Characterisation of a laminarin sulphate which inhibits basic fibroblast growth factor binding and endothelial cell proliferation.
We have evaluated a series of laminarin sulphates with different degrees of sulphation (0.3-2.3) as antagonists of basic fibroblast growth factor (bFGF) and as inhibitors of the bFGF-dependent endothelial cell line FBHE. Inhibition of binding of bFGF by the laminarin sulphates increased with increasing degree of sulphation. Binding of bFGF to low affinity sites on BHK cells was inhibited more strongly than binding to high affinity sites. IC50 values for inhibition of binding to low and high affinity sites by the most highly sulphated laminarin sulphate (LAM S5; degree of sulphation 2.31) were 12 +/- 8 micrograms/ml and 69 +/- 66 micrograms/ml, respectively. LAM S5 dissociated bFGF from low affinity sites on BHK cells but not from high affinity sites. LAM S5 increased the electrophoretic mobility of bFGF indicating that LAM S5 binds directly to bFGF. LAM S5 reduced uptake of bFGF by FBHE cells by 67%. Increasing the degree of sulphation of laminarin sulphates increased the inhibition of bFGF-stimulated DNA synthesis of the endothelial cell line FBHE (IC50 for LAM S5 approx. 1 microgram/ml). There was no inhibition of DNA synthesis of FBHE cells by LAM S5 in the presence of 1 microgram/ml bFGF indicating that bFGF antagonism is involved in the anti-proliferative activity of this compound. LAM S5 may be of value against diseases associated with bFGF-dependent cell proliferation.


Benzylaminoethyureido-Tailed Benzenesulfonamides: Design, Synthesis, Kinetic and X-ray Investigations on Human Carbonic Anhydrases.
A drug design strategy of carbonic anhydrase inhibitors (CAIs) belonging to sulfonamides incorporating ureidoethylaminobenzyl tails is presented. A variety of substitution patterns on the ring and the tails, located on para- or meta- positions with respect to the sulfonamide warheads were incorporated in the new compounds. Inhibition of human carbonic anhydrases (hCA) isoforms I, II, IX and XII, involving various pathologies, was assessed with the new compounds. Selective inhibitory profile towards hCA II was observed, the most active compounds being low nM inhibitors (KIs of 2.8-9.2 nM, respectively). Extensive X-ray crystallographic analysis of several sulfonamides in an adduct with hCA I allowed an in-depth understanding of their binding mode and to lay a detailed structure-activity relationship.


Characterization of putative multidrug resistance transporters of the major facilitator-superfamily expressed in Salmonella Typhi.
Multidrug resistance mediated by efflux pumps is a well-known phenomenon in infectious bacteria. Although much work has been carried out to characterize multidrug efflux pumps in Gram-negative and Gram-positive bacteria, such information is still lacking for many deadly pathogens. The aim of this study was to gain insight into the substrate specificity of previously uncharacterized transporters of Salmonella Typhi to identify their role in the development of multidrug resistance. S. Typhi genes encoding putative members of the major facilitator superfamily were cloned and expressed in the drug-hypersensitive Escherichia coli strain KAM42, and tested for transport of 25 antibacterial compounds, including representative antibiotics of various classes, antiseptics, dyes and detergents. Of the 15 tested putative transporters, STY0901, STY2458 and STY4874 exhibited a drug-resistance phenotype. Among these, STY4874 conferred resistance to at least ten of the tested antimicrobials: ciprofloxacin, norfloxacin, levofloxacin, kanamycin, streptomycin, gentamycin, nalidixic acid, chloramphenicol, ethidium bromide, and acriflavine, including fluoroquinolone antibiotics, which were drugs of choice to treat S. Typhi infections. Cell-based functional studies using ethidium bromide and acriflavine showed that STY4874 functions as a H(+)-dependent exporter. These results suggest that STY4874 may be an important drug target, which can now be tested by studying the susceptibility of a STY4874-deficient S. Typhi strain to antimicrobials.


The continuing enigma of the fetal echogenic intracardiac focus in prenatal ultrasound.
The present article aims to review recent literature about intracardiac echogenic foci (ICEFs), with special emphasis in potential causes, relationships with aneuploidy and cardiac function. The presence of an ICEF may be considered as a soft marker that may be of interest only in high-risk populations for chromosomal abnormalities. In cases of isolated ICEF in euploid fetuses there is not evidence of an altered cardiac function and a detailed echocardiogram is not recommended as long as the second trimester scan is normal. Cases of fetal myocardial diffuse or multiple calcifications may be related to inflammatory and hypoxic changes in fetal heart, but the relationships between these conditions and isolated ICEF remain unclear. The origin of ICEF remains unclear. It is considered a normal developmental variant, but either inflammatory or hypoxic processes could be involved in its appearance. In low-risk populations for aneuploidy, the presence of an ICEF is not an indication for invasive procedures. In high-risk populations this depends on the previous calculated risk. In euploid fetuses, cardiac function is not significantly altered but further studies in specific populations may be justified. Fetuses with diffuse myocardial calcification or multiple foci may need further investigations.


Establishment and validation of early prediction model for hypertriglyceridemic severe acute pancreatitis.
The prevalence of hypertriglyceridaemia-induced acute pancreatitis (HTG-AP) is increasing due to improvements in living standards and dietary changes. However, currently, there is no clinical multifactor scoring system specific to HTG-AP. This study aimed to screen the predictors of HTG-SAP and combine several indicators to establish and validate a visual model for the early prediction of HTG-SAP. The clinical data of 266 patients with HTG-SAP were analysed. Patients were classified into severe (N = 42) and non-severe (N = 224) groups according to the Atlanta classification criteria. Several statistical analyses, including one-way analysis, least absolute shrinkage with selection operator (LASSO) regression model, and binary logistic regression analysis, were used to evaluate the data. The univariate analysis showed that several factors showed no statistically significant differences, including the number of episodes of pancreatitis, abdominal pain score, and several blood diagnostic markers, such as lactate dehydrogenase (LDH), serum calcium (Ca2+), C-reactive protein (CRP), and the incidence of pleural effusion, between the two groups (P < 0.000). LASSO regression analysis identified six candidate predictors: CRP, LDH, Ca2+, procalcitonin (PCT), ascites, and Balthazar computed tomography grade. Binary logistic regression multivariate analysis showed that CRP, LDH, Ca2+, and ascites were independent predictors of HTG-SAP, and the area under the curve (AUC) values were 0.886, 0.893, 0.872, and 0.850, respectively. The AUC of the newly established HTG-SAP model was 0.960 (95% confidence interval: 0.936-0.983), which was higher than that of the bedside index for severity in acute pancreatitis (BISAP) score, modified CT severity index, Ranson score, and Japanese severity score (JSS) CT grade (AUC: 0.794, 0.796, 0.894 and 0.764, respectively). The differences were significant (P < 0.01), except for the JSS prognostic indicators (P = 0.130). The Hosmer-Lemeshow test showed that the predictive results of the model were highly consistent with the actual situation (P > 0.05). The decision curve analysis plot suggested that clinical intervention can benefit patients when the model predicts that they are at risk for developing HTG-SAP. CRP, LDH, Ca2+, and ascites are independent predictors of HTG-SAP. The prediction model constructed based on these indicators has a high accuracy, sensitivity, consistency, and practicability in predicting HTG-SAP.


Cotranscriptional association of mRNA export factor Yra1 with C-terminal domain of RNA polymerase II.
The unique C-terminal domain (CTD) of RNA polymerase II, composed of tandem heptad repeats of the consensus sequence YSPTSPS, is subject to differential phosphorylation throughout the transcription cycle. Several RNA processing factors have been shown to bind the phosphorylated CTD and use it to localize to nascent pre-mRNA during transcription. In Saccharomyces cerevisiae, the mRNA export protein Yra1 (ALY/RNA export factor in metazoa) cotranscriptionally associates with mRNA and delivers it to the nuclear pore complex for export to the cytoplasm. Here we report that Yra1 directly binds in vitro the hyperphosphorylated form of the CTD characteristic of elongating RNA polymerase II and contains a phospho-CTD-interacting domain within amino acids 18-184, which also include an "RNA recognition motif" (RRM) (residues 77-184). Using UV cross-linking, we showed that the RRM alone binds RNA, although a larger segment extending to the C terminus (amino acids 77-226) displayed stronger RNA binding activity. Although the RRM is implicated in both RNA and CTD binding, RRM point mutations separated these two functions. Both functions are important in vivo as RNA binding-defective or CTD binding-defective versions of Yra1 engendered growth and mRNA export defects. We also report the construction and characterization of a useful new temperature-sensitive YRA1 allele (R107A/F126A). Using ChIP, we demonstrated that removing the N-terminal 76 amino acids of Yra1 (all of the phospho-CTD-interacting domain up to the RRM) results in a 10-fold decrease in Yra1 recruitment to genes during elongation. These results indicate that the phospho-CTD is likely involved directly in the cotranscriptional recruitment of Yra1.


Pregnancy outcomes after conservative surgical management of ovarian neoplasms treated at a single institution.
Conservative surgical management of ovarian neoplasms can result in favorable oncologic and obstetric outcomes. All reproductive-age women (18-45 years old) with stage IA to stage IIC ovarian neoplasms (N = 161) were retrospectively identified from a single institution's tumor registry between 1990 and 2007. Operative, pathological, outpatient, and delivery records were reviewed to confirm histological findings, stage, extent of surgical resection, adjuvant treatment, oncologic outcomes, and pregnancy outcomes. Women who underwent conservative surgical management for ovarian neoplasms (n = 61 [37.9%]) were identified, including those with low malignant potential (LMP, n = 36), epithelial (n = 12), germ cell (n = 6), and sex cord (n = 7) tumors. Thirteen women conceived 23 pregnancies, producing a pregnancy rate of 25.0% overall and of 68.4% for those attempting conception. Women with LMP tumors conceived most pregnancies and had the highest number of antenatal complications. Of those receiving adjuvant chemotherapy (n = 8), 12.5% were able to conceive after their treatment and with no reported congenital anomalies. Pregnancy after a diagnosis of ovarian neoplasm did not impact disease recurrence (0% vs 7.7%, P = 0.56) or survival (100% vs 100%, P = 1.0). Conservative surgery may be an acceptable option for reproductive-age women with early-stage ovarian neoplasms. We report pregnancy and oncologic outcomes for a cohort of women managed conservatively for LMP, epithelial, germ cell, and sex cord ovarian neoplasms.


