Carcinoma ex basal cell adenoma of the parotid gland: A report of an extremely rare case.
Malignant non-basaloid tumors that arise from basal cell adenoma (BCA) are extremely rare. The patient was a 72-year-old Japanese male, who had noticed swelling of the left parotid region 21 years ago. A superficial lobectomy was performed. About 60% of the tumor was made up of cribriform and trabecular tissue composed of basaloid cells, which exhibited mild atypia and nuclear expression of β-catenin. This portion of the tumor was considered to be a BCA. In the other part of the tumor, the proliferation of large eosinophilic atypical cells, most of which formed intraductal structures, was observed. These tumor cells displayed cellular atypia, and some of them formed Roman bridge structures or contributed to intracapsular invasion. Immunohistochemically, these cells were positive for cytokeratin 7, gross cystic disease fluid proten-15 (GCDFP-15), androgen receptor (AR), and mammaglobin (MMG) and exhibited a high Ki-67 labeling index. So, this portion of the tumor was considered to be a salivary duct carcinoma (SDC). The tumor's final diagnosis was SDC ex BCA (intracapsular type), which is extremely rare. GCDFP-15, AR, MMG, and Ki-67 are useful immunohistochemical markers for diagnosing SDC ex BCA.


New insights into extracellular vesicle biogenesis and function.
It is becoming increasingly evident that most cell types are capable of forming and releasing multiple distinct classes of membrane-enclosed packages, referred to as extracellular vesicles (EVs), as a form of intercellular communication. Microvesicles (MVs) represent one of the major classes of EVs and are formed by the outward budding of the plasma membrane. The second major class of EVs, exosomes, are produced as components of multivesicular bodies (MVBs) and are released from cells when MVBs fuse with the cell surface. Both MVs and exosomes have been shown to contain proteins, RNA transcripts, microRNAs and even DNA that can be transferred to other cells and thereby trigger a broad range of cellular activities and biological responses. However, EV biogenesis is also frequently de-regulated in different pathologies, especially cancer, where MVs and exosomes have been suggested to promote tumor cell growth, therapy resistance, invasion and even metastasis. In this Review, we highlight some of the recent advances in this rapidly emerging and exciting field of cell biology, focusing on the underlying mechanisms that drive MV and exosome formation and release, with a particular emphasis on how EVs potentially impact different aspects of cancer progression and stem cell biology.


Elbasvir/Grazoprevir for HCV Infection in Russia: A Randomized Trial.
Hepatitis C virus (HCV) infection is a major healthcare concern in Russia, where almost 5 million individuals are viremic. Elbasvir/grazoprevir is a fixed-dose combination therapy for the treatment of HCV genotype 1 and genotype 4 infection. The present analysis aimed to assess the safety and efficacy of elbasvir/grazoprevir in individuals with HCV infection enrolled at Russian study sites in the C-CORAL study. C-CORAL (Protocol PN-5172-067; NCT02251990) was a Phase 3, placebo-controlled, double-blind study conducted throughout Asia and Russia. Treatment-naive participants with chronic HCV infection were randomly assigned to receive immediate or deferred treatment with elbasvir 50 mg/grazoprevir 100 mg once daily for 12 weeks. Participants in the immediate-treatment group received elbasvir/grazoprevir for 12 weeks, and those in the deferred-treatment group received placebo for 12 weeks, followed by open-label elbasvir/grazoprevir for 12 weeks. The primary endpoint was sustained virologic response at 12 weeks after completion of therapy (SVR12). One hundred and nineteen Russian participants were randomized (immediate-treatment group, n=88; deferred-treatment group, n=31). Most participants were white (99%) with HCV genotype 1b infection (97%) and mild-to-moderate (F0-F2) fibrosis (70%). SVR12 was achieved by 98.9% participants in the immediate-treatment group and by 100% of those receiving deferred elbasvir/grazoprevir in the deferred-treatment group. One participant relapsed with nonstructural protein 5A (NS5A) L28M and Y93H resistance-associated substitutions at baseline and at time of failure. Drug-related adverse events were reported by 19% of participants receiving elbasvir/grazoprevir in the immediate-treatment group and by 16% of those receiving placebo in the deferred-treatment group. No serious adverse event or deaths occurred, and no participant discontinued treatment owing to an adverse event. Elbasvir/grazoprevir for 12 weeks was highly effective in treatment-naive Russian individuals with HCV genotype 1b infection.


