarin-related bleeding is a major reason for hospitalisation for adverse drug effects. Warfarin is cheap but therapeutic monitoring increases treatment costs. Novel oral anticoagulants (NOACs) have more rapid onset and offset of action than warfarin, and more predictable dosing requirements. To determine the best oral anticoagulant/s for prevention of stroke in AF and for primary prevention, treatment and secondary prevention of VTE. Four systematic reviews, network meta-analyses (NMAs) and cost-effectiveness analyses (CEAs) of randomised controlled trials. Hospital (VTE primary prevention and acute treatment) and primary care/anticoagulation clinics (AF and VTE secondary prevention). Patients eligible for anticoagulation with warfarin (stroke prevention in AF, acute treatment or secondary prevention of VTE) or LMWH (primary prevention of VTE). NOACs, warfarin and LMWH, together with other interventions (antiplatelet therapy, placebo) evaluated in the evidence network. Efficacy Stroke, symptomatic VTE, symptomatic deep-vein thrombosis and symptomatic pulmonary embolism. Safety Major bleeding, clinically relevant bleeding and intracranial haemorrhage. We also considered myocardial infarction and all-cause mortality and evaluated cost-effectiveness. MEDLINE and PREMEDLINE In-Process & Other Non-Indexed Citations, EMBASE and The Cochrane Library, reference lists of published NMAs and trial registries. We searched MEDLINE and PREMEDLINE In-Process & Other Non-Indexed Citations, EMBASE and The Cochrane Library. The stroke prevention in AF review search was run on the 12 March 2014 and updated on 15 September 2014, and covered the period 2010 to September 2014. The search for the three reviews in VTE was run on the 19 March 2014, updated on 15 September 2014, and covered the period 2008 to September 2014. Two reviewers screened search results, extracted and checked data, and assessed risk of bias. For each outcome we conducted standard meta-analysis and NMA. We evaluated cost-effectiveness using discrete-time Markov models. Apixaban (Eliquis®, Bristol-Myers Squibb, USA; Pfizer, USA) [5 mg bd (twice daily)] was ranked as among the best interventions for stroke prevention in AF, and had the highest expected net benefit. Edoxaban (Lixiana®, Daiichi Sankyo, Japan) [60 mg od (once daily)] was ranked second for major bleeding and all-cause mortality. Neither the clinical effectiveness analysis nor the CEA provided strong evidence that NOACs should replace postoperative LMWH in primary prevention of VTE. For acute treatment and secondary prevention of VTE, we found little evidence that NOACs offer an efficacy advantage over warfarin, but the risk of bleeding complications was lower for some NOACs than for warfarin. For a willingness-to-pay threshold of > £5000, apixaban (5 mg bd) had the highest expected net benefit for acute treatment of VTE. Aspirin or no pharmacotherapy were likely to be the most cost-effective interventions for secondary prevention of VTE: our results suggest that it is not cost-effective to prescribe NOACs or warfarin for this indication. NOACs have advantages over warfarin in patients with AF, but we found no strong evidence that they should replace warfarin or LMWH in primary prevention, treatment or secondary prevention of VTE. These relate mainly to shortfalls in the primary data: in particular, there were no head-to-head comparisons between different NOAC drugs. Calculating the expected value of sample information to clarify whether or not it would be justifiable to fund one or more head-to-head trials. This study is registered as PROSPERO CRD42013005324, CRD42013005331 and CRD42013005330. The National Institute for Health Research Health Technology Assessment programme.


CRISPR/Cas9-mediated efficient targeted mutagenesis in Chardonnay (Vitis vinifera L.).
The type II clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 system (CRISPR/Cas9) has been successfully applied to edit target genes in multiple plant species. However, it remains unknown whether this system can be used for genome editing in grape. In this study, we described genome editing and targeted gene mutation in 'Chardonnay' suspension cells and plants via the CRISPR/Cas9 system. Two single guide RNAs (sgRNAs) were designed to target distinct sites of the L-idonate dehydrogenase gene (IdnDH). CEL I endonuclease assay and sequencing results revealed the expected indel mutations at the target site, and a mutation frequency of 100% was observed in the transgenic cell mass (CM) as well as corresponding regenerated plants with expression of sgRNA1/Cas9. The majority of the detected mutations in transgenic CM were 1-bp insertions, followed by 1- to 3-nucleotide deletions. Off-target activities were also evaluated by sequencing the potential off-target sites, and no obvious off-target events were detected. Our results demonstrated that the CRISPR/Cas9 system is an efficient and specific tool for precise genome editing in grape.


Involvement of nitric oxide in the elimination of a transient retinotectal projection in development.
The adult pattern of axonal connections from the eye to the brain arises during development through the refinement of a roughly ordered set of connections. In the chick visual system, refinement normally results in the loss of the ipsilateral retinotectal connections. Inhibition of nitric oxide synthesis reduced the loss of these transient connections. Because nitric oxide is expressed by tectal cells with which retinal axons connect and because reduction of nitric oxide synthesis by tectal cells resulted in a change in the connections of retinal axons, nitric oxide probably serves as a messenger from tectal cells back to retinal axons during development.


Clinico-pathological features of erythema nodosum leprosum: A case-control study at ALERT hospital, Ethiopia.
Leprosy reactions are a significant cause of morbidity in leprosy population. Erythema nodosum leprosum (ENL) is an immunological complication affecting approximately 50% of patients with lepromatous leprosy (LL) and 10% of borderline lepromatous (BL) leprosy. ENL is associated with clinical features such as skin lesions, neuritis, arthritis, dactylitis, eye inflammation, osteitis, orchitis, lymphadenitis and nephritis. ENL is treated mainly with corticosteroids and corticosteroids are often required for extended periods of time which may lead to serious adverse effects. High mortality rate and increased morbidity associated with corticosteroid treatment of ENL has been reported. For improved and evidence-based treatment of ENL, documenting the systems affected by ENL is important. We report here the clinical features of ENL in a cohort of patients with acute ENL who were recruited for a clinico-pathological study before and after prednisolone treatment. A case-control study was performed at ALERT hospital, Ethiopia. Forty-six LL patients with ENL and 31 non-reactional LL matched controls were enrolled to the study and followed for 28 weeks. Clinical features were systematically documented at three visits (before, during and after predinsolone treatment of ENL cases) using a specifically designed form. Skin biopsy samples were obtained from each patient before and after treatment and used for histopathological investigations to supplement the clinical data. Pain was the most common symptom reported (98%) by patients with ENL. Eighty percent of them had reported skin pain and more than 70% had nerve and joint pain at enrolment. About 40% of the patients developed chronic ENL. Most individuals 95.7% had nodular skin lesions. Over half of patients with ENL had old nerve function impairment (NFI) while 13% had new NFI at enrolment. Facial and limb oedema were present in 60% patients. Regarding pathological findings before treatment, dermal neutrophilic infiltration was noted in 58.8% of patients with ENL compared to 14.3% in LL controls. Only 14.7% patients with ENL had evidence of vasculitis at enrolment. In our study, painful nodular skin lesions were present in all ENL patients. Only 58% patients had dermal polymorphonuclear cell infiltration showing that not all clinically confirmed ENL cases have neutrophilic infiltration in lesions. Very few patients had histological evidence of vasculitis. Many patients developed chronic ENL and these patients require inpatient corticosteroid treatment for extended periods which challenges the health service facility in resource poor settings, as well as the patient's quality of life.