Neonatal bleeding in transgenic mice expressing urokinase-type plasminogen activator.
Spontaneous intestinal and intra-abdominal bleeding was observed in a high percentage of newborn transgenic mice carrying the murine urokinase-type plasminogen activator (uPA) gene linked to the albumin enhancer/promoter. These hemorrhagic events were directly related to transgene expression in the liver and the development of high plasma uPA levels. Two lines were established from surviving founder mice that displayed multigenerational transmission of the bleeding phenotype. Fatal hemorrhaging developed between 3 and 84 hr after birth in about half of the transgenic offspring of these lines; transgenic pups that did not bleed nevertheless passed the phenotype to their young. The phenotypic variability could not be explained by differences in transgene expression. All transgenic neonates were severely hypofibrinogenemic and displayed loss of clotting function that extended beyond the risk period for bleeding. These mice provide a means of studying the pathophysiology of plasminogen hyperactivation and evaluating therapeutic protocols designed to prevent bleeding.


Racial Differences in DNA-Methylation of CpG Sites Within Preterm-Promoting Genes and Gene Variants.
Objective To evaluate the role DNA methylation may play in genes associated with preterm birth for higher rates of preterm births in African-American women. Methods Fetal cord blood samples from births collected at delivery and maternal demographic and medical information were used in a cross-sectional study to examine fetal DNA methylation of genes implicated in preterm birth among black and non-black infants. Allele-specific DNA methylation analysis was performed using a methylation bead array. Targeted maximum likelihood estimation was applied to examine the relationship between race and fetal DNA methylation of candidate preterm birth genes. Receiver-operating characteristic analyses were then conducted to validate the CpG site methylation marker within the two racial groups. Bootstrapping, a method of validation and replication, was employed. Results 42 CpG sites were screened within 20 candidate gene variants reported consistently in the literature as being associated with preterm birth. Of these, three CpG sites on TNFAIP8 and PON1 genes (corresponding to: cg23917399; cg07086380; and cg07404485, respectively) were significantly differentially methylated between black and non-black individuals. The three CpG sites showed lower methylation status among infants of black women. Bootstrapping validated and replicated results. Conclusion for Practice Our study identified significant differences in levels of methylation on specific genes between black and non-black individuals. Understanding the genetic/epigenetic mechanisms that lead to preterm birth may lead to enhanced prevention strategies to reduce morbidity and mortality by eventually providing a means to identify individuals with a genetic predisposition to preterm labor.


Dislocation of a primary total hip arthroplasty is more common in patients with a lumbar spinal fusion.
Lumbar fusion is known to reduce the variation in pelvic tilt between standing and sitting. A flexible lumbo-pelvic unit increases the stability of total hip arthroplasty (THA) when seated by increasing anterior clearance and acetabular anteversion, thereby preventing impingement of the prosthesis. Lumbar fusion may eliminate this protective pelvic movement. The effect of lumbar fusion on the stability of total hip arthroplasty has not previously been investigated. The Medicare database was searched for patients who had undergone THA and spinal fusion between 2005 and 2012. PearlDiver software was used to query the database by the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) procedural code for primary THA and lumbar spinal fusion. Patients who had undergone both lumbar fusion and THA were then divided into three groups: 1 to 2 levels, 3 to 7 levels and 8+ levels of fusion. The rate of dislocation in each group was established using ICD-9-CM codes. Patients who underwent THA without spinal fusion were used as a control group. Statistical significant difference between groups was tested using the chi-squared test, and significance set at p < 0.05. At one-year follow-up, 14 747 patients were found to have had a THA after lumbar spinal fusion (12 079 1 to 2 levels, 2594 3 to 7 levels, 74 8+ levels). The control group consisted of 839 004 patients. The dislocation rate in the control group was 1.55%. A higher rate of dislocation was found in patients with a spinal fusion of 1 to 2 levels (2.96%, p < 0.0001) and 3 to 7 levels (4.12%, p < 0.0001). Patients with 3 to 7 levels of fusion had a higher rate of dislocation than patients with 1 to 2 levels of fusion (odds ratio (OR) = 1.60, p < 0.0001). When groups were matched for age and gender to the unfused cohort, patients with 1 to 2 levels of fusion had an OR of 1.93 (95% confidence interval (CI) 1.42 to 2.32, p < 0.001), and those with 3 to 7 levels of fusion an OR of 2.77 (CI 2.04 to 4.80, p < 0.001) for dislocation. Patients with a previous history of lumbar spinal fusion have a significantly higher rate of dislocation of their THA than age- and gender-matched patients without a lumbar spinal fusion. Cite this article: Bone Joint J 2017;99-B:585-91.


cdk1- and cdk2-mediated phosphorylation of MyoD Ser200 in growing C2 myoblasts: role in modulating MyoD half-life and myogenic activity.
We have examined the role of protein phosphorylation in the modulation of the key muscle-specific transcription factor MyoD. We show that MyoD is highly phosphorylated in growing myoblasts and undergoes substantial dephosphorylation during differentiation. MyoD can be efficiently phosphorylated in vitro by either purified cdk1-cyclin B or cdk1 and cdk2 immunoprecipitated from proliferative myoblasts. Comparative two-dimensional tryptic phosphopeptide mapping combined with site-directed mutagenesis revealed that cdk1 and cdk2 phosphorylate MyoD on serine 200 in proliferative myoblasts. In addition, when the seven proline-directed sites in MyoD were individually mutated, only substitution of serine 200 to a nonphosphorylatable alanine (MyoD-Ala200) abolished the slower-migrating hyperphosphorylated form of MyoD, seen either in vitro after phosphorylation by cdk1-cyclin B or in vivo following overexpression in 10T1/2 cells. The MyoD-Ala200 mutant displayed activity threefold higher than that of wild-type MyoD in transactivation of an E-box-dependent reporter gene and promoted markedly enhanced myogenic conversion and fusion of 10T1/2 fibroblasts into muscle cells. In addition, the half-life of MyoD-Ala200 protein was longer than that of wild-type MyoD, substantiating a role of Ser200 phosphorylation in regulating MyoD turnover in proliferative myoblasts. Taken together, our data show that direct phosphorylation of MyoD Ser200 by cdk1 and cdk2 plays an integral role in compromising MyoD activity during myoblast proliferation.


An analysis of practice-level mortality data to inform a health needs assessment.
The utility of practice death registers has been indicated but, in the wake of the recent Harold Shipman case in the United Kingdom, the value of individual practice-level analysis has been questioned. To assess the value of analysing practice-level mortality data to inform health needs assessment. Comparative analyses of mortality. Two large practices, an inner-city study practice, and a reference practice in a medium-sized town. All premature deaths (aged one to 74 years) during 1994-1998 at the study practice (n = 170), and reference practice (n = 340), were identified. Cause-specific standardised mortality ratios (SMRs) were calculated using national reference data. The proportions of the total number of years of life lost (YLL) up to age 75 years associated with alcoholism, drug dependency, and severe mental illness were calculated and a comparison between practices was made, using standardised proportional mortality methods. Significantly raised SMRs for the study practice were lung cancer (SMR = 234), digestive system diseases (SMR = 362), and injuries and poisonings (SMR = 180). Having standardised for age, there were nearly four times as many YLLs in the study practice population associated with a history of alcoholism, and over three times as many associated with drug dependency, compared with the reference practice. Mortality analyses can provide useful insights for informing needs assessment in an individual practice. Small number problems may occur with smaller practice populations, but collation of data at PCG/T level also has potential utility. The study reinforces the argument that practices need to set up and maintain complete and accurate death registers.


STATURE THROUGHOUT THE WORLD.
Tropic jungle life has an influence that decreases stature, and so has Arctic cold and waste. In each there is difficulty in procuring proper food, and discomfort in the extreme. The active life of the temperate zone with its comforts and abundant food supply produces the tallest statures. The greatest extremes of small stature are found among the Negrillos of Central Africa and the Aymaras of Central South America, in the jungles of excessive heat and poor food supply. Next to these come the Eskimos, Lapps and Siberians, with ice and excessive cold and poor food supply. On the other hand, the littoral and southern Baltic regions in Europe, the western part of Asia, eastern Africa, and the plains and pampas of the Americas with their active life, abundant food supply and temperate climate produce the tallest statures. Certain stocks may move into areas for which they are not fitted and remain for a time, and such conditions exist throughout the world to-day where recent movements of peoples have taken place, but ultimately there is a survival of the stock best fitted for the environment, and the unfitted stocks disappear by amalgamation, eradication or dispersal. Sea areas and probably sea food have an influence in reducing stature. The present Mediterranean peoples and the primordial British have small statures and so does Japan, yet they came from taller continental stocks. The Central Americans and Fuegians are smaller than the continental peoples near-by. The Malays and southern Asiaties are smaller than the peoples of the interior of the continent. Other instances might be cited. There is some evidence that the seaboard statures of the United States are less than those of the interior, but other factors enter here. Looked at in its broadest sense, environment molds the individual, selection retains the fittest under different environments, and heredity carries on the results.


Laboratory findings, histopathology, and immunophenotype of lymphoma in domestic ferrets.
Lymphoma is a common tumor in ferrets, but anatomic distribution, histomorphology, immunophenotype, laboratory abnormalities, and response to chemotherapy are incompletely defined. In this study, lymphoma was diagnosed by histopathology of tumor tissue in 29 ferrets ranging in age from 0.8 to 8.5 years, including 12 males and 17 females. Tumors involved the viscera of the abdominal cavity (n = 11), thoracic cavity (n = 1), or abdominal and thoracic cavities (n = 7); the skin (n = 2); or the viscera of both body cavities plus other sites (n = 8). Microscopically, all tumors had diffuse architecture. Assessment by histomorphology and immunophenotype classified tumors as peripheral T-cell lymphoma (n = 17), anaplastic large T-cell lymphoma (n = 5), anaplastic large B-cell lymphoma (n = 4), diffuse large B-cell lymphoma (n = 1), and Hodgkin-like lymphoma (n = 2). Cytologic evaluation of tumor tissue was diagnostic in 11 of 13 cases. Twenty-two of 27 ferrets had anemia, 2 had leukemia, and 5 were neutropenic. Common comorbid disorders were adrenal disease (n = 27) and insulinoma (n = 6). Tumors most frequently involved mesenteric lymph nodes, while enlargement of peripheral lymph nodes was uncommon (n = 3). Ferrets with Hodgkin-like lymphoma had massive enlargement of single lymph nodes. Mean survival of ferrets not immediately euthanized was 5.0 months (T-cell lymphoma) and 8.4 months (B-cell lymphoma). Ferrets treated with chemotherapy survived an average of 4.3 months (T-cell lymphoma, n = 9) or 8.8 months (B-cell lymphoma, n = 4). Results indicate that lymphomas in ferrets most commonly affect abdominal viscera, may be amenable to cytologic diagnosis, are frequently associated with anemia and, in some cases, may be chemosensitive, resulting in relatively long survival times.