Effect of artemisinin on improving islet function in rats fed with maternal high-fat diet.
Maternal high-fat diet (HFD) is a detrimental factor in developing glucose intolerance, obesity, and islet dysfunction. However, the effect of artemisinin on maternal HFD and whether it is related to the alterations of islet function is seldom studied since artemisinin treatments not only attenuate insulin resistance (IR) and restore islet ß cell function in Diabetes mellitus type 2. Female rats were randomly fed a HFD (45% kcal from fat), HFD + artemisinin, or a regular chow diet (RCD) before pregnancy and during gestation. Glucose metabolism and the β cell phenotypes were assessed. Maternal HFD increased islet load in female rats, proliferation of pancreatic β cells, increased insulinogen, and decreased insulin secretion response to high glucose stimulation with delayed insulin release, increased fasting glucose, and glucose area under the curve compared with the general diet group. HFD inhibited expression of Foxo1 and PAX6 in female rats. Under the effect of both HFD and pregnancy, islet load was further increased, insulinogen was further increased, and fasting insulin level and fasting glucose were higher than RCD fed general-pregnancy group. ALDH1a3 transdifferentiation and PAX6, Foxo1, and PDX1 expression were increased in islets of high-fat pregnant rats. When adding artemisinin in HFD treated pregnant rats, islet function was significantly improved. Intervention with artemisinin in maternal HFD resulted in reduced islet size, decreased number of β-cells and improved islet microcirculation, insulin processing shear process, decreased insulinogen/insulin ratio, and restored islet function through increased expression of PC1/3.


Chronic tension-type headache: an investigation of the appraisal process.
The few existing studies that have explicitly examined appraisal of stress in tension-type headache sufferers have found that these individuals perceive events in a more negative manner than headache-free controls. However, can it necessarily be assumed that they perceive events in a less accurate manner than headache-free controls? To answer this question, sixty chronic tension-type headache sufferers and 60 controls engaged in a 40-trial contingency learning task in which they had to estimate the amount of control they could exert. Results provide little support for the notion that the cognitive appraisal of tension-type headache sufferers is more accurate than that of headache-free individuals. However, differences were found in perceived control when behavioral involvement was considered. This result is discussed in terms of learned helplessness theory.


Low-dose lipopolysaccharide inducing continuous and obvious increase in urinary protein in hyperglycemic rats and the underlying mechanism.
Proteinuria is an important hallmark of diabetic nephropathy models, however it takes a long time for the proteinuria and is not stable. Therefore, low-dose lipopolysaccharide (LPS) was investigated in this work to induce rapid and stable proteinuria in hyperglycemic rats and the underlying mechanism was studied. Hyperglycemia rats was induced by high-fat feeding combined with intraperitoneal injection of streptozotocin (STZ). After 21 days, the model rats received a subinjury dose of 0.8 mg / kg LPS intraperitoneally (i.p.). We detected related biochemical indexes at different time periods after LPS injection and examined the expression of glomerular podocyte-associated proteins. Simultaneously, we measured expression of inflammatory factors, apoptotic proteins and albumin (ALB) in the renal cortex and renal medulla, respectively. PAS (Periodic Acid Schiff) staining was used to observe renal pathology. After LPS injection, urinary microalbumin (umALB) increased significantly and lasted longer. The expression of Nephrin, Podocin and necroptosis factor kappa B (NF-κB) in rennal cortex and Interleukin 18 (IL-18), Caspase-1, NF-κB and ALB in the renal medulla was significantly changed. Pathologically, the glomerular basement membrane was observed to be significantly thickened, the renal tubules were dilated, and the epithelial cells fell off in a circle. LPS promoted the continuous increase in urinary microalbumin in hyperglycemic rats, which was related to the damage to the glomerular basement membrane and renal tubular epithelial cells and to the inflammatory reaction in the kidney involved in NF-κB signaling, and this pathological damage can help to establish a stable model of diabetic nephropathy with increased proteinuria.