Accounting for intensity variation in image analysis of large-scale multiplexed clinical trial datasets.
Multiplex immunofluorescence (mIF) imaging can provide comprehensive quantitative and spatial information for multiple immune markers for tumour immunoprofiling. However, application at scale to clinical trial samples sourced from multiple institutions is challenging due to pre-analytical heterogeneity. This study reports an analytical approach to the largest multi-parameter immunoprofiling study of clinical trial samples to date. We analysed 12,592 tissue microarray (TMA) spots from 3,545 colorectal cancers sourced from more than 240 institutions in two clinical trials (QUASAR 2 and SCOT) stained for CD4, CD8, CD20, CD68, FoxP3, pan-cytokeratin, and DAPI by mIF. TMA slides were multi-spectrally imaged and analysed by cell-based and pixel-based marker analysis. We developed an adaptive thresholding method to account for inter- and intra-slide intensity variation in TMA analysis. Applying this method effectively ameliorated inter- and intra-slide intensity variation improving the image analysis results compared with methods using a single global threshold. Correlation of CD8 data derived by our mIF analysis approach with single-plex chromogenic immunohistochemistry CD8 data derived from subsequent sections indicates the validity of our method (Spearman's rank correlation coefficients ρ between 0.63 and 0.66, p ≪ 0.01) as compared with the current gold standard analysis approach. Evaluation of correlation between cell-based and pixel-based analysis results confirms equivalency (ρ > 0.8, p ≪ 0.01, except for CD20 in the epithelial region) of both analytical approaches. These data suggest that our adaptive thresholding approach can enable analysis of mIF-stained clinical trial TMA datasets by digital pathology at scale for precision immunoprofiling.


Anaerobic ammonium oxidation (anammox) in Chesapeake Bay sediments.
Anaerobic ammonium oxidation (anammox) has recently been recognized as a pathway for the removal of fixed N from aquatic ecosystems. However, the quantitative significance of anammox in estuarine sediments is variable, and measurements have been limited to a few estuaries. We measured anammox and conventional denitrification activities in sediments along salinity gradients in the Chesapeake Bay and two of its sub-estuaries, the Choptank River and Patuxent River. Homogenized sediments were incubated with (14/15)N amendments of NH4+, NO3-, and NO2- to determine relative activities of anammox and denitrification. The percent of N2 production due to anammox (ra%) ranged from 0 to 22% in the Chesapeake system, with the highest ra% in the freshwater portion of the main stem of upper Chesapeake Bay, where water column NO3- concentrations are consistently high. Intermediate levels of relative anammox (10%) were detected at locations corresponding to tidal freshwater and mesohaline locations in the Choptank River, whereas anammox was not detected in the tidal freshwater location in the Patuxent River. Anammox activity was also not detected in the seaward end of Chesapeake Bay, where water column No3- concentrations are consistently low. The ra% did not correlate with NH4+ accumulation rate in anoxic sediment incubations, but ra% was related to water column NO3- concentrations and salinity. Anammox bacterial communities were also examined by amplifying DNA extracted from the upper Chesapeake Bay sediment with polymerase chain reaction (PCR) primers that are specific for 16S rRNA genes of anammox organisms. A total of 35 anammox-like sequences were detected, and phylogenetic analysis grouped the sequences in two distinct clusters belonging to the Candidatus "Scalindua" genus.


Comparing apples and oranges: using reward-specific and reward-general subjective value representation in the brain.
The ability of human subjects to choose between disparate kinds of rewards suggests that the neural circuits for valuing different reward types must converge. Economic theory suggests that these convergence points represent the subjective values (SVs) of different reward types on a common scale for comparison. To examine these hypotheses and to map the neural circuits for reward valuation we had food and water-deprived subjects make risky choices for money, food, and water both in and out of a brain scanner. We found that risk preferences across reward types were highly correlated; the level of risk aversion an individual showed when choosing among monetary lotteries predicted their risk aversion toward food and water. We also found that partially distinct neural networks represent the SVs of monetary and food rewards and that these distinct networks showed specific convergence points. The hypothalamic region mainly represented the SV for food, and the posterior cingulate cortex mainly represented the SV for money. In both the ventromedial prefrontal cortex (vmPFC) and striatum there was a common area representing the SV of both reward types, but only the vmPFC significantly represented the SVs of money and food on a common scale appropriate for choice in our data set. A correlation analysis demonstrated interactions across money and food valuation areas and the common areas in the vmPFC and striatum. This may suggest that partially distinct valuation networks for different reward types converge on a unified valuation network, which enables a direct comparison between different reward types and hence guides valuation and choice.


The arginine finger domain of ExoT contributes to actin cytoskeleton disruption and inhibition of internalization of Pseudomonas aeruginosa by epithelial cells and macrophages.
Pseudomonas aeruginosa, an important nosocomial pathogen of humans, expresses a type III secretion system that is required for virulence. Previous studies demonstrated that the lung-virulent strain PA103 has the capacity to be either cytotoxic or invasive. Analyses of mutants suggest that PA103 delivers a negative regulator of invasion, or anti-internalization factor, to host cells via a type III secretion system. In this work we show that the type III secreted protein ExoT inhibits the internalization of PA103 by polarized epithelial cells (Madin-Darby canine kidney cells) and J774.1 macrophage-like cells. ExoS, which is closely related to ExoT but has additional ADP-ribosylating activity, can substitute for ExoT as an anti-internalization factor. ExoT contains a signature arginine finger domain found in GTPase-activating proteins. Mutation of the conserved arginine in ExoT diminished its anti-internalization activity and altered its ability to disrupt the actin cytoskeleton. Cell fractionation experiments showed that ExoT is translocated into host cells and that mutation of the arginine finger did not disrupt translocation. In a mouse model of acute pneumonia, PA103DeltaUDeltaT reached the lungs as efficiently as PA103DeltaU but showed reduced colonization of the liver. This finding suggests that the ability to resist internalization may be important for virulence in vivo.