Resistance, tolerance and environmental transmission dynamics determine host extinction risk in a load-dependent amphibian disease.
While disease-induced extinction is generally considered rare, a number of recently emerging infectious diseases with load-dependent pathology have led to extinction in wildlife populations. Transmission is a critical factor affecting disease-induced extinction, but the relative importance of transmission compared to load-dependent host resistance and tolerance is currently unknown. Using a combination of models and experiments on an amphibian species suffering extirpations from the fungal pathogen Batrachochytrium dendrobatidis (Bd), we show that while transmission from an environmental Bd reservoir increased the ability of Bd to invade an amphibian population and the extinction risk of that population, Bd-induced extinction dynamics were far more sensitive to host resistance and tolerance than to Bd transmission. We demonstrate that this is a general result for load-dependent pathogens, where non-linear resistance and tolerance functions can interact such that small changes in these functions lead to drastic changes in extinction dynamics.


[The development of the medical school in Brazil].
Since its beginnings in 1808, medical instruction in Brazil has evolved and changed as Brazilian society has done. Medical schools, at first a concern of government alone, between 1931 and 1960 began to attract private enterprise, which established six of the 19 schools founded during those years. Since then the share of the private sector has grown, and there was a veritable explosion between 1966 and 1970, when 30 new medical schools were set up, 25 of them in the private sector (more than half of the total of 76 schools in operation today are in the private sector). The authors note that the increase in the number of medical schools and courses has not made them less selective, diminished the importance of the government-operated schools--which are still the leading institutions--or made it more certain that the education imparted in the private schools is of satisfactory quality. The medical schools are clustered chiefly in the southeast, and they are partial to the industrial urban setting, especially the Rio-São Paulo axis (where half of the country's physicians are concentrated), which makes for yet another severe imbalance among regions.


Determination of aromatic hydrotropic drugs in pharmaceutical preparations by the surfactant-binding degree method.
An aggregation parameter-based analytical approach, the surfactant-dye binding degree (SDBD) method, was used, for the first time, to determine aromatic hydrotropic compounds. The anionic dye Coomassie Brilliant Blue G (CBBG) was used as inductor of didodecyldimethylammonium bromide (DDABr) aggregates, whose formation was monitored from changes in the spectral features of the dye. Interactions between hydrotrope and DDABr molecules resulted in a decrease of the degree of binding of the cationic surfactant to CBBG, which was proportional to the concentration of hydrotrope in the aqueous solution. The CBBG-DDABr-hydrotrope chemical system was found to fit to the mathematical expression previously derived for the determination of amphiphilic compounds. The hydrotrope-surfactant bond strength determined the sensitivity achieved for the determination of hydrotropic compounds, which was highly dependent on the molecular structure of the analyte. The high precision (the relative standard deviation for 7 mg l(-1) of salicylic acid was 0.8%), rapidity (measurements were performed in a few minutes) and low cost (in both instrumentation and reactants) of the proposed method, made it especially suitable for quality control. The practical analytical applicability of the SDBD method for the control of hydrotropic drugs in pharmaceutical preparations was demonstrated by quantifying salicylic acid and acetyl salicylic acid in liquid (solutions) and solid (tablets, granulates, unguents, gels and creams) samples, which were directly analyzed after dissolution of the samples.


Enzymatic determination of itoic acid, a Bacillus subtilis siderophore, and 2,3-dihydroxybenzoic acid.
A specific enzymatic method to determine the amounts of itoic acid, a Bacillus subtilis siderophore, and 2,3-dihydroxybenzoic acid (2,3-DHBA) was devised. A sample was incubated first with hippurate hydrolase and then with 2,3-DHBA-3,4-dioxygenase. Itoic acid was estimated from the increase in A374. The incubation with the first enzyme was omitted for the determination of 2,3-DHBA.


The herpes simplex virus 1 UL 17 gene is required for localization of capsids and major and minor capsid proteins to intranuclear sites where viral DNA is cleaved and packaged.
In nuclei of cells infected with herpes simplex virus (HSV), synthesized viral DNA accumulates as concatamers that are cleaved into genomic lengths and inserted into preformed capsids. Whereas newly replicated DNA and enzymes required for DNA synthesis accumulate in sites of infected cell nuclei termed replication compartments, the intranuclear site of DNA cleavage and packaging is currently controversial. DNA packaging requires the UL6, UL15, UL17, UL25, UL28, UL32, and UL33 genes in addition to the major capsid proteins. Using confocal immunofluorescence microscopy, it was observed that in > 95% of HEp-2 cells fixed at late times after infection with wild-type HSV-1, capsids, major capsid proteins ICP5 and ICP35, and the UL6-encoded minor capsid protein localized in DNA replication compartments. These data support the hypothesis that capsid assembly and DNA cleavage/packaging normally occur in HEp-2 cell replication compartments. In contrast, cells infected with a viral mutant lacking functional UL17 contained antigenically dense nuclear aggregates that stained with ICP35, ICP5, and capsid specific antibodies. Cells infected with the UL17 mutant virus also displayed UL6-specific fluorescence in a diffuse pattern at the nuclear periphery in regions not containing ICP35 and ICP5. Displacement of ICP35 from replication compartments was not observed in cells infected with cleavage/packaging mutants lacking UL28 and UL33. We conclude that the UL17 gene is required for correct targeting of capsids and major and minor capsid proteins to the DNA replication compartment of HEp-2 cells and deduce that this targeting reflects one functional role of UL17 in viral DNA cleavage and packaging.


Ischemic pre-conditioning preserves brain mitochondrial functions during the middle cerebral artery occlusion in rat.
It is well known that a brief period of ischemia increases tolerance to a subsequent severe ischemic episode. In the present study, bilateral carotid arteries occlusion (BCAO) was applied as pre-conditioning to testify whether this kind of ischemia could preserve the function of mitochondria with the impairment induced by middle cerebral artery occlusion (MCAO) in brain. The activities of respiratory enzyme complex I to IV, mitochondria swelling, membrane potential, and membrane fluidity were investigated. The results showed that the percentage of infarct area decreased greatly due to the ischemic pre-conditioning (IP) revealing the preventive effect of IP on infarct size. The activities of respiratory enzyme complex III, IV were effectively preserved (p < 0.05, p < 0.05) compared with MCAO group through ischemic pre-conditioning. Mitochondrial swelling and membrane fluidity were protected by IP, and also an increased trend was found in membrane potential, which indicated that the integrity of mitochondrial membrane was maintained. It suggested that the function of mitochondrial energy metabolism in brain ischemia was effectively protected by this kind of pre-conditioning.


[Summary of the Dutch College of General Practitioners' practice guideline 'Dizziness'].
In the Dutch College of General Practitioners' practice guideline entitled 'Dizziness', a distinction is made between two types of dizziness. The first is vertigo which is characterised by a sense of spinning or movement and which is caused by problems with the vestibular organ. The second is lightheadedness or the feeling that one is going to faint; the cause of this problem is found outside the vestibular organ. In making the diagnosis, history-taking and a limited physical examination is usually sufficient. Providing the patient with information plays an important role in the treatment. Medication for dizziness is not advised. In case of nausea and vomiting, specific medication can be prescribed to treat these symptoms. Referral to secondary care is indicated if there are signs of a serious condition or if additional investigations are required.


Employment Status and Sick Leave After First-Time Implantable Cardioverter Defibrillator Implantation: Results From the COPE-ICD Trial.
In the Copenhagen Outpatient Programme-Implantable Cardioverter Defibrillator (COPE-ICD) Trial, a positive effect from a cost-saving, comprehensive cardiac rehabilitation program was found on physical and mental health in patients with ICDs. In the context of the COPE-ICD Trial, the aims of this study is to investigate (a) employment status after ICD implantation, (b) the number of sick days related to ICD implantation, (c) differences in employment status and sick days between rehabilitation and usual care groups, and (d) predictors of employment status and sick leave. Patients with first-time ICD implantation were randomized to comprehensive cardiac rehabilitation or usual care. One year after ICD implantation, patients answered questions about employment status and sick leave. Differences were tested using the Student t test and the χ test. Predictors of employment status and sick leave were tested using logistic regression and linear regression models. A total of 196 patients were randomized. The questionnaire was completed by 138 patients (70%). In total, 47% had worked before ICD implantation. After 1 year, 81% were still working and their mean (SD) number of sick days was 33.8 (58.3). Age 60 years or younger and secondary ICD indication were predictors of working after 1 year. Patients with secondary ICD indication had more sick days and patients who were not married had fewer sick days. Most patients who worked before ICD implantation returned to work after the ICD was placed. Those who were married and had an ICD for secondary prevention took more sick days after the ICD than did those without these characteristics. Those who were younger and have a secondary indication ICD were more likely to be working 1 year after ICD implantation.


Cyclin C Regulated Oxidative Stress Responsive Transcriptome in Mus musculus Embryonic Fibroblasts.
The transcriptional changes that occur in response to oxidative stress help direct the decision to maintain cell viability or enter a cell death pathway. Cyclin C-Cdk8 is a conserved kinase that associates with the RNA polymerase II Mediator complex that stimulates or represses transcription depending on the locus. In response to oxidative stress, cyclin C, but not Cdk8, displays partial translocation into the cytoplasm. These findings open the possibility that cyclin C relocalization is a regulatory mechanism governing oxidative stress-induced transcriptional changes. In the present study, the cyclin C-dependent transcriptome was determined and compared to transcriptional changes occurring in oxidatively stressed Mus musculus embryonic fibroblasts. We observed a similar number (∼2000) of genes up or downregulated in oxidatively stressed cells. Induced genes include cellular repair/survival factors while repressed loci were generally involved in proliferation or differentiation. Depleting cyclin C in unstressed cells produced an approximately equal number of genes (∼2400) that were repressed by, or whose transcription required, cyclin C. Consistent with the possibility that cyclin C nuclear release contributes to transcriptional remodeling in response to oxidative stress, we found that 37% cyclin C-dependent genes were downregulated following stress. Moreover, 20% of cyclin C- repressed genes were induced in response to stress. These findings are consistent with a model that cyclin C relocalization to the cytoplasm, and corresponding inactivation of Cdk8, represents a regulatory mechanism to repress and stimulate transcription of stress-responsive genes.


Ring-opening polymerization of cyclohexene oxide using aluminum amine-phenolate complexes.
Remarkably active (down to 0.001% Al) catalysts for ring-opening polymerization of cyclohexene oxide under neat reaction conditions are reported. High molecular weight polymers with uniform dispersity are produced. Kinetic data from NMR studies and MALDI-TOF MS data of the polymers provide some mechanistic insight.