Diagnostic and prognostic impact of cytochemically assessed nuclear DNA contents in human adenocarcinomas of the mammary gland.
Cytochemical assessments of the nuclear DNA contents in carcinomas of the breast can be used for both prognostic and diagnostic purposes. Two main techniques are currently being used, viz. flow cytometry (FCM) and microspectrophotometry (MSP). An account of their advantages and disadvantages is given. In addition, an old, rather crude, cytophotometric technique can be used for histopathological sections of paraffin-embedded specimens. The principal sampling procedures are fine-needle aspiration biopsy and the so-called imprint technique, where the specimens are made from the cut surface of the freshly excised operation specimen. Paraffin-embedded histopathological material can also be used, applying a newly developed MSP procedure, where isolated nuclei from deparaffinized/disintegrated specimens are analyzed. Most important is that intact cell nuclei, representative for the whole tumour nodule, can be obtained. The simultaneous use of FCM and MSP is also of utmost importance for the reliability in the interpretation of the results. Then, a kind of "DNA malignancy grading" is obtained that in several investigations has proven itself to be an excellent prognosticating tool that can be used for making an adequate choice of therapy for the individual patient. The diagnostic value of the results of the cytochemically assessed nuclear DNA distribution patterns is not so high as the prognostic one. Tumours with a diploid type of nuclear DNA content can be found both among benign and malignant neoplasms. However, a neoplasm with an aneuploid DNA distribution pattern can almost certainly be considered highly malignant.


Serum complement levels and perinatal outcome in pregnancies complicated by systemic lupus erythematosus.
Previous reports suggest an association between maternal serum complement component (C3 and C4) levels and perinatal outcome. Eighteen women with systemic lupus erythematosus (21 pregnancies) were prospectively studied. C3 and C4 levels were measured monthly unless more frequent data were needed for acute medical management. Perinatal outcome was then correlated with both maternal clinical and serologic status before and during pregnancy. Twelve pregnancies were carried to term, 11 of which began in clinical remission. Eight of these 12 had normal C3 and C4 levels at the onset of pregnancy, and 11 were normal at delivery. Four pregnancies were delivered prematurely (with one neonatal death from septic meningitis); all began pregnancy in clinical remission but three had low C3 and C4 levels before delivery. Three pregnancies beginning in clinical remission ended in spontaneous first- or early second-trimester abortions; all had low C3 and C4 levels before aborting. Normal complement component levels were observed in the remaining two pregnancies that were terminated electively; they had no evidence of fetal or placental abnormality. The correlation of maternal serum complement levels and pregnancy outcome in this series suggests that these parameters may help in the assessment of fetal as well as maternal prognosis in lupus-complicated gestations.


Report of a gingival "surgical cyst" developing secondarily to a subepithelial connective tissue graft.
The subepithelial connective tissue graft is a valuable technique used to cover exposed root surfaces or to increase the zone of attached gingiva. Cysts, such as the gingival cyst of the adult, are epithelial-lined cysts which may arise from heterotropic glandular tissue, a proliferating rete peg, remnants of the dental lamina, or traumatic implantation of epithelial tissue. In this paper, a case report is described of a "surgical cyst" or cystic morphogenesis of surgically implanted epithelium developing secondarily to a subepithelial connective tissue graft.