Dose-dependent effect of Bisphenol-A on insulin signaling molecules in cardiac muscle of adult male rat.
Environmental contaminant, Bisphenol-A (BPA) is a xenoestrogen, an essential component used for the production of two classes of polymers such as polycarbonate and epoxy resin which disrupts the normal endocrine function. BPA has intense effects on mice endocrine pancreas, an essential tissue involved in glucose metabolism. It disrupts pancreatic β-cell insulin content, induces hyperinsulinemia and insulin resistance in male rats. Cardiac muscle is an insulin responsive organ and insulin has direct effects on glucose transport. The present study was designed to assess the effect of BPA on insulin signaling molecules in the cardiac muscle of adult male Wistar rat. Adult male Wistar rats (200-250 g) were selected and divided into following groups: Group 1: Control (vehicle treated), Group 2: Rats treated with 10 mg BPA/kg b.wt./day for 30 days orally, Group 3: Rats treated with 100 mg BPA/kg b.wt./day for 30 days orally, Group 4: Rats treated with 400 mg BPA/kg b.wt./day for 30 days orally. IR (insulin receptor) and pIRTyr1162 proteins were significantly decreased in the high dose group (400 mg). There was no change in IRS1 (insulin receptor substrate-1) and Akt proteins. Whereas, a decrease in pIRS1Tyr632 (100 mg and 400 mg), pAkt Ser473 (400 mg) and GLUT4 (glucose transporter 4) (cytosolic and plasma membrane) proteins was observed which may affect the cardiovascular function. It is concluded that BPA exposure has adverse effect on cardiac insulin signal transduction which may affect its function.


Physical activity and cognitive function in a cross-section of younger and older community-dwelling individuals.
Previous reports have indicated a small, positive relationship between physical activity and cognition. However, the majority of research has focused on older adults, with few studies examining this relationship during earlier periods of the life span. This study examined the relationship of physical activity to cognition in a cross-section of 241 community-dwelling individuals 15-71 years of age with a task requiring variable amounts of executive control. Data were analyzed with multiple regression, which controlled for age, sex, and IQ. Participants reported their physical activity behavior and were tested for reaction time (RT) and response accuracy on congruent and incongruent conditions of a flanker task, which manipulates interference control. After controlling for confounding variables, an age-related slowing of RT was observed during both congruent and incongruent flanker conditions. However, physical activity was associated with faster RT during these conditions, regardless of age. Response accuracy findings indicated that increased physical activity was associated with better performance only during the incongruent condition for the older cohort. Findings suggest that physical activity may be beneficial to both general and selective aspects of cognition, particularly among older adults.


Isolation and identification of African horsesickness virus from naturally infected dogs in Upper Egypt.
African horsesickness virus was isolated from blood samples of street dogs in Aswan Province in Arab Republic of Egypt. Of six isolated "dog strain" African horsesickness viruses, three viruses designated D2, D6 and D10 have been identified as type 9 African horsesickness virus. Methods of isolation, tissue culture adaptation, serological indentification and typing are described. Horses experimentally infected with dog viruses showed febrile reaction and characteristic clinical and pathological signs of African horsesickness. Reisolation of African horsesickness virus type 9 was achieved from the horses during serial passages.


Macular function assessed by microperimetry in patients with enhanced S-cone syndrome.
Enhanced S-cone syndrome (ESCS), also known as Goldmann-Favre syndrome, is a progressive retinal degeneration that frequently presents with night blindness and nummular pigment clumping around the vascular arcades and is caused by recessive mutations in the photoreceptor-specific NR2E3 transcription factor. A unique feature of this disease is the development of retinoschisis of the macula. This study used fine anatomic and functional assessments within this region to determine whether the loss of retinal function was due to progressive schisis or a primary photoreceptor loss, similar to other rod-cone dystrophies. Cross-sectional, prospective study. Nine probands (n=18 eyes) and 3 controls (n=6 eyes) were studied at Moorfields Eye Hospital in London, United Kingdom. Histories were obtained and visual acuity was measured using Early Treatment Diabetic Retinopathy Study protocol. Autofluorescence (AF), fundus photography, and spectral domain optical coherence tomography (OCT) imaging were co-registered to detailed microperimetry (Nidek MP1; NAVIS software version 1.7.2; Nidek Technologies, Padova, Italy) data for statistical analysis. Retinal sensitivity (decibels) in a customized test grid of the macula; retinal structure assessed with OCT and AF. Patients were divided into 3 cohorts roughly based on life span and documentation of schisis: (1) no schisis, childhood; (2) macular schisis, young adults; (3) resolved schisis, older adults. Retinal sensitivity was significantly attenuated in those with schisis and did not recover in those whose schisis had resolved despite retinal thickness comparable to that of controls. All probands exhibited loss of AF peripherally (and corresponding loss of retinal sensitivity), but there was relative preservation of AF within the macula. Development of macular retinoschisis in ESCS is an important feature of the disease and contributes to attenuated retinal sensitivity that persists after resolution of retinoschisis. The central macula appears to be compromised more by foveoschisis than photoreceptor loss. In contrast, the peripheral retina (ordinarily a rod-rich region) is affected early in the disease process and degenerates rapidly because of photoreceptor loss. Thus, 2 distinct mechanisms of retinal degeneration may exist in ESCS, corresponding to regions of the retina that may experience either normal or abnormal photoreceptor development.


Specialty choice in UK junior doctors: is psychiatry the least popular specialty for UK and international medical graduates?
In the UK and many other countries, many specialties have had longstanding problems with recruitment and have increasingly relied on international medical graduates to fill junior and senior posts. We aimed to determine what specialties were the most popular and desirable among candidates for training posts, and whether this differed by country of undergraduate training. We conducted a database analysis of applications to Modernising Medical Careers for all training posts in England in 2008. Total number of applications (as an index of popularity) and applications per vacancy (as an index of desirability) were analysed for ten different specialties. We tested whether mean consultant incomes correlated with specialty choice. In, 2008, there were 80,949 applications for specialty training in England, of which 31,434 were UK graduates (39%). Among UK medical graduates, psychiatry was the sixth most popular specialty (999 applicants) out of 10 specialty groups, while it was fourth for international graduates (5,953 applicants). Among UK graduates, surgery (9.4 applicants per vacancy) and radiology (8.0) had the highest number of applicants per vacancy and paediatrics (1.2) and psychiatry (1.1) the lowest. Among international medical graduates, psychiatry had the fourth highest number of applicants per place (6.3). Specialty popularity for UK graduates was correlated with predicted income (p = 0.006). Based on the number of applicants per place, there was some consistency in the most popular specialties for both UK and international medical graduates, but there were differences in the popularity of psychiatry. With anticipated decreases in the number of new international medical graduates training in the UK, university departments and professional associations may need to review strategies to attract more UK medical graduates into certain specialties, particularly psychiatry and paediatrics.