Regulation of RNA editing by RNA-binding proteins in human cells.
Adenosine-to-inosine (A-to-I) editing, mediated by the ADAR enzymes, diversifies the transcriptome by altering RNA sequences. Recent studies reported global changes in RNA editing in disease and development. Such widespread editing variations necessitate an improved understanding of the regulatory mechanisms of RNA editing. Here, we study the roles of >200 RNA-binding proteins (RBPs) in mediating RNA editing in two human cell lines. Using RNA-sequencing and global protein-RNA binding data, we identify a number of RBPs as key regulators of A-to-I editing. These RBPs, such as TDP-43, DROSHA, NF45/90 and Ro60, mediate editing through various mechanisms including regulation of ADAR1 expression, interaction with ADAR1, and binding to Alu elements. We highlight that editing regulation by Ro60 is consistent with the global up-regulation of RNA editing in systemic lupus erythematosus. Additionally, most key editing regulators act in a cell type-specific manner. Together, our work provides insights for the regulatory mechanisms of RNA editing.


[Sequencing and analysis of the complete mitochondrial genome of Remiz consobrinus].
The complete mitochondrial genome sequence of Remiz consobrinus was determined using long PCR and conserved primers walking approaches. Based on the results of assembling and annotation, the structure, sequence composition and codon usage of the genome protein-coding genes were analyzed, and the prediction of the secondary structure of 22 tRNA and 2 rRNA, the control region structure, and the phylogeny were also conducted, which provided new information for phylogenetic studies of passerine birds. The entire mitochondrial genome of Remiz consobrinus was 16 737 bp in length, the accession number was KC463856 and the content of A, T, C, and G were 27.8%, 21.5%, 35.4%, and 15.3%, respectively. The genome harbored the same gene order with that of other birds, and contained 13 protein coding genes (PCGs), 22 tRNA, 2 rRNA, and a non-coding control region. There were 77 bp intergenic intervals between 18 pair genes and 30 bp overlaps between 7 pair genes. Except for ND3 gene, which used ATT as the initiation codon, all other PCGs started with the typical ATG codon. Except for COIII and ND4, which used incomplete termination codon T, the other 11 PCGs used standard TAA, TAG, AGA or AGG as termination codons. The tRNAs all formed typical cloverleaf secondary structure, except for tRNASer-AGN, which lost the DHU arm in its structure. A total of 27 base mismatches appeared, with 19 common G-U mismatches. The predicted secondary structure of SrRNA and LrRNA contained 3 domains with 47 helices and 6 domains with 60 helices, respectively. Besides F-box, D-box, C-box, and B-box, Bird similarity-box and CSB1-box were also found in the control region of Remiz consobrinus, as found in other bird species. Our results suggest Remizidae as a separate family. The monophyly of Sylviidae and Remizidae was supported.


Ceftaroline fosamil, a cephalosporin derivative for the potential treatment of MRSA infection.
Forest Laboratories (following its acquisition of Cerexa Inc, a spin-out from Peninsula Pharmaceuticals Inc), under license from Takeda, is developing ceftaroline fosamil, an N-phosphono prodrug of ceftaroline, a derivative of a fourth-generation cephalosporin, for the potential treatment of methicillin-resistant Staphylococcus aureus infection. Phase III clinical trials evaluating ceftaroline fosamil for community acquired pneumonia and complicated skin and skin structure infections are underway.


Recurrence, death, and secondary malignancy after ovarian conservation for young women with early-stage low-grade endometrial cancer.
To examine the association between ovarian conservation and oncologic outcome in surgically-treated young women with early-stage, low-grade endometrial cancer. This multicenter retrospective study examined women aged <50 with stage I grade 1-2 endometrioid endometrial cancer who underwent primary surgery with hysterectomy from 2000 to 2014 (US cohort n = 1196, and Japan cohort n = 495). Recurrence patterns, survival, and the presence of a metachronous secondary malignancy were assessed based on ovarian conservation versus oophorectomy. During the study period, the ovarian conservation rate significantly increased in the US cohort from 5.4% to 16.4% (P = 0.020) whereas the rate was unchanged in the Japan cohort (6.3-8.7%, P = 0.787). In the US cohort, ovarian conservation was not associated with disease-free survival (hazard ratio [HR] 0.829, 95% confidence interval [CI] 0.188-3.663, P = 0.805), overall survival (HR not estimated, P = 0.981), or metachronous secondary malignancy (HR 1.787, 95% CI 0.603-5.295, P = 0.295). In the Japan cohort, ovarian conservation was associated with decreased disease-free survival (HR 5.214, 95% CI 1.557-17.464, P = 0.007) and an increased risk of a metachronous secondary malignancy, particularly ovarian cancer (HR 7.119, 95% CI 1.349-37.554, P = 0.021), but was not associated with overall survival (HR not estimated, P = 0.987). Ovarian recurrence or metachronous secondary ovarian cancer occurred after a median time of 5.9 years, and all cases were salvaged. Our study suggests that adoption of ovarian conservation in young women with early-stage low-grade endometrial cancer varies by population. Ovarian conservation for young women with early-stage, low-grade endometrial cancer may be potentially associated with increased risks of ovarian recurrence or metachronous secondary ovarian cancer in certain populations; nevertheless, ovarian conservation did not negatively impact overall survival.


[Amiloride sensitive sodium channels (ENaC) and their regulation by proteases].
ENaC is a channel that mediates entry of Na+ from the luminal fluid into the cells in many reabsorbing epithelia and it is also expressed in human placenta. ENaC is crucial in the control of electrolyte and extracellular volume homeostasis. ENaC is regulated by several hormones, including aldosterone and blocked by amiloride and its analogs. ENaC channels are composed by three homologous subunits, α, β and γ that form the pore where Na ions are transported. Two factors regulate the activity of ENaC channels: 1) the number of channels inserted in the membrane and 2) the open probability of the channels or time that the channel is open. The number of channels is the result of a balance between the synthesis and degradation of ENaC channels. The open probability depends on the proteolysis of specific segments in the α and γ subunits of ENaC by multiple proteases inside of the cell or in the extracellular space. Among the most studied proteases are furin, prostasin, elastase, plasmin and trypsin. There are endogenous substances that block the activity of these proteases such as aprotinin, bikunin and nexin-1 and the expression of both, proteases and their inhibitors are controlled by the rate of Na+ movement, aldosterone and TFG-β levels. In this work we present some examples of this regulation and the potential role that this process may play under normal and pathological conditions such as cystic fibrosis, kidney diseases and hypertension.


The influence of foot position on lower leg muscle activity during a heel raise exercise measured with fine-wire and surface EMG.
Exercises for lower leg muscles are important to improve function. To examine the influence of foot position on lower leg muscle activity during heel raises. Cross-sectional laboratory study. Laboratory. Fourteen healthy men participated in this study. The muscle activity levels of the tibialis posterior (TP), peroneus longus (PL), flexor digitorum longus (FDL) and medial gastrocnemius (MG) were measured. The heel raises consisted of three foot positions: 1) neutral, 2) 30° abduction, and 3) 30° adduction. The EMG data for five repetitions of each foot position were normalized to maximum voluntary contraction. One-way repeated measure ANOVA was employed for statistical analysis. The muscle activity level of TP, PL and FDL was significantly different between the three foot positions during the heel raises. TP and FDL showed the highest activity level in 30° foot adduction while PL demonstrated the highest activity level in 30° foot abduction. Heel raises with 30° foot adduction and abduction positions can change lower leg muscle activity; These findings suggest that altering foot posture during the heel raise exercise may benefit patients with impaired TP, PL or FDL function.


Immune Phenotyping of Patients With Acute Vogt-Koyanagi-Harada Syndrome Before and After Glucocorticoids Therapy.
Previous studies have established that disturbed lymphocytes are involved in the pathogenesis of Vogt-Koyanagi-Harada (VKH) syndrome. Accordingly, glucocorticoids (GCs), with their well-recognized immune-suppressive function, have been widely used for treatment of VKH patients with acute relapses. However, the systemic response of diverse immune cells to GC therapy in VKH is poorly characterized. To address this issue, we analyzed immune cell subpopulations and their phenotype, as well as cytokine profiles in peripheral blood from VKH patients (n=25) and health controls (HCs, n=21) by flow cytometry and luminex technique, respectively. For 16 patients underwent GC therapy (methylprednisolone, MP), the aforementioned measurements as well as the transcriptome data from patients before and after one-week's GC therapy were also compared to interrogate the systemic immune response to GC therapy. Lymphocyte composition in the blood was different in VKH patients and HCs. VKH patients had significantly higher numbers of T cells with more activated, polarized and differentiated phenotype, more unswitched memory B cells and monocytes, as compared to HCs. MP treatment resulted in decreased frequencies of T cells and NK cells, inhibited NK cell activation and T cell differentiation, and more profoundly, a marked shift in the distribution of monocyte subsets. Collectively, our findings suggest that advanced activation and differentiation, as well as dysregulated numbers of peripheral lymphocytes are the major immunological features of VKH, and GC therapy with MP not only inhibits T cell activation directly, but also affects monocyte subsets, which might combinatorically result in the inhibition of the pathogenic immune response.