Trichostatin A induces bladder cancer cell death via intrinsic apoptosis at the early phase and Sp1‑survivin downregulation at the late phase of treatment.
Histone deacetylase (HDAC) inhibitors have been widely shown to result in cancer cell death. The present study investigated the mechanisms underlying the antitumor effects of the phytochemical trichostatin A (TSA), a classic pan-HDAC inhibitor, in 5,637 urinary bladder cancer cells. It was found that TSA caused cell cycle arrest at the G2/M and G1 phase accompanied by reduced expression of cyclin D1 and upregulated induction of p21. In addition, TSA induced morphological changes, reduced cell viability and apoptotic cell death in 5,637 cells through caspase-3 activation followed by PARP cleavage. The loss of mitochondrial membrane potential (MMP) indicated that TSA induced apoptosis in 5,637 cells through the intrinsic mitochondrial pathway. TSA significantly suppressed Akt activity at 12 h after treatment, suggesting that the apoptosis in the early phase was mediated by Akt inhibition. In addition, the protein level of transcription factor Sp1 was decreased at 24 h after TSA treatment, which likely led to the downregulation of survivin gene expression, and then contributed to the antitumor activity of TSA. Taken together, the present study delineated that TSA-induced growth inhibition and apoptosis in 5,637 cells was associated with pAKT inhibition and MMP loss at the early phase, followed by downregulation of Sp1 and survivin at the late phase of treatment.


Swift-Hohenberg model for magnetoconvection.
A model system of partial differential equations in two dimensions is derived from the three-dimensional equations for thermal convection in a horizontal fluid layer in a vertical magnetic field. The model consists of an equation of Swift-Hohenberg type for the amplitude of convection, coupled to an equation for a large-scale mode representing the local strength of the magnetic field. The model facilitates both analytical and numerical studies of magnetoconvection in large domains. In particular, we investigate the phenomenon of flux separation, where the domain divides into regions of strong convection with a weak magnetic field and regions of weak convection with a strong field. Analytical predictions of flux separation based on weakly nonlinear analysis are extended into the fully nonlinear regime through numerical simulations. The results of the model are compared with simulations of the full three-dimensional magnetoconvection problem.


Effect of extracellular manganese on amylase release from dispersed pancreatic acini.
In dispersed acini from guinea pig pancreas, adding extracellular manganese increased amylase release. A significant effect could be detected with 0.25 mM manganese, and maximal stimulation occurred with 1 mM manganese. When manganese was added, the rate of amylase release did not change during the first 20 min of incubation and then gradually increased to a new steady state by 80 min, which with 1 mM manganese represented a fourfold increase in the rate of enzyme release. Extracellular manganese inhibited the stimulation of amylase release caused by those secretagogues that mobilize cellular calcium but augmented the stimulation caused by those secretagogues whose actions are mediated by cellular cAMP. The mechanism by which manganese altered stimulated amylase secretion differed from the mechanism by which manganese stimulated basal amylase release because the change in stimulated release was maximal within 10 min, whereas the change in basal release did not occur until after 20 min. The actions of manganese on secretagogue-stimulated amylase release were not attributable to manganese-induced changes in secretagogue-stimulated calcium outflux or cAMP and, instead, appear to result from actions of manganese on one of the later steps in the mechanisms for stimulating the secretory process.


[Clinical Guidelines 2.0: Translation of Tables 2 and 3 and Apendix 1 of Schünemann HJ et al. CMAJ. 2014; 186:E123-42].
Clinical Practice Guidelines are instruments to support decision to improve the quality of clinical care. An expert group from McMaster University (Canada) has developed - from high-quality literature sources - a guidance on the practical steps for their development, dissemination, implementation and evaluation. This is the 1st time anyone seeks to bring together in one document all information regarding the Clinical Practice Guidelines. Due to the interest of this paper, the Centre for Evidence Based Medicine at the University of Lisbon School of Medicine contacted the authors of the article and the journal where it was published (the Canadian Medical Association Journal) in order to translate the most relevant parts of the article (including the practice tables), which was agreed. This guide should be useful to those who, being interested in the development, dissemination and implementation of Clinical Practice Guidelines, want to ensure their intrinsic quality based on relevant and updated evidence.