Reliability of the diagnostic tests for Cushing's syndrome performed in a tertiary referral center.
The study aimed to retrospectively evaluate the reliability of the diagnostic and location tests in Cushing's Syndrome (CS). Eighty-seven patients diagnosed with CS between 1995 and 2007 by Endocrinology Metabolism Department of Cerrahpasa Medical School were included in the study. The control group consisted of 91 patients who presented to the outpatient clinic because of obesity. The diagnostic tests were as follows: 1 mg dexamethasone suppression test (DST), 24-h urinary free cortisol (UFC), midnight cortisol level (MCL), ACTH level and overnight 8 mg DST. The sensitivity and specificity of UFC were 81 and 66 % respectively for the cut-off point of 50 μg/day, whereas they were 64 and 76 % respectively for the cut-off point of 100 μg/day. For the cut-off value of 1.8/μg/dL for MCL and 1 mDST, the sensitivity rates were 100 and 98 %, while the specificity rates were 88 and 33 %, respectively. Among the location tests, the sensitivity and specificity of ACTH under 10 pg/mL for adrenal CS were 92 and 94 % respectively. The sensitivity and specificity of ACTH higher than 30 pg/mL for ACTH-dependent CS were 69 and 100 % respectively. The sensitivity rates of 8 mg DST for 50 and 60 % suppressions were 83 and 79 % respectively, whereas the specificity rates were 75 and 88 % respectively. 1 mg DST (cut-off <1.8 μg/dL) and UFC (50 μg/24 h) are appropriate tests for screening CS. Overnight 8 mg DST with 60 % suppression for Cushing's Disease (CD) and ACTH levels <10 pg/mL for adrenal CS, ACTH levels >30 pg/mL for ACTH dependency were identified as the best tests for the differential diagnosis of the subtypes.


Predictors of mortality in childhood burns: an 8-year review.
This study aims to analyze the epidemiological characteristics and predictors of mortality from burn injuries in childhood patients admitted to our hospital during an eight-year period. The medical records of acute childhood burn patients were reviewed retrospectively. All variables thought to be associated with mortality were entered in a multiple binary logistic regression model (method = stepwise). The magnitude of risk was measured by the odds ratio, and the 95% confidence interval was estimated. A total of 2269 acute childhood burn patients were admitted during the study period. A total of 86 (3.8%) children died due to burn injuries. Deaths were seen 1.849 times more in males than in females. According to the 1%-10% total body surface area (TBSA) burned group, mortality occurred 121.116 times more in the >41% TBSA burned group. Most burn injuries can be avoided by keeping children away from hazardous and dangerous environments. Also, requiring a multidisciplinary management in these patients, quality of care services given by physicians and nurses certainly will create a positive impact on patients' outcomes.


Outcomes of apixaban vs. warfarin by type and duration of atrial fibrillation: results from the ARISTOTLE trial.
It is uncertain whether the benefit from apixaban varies by type and duration of atrial fibrillation (AF). A total of 18 201 patients with AF [2786 (15.3%) with paroxysmal and 15 412 (84.7%) with persistent or permanent] were randomized to apixaban or warfarin. In this pre-specified secondary analysis, we compared outcomes and treatment effect of apixaban vs. warfarin by AF type and duration. The primary efficacy endpoint was a composite of ischaemic or haemorrhagic stroke or systemic embolism. The secondary efficacy endpoint was all-cause mortality. There was a consistent reduction in stroke or systemic embolism (P for interaction = 0.71), all-cause mortality (P for interaction = 0.75), and major bleeding (P for interaction = 0.50) with apixaban compared with warfarin for both AF types. Apixaban was superior to warfarin in all studied endpoints, regardless of AF duration at study entry (P for all interactions >0.13). The rate of stroke or systemic embolism was significantly higher in patients with persistent or permanent AF than patients with paroxysmal AF (1.52 vs. 0.98%; P = 0.003, adjusted P = 0.015). There was also a trend towards higher mortality in patients with persistent or permanent AF (3.90 vs. 2.81%; P = 0.0002, adjusted P = 0.066). The risks of stroke, mortality, and major bleeding were lower with apixaban than warfarin regardless of AF type and duration. Although the risk of stroke or systemic embolism was lower in paroxysmal than persistent or permanent AF, apixaban is an attractive alternative to warfarin in patients with AF and at least one other risk factor for stroke, regardless of the type or duration of AF.


Ovarian cycle-specific regulation of adipose tissue lipid storage by testosterone in female nonhuman primates.
Previous studies in rodents and humans suggest that hyperandrogenemia causes white adipose tissue (WAT) dysfunction in females, although the underlying mechanisms are poorly understood. In light of the differences in the length of the ovarian cycle between humans and rodents, we used a nonhuman primate model to elucidate the effects of chronic hyperandrogenemia on WAT function in vivo. Female rhesus macaques implanted with testosterone capsules developed insulin resistance and altered leptin secretion on a high-fat, Western-style diet. In control visceral WAT, lipolysis and hormone-sensitive lipase expression were upregulated during the luteal phase compared with the early follicular (menses) phase of the ovarian cycle. Hyperandrogenemia attenuated elevated lipolysis and hormone-sensitive lipase activity in visceral WAT during the luteal phase but not during menses. Under control conditions, insulin-stimulated Akt and Erk activation and fatty acid uptake in WAT were not significantly affected by the ovarian cycle. In contrast, testosterone treatment preferentially increased fatty acid uptake and insulin signaling at menses. The fatty acid synthase and glucose transporter-4 genes were upregulated by testosterone during the luteal phase. In summary, this study reveals ovarian stage-specific fluctuations in adipocyte lipolysis and suggests that male sex hormones increase and female sex hormones decrease lipid storage in female WAT.