Factors associated with soil-transmitted helminths infection in Benin: Findings from the DeWorm3 study.
Despite several years of school-based MDA implementation, STH infections remain an important public health problem in Benin, with a country-wide prevalence of 20% in 2015. The DeWorm3 study is designed to assess the feasibility of using community-based MDA with albendazole to interrupt the transmission of STH, through a series of cluster-randomized trials in Benin, India and Malawi. We used the pre-treatment baseline survey data to describe and analyze the factors associated with STH infection in Comé, the study site of the DeWorm3 project in Benin. These data will improve understanding of the challenges that need to be addressed in order to eliminate STH as a public health problem in Benin. Between March and April 2018, the prevalence of STH (hookworm spp., Ascaris and Trichuris trichiura) was assessed by Kato-Katz in stool samples collected from 6,153 residents in the community of Comé, Benin using a stratified random sampling procedure. A standardized survey questionnaire was used to collect information from individual households concerning factors potentially associated with the presence and intensity of STH infections in pre-school (PSAC, aged 1-4), school-aged children (SAC, aged 5-14) and adults (aged 15 and above). Multilevel mixed-effects models were used to assess associations between these factors and STH infection. The overall prevalence of STH infection was 5.3%; 3.2% hookworm spp., 2.1% Ascaris lumbricoides and 0.1% Trichuris. Hookworm spp. were more prevalent in adults than in SAC (4.4% versus 2.0%, respectively; p = 0.0001) and PSAC (4.4% versus 1.0%, respectively; p<0.0001), whilst Ascaris lumbricoides was more prevalent in SAC than in adults (3.0% versus 1.7%, respectively; p = 0.004). Being PSAC (adjusted Odds Ratio (aOR) = 0.2, p< 0.001; adjusted Infection Intensity Ratio (aIIR) = 0.1, p<0.001) or SAC (aOR = 0.5, p = 0.008; aIIR = 0.3, p = 0.01), being a female (aOR = 0.6, p = 0.004; aIIR = 0.3, p = 0.001), and having received deworming treatment the previous year (aOR = 0.4, p< 0.002; aIIR = 0.2, p<0.001) were associated with a lower prevalence and intensity of hookworm infection. Lower income (lowest quintile: aOR = 5.0, p<0.001, 2nd quintile aOR = 3.6, p = 0.001 and 3rd quintile aOR = 2.5, p = 0.02), being a farmer (aOR = 1.8, p = 0.02), medium population density (aOR = 2.6, p = 0.01), and open defecation (aOR = 0.5, p = 0.04) were associated with a higher prevalence of hookworm infection. Lower education-no education, primary or secondary school- (aIIR = 40.1, p = 0.01; aIIR = 30.9, p = 0.02; aIIR = 19.3, p = 0.04, respectively), farming (aIIR = 3.9, p = 0.002), natural flooring (aIIR = 0.2, p = 0.06), peri-urban settings (aIIR = 6.2, 95%CI 1.82-20.90, p = 0.003), and unimproved water source more than 30 minutes from the household (aIIR = 13.5, p = 0.02) were associated with a higher intensity of hookworm infection. Improved and unshared toilet was associated with lower intensity of hookworm infections (aIIR = 0.2, p = 0.01). SAC had a higher odds of Ascaris lumbricoides infection than adults (aOR = 2.0, p = 0.01) and females had a lower odds of infection (aOR = 0.5, p = 0.02). Hookworm spp. are the most prevalent STH in Comé, with a persistent reservoir in adults that is not addressed by current control measures based on school MDA. Expanding MDA to target adults and PSAC is necessary to substantially impact population prevalence, particularly for hookworm. ClinicalTrials.gov NCT03014167.


Upregulation of gp91phox subunit of NAD(P)H oxidase contributes to erectile dysfunction caused by long-term nitric oxide inhibition in rats: reversion by regular physical training.
To test the hypothesis that glyco protein 91phox (gp91(phox)) subunit of nicotinamide adenine dinucleotide phosphate [NAD(P)H] oxidase is a fundamental target for physical activity to ameliorate erectile dysfunction (ED). Vascular risk factors are reported to contribute to ED. Regular physical exercise prevents cardiovascular diseases by increasing nitric oxide (NO) production and/or decreasing NO inactivation. Male Wistar rats received the NO synthesis inhibitor N(omega)-nitro-l-arginine methyl ester (l-NAME) for 4 weeks, after which animals were submitted to a run training program for another 4 weeks. Erectile functions were evaluated by in vitro cavernosal relaxations and intracavernous pressure measurements. Expressions of gp91(phox) subunit and neuronal nitric oxidase synthase in erectile tissue, as well as superoxide dismutase activity and nitrite/nitrate (NO(x)) levels were determined. The in vitro acetylcholine- and electrical field stimulation-induced cavernosal relaxations, as well as the increases in intracavernous pressure were markedly reduced in sedentary rats treated with l-NAME. Run training significantly restored the impaired cavernosal relaxations. No alterations in the neuronal nitric oxidase synthase protein expression (and its variant penile neuronal nitric oxidase synthase) were detected. A reduction of NO(x) levels and superoxide dismutase activity was observed in l-NAME-treated animals, which was significantly reversed by physical training. Gene expression of subunit gp91(phox) was enhanced by approximately 2-fold in erectile tissue of l-NAME-treated rats, and that was restored to basal levels by run training. Our study shows that ED seen after long-term l-NAME treatment is associated with gp91(phox) subunit upregulation and decreased NO bioavailability. Exercise training reverses the increased oxidative stress in NO-deficient rats, ameliorating the ED.


Using fuzzy methods to model nearest neighbor rules.
The basic principle used in the construction of nearest-neighbor models is discussed. The induced ordered weighted averaging (IOWA) operators are shown to provide a useful formal structure for building nearest-neighbor models. A methodology for learning IOWA operator nearest-neighbor models is described. Various types of nearest-neighbor rules are investigated, including those based on a linguistic specification. The situation in which the value of interest lies in an ordinal set is also considered. It is shown that the weighted median provides a useful tool for constructing nearest-neighbor rules in this case.


An ERP correlate of metrical stress in spoken word recognition.
Rhythmic properties of spoken language such as metrical stress, that is, the alternation of strong and weak syllables, are important in speech recognition of stress-timed languages such as Dutch and English. Nineteen subjects listened passively to or discriminated actively between sequences of bisyllabic Dutch words, which started with either a weak or a strong syllable. Weak-initial words, which constitute 12% of the Dutch lexicon, evoked more negativity than strong-initial words in the interval between P2 and N400 components of the auditory event-related potential. This negativity was denoted as N325. The N325 was larger during stress discrimination than during passive listening. N325 was also larger when a weak-initial word followed a sequence of strong-initial words than when it followed words with the same stress pattern. The latter difference was larger for listeners who performed well on stress discrimination. It was concluded that the N325 is probably a manifestation of the extraction of metrical stress from the acoustic signal and its transformation into task requirements.


HNF1B controls proximal-intermediate nephron segment identity in vertebrates by regulating Notch signalling components and Irx1/2.
The nephron is a highly specialised segmented structure that provides essential filtration and resorption renal functions. It arises by formation of a polarised renal vesicle that differentiates into a comma-shaped body and then a regionalised S-shaped body (SSB), with the main prospective segments mapped to discrete domains. The regulatory circuits involved in initial nephron patterning are poorly understood. We report here that HNF1B, a transcription factor known to be involved in ureteric bud branching and initiation of nephrogenesis, has an additional role in segment fate acquisition. Hnf1b conditional inactivation in murine nephron progenitors results in rudimentary nephrons comprising a glomerulus connected to the collecting system by a short tubule displaying distal fates. Renal vesicles develop and polarise normally but fail to progress to correctly patterned SSBs. Major defects are evident at late SSBs, with altered morphology, reduction of a proximo-medial subdomain and increased apoptosis. This is preceded by strong downregulation of the Notch pathway components Lfng, Dll1 and Jag1 and the Irx1/2 factors, which are potential regulators of proximal and Henle's loop segment fates. Moreover, HNF1B is recruited to the regulatory sequences of most of these genes. Overexpression of a HNF1B dominant-negative construct in Xenopus embryos causes downregulation specifically of proximal and intermediate pronephric segment markers. These results show that HNF1B is required for the acquisition of a proximo-intermediate segment fate in vertebrates, thus uncovering a previously unappreciated function of a novel SSB subcompartment in global nephron segmentation and further differentiation.


Technology evaluation: Theratope, Biomira Inc.
Active specific immunotherapy, the use of 'vaccines' to stimulate therapeutic tumor antigen-specific immune responses, holds promise as a complementary approach to chemotherapy, radiation and surgery for the treatment of patients with cancers that have a high risk of relapse or progressive disease. Important components of an agent used for active immunotherapy are immunogens in the form of tumor-associated antigen(s) and an adjuvant or carrier molecule to promote presentation of the antigen to the immune effector cells. Possible antigens include tumor-expressed proteins or carbohydrate structures such as the glycoprotein mucin and its epitopes. The Theratope vaccine, consisting of a synthetic mimic of the mucin-associated glycan epitope STn conjugated to the carrier molecule keyhole limpet hemocyanin, has been developed for immunizing patients with mucin-expressing tumors. In murine and human studies, the vaccine has been shown to stimulate anti-STn antibodies and mucin-specific T-cell responses. The immune response is augmented by pretreatment with intravenous cyclophosphamide that serves to inhibit suppressor T-cells. Phase II studies suggested a survival benefit for breast cancer patients who received the Theratope vaccine after intravenous cyclophosphamide. A multinational phase III study testing the Theratope vaccine in patients with metastatic breast cancer who have had a clinical response or stability of disease is ongoing. Other malignancies for which the vaccine may be applicable include ovarian and gastrointestinal cancers.


Characterization of Canine parvovirus 2 variants circulating in Greece.
The aim of the present study was to characterize Canine parvovirus 2 (CPV-2) variants currently circulating in Greece. Between March 2008 and March 2009, 167 fecal samples were collected from diarrheic dogs from different regions of Greece. Canine parvovirus 2 was detected by standard polymerase chain reaction, whereas minor groove binder probe assays were used to distinguish genetic variants and discriminate between vaccine and field strains. Of 84 CPV-2-positive samples, 81 CPV-2a, 1 CPV-2b, and 2 CPV-2c were detected. Vaccine strains were not detected in any sample. Sequence analysis of the VP2 gene of the 2 CPV-2c viruses revealed up to 100% amino acid identity with the CPV-2c strains previously detected in Europe. The results indicated that, unlike other European countries, CPV-2a remains the most common variant in Greece, and that the CPV-2c variant found in Europe is also present in Greece.


In utero and peripubertal metals exposure in relation to reproductive hormones and sexual maturation and progression among girls in Mexico City.
There is increasing evidence that several metals are endocrine disrupting chemicals (EDCs). In utero development and adolescence are critical windows of susceptibility to EDC exposure. With the exception of a few heavy metals, few human studies have evaluated the impact of metal exposure on pubertal development. Our aim was to investigate measures of in utero and peripubertal metal exposure in relation to reproductive hormone levels and sexual maturation and progression among girls from the Early Life Exposure in Mexico to Environmental Toxicants (ELEMENT) cohorts. We measured urinary concentrations of aluminum (Al), arsenic (As), barium (Ba), cadmium (Cd), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), nickel (Ni), antimony (Sb), selenium (Se), and zinc (Zn) in samples collected from women during their third trimester of pregnancy and from their female children at 8-13 years (n = 132). We measured serum testosterone, estradiol, dehydroepiandrosterone sulfate (DHEA-S), inhibin B, and sex hormone-binding globulin (SHBG) at age 8-13, and assessed Tanner stages for sexual maturation (breast, pubic hair development, and menarche status), at two time points (8-13, 14-18 years). We used linear regression to independently examine in utero and peripubertal metal concentrations as predictors of peripubertal hormones. In a longitudinal analysis using generalized estimation equations, we evaluated Tanner stage and menarche progression in relation to individual in utero and peripubertal metal concentrations. We found that higher in utero Zn was associated with increased inhibin B. Several metals at 8-13 years were associated with higher DHEA-S and estradiol, while Ni was positively but Cu was negatively associated with testosterone. In utero Ni, Al, and Cd were associated with slower progression of breast development after adjustment for child age and BMI z-score. For example, an IQR increase in in utero Al exposure was associated with 0.82 times lower odds of progressing to a higher Tanner stage for breast development per year (95% CI: 0.68, 0.99). Peripubertal concentrations of Ba and Al were also associated with being at a higher pubic hair Tanner stage and menarche at 8-13, but lower odds of progressing to the next stage at 14-18 years. We used Bayesian kernel machine regression (BKMR) to model the joint effect of multiple metals while accounting for correlated exposures, as well as potential non-linear relationships between metals and outcomes of interest, which yielded results similar to individual analyses. These findings suggest that female reproductive development may be vulnerable to the effects of metal exposure, and using both Tanner stages and hormone levels may provide clues about underlying mechanisms in two sensitive periods of development.