Direct optical detection of viral nucleoprotein binding to an anti-influenza aptamer.
We have demonstrated label-free optical detection of viral nucleoprotein binding to a polyvalent anti-influenza aptamer by monitoring the surface-enhanced Raman (SERS) spectra of the aptamer-nucleoprotein complex. The SERS spectra demonstrated that selective binding of the aptamer-nucleoprotein complex could be differentiated from that of the aptamer alone based solely on the direct spectral signature for the aptamer-nucleoprotein complex. Multivariate statistical methods, including principal components analysis, hierarchical clustering, and partial least squares, were used to confirm statistically significant differences between the spectra of the aptamer-nucleoprotein complex and the spectra of the unbound aptamer. Two separate negative controls were used to evaluate the specificity of binding of the viral nucleoproteins to this aptamer. In both cases, no spectral changes were observed that showed protein binding to the control surfaces, indicating a high degree of specificity for the binding of influenza viral nucleoproteins only to the influenza-specific aptamer. Statistical analysis of the spectra supports this interpretation. AFM images demonstrate morphological changes consistent with formation of the influenza aptamer-nucleoprotein complex. These results provide the first evidence for the use of aptamer-modified SERS substrates as diagnostic tools for influenza virus detection in a complex biological matrix.


Bordetella parapertussis infection in children: epidemiology, clinical symptoms, and molecular characteristics of isolates.
The clinical trial conducted in Italy to evaluate the efficacy of acellular pertussis vaccines provided an opportunity to estimate the frequency of clinical infections with Bordetella parapertussis and to compare the clinical characteristics of children suffering from Bordetella pertussis illness with those of children with B. parapertussis illness. This study dealt with 76 B. parapertussis infections diagnosed from a population of 15,601 children participating in the follow-up of suspected cases of pertussis. An overall incidence of 2.1 cases of laboratory-confirmed parapertussis per 1,000 person-years was observed. Children affected by B. parapertussis infections showed a less severe clinical picture both in the duration of symptoms and in the percentage of patients affected, even when compared with vaccinated children with pertussis. To characterize the isolated strains, we performed assays for susceptibility to erythromycin and sulfamethoxazole-trimethoprim, and we examined the genomic DNAs by pulsed-field gel electrophoresis. The results showed a high degree of genetic stability among B. parapertussis strains regardless of time of collection and geographical distribution.


Detoxification of Fusaric Acid by the Soil Microbe Mucor rouxii.
Fusarium oxysporum f. sp. vasinfectum race 4 (VCG0114), which causes root rot and wilt of cotton (Gossypium hirsutum and G. barbadense), has been identified recently for the first time in the western hemisphere in certain fields in the San Joaquin Valley of California. This pathotype produces copious quantities of the plant toxin fusaric acid (5-butyl-2-pyridinecarboxylic acid) compared to other isolates of F. oxysporum f. sp. vasinfectum (Fov) that are indigenous to the United States. Fusaric acid is toxic to cotton plants and may help the pathogen compete with other microbes in the soil. We found that a laboratory strain of the fungus Mucor rouxii converts fusaric acid into a newly identified compound, 8-hydroxyfusaric acid. The latter compound is significantly less phytotoxic to cotton than the parent compound. On the basis of bioassays of hydroxylated analogues of fusaric acid, hydroxylation of the butyl side chain of fusaric acid may affect a general detoxification of fusaric acid. Genes that control this hydroxylation may be useful in developing biocontrol agents to manage Fov.


Combination antiretroviral therapy including ritonavir in children infected with human immunodeficiency.
To assess the efficacy of combination therapy that includes ritonavir in HIV-1 infected children. A monocentric retrospective study. Twenty-two children with a minimum follow-up of 6 months under triple therapy including ritonavir were analysed for treatment efficacy. At entry, all the patients were protease inhibitor naive and all but two had received previous antiretroviral therapy during a median period of 5 years. Their initial median CD4+ lymphocyte count and viral load were 121 x 10(6)/l and 5.08 log10 copies/ml, respectively. Clinical and biological evaluation included clinical assessment every 6 weeks and determination of CD4 cell count and HIV-RNA concentration every 3 months. Median length of follow-up on triple therapy was 15 months (range: 7-21 months). Neither progression in the CDC classification nor death occurred. No significant change in mean weight SD scores was noted when baseline values were compared with values obtained after 1 year of triple therapy. Median CD4 count increases were of 210 x 10(6)/l, 415 x 10(6)/l, and 472 x 10(6)/l cells at 6, 12, and 18 months, respectively. Among the patients baseline characteristics, neither age nor initial CD4 cells count influenced the magnitude of immunologic improvement. There were median decreases of 1.14, 0.95, and 1.5 log10 per ml of plasma in the concentration of viral RNA at 6, 12, and 18 months respectively. Seven patients maintained an undetectable viral load when under treatment. The introduction of at least one new reverse transcriptase inhibitor at the initiation of triple therapy correlated significantly with a greater viral suppression. Despite variable viral response, antiretroviral-experienced HIV-infected children demonstrated a substantial CD4 cell increase during a median period of 15 months of ritonavir containing combination therapy.