Total laparoscopic Roux-en-Y cholangiojejunostomy for the treatment of biliary disease.
Roux-en-Y cholangiojejunostomy (RCJS) has been widely used in biliary bypass surgeries, but in most reported literature, an assisted miniincision was needed, and studies reporting total laparoscopic Roux-en-Y cholangiojejunostomy (TLRCJS) are rare. The goal of this study was to investigate how to treat hepatic portal bile duct diseases and perform jejunojejunostomy and cholangiojejunostomy totally laparoscopically. We evaluated the feasibility of TLRCJS in treating biliary tract diseases. TLRCJS were performed in 103 patients from January 2000 to August 2011. There were 28 cases of recurrent choledocholithiasis combined with stricture of the common bile duct (CBD) after several stone extractions, 3 patients with iatrogenic bile duct injury, 24 patients with choledochal cyst, 36 patients with hepatic portal cholangiocarcinoma, and 12 patients with cancer of the pancreatic head and periampullary cancer. All surgeries were performed through 5 trocars. First, laparoscopic surgery on the CBD was performed according to the original disease. The CBD was opened and stones were extracted in choledocholithiasis patients. In iatrogenic injury patients, strictured CBD was resected and repaired. Dilated CBD or choledochal cyst with tumor was transected. In patients with malignant jaundice, the CBD was opened longitudinally. At the same time, the bile duct was prepared for cholangiojejunostomy. Second, the positions of the laparoscope and surgeons were altered. The jejunal mesentery and jejunum were transected, and side-to-side jejunojejunostomy (JJS) was performed. The laparoscope and surgeon positions were exchanged again; the Rouxen-Y biliary limb was lifted close to the residual bile duct; and side-to-side or end-to-side choledochojejunostomy (CJS) was performed. Finally, an abdominal drainage tube was placed. All the surgeries were performed successfully. The diameter of the residual bile duct ranged from 0.4 to 3.2 cm (average, 0.9 cm). Three patients had postoperative bile leakage and were treated from 1 week to approximately 1 month with abdominal drainage. Postoperative intraperitoneal hemorrhage and stress ulcer of the stomach occurred in 2 patients with biliary tract injury combined with obstructive jaundice. One with intraperitoneal hemorrhage was cured by another laparoscopic surgery. The other patient was cured after 2 days of abdominal drainage, antacids, and hemostatic drug therapy. The follow-up duration of 95 patients was 4 to 93 months (average, 48.3 months). The follow-up rate was 92.2% (95/103). Patients with cancer died of metastasis or cachexia during 14-month follow-up with no postoperative complication. Reflux cholangitis occurred in 3 patients 2, 3, and 5 years after the operation, respectively. No anastomotic stricture or other complication was found in other patients during the follow-up. TLRCJS is the best and first choice for patients with biliary tract diseases that need biliary-jejunal anastomosis. But it is essential that the surgeon has proficiency in laparoscopic surgeries.


Variability of activity patterns across mood disorders and time of day.
Few actigraphy studies in mood disorders have simultaneously included unipolar (UP) and bipolar (BD) depression or BD mixed states as a separate subgroup from mania. This study compared objectively measured activity in UP, BD depression, mania and mixed states and examined if patterns differed according to time of day and/or diagnostic group. Eighty -eight acutely admitted inpatients with mood disorders (52 UP; 18 mania; 12 BD depression; 6 mixed states) underwent 24 hours of actigraphy monitoring. Non-parametric analyses were used to compare median activity level over 24 h (counts per minute), two time series (64-min periods of continuous motor activity) in the morning and evening, and variability in activity across and within groups. There was no between-group difference in 24-h median level of activity, but significant differences emerged between BD depression compared to mania in the active morning period, and between UP and mania and mixed states in the active evening period. Within-group analyses revealed that UP cases showed several significant changes between morning and evening activity, with fewer changes in the BD groups. Mean activity over 24 hours has limited utility in differentiating UP and BD. In contrast, analysis of non-linear variability measures of activity at different times of day could help objectively distinguish between mood disorder subgroups. ClinicalTrials.gov Identifier: NCT01415323 , first registration July 6, 2011.


Allelic and locus heterogeneity in inherited venous malformations.
Venous malformations are low-flow vascular lesions consisting of disorganized thin-walled vascular channels. These can occur sporadically but also as an autosomal dominant condition termed venous malformations, cutaneous and mucosal (VMCM; OMIM 600195). In two large unrelated kindreds mapping to chromosome 9, the identical R849W missense mutation was identified in the first kinase domain of Tie2, an endothelial cell-specific receptor tyrosine kinase. We report here the identification of four new kindreds with inherited venous malformations. Unlike the initial two families described, these four families demonstrate allelic and locus heterogeneity. In one of these families, the R849W mutation co-segregates with the disease phenotype. Three other families with venous malformations lack this mutation. One of these families is linked to markers near TIE2 on chromosome 9. In this family, we identified a novel mutation within the first kinase domain of Tie2 resulting in a Y897S change. Results from COS-1 cell transfections using expression constructs containing either the R849W or the Y897S mutation suggest that the receptors containing either mutation show ligand-independent hyperphosphorylation. These results suggest a gain-of-function mechanism for development of venous malformations in these families. Of the two remaining families, one excludes linkage to the TIE2 locus, establishing the existence of at least one additional locus for dominantly inherited venous malformations.


Machine learning for the New York City power grid.
Power companies can benefit from the use of knowledge discovery methods and statistical machine learning for preventive maintenance. We introduce a general process for transforming historical electrical grid data into models that aim to predict the risk of failures for components and systems. These models can be used directly by power companies to assist with prioritization of maintenance and repair work. Specialized versions of this process are used to produce 1) feeder failure rankings, 2) cable, joint, terminator, and transformer rankings, 3) feeder Mean Time Between Failure (MTBF) estimates, and 4) manhole events vulnerability rankings. The process in its most general form can handle diverse, noisy, sources that are historical (static), semi-real-time, or realtime, incorporates state-of-the-art machine learning algorithms for prioritization (supervised ranking or MTBF), and includes an evaluation of results via cross-validation and blind test. Above and beyond the ranked lists and MTBF estimates are business management interfaces that allow the prediction capability to be integrated directly into corporate planning and decision support; such interfaces rely on several important properties of our general modeling approach: that machine learning features are meaningful to domain experts, that the processing of data is transparent, and that prediction results are accurate enough to support sound decision making. We discuss the challenges in working with historical electrical grid data that were not designed for predictive purposes. The "rawness" of these data contrasts with the accuracy of the statistical models that can be obtained from the process; these models are sufficiently accurate to assist in maintaining New York City's electrical grid.