Use of electrical bone stimulation in spinal fusion.
Spinal fusion is commonly done to manage deformity, restore stability, and eliminate excessive motion at specific spinal levels. Pseudarthrosis limits the clinical success of spinal fusion. Three types of electrical stimulation, which is used to manage non-union in long bones, recently have been applied in an attempt to enhance the rate of spinal fusion. Direct current electrical stimulation is internal and thus eliminates dependence on patient compliance. Pulsed electromagnetic fields and capacitively coupled electrical stimulation are external techniques that require patient compliance but do not have the increased risk associated with implantable devices. Firm conclusions about efficacy are difficult to establish because of inconsistencies in both determining a reliable, reproducible end point for fusion and in incorporating the effect of patient parameters. Most data indicate a positive effect for use of direct current stimulation, but further studies are necessary to determine its appropriateness as an adjuvant to spinal fusion.


A comparison of Bt transgene, hybrid background, water stress, and insect stress effects on corn leaf and ear injury and subsequent yield.
Experimentally manipulated water and insect stresses were applied to field-grown corn with different Bacillus thuringiensis (Bt) transgenes and no Bt transgenes, and different nontransgenic hybrid backgrounds (2011 and 2012, Corpus Christi, TX). Differences in leaf injury, ear injury, and yield were detected among experimental factors and their interactions. Under high and low water stress, injury from noctuid larvae (Lepidoptera: Noctuidae) on leaves during vegetative growth (primarily from fall armyworm, Spodoptera frugiperda J.E. Smith) and on developing ears (primarily from corn earworm, Helicoverpa zea [Boddie]) was lowest on more recent releases of Bt hybrids (newer Bt hybrids) expressing Cry1A.105+Cry2Ab2 and Cry 3Bb1, compared with earlier Bt hybrids (older Bt hybrids) expressing Cry1Ab and Cry3Bb1 and non-Bt hybrids. High water stress led to increased leaf injury under substantial fall armyworm feeding pressure in 2011 (as high as 8.7 on a 1-9 scale of increasing injury). In contrast, ear injury by corn earworm (as high as 20 cm(2) of surface area of injury) was greater in low water stress conditions. Six hybrid backgrounds did not influence leaf injury, while ear injury differences across hybrid backgrounds were detected for non-Bt and older Bt hybrid versions. While newer Bt hybrids expressing Cry1A.105+Cry2Ab2 and Cry 3Bb1 had consistent low leaf injury and high yield and low but less consistent ear injury across six hybrid backgrounds, water stress was a key factor that influenced yield. Bt transgenes played a more variable and lesser role when interacting with water stress to affect yield. These results exemplify the interplay of water and insect stress with plant injury and yield, their interactions with Bt transgenes, and the importance of these interactions in considering strategies for Bt transgene use where water stress is common.


Mode of delivery in pregnancies with premature rupture of membranes at or before term following induction of labor with vaginal prostaglandin E2.
Our aim was to evaluate the mode of delivery in pregnancies complicated with premature rupture of the membranes (PROM) at or before term following induction of labor with vaginal application of prostaglandin E2 (PGE2), and to identify possible predictors leading to cesarean section (CS). The study sample consisted of 220 women with term-PROM who did not enter spontaneous labor after 24 hours of expectant management and 42 with preterm (P)-PROM, who underwent labor induction with vaginal PGE2. Findings were compared with 115 women admitted for elective induction of labor, and 510 women with normal spontaneous-onset labor. There were no between-group differences in gravidity, parity, nulliparity rate, or number of PGE2 tablets used. Women with P-PROM were admitted at 34.6 +/- 2.4 weeks gestation, and delivered at 36.1 +/- 1.6 weeks (mean interval, 5.0 +/- 7.9 days; range, 0.5 to 40 days). The rate of CS was different only between the term-PROM (18.6%) and spontaneous onset of labor (9%) groups (odds ratio, 2.08; 95% confidence interval, 1.4 to 3.4). The rate of low 5-minute Apgar score (< 7) was similar in all groups, but the incidence of nonreassuring fetal heart rate pattern leading to CS was significantly lower in the PROM groups. A logistic regression model and forward likelihood analysis in the PROM groups yielded parity, higher number of PGE2 tablets used, and higher birth weight as independent and significant variables associated with increased risk of CS. Using our management protocol, 36.4% and 20.4% of women with term-PROM and P-PROM, respectively, required labor induction with PGE2, which was successful in more than 80% of cases, with no apparent serious maternal or fetal complications.


Aspergillus flavus: the major producer of aflatoxin.
SUMMARY Aspergillus flavus is an opportunistic pathogen of crops. It is important because it produces aflatoxin as a secondary metabolite in the seeds of a number of crops both before and after harvest. Aflatoxin is a potent carcinogen that is highly regulated in most countries. In the field, aflatoxin is associated with drought-stressed oilseed crops including maize, peanut, cottonseed and tree nuts. Under the right conditions, the fungus will grow and produce aflatoxin in almost any stored crop seed. In storage, aflatoxin can be controlled by maintaining available moisture at levels below that which will support growth of A. flavus. A number of field control measures are being utilized or explored, including: modification of cultural practices; development of resistant crops through molecular and proteomic techniques; competitive exclusion using strains that do not produce aflatoxin; and development of field treatments that would block aflatoxin production. Aspergillus flavus Link (teleomorph unknown) kingdom Fungi, phyllum Ascomycota, order Eurotiales, class Eurotiomycetes, family Trichocomaceae, genus Aspergillus, species flavus. Aspergillus flavus has a broad host range as an opportunistic pathogen/saprobe. It is an extremely common soil fungus. The major concern with this fungus in agriculture is that it produces highly carcinogenic toxins called aflatoxins which are a health hazard to animals. In the field, A. flavus is predominantly a problem in the oilseed crops maize, peanuts, cottonseed and tree nuts. Under improper storage conditions, A. flavus is capable of growing and forming aflatoxin in almost any crop seed. It also is a pathogen of animals and insects. In humans it is predominantly an opportunistic pathogen of immunosuppressed patients. http://www.aspergillusflavus.org, http://www.aflatoxin.info/health.asp, plantpathology.tamu.edu/aflatoxin, http://www.aspergillus.org.uk.


Head covering - a major modifiable risk factor for sudden infant death syndrome: a systematic review.
Some victims of sudden infant death syndrome (SIDS) are found with their heads covered with bedclothes, but the significance of this is uncertain. The aim of this review is to describe the prevalence of head covering, the magnitude of the risk and how far the suggested causal mechanisms agree with current epidemiological evidence. Systematic review of population-based age-matched controlled studies. Controlled observations of head covering for the final sleep were found in 10 studies. The pooled prevalence in SIDS victims was 24.6% (95% CI 22.3% to 27.1%) compared to 3.2% (95% CI 2.7% to 3.8%) among controls. The pooled univariate odds ratio (OR) was 9.6 (95% CI 7.9 to 11.7) and the pooled adjusted OR from studies mainly conducted after the fall in SIDS rate was 16.9 (95% CI 12.6 to 22.7). The risk varied in strength but was significant across all studies. In a quarter of cases and controls head covering had occurred at least once previously (pooled adjusted OR = 1.1; 95% CI 0.9 to 1.4). The population attributable risk (27.1%; 95% CI 24.7% to 29.4%) suggests avoiding head covering might reduce SIDS deaths by more than a quarter. The epidemiological evidence does not fully support postulated causal mechanisms such as hypoxia, hypercapnoea and thermal stress, but neither does it support the idea that head covering is part of some terminal struggle. Head covering is a major modifiable risk factor associated with SIDS deaths and parental advice to avoid this situation should be emphasised.


Pathogenesis and pathology of hematogenous infections of the fetus and newborn.
The earlier infections occur in intrauterine life the more severe are they. When the infection develops during embryogenesis, the lesions are much more serious, sometimes causing disruptions (malformations). At this time the organs are not completely formed and microorganisms may interfere with organogenesis to such an extent that the development of the functions necessary for viability become impaired. Infection acquired in utero may result in resorption of the embryo, abortion, stillbirth, neonatal death, intrauterine growth retardation (IUGR), or prematurity. The infected newborns commonly are apparently normal at birth but they may develop a late onset disease. Otherwise, neonates presenting symptomatology may derelop untoward sequelae.


Importance of vagus nerves in duodenal acid neutralization in anesthetized pigs.
During the cephalic phase of gastric acid secretion, vagally mediated synchronous stimulation of bicarbonate provides protection against the acid. The purpose of this study was to determine simultaneously the effect of electrical vagal stimulation (EVS) on pancreatic, hepatic, and duodenal mucosal bicarbonate secretion, thereby estimating their relative importance in vagally induced duodenal acid neutralization. Splanchnicotomy increased vagally induced pancreaticobiliary bicarbonate secretion, whereas duodenal mucosal bicarbonate secretion was unchanged. After splanchnicotomy, EVS (10 ms, 15 mA, 12 Hz) significantly increased pancreatic bicarbonate secretion (0-4.17 mmol/h), hepatic bicarbonate secretion (0.16 to 0.22 mmol/h), and duodenal mucosal bicarbonate secretion (0.17 to 0.31 mmol/h). Pancreaticobiliary bicarbonate secretion was atropine resistant, whereas vagally induced duodenal mucosal bicarbonate secretion was diminished by atropine (2.0 mg/kg). After splanchnicotomy, EVS (10 ms, 15 mA, 12 Hz) had no effect on portal plasma concentration of secretin, whereas vasoactive intestinal peptide was increased (14-29 pM). EVS at 12 Hz with varying duration (3 or 10 ms) and amplitude (3-50 mA) had no further effect on the bicarbonate secretion from the three organs. In addition, biliary [14C]mannitol clearance was shown not to be a reliable marker of canalicular bile secretion in pigs. These results suggest that in the anesthetized pig 1) vagal stimulation is only of minor importance to hepatic bicarbonate secretion; 2) vagal stimulation activates pancreatic bicarbonate secretion through both cholinergic muscarinic and noncholinergic transmission; and 3) vagal stimulation induces duodenal mucosal bicarbonate secretion mainly through cholinergic muscarinic transmission. In conclusion, these results suggest that only pancreatic and duodenal bicarbonate production play a role in vagally induced duodenal acid neutralization.


Integrated analysis of transcriptomic and proteomic data reveals novel regulators of soybean (Glycine max) hypocotyl development.
Hypocotyl elongation directly affects the seedling establishment and soil-breaking after germination. In soybean (Glycine max ), the molecular mechanisms regulating hypocotyl development remain largely elusive. To decipher the regulatory landscape, we conducted proteome and transcriptome analysis of soybean hypocotyl samples at different development stages. Our results showed that during hypocotyl development, many proteins were with extreme high translation efficiency (TE) and may act as regulators. These potential regulators include multiple peroxidases and cell wall reorganisation related enzymes. Peroxidases may produce ROS including H2 O2 . Interestingly, exogenous H2 O2 application promoted hypocotyl elongation, supporting peroxidases as regulators of hypocotyl development. However, a vast variety of proteins were shown to be with dramatically changed TE during hypocotyl development, including multiple phytochromes, plasma membrane intrinsic proteins (PIPs) and aspartic proteases. Their potential roles in hypocotyl development were confirmed by that ectopic expression of GmPHYA1 and GmPIP1-6 in Arabidopsis thaliana affected hypocotyl elongation. In addition, the promoters of these potential regulatory genes contain multiple light/gibberellin/auxin responsive elements, while the expression of some members in hypocotyls was significantly regulated by light and exogenous auxin/gibberellin. Overall, our results revealed multiple novel regulatory factors of soybean hypocotyl elongation. Further research on these regulators may lead to new approvals to improve soybean hypocotyl traits.


Combined-modality therapy with gemcitabine and radiotherapy as a bladder preservation strategy: results of a phase I trial.
We conducted a phase I trial of gemcitabine given twice weekly with concurrent radiotherapy in patients with muscle-invasive bladder cancer. Eligible patients underwent maximal transurethral resection of their bladder tumors followed by twice-weekly infusion of gemcitabine with 2 Gy/d concurrent radiotherapy to the bladder, for a total of 60 Gy over 6 weeks. The starting dose of gemcitabine was 10 mg/m(2) with subsequent dose levels of 20, 27, 30, and 33 mg/m(2). The primary end point was the determination of the maximum-tolerated dose (MTD) of twice weekly gemcitabine with concurrent radiotherapy. Secondary end points included assessment of toxicity associated with combined-modality therapy and initial assessment of the rate of bladder preservation. Twenty-four patients were enrolled and 23 were assessable for toxicity and response. No significant toxicity was demonstrated at the 10 or 20 mg/m(2) twice-weekly doses. Dose-limiting toxicity (DLT) occurred in two of three patients treated at 33 mg/m(2). Intermediate dose levels of 27 and 30 mg/m(2) were then evaluated. The MTD of gemcitabine was 27 mg/m(2). The DLT was systemic, manifested as an elevation in liver function tests, malaise, and edema. Fifteen of 23 patients (65%) are alive with bladders intact and no evidence of recurrent disease at a median follow-up of 43 months. Twice-weekly gemcitabine with concurrent radiotherapy at 2 Gy/d to a total dose of 60 Gy is well-tolerated. The MTD of gemcitabine is 27 mg/m(2). There is a high rate of bladder preservation in this selected group of patients.


Comparative insights into the mechanisms of renal organic anion and cation secretion.
Comparative models have played a major role in defining the mechanisms that enable vertebrate proximal tubules to transport organic anions and cations from the peritubular interstitium to the urine. The unique advantages of these models and their contributions to our understanding of organic anion and cation transport mechanisms are summarized here. Recent studies of the organic anion transport system suggest that transport is coupled to metabolic energy via indirect coupling to the sodium gradient. Organic anions enter the cell across the basolateral membrane in exchange for alpha-ketoglutarate (alpha-KG), and the alpha-KG is returned to the interior via Na-alpha-KG cotransport. Indirect coupling to Na has been demonstrated in both isolated membranes and intact renal epithelial cells of species ranging from marine crustaceans to mammals. This mechanism was shown to drive not only cellular accumulation but also secretory transepithelial fluxes of organic anions. Luminal exit of secreted organic anions appears to be carrier mediated but is, at present, poorly understood, with mediated potential-driven efflux and anion exchange-driven efflux implicated in some species. As for organic anions, the renal clearance of some organic cations approaches the renal plasma flow. Although there is considerable variation in the handling of specific substrates between species, the basic properties of organic cation transport include carrier-mediated potential-driven uptake at the basolateral membrane, intracellular sequestration that reduces the free concentration of the cation, and luminal exit by organic cation-proton exchange. Reabsorptive transport is also observed for some organic cations, but its mechanisms and driving forces are not well understood.


Social physique anxiety in postmenopausal women.
Social physique anxiety (SPA), the degree to which individuals become anxious when others observe their bodies, is an important concept for postmenopausal women because it may be directly related to their physical activity behavior. Women with high levels of social physique anxiety may not participate in health-enhancing physical activity. Therefore, the main purpose of this study was to examine SPA in postmenopausal women relative to leisure time physical activity (LTPA, activities such as exercise, recreation, and sports). Other factors, such as percent body fat, body fat distribution, age, and hormone replacement therapy status were also examined for their relationship to SPA. Women who expended < or = 500 kcal.wk-1 in LTPA had significantly higher SPA than women who expended > or = 2,000 kcal.wk-1, independent of percent body fat. Women with more than 37.5% body fat had significantly higher SPA than those with less than 37.5% body fat. Women with upper BFD (waist-to-hip ratio or WHR > 0.85) had higher SPA than women with lower BFD (WHR < 0.75). There were no differences in SPA relative to age or HRT status. It appears that a sedentary lifestyle, high percent body fat, and upper body fat distribution are associated with increased SPA in postmenopausal women, thus health promotion professionals should be aware of these concerns when developing physical activity interventions for postmenopausal women.


[Purification and survival of retinal ganglion cell].
We had used a specific anti-Thy 1.1 antibody binding method and a nylonmembrane sieve method to isolate and purify retinal ganglion cells from neonatal rats in order to compare the effect of tectal extract on these purified cells retinal ganglion cells. Isolated retinal cell suspension with retinal ganglion cells retrograde-prelabelled with Fast Blue were seeded on culture dishes coated with the specific anti-Thy 1.1 antibody for 30 minutes before nonadherent cells were removed. The percentage purity of the adherent retinal ganglion cells determined microscopically to be 95%. However, the percentage purity of the Fast Blue-labelled retinal ganglion cells recovered using the nylon membrane of pore size 15 microns was only 60 +/- 5%. Retinal ganglion cells purified by both methods could survive and grow into large, active neurons with neurite outgrowths in the presence of tectal extract. A MTT colorimetric microassay was used to quantify the survival growth activity of these purified retinal ganglion cells after culture for 24 hours. The result showed that the optical density ratio (+Te/-Te) of the retinal ganglion cells purified by anti-Thy 1.1 antibody binding method was 12.3 (0.111/0.009) and by the nylon membrane method was 6.4 (0.102/0.016), and the optical density ratio of the non-purified retinal cells was 3.8 (0.095/0.025), p less than 0.01 for all 3 sets of results. It was concluded that in the absence of other cells, the purified retinal ganglion cells responded specifically to the trophic activity in tectal extract, the purer the retinal ganglion cells and the clearer the effect.


Uncovering Intramolecular π-Type Hydrogen Bonds in Solution by NMR Spectroscopy and DFT Calculations.
Reaction between the phosphinito bridged diplatinum species [(PHCy2 )Pt(μ-PCy2 ){κ(2) P,O-μ-P(O)Cy2 }Pt(PHCy2 )](Pt-Pt) (1), and (trimethylsilyl)acetylene at 273 K affords the σ-acetylide complex [(PHCy2 )(η(1) -Me3 SiC≡C)Pt(μ-PCy2 )Pt(PHCy2 ){κP-P(OH)Cy2 }](Pt-Pt) (2) featuring an intramolecular π-type hydrogen bond. Scalar and dipolar couplings involving the POH proton were detected by 2D NMR experiments. Relativistic DFT calculations of the geometry, relative energy, and NMR properties of model systems of 2 confirmed the structural assignment and allowed the energy of the π-type hydrogen bond to be estimated (ca. 22 kJ mol(-1) ).


Extratemporal atrophy in patients with complex partial seizures of left temporal origin.
Total cerebral, temporal lobe, hippocampal, caudate, and lenticular nuclei volumes were quantified from magnetic resonance images of 21 patients with left temporal lobe epilepsy and medically intractable complex partial seizures. These regional brain volumes were compared with the same measures in 19 controls. No significant differences in total cerebral, left temporal lobe, right temporal lobe, or total temporal lobe volumes were found. As expected, left hippocampal volumes were significantly smaller in the patients with epilepsy than in control subjects. The left hippocampus-to-right hippocampus volume ratio was significantly lower in patients than in control subjects. In addition to left hippocampal volumes, mean left thalamic, left caudate, and bilateral lenticular volumes were significantly smaller in the patients with epilepsy than in control subjects. The left-to-right thalamic volume ratio was also significantly lower in the patients with epilepsy compared with control subjects, but there were no significant group differences in caudate or lenticular ratios. These results show that medically intractable temporal lobe epilepsy is associated with volume loss in brain structures outside the presumably involved hippocampus. The pathophysiological significance of our findings is uncertain. They could be related to the underlying cause of the disorder. However, volume loss also may reflect damage due to involvement of these structures in recurrent seizure activity.


Melatonin sensitizes human breast cancer cells to ionizing radiation by downregulating proteins involved in double-strand DNA break repair.
Radiation and adjuvant endocrine therapy are nowadays considered a standard treatment option after surgery in breast cancer. Melatonin exerts oncostatic actions on human breast cancer cells. In the current study, we investigated the effects of a combination of radiotherapy and melatonin on human breast cancer cells. Melatonin (1 mm, 10 μm and 1 nm) significantly inhibited the proliferation of MCF-7 cells. Radiation alone inhibited the MCF-7 cell proliferation in a dose-dependent manner. Pretreatment of breast cancer cells with melatonin 1 wk before radiation led to a significantly greater decrease of MCF-7 cell proliferation compared with radiation alone. Melatonin pretreatment before radiation also decreased G2 -M phase arrest compared with irradiation alone, with a higher percentage of cells in the G0 -G1 phase and a lower percentage of cells in S phase. Radiation alone diminished RAD51 and DNA-protein kinase (PKcs) mRNA expression, two main proteins involved in double-strand DNA break repair. Treatment with melatonin for 7 days before radiation led to a significantly greater decrease in RAD51 and DNA-PKcs mRNA expression compared with radiation alone. Our findings suggest that melatonin pretreatment before radiation sensitizes breast cancer cells to the ionizing effects of radiation by decreasing cell proliferation, inducing cell cycle arrest and downregulating proteins involved in double-strand DNA break repair. These findings may have implications for designing clinical trials using melatonin and radiotherapy.