Structural characterization of the tetrameric form of the major cat allergen Fel d 1.
Felis domesticus allergen 1(Fel d 1) is a 35 kDa tetrameric glycoprotein formed by two heterodimers which elicits IgE responses in 95% of patients with allergy to cat. We have previously established in vitro conditions for the appropriate folding of recombinant Fel d 1 using a direct linkage of chain 1 to chain 2 (construct Fel d 1 (1+2)) and chain 2 to chain 1 (construct Fel d 1 (2+1)). Although the crystal structure of Fel d 1 (2+1) revealed a striking structural similarity to that of uteroglobin, a steroid-inducible cytokine-like molecule with anti-inflammatory and immunomodulatory properties, no functional tetrameric form of Fel d 1 could be identified. Here we present the crystal structure of the Fel d 1 (1+2) tetramer at 1.6 A resolution. Interestingly, the crystal structure of tetrameric Fel d 1 reveals two different calcium-binding sites. Symmetrically positioned on each side of the Fel d 1 tetramer, the external Ca(2+)-binding sites correspond to a putative Ca(2+)-binding site previously suggested for uteroglobin. The second Ca(2+)-binding site lies within the dimerization interface, stabilizing the formation of the Fel d 1 tetramer, and inducing important local conformational changes that directly govern the shape of two water-filled cavities. The crystal structure suggests a potential portal for an unknown ligand. Alternatively, the two cavities could be used by the allergen as a conditional inner space allowing for the spatial rearrangement of centrally localized side-chains, such as Asp130, without altering the overall fold of the molecule. The striking structural similarity of the major cat allergen to uteroglobin, coupled to the identification in the present study of a common Ca(2+)-binding site, let us speculate that Fel d 1 could provoke an allergic response through the modulation of phospholipase A2, by sequestering Ca ions in a similar manner as previously suggested for uteroglobin.


Characterization of profilin polymorphism in pollen with a focus on multifunctionality.
Profilin, a multigene family involved in actin dynamics, is a multiple partners-interacting protein, as regard of the presence of at least of three binding domains encompassing actin, phosphoinositide lipids, and poly-L-proline interacting patches. In addition, pollen profilins are important allergens in several species like Olea europaea L. (Ole e 2), Betula pendula (Bet v 2), Phleum pratense (Phl p 12), Zea mays (Zea m 12) and Corylus avellana (Cor a 2). In spite of the biological and clinical importance of these molecules, variability in pollen profilin sequences has been poorly pointed out up until now. In this work, a relatively high number of pollen profilin sequences have been cloned, with the aim of carrying out an extensive characterization of their polymorphism among 24 olive cultivars and the above mentioned plant species. Our results indicate a high level of variability in the sequences analyzed. Quantitative intra-specific/varietal polymorphism was higher in comparison to inter-specific/cultivars comparisons. Multi-optional posttranslational modifications, e.g. phosphorylation sites, physicochemical properties, and partners-interacting functional residues have been shown to be affected by profilin polymorphism. As a result of this variability, profilins yielded a clear taxonomic separation between the five plant species. Profilin family multifunctionality might be inferred by natural variation through profilin isovariants generated among olive germplasm, as a result of polymorphism. The high variability might result in both differential profilin properties and differences in the regulation of the interaction with natural partners, affecting the mechanisms underlying the transmission of signals throughout signaling pathways in response to different stress environments. Moreover, elucidating the effect of profilin polymorphism in adaptive responses like actin dynamics, and cellular behavior, represents an exciting research goal for the future.


Specific recognition of bovine serum albumin using superparamagnetic molecularly imprinted nanomaterials prepared by two-stage core-shell sol-gel polymerization.
A novel type of uniform magnetic imprinted nanomaterial for the recognition of bovine serum albumin (BSA) was prepared by anchoring MIP shells on the surface of silica deposited Fe3O4 NPs via a surface imprinting process and two-stage core-shell sol-gel polymerization. The resulting magnetic nanomaterials were characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, and a vibrating sample magnetometer (VSM). The measurements indicated that the as-synthesized nanospheres exhibited good dispersion, high crystallinity, and satisfactory superparamagnetic properties. Moreover, the obtained Fe3O4@BSA-MIPs had a high saturation magnetization (43.82 emu g-1), which allowed them to be easily separated from solution by means of an external magnetic field. The thickness of the imprinted polymer layer was approximately 5 nm, which would be effective for the mass transport between the solution and the surface of Fe3O4@BSA-MIPs. The kinetic adsorption experiment showed that the imprinted nanomaterials could reach equilibrium within 15 min and be well described by the second-order kinetics model, indicating chemical adsorption might be the rate-limiting step. Meanwhile, the imprinting factor and selectivity coefficient of the Fe3O4@BSA-MIPs were as high as 16.4 and 4.65, displaying excellent selectivity towards BSA. In addition, the resulting imprinted polymers were without obvious deterioration after ten adsorption-desorption cycles and different batches of which exhibited excellent reproducibility. Successful application in the selective separation and enrichment of BSA from a bovine blood sample and good recovery after a reasonably mild elution suggested that the Fe3O4@BSA-MIPs could specifically capture BSA from a real complex matrix and had potential value in practical applications.


Hypothalamic-pituitary-testicular axis disruptions in older men are differentially linked to age and modifiable risk factors: the European Male Aging Study.
The cause of declining testosterone (T) in aging men and their relationships with risk factors are unclear. The objective of the study was to investigate the relationships between lifestyle and health with reproductive hormones in aging men. This was a baseline cross-sectional survey on 3200 community-dwelling men aged 40-79 yr from a prospective cohort study in eight European countries. Four predictors were associated with distinct modes of altered function: 1) age: lower free T (FT; -3.12 pmol/liter.yr, P < 0.001) with raised LH, suggesting impaired testicular function; 2) obesity: lower total T (TT; -2.32 nmol/liter) and FT (-17.60 pmol/liter) for body mass index (BMI; > or = 25 to < 30 kg/m(2)) and lower TT (-5.09 nmol/liter) and FT (-53.72 pmol/liter) for BMI 30 kg/m(2) or greater (P < 0.001-0.01, referent: BMI < 25 kg/m(2)) with unchanged/decreased LH, indicating hypothalamus/pituitary dysfunction; 3) comorbidity: lower TT (-0.80 nmol/liter, P < 0.01) with unchanged LH in younger men but higher LH in older men; and 4) smoking: higher SHBG (5.96 nmol/liter, P < 0.001) and LH (0.77 U/liter, P < 0.01) with increased TT (1.31 nmol/liter, P < 0.001) but not FT, compatible with a resetting of T-LH-negative feedback due to elevated SHBG. Complex multiple alterations in the hypothalamic-pituitary-testicular axis function exist in aging men against a background of progressive age-related testicular impairment. These changes are differentially linked to specific risk factors. Some risk factors operate independently of but others interact with age, in contributing to the T decline. These potentially modifiable risk factors suggest possible preventative measures to maintain T during aging in men.