Molecular Epidemiology of Dengue in Panama: 25 Years of Circulation.
Dengue virus (DENV) is the most prevalent arbovirus in terms of human public health importance globally. In addition to DENV epidemiological surveillance, genomic surveillance may help investigators understand the epidemiological dynamics, geographic distribution, and temporal patterns of DENV circulation. Herein, we aimed to reconstruct the molecular epidemiology and phylogeny of DENV in Panama to connect the epidemiological history of DENV dispersal and circulation in Latin America. We retrospectively analyzed the epidemiological data obtained during 25 years of DENV surveillance in Panama. DENV was reintroduced in Panama in 1993 after a 35 year absence of autochthonous transmission. The increase in the number of total dengue cases has been accompanied by an increase in severe and fatal cases, with the highest case fatality rate recorded in 2011. All four serotypes were detected in Panama, which is characterized by serotype replacement and/or co-circulation of multiple serotypes. Phylogenetic analysis of datasets collected from envelope (E) gene sequences obtained from viruses isolated from human sera demonstrated that circulating viruses were highly diverse and clustered in distinct clades, with co-circulation of clades from the same genotype. Our analyses also suggest that Panamanian strains were related to viruses from different regions of the Americas, suggesting a continuous exchange of viruses within the Americas.


Laparoscopic splenectomy for splenic hamartoma: Case management and clinical consequences.
Splenic hamartoma is a rare benign tumor, and although minimally invasive surgery may be suitable for this condition, there have only been 2 previous reports of laparoscopic surgery. Here we report the third case of splenic hamartoma managed by laparoscopic splenectomy. A 37-year-old male was incidentally diagnosed by abdominal ultrasonography with a hypoechoic mass measuring 2.5 cm × 2.4 cm in the spleen. Color Doppler sonography showed multiple flow signals within the mass and contrast-enhanced computed tomography revealed strong enhancement of the lesion. On T1- and T2-weighted magnetic resonance images, the splenic mass was demonstrated as isointense and hyperintense respectively. Although a malignant tumor could not be ruled out, a hand-assisted laparoscopic splenectomy was performed because the splenic mass was limited in size and had not invaded adjacent organs. The pathological diagnosis was splenic hamartoma. The postoperative course was uneventful and the patient was discharged by the seventh postoperative day. Although splenic hamartomas have some specific imaging features, more reports and analyses of these cases are required to increase the reliability of the diagnosis and management. Hand-assisted laparoscopic splenectomy may play a pivotal role in the postoperative diagnosis and management of this condition.


Gut dysbiosis, defective autophagy and altered immune responses in neurodegenerative diseases: Tales of a vicious cycle.
The human microbiota comprises trillions of symbiotic microorganisms and is involved in regulating gastrointestinal (GI), immune, nervous system and metabolic homeostasis. Recent observations suggest a bidirectional communication between the gut microbiota and the brain via immune, circulatory and neural pathways, termed the Gut-Brain Axis (GBA). Alterations in gut microbiota composition, such as seen with an increased number of pathobionts and a decreased number of symbionts, termed gut dysbiosis or microbial intestinal dysbiosis, plays a prominent role in the pathogenesis of central nervous system (CNS)-related disorders. Clinical reports confirm that GI symptoms often precede neurological symptoms several years before the development of neurodegenerative diseases (NDDs). Pathologically, gut dysbiosis disrupts the integrity of the intestinal barrier leading to ingress of pathobionts and toxic metabolites into the systemic circulation causing GBA dysregulation. Subsequently, chronic neuroinflammation via dysregulated immune activation triggers the accumulation of neurotoxic misfolded proteins in and around CNS cells resulting in neuronal death. Emerging evidence links gut dysbiosis to the aggravation and/or spread of proteinopathies from the peripheral nervous system to the CNS and defective autophagy-mediated proteinopathies. This review summarizes the current understanding of the role of gut microbiota in NDDs, and highlights a vicious cycle of gut dysbiosis, immune-mediated chronic neuroinflammation, impaired autophagy and proteinopathies, which contributes to the development of neurodegeneration in Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis and frontotemporal lobar degeneration. We also discuss novel therapeutic strategies targeting the modulation of gut dysbiosis through prebiotics, probiotics, synbiotics or dietary interventions, and faecal microbial transplantation (FMT) in the management of NDDs.


Assessing implementation fidelity in the First Episode Rapid Early Intervention for Eating Disorders service model.
The First Episode Rapid Early Intervention for Eating Disorders (FREED) service model is associated with significant reductions in wait times and improved clinical outcomes for emerging adults with recent-onset eating disorders. An understanding of how FREED is implemented is a necessary precondition to enable an attribution of these findings to key components of the model, namely the wait-time targets and care package. This study evaluated fidelity to the FREED service model during the multicentre FREED-Up study. Participants were 259 emerging adults (aged 16-25 years) with an eating disorder of <3 years duration, offered treatment through the FREED care pathway. Patient journey records documented patient care from screening to end of treatment. Adherence to wait-time targets (engagement call within 48 h, assessment within 2 weeks, treatment within 4 weeks) and care package, and differences in adherence across diagnosis and treatment group were examined. There were significant increases (16-40%) in adherence to the wait-time targets following the introduction of FREED, irrespective of diagnosis. Receiving FREED under optimal conditions also increased adherence to the targets. Care package use differed by component and diagnosis. The most used care package activities were psychoeducation and dietary change. Attention to transitions was less well used. This study provides an indication of adherence levels to key components of the FREED model. These adherence rates can tentatively be considered as clinically meaningful thresholds. Results highlight aspects of the model and its implementation that warrant future examination.


Recommended medical and non-medical factors to assess military preventable deaths: subject matter experts provide valuable insights.
Historically, there has been variability in the methods for determining preventable death within the US Department of Defense. Differences in methodologies partially explain variable preventable death rates ranging from 3% to 51%. The lack of standard review process likely misses opportunities for improvement in combat casualty care. This project identified recommended medical and non-medical factors necessary to (1) establish a comprehensive preventable death review process and (2) identify opportunities for improvement throughout the entire continuum of care. This qualitative study used a modified rapid assessment process that includes the following steps: (1) identification and recruitment of US government subject matter experts (SMEs); (2) multiple cycles of data collection via key informant interviews and focus groups; (3) consolidation of information collected in these interviews; and (4) iterative analysis of data collected from interviews into common themes. Common themes identified from SME feedback were grouped into the following subject areas: (1) prehospital, (2) in-hospital and (3) forensic pathology. Medical recommendations for military preventable death reviews included the development, training, documentation, collection, analysis and reporting of the implementation of the Tactical Combat Casualty Care Guidelines, Joint Trauma System Clinical Practice Guidelines and National Association of Medical Examiners autopsy standards. Non-medical recommendations included training, improved documentation, data collection and analysis of non-medical factors needed to understand how these factors impact optimal medical care. In the operational environment, medical care must be considered in the context of non-medical factors. For a comprehensive preventable death review process to be sustainable in the military health system, the process must be based on an appropriate conceptual framework implemented consistently across all military services.


Land availability for biofuel production.
Marginal agricultural land is estimated for biofuel production in Africa, China, Europe, India, South America, and the continental United States, which have major agricultural production capacities. These countries/regions can have 320-702 million hectares of land available if only abandoned and degraded cropland and mixed crop and vegetation land, which are usually of low quality, are accounted. If grassland, savanna, and shrubland with marginal productivity are considered for planting low-input high-diversity (LIHD) mixtures of native perennials as energy crops, the total land availability can increase from 1107-1411 million hectares, depending on if the pasture land is discounted. Planting the second generation of biofuel feedstocks on abandoned and degraded cropland and LIHD perennials on grassland with marginal productivity may fulfill 26-55% of the current world liquid fuel consumption, without affecting the use of land with regular productivity for conventional crops and without affecting the current pasture land. Under the various land use scenarios, Africa may have more than one-third, and Africa and Brazil, together, may have more than half of the total land available for biofuel production. These estimations are based on physical conditions such as soil productivity, land slope, and climate.


Functional architecture of the cell's nucleus in development, aging, and disease.
In eukaryotes, the function of the cell's nucleus has primarily been considered to be the repository for the organism's genome. However, this rather simplistic view is undergoing a major shift, as it is increasingly apparent that the nucleus has functions extending beyond being a mere genome container. Recent findings have revealed that the structural composition of the nucleus changes during development and that many of these components exhibit cell- and tissue-specific differences. Increasing evidence is pointing to the nucleus being integral to the function of the interphase cytoskeleton, with changes to nuclear structural proteins having ramifications affecting cytoskeletal organization and the cell's interactions with the extracellular environment. Many of these functions originate at the nuclear periphery, comprising the nuclear envelope (NE) and underlying lamina. Together, they may act as a "hub" in integrating cellular functions including chromatin organization, transcriptional regulation, mechanosignaling, cytoskeletal organization, and signaling pathways. Interest in such an integral role has been largely stimulated by the discovery that many diseases and anomalies are caused by defects in proteins of the NE/lamina, the nuclear envelopathies, many of which, though rare, are providing insights into their more common variants that are some of the major issues of the twenty-first century public health. Here, we review the contributions that mouse mutants have made to our current understanding of the NE/lamina, their respective roles in disease and the use of mice in developing potential therapies for treating the diseases.


Progress in diagnosis and treatment of difficult-to-treat asthma in children.
At present, medications containing inhaled corticosteroids (ICS-containing) are the keystones of asthma treatment. The majority of asthmatic children can significantly improve clinical outcomes with little worsening by standardized inhaled glucocorticoid treatment, but there is still a small proportion of children who are unable to achieve good symptom control even after the maximum standardized treatment, known as 'children with difficult-to-treat asthma (DA)'. The high heterogeneity of DA makes therapy challenging and expensive, which poses a serious risk to children's health and makes it extremely difficult for clinical physicians to accurately identify and treat children with DA. This article reviews the definition, evaluation, and treatment of this asthma in order to provide a reference for optimal clinical decision-making.