The protein kinase IKKepsilon regulates energy balance in obese mice.
Obesity is associated with chronic low-grade inflammation that negatively impacts insulin sensitivity. Here, we show that high-fat diet can increase NF-kappaB activation in mice, which leads to a sustained elevation in level of IkappaB kinase epsilon (IKKepsilon) in liver, adipocytes, and adipose tissue macrophages. IKKepsilon knockout mice are protected from high-fat diet-induced obesity, chronic inflammation in liver and fat, hepatic steatosis, and whole-body insulin resistance. These mice show increased energy expenditure and thermogenesis via enhanced expression of the uncoupling protein UCP1. They maintain insulin sensitivity in liver and fat, without activation of the proinflammatory JNK pathway. Gene expression analyses indicate that IKKepsilon knockout reduces expression of inflammatory cytokines, and changes expression of certain regulatory proteins and enzymes involved in glucose and lipid metabolism. Thus, IKKepsilon may represent an attractive therapeutic target for obesity, insulin resistance, diabetes, and other complications associated with these disorders.


When mRNA translation meets decay.
Messenger RNA (mRNA) translation and mRNA degradation are important determinants of protein output, and they are interconnected. Previously, it was thought that translation of an mRNA, as a rule, prevents its degradation. mRNA surveillance mechanisms, which degrade mRNAs as a consequence of their translation, were considered to be exceptions to this rule. Recently, however, it has become clear that many mRNAs are degraded co-translationally, and it has emerged that codon choice, by influencing the rate of ribosome elongation, affects the rate of mRNA decay. In this review, we discuss the links between translation and mRNA stability, with an emphasis on emerging data suggesting that codon optimality may regulate mRNA degradation.


Regulation of human mononuclear phagocyte migration by cell surface-binding proteins for advanced glycation end products.
Nonenzymatic glycation of proteins occurs at an accelerated rate in diabetes and can lead to the formation of advanced glycation end products of proteins (AGEs), which bind to mononuclear phagocytes (MPs) and induce chemotaxis. We have isolated two cell surface-associated binding proteins that mediate the interaction of AGEs with bovine endothelial cells. One of these proteins is a new member of the immunoglobulin superfamily of receptors (termed receptor for AGEs or RAGE); and the second is a lactoferrin-like polypeptide (LF-L). Using monospecific antibodies to these two AGE-binding proteins, we detected immunoreactive material on Western blots of detergent extracts from human MPs. Radioligand-binding studies demonstrated that antibody to the binding proteins blocked 125I-AGE-albumin binding and endocytosis by MPs. Chemotaxis of human MPs induced by soluble AGE-albumin was prevented in a dose-dependent manner by intact antibodies raised to the AGE-binding proteins, F(ab')2 fragments of these antibodies and by soluble RAGE. When MP migration in response to N-formyl-Met-Leu-Phe was studied in a chemotaxis chamber with AGE-albumin adsorbed to the upper surface of the chamber membrane, movement of MPs to the lower compartment was decreased because of interaction of the glycated proteins with RAGE and LF-L on the cell surface. The capacity of AGEs to attract and retain MPs was shown by implanting polytetrafluoroethylene (PTFE) mesh impregnated with AGE-albumin into rats: within 4 d a florid mononuclear cell infiltrate was evident in contrast to the lack of a significant cellular response to PTFE with adsorbed native albumin. These data indicate that RAGE and LF-L have a central role in the interaction of AGEs with human mononuclear cells and that AGEs can serve as a nidus to attract MPs in vivo.


The association between psychostimulant use in pregnancy and adverse maternal and neonatal outcomes: results from a distributed analysis in two similar jurisdictions.
Conflicting evidence suggests a possible association between use of prescribed psychostimulants during pregnancy and adverse perinatal outcomes. We conducted population-based cohort studies including pregnancies conceived between April 2002 and March 2017 (Ontario, Canada; N = 554 272) and January 2003 to April 2011 [New South Wales (NSW), Australia; N = 139 229]. We evaluated the association between exposure to prescription amphetamine, methylphenidate, dextroamphetamine or lisdexamfetamine during pregnancy and pre-eclampsia, placental abruption, preterm birth, low birthweight, small for gestational age and neonatal intensive care unit admission. We used inverse probability of treatment weighting based on propensity scores to balance measured confounders between exposed and unexposed pregnancies. Additionally, we restricted the Ontario cohort to social security beneficiaries where supplementary confounder information was available. In Ontario and NSW respectively, 1360 (0.25%) and 146 (0.10%) pregnancies were exposed to psychostimulants. Crude analyses indicated associations between exposure and nearly all outcomes [OR range 1.15-2.16 (Ontario); 0.97-2.20 (NSW)]. Nearly all associations were attenuated after weighting. Pre-eclampsia was the exception: odds remained elevated in the weighted analysis of the Ontario cohort (OR 2.02, 95% CI 1.42-2.88), although some attenuation occurred in NSW (weighted OR 1.50, 95% CI 0.77-2.94) and upon restriction to social security beneficiaries (weighted OR 1.24, 95% CI 0.64-2.40), and confidence intervals were wide. We observed higher rates of outcomes among exposed pregnancies, but the attenuation of associations after adjustment and likelihood of residual confounding suggests psychostimulant exposure is not a major causal factor for most measured outcomes. Our findings for pre-eclampsia were inconclusive; exposed pregnancies may benefit from closer monitoring.


Genomic analysis of Epstein-Barr virus in nasal and peripheral T-cell lymphoma: a comparison with nasopharyngeal carcinoma in an endemic area.
The Epstein-Barr virus (EBV) is prevalent in nasal and peripheral T-cell lymphoma (NPTL) in Taiwan, where nasopharyngeal carcinoma (NPC) is endemic. In order to understand the pathogenesis of these two malignancies in this endemic area, genomic analysis of EBV in NPTL with comparison to NPC is important. We investigated the EBV subtype (types A and B), BamH-I "f" variant, and the Xho-I site mutant of the latent membrane protein-1 (LMP-1) gene in 19 cases of EBV-associated NPTL and in 30 cases of NPC. EBV DNA from three patients with infectious mononucleosis (IM) was simultaneously studied as representative of normal healthy carriers. Similar to NPC and IM, the EBV in NPTL was found to belong to the type A strain in the majority (18 of 19) of cases by analyzing the 3' divergence of EBNA-2 genes. The extra restriction enzyme site in the BamHI-F region ("f" variant) of EBV DNA was frequently (15 of 30) demonstrated in NPC, but only rarely (1 of 19) was it detected in NPTL and IM (0 of 3). The Xho-I site mutant of the LMP-1 gene previously characterized in Chinese NPC also prevailed in NPTL and IM with an identical nucleotide sequence. No correlation exists between the EBV subtype and its variants. In conclusion, type A EBV is prevalent in Taiwanese NPTL, a finding much distinct from the dominance of type B virus in nonendemic European patients. The EBV genomes in NPTL are closely similar to those in IM or normal healthy carriers, but are distinct from NPC for the infrequency of the "f" variant. The prevalence of the LMP-1 mutant in this endemic region suggests that this EBV strain may confer a growth advantage role in the pathogenesis of these EBV-associated diseases. The rarity of the "f" variant in NPTL and its high frequency in NPC may explain the differential tumorigenesis of different EBV strains.


Proglycogen, macroglycogen, glucose, and glucose-6-phosphate concentrations in skeletal muscles of horses with polysaccharide storage myopathy performing light exercise.
To determine concentrations of proglycogen (PG), macroglycogen (MG), glucose, and glucose-6-phosphate (G-6-P) in skeletal muscle of horses with polysaccharide storage myopathy (PSSM) before and after performing light submaximal exercise. 6 horses with PSSM and 4 control horses. Horses with PSSM completed repeated intervals of 2 minutes of walking followed by 2 minutes of trotting on a treadmill until muscle cramping developed. Four untrained control horses performed a similar exercise test for up to 20 minutes. Serum creatine kinase (CK) activity was measured before and 4 hours after exercise. Concentrations of total glycogen (G(t)), PG, MG, G-6-P, free glucose, and lactate were measured in biopsy specimens of gluteal muscle obtained before and after exercise. Mean serum CK activity was 26 times higher in PSSM horses than in control horses after exercise. Before exercise, muscle glycogen concentrations were 1.5, 2.2, and 1.7 times higher for PG, MG, and G(t), respectively, in PSSM horses, compared with concentrations in control horses. No significant changes in G(t), PG, MG, G-6-P, and lactate concentrations were detected after exercise. However, free glucose concentrations in skeletal muscle increased significantly in PSSM horses after exercise. Analysis of the results suggests that glucose uptake in skeletal muscle is augmented in horses with PSSM after light exercise. There is excessive storage of PG and MG in horses with PSSM, and high concentrations of the 2 glycogen fractions may affect functional interactions between glycogenolytic and glycogen synthetic enzymes and glycosomes.


Boll weevil (Coleoptera: Curculionidae) survival through the seed cotton cleaning process in the cotton gin.
There is concern that gins located in boll weevil, Anthonomus grandis grandis Boheman, eradication zones may become points of reintroduction when they process cotton grown in a neighboring infested area. We estimated boll weevil survival through two typical machine sequences used in commercial cotton gins to clean and dry the seed cotton in advance of the gin stand, as well as separately through two incline cylinder cleaners or one or two tower dryers operating at different temperatures. Large numbers of laboratory-reared adult boll weevils were marked with fluorescent powder, fed into the test system, and recovered with the assistance of blacklights. We found no evidence of survival through the seed cotton cleaning systems even when the dryers were not heated, or when passed separately through the two incline cleaners alone. Upper confidence limits (95%) were calculated for the observed zero recoveries based on sample size and the binomial distribution, and these represent the statistical worst-case (i.e., highest) survival potential. Survival through heated tower dryers declined rapidly to zero at higher temperatures, especially when two dryers were running. Although we conclude that the potential for survival of weevils in the seed cotton to the gin stand is zero or close to zero, a small percentage of live weevils was recovered in the green boll/rock trap, which may represent the greatest threat of reintroduction at the gin. Escape of live weevils with the gin trash is also possible, and studies addressing this issue will be presented elsewhere.


Genetic redundancy of senescence-associated transcription factors in Arabidopsis.
Leaf senescence is a genetically programmed process that constitutes the last stage of leaf development, and involves massive changes in gene expression. As a result of the intensive efforts that have been made to elucidate the molecular genetic mechanisms underlying leaf senescence, 184 genes that alter leaf senescence phenotypes when mutated or overexpressed have been identified in Arabidopsis thaliana over the past two decades. Concurrently, experimental evidence on functional redundancy within senescence-associated genes (SAGs) has increased. In this review, we focus on transcription factors that play regulatory roles in Arabidopsis leaf senescence, and describe the relationships among gene duplication, gene expression level, and senescence phenotypes. Previous findings and our re-analysis demonstrate the widespread existence of duplicate SAG pairs and a correlation between gene expression levels in duplicate genes and senescence-related phenotypic severity of the corresponding mutants. We also highlight effective and powerful tools that are available for functional analyses of redundant SAGs. We propose that the study of duplicate SAG pairs offers a unique opportunity to understand the regulation of leaf senescence and can guide the investigation of the functions of redundant SAGs via reverse genetic approaches.


Anesthetic management of patients undergoing extrapleural pneumonectomy for mesothelioma.
Extrapleural pneumonectomy is a radical and aggressive surgery that presents a great challenge to the thoracic anesthesiologist. This surgery is performed routinely by only a few centers in the world and this review represents our institution's experience in anesthetic care. Prominent among the developing multimodal treatment options is the combination of extrapleural pneumonectomy with intraoperative intracavitary hyperthermic chemotherapy. Outcome survival benefits have recently been demonstrated for the less completely cytoreductive pleurectomy procedure when combined with intraoperative intracavitary hyperthermic chemotherapy and trials are well under way for extrapleural pneumonectomy plus intraoperative intracavitary hyperthermic chemotherapy. Anesthetic management of extrapleural pneumonectomy is further impacted by these developments. Anesthetic management importantly contributes to containment of the perioperative complications of extrapleural pneumonectomy. An appreciation of the technical aspects and physiologic disruptions associated with extrapleural pneumonectomy is critical to effective management. While data on this relatively uncommon surgical procedure are scarce, some referral centers have accumulated extensive experience. This review summarizes relevant surgical aspects and anesthetic insights from the Brigham and Women's Hospital experience. Included are the anesthetic implications of intraoperative intracavitary hyperthermic chemotherapy in combination with extrapleural pneumonectomy - an emerging therapeutic option in the treatment of malignant pleural mesothelioma.


Change in enzyme activity and ribonucleic acid concentration within the epidermal cell of the rat during the growth stage of the hair cycle.
The rat hair cycle produces rapid anatomical and physiological changes in the entire skin. By the 4th day of the cycle, the epidermis has doubled in thickness. The present study shows that, during this proliferative period, there is a correlation between the biological state (growth) and the molecular composition (enzyme concentrations) of the epidermal cell. In proliferated 4th day epidermis there has occurred not only an increase in the number of viable cells but also an increase in succinic dehydrogenase and cytochrome oxidase activity and ribonucleic acid (RNA) content in the viable epidermal cell. Moreover, RNA content and enzyme activities appear to be correlated. The significance of these findings is discussed in the light of recent evidence supporting the relationship between RNA and enzyme synthesis and the role of hormones in the induction of enzyme adaptation.