Adolescent development and the measurement of juvenile psychopathy.
Researchers are currently seeking antecedents of psychopathy among children and adolescents. Instruments developed for this purpose are likely to be used in the future in ways that may have serious implications for youths in delinquency and criminal cases. In this regard, and because the concept and measurement of psychopathy have been developed in reference to adults, there is reason to be concerned about potential developmental sources of false positives when measuring psychopathic traits in juvenile offenders. We provide the basis for our concern, as well as standards regarding the types of research evidence that should be provided before psychopathy measures are accepted for use in juvenile forensic assessments.


Molecular mechanisms of sleep and wakefulness.
Major questions on the biology of sleep include the following: what are the molecular functions of sleep; why can wakefulness only be sustained for defined periods before there is behavioral impairment; what genes contribute to the individual differences in sleep and the response to sleep deprivation? Behavioral criteria to define sleep have facilitated identification of sleep states in a number of different model systems: Drosophila, zebrafish, and Caenorhabditis elegans. Each system has unique strengths. Studies in these model systems are identifying conserved signaling mechanisms regulating sleep that are present in mammals. For example, the PKA-CREB signaling mechanism promotes wakefulness in Drosophila, mice, and C. elegans. Microarray studies indicate that genes whose expression is upregulated during sleep are involved in macromolecule biosynthesis (proteins, lipids [including cholesterol], heme). Thus, a key function of sleep is likely to be macromolecule synthesis. Moreover, in all species studied to date, there is upregulation of the molecular chaperone BiP with extended wakefulness. Sleep deprivation leads to cellular ER stress in brain and the unfolded protein response. Identification of genes regulating sleep has the potential for translational studies to elucidate the genetics of sleep and response to sleep deprivation in humans.


Conjugate heptavalent pneumococcal vaccine outcome improvements.
Heptavalent conjugate pneumococcal vaccine (PCV7) is now routinely administered to infants. Medical records from a pediatric outpatient clinic were used to identify two study groups: 1) those born between January 1995-May 1997(controls), 2) those born between April 2000--May 2001 (PCV7 group). The PCV7 group showed a lower incidence of otitis media episodes, clinic visits for fever and respiratory symptoms, antibiotic prescriptions, ceftriaxone use, and hospitalizations (singled sided probabilities).


Osteopathic Evaluation of Urinary Retention Caused by Atypical Presentation of Invasive Cervical Cancer Mimicking Primary Urothelial Tumor.
Cancer of the uterine cervix is the third most common gynecologic cancer diagnosis and cause of death among gynecologic cancers in the United States. Cervical cancer is frequently asymptomatic until it reaches a locally advanced stage. The authors present a case of urinary retention caused by an atypical presentation of invasive cervical cancer mimicking primary urothelial tumor. The patient was evaluated by a multidisciplinary team, with osteopathic structural examination, intraoperative examination, surgical resection, and histopathologic analysis.


Next-Generation Digital Biomarkers for Tuberculosis and Antibiotic Stewardship: Perspective on Novel Molecular Digital Biomarkers in Sweat, Saliva, and Exhaled Breath.
The internet of health care things enables a remote connection between health care professionals and patients wearing smart biosensors. Wearable smart devices are potentially affordable, sensitive, specific, user-friendly, rapid, robust, lab-independent, and deliverable to the end user for point-of-care testing. The datasets derived from these devices are known as digital biomarkers. They represent a novel patient-centered approach to collecting longitudinal, context-derived health insights. Adding automated, analytical smartphone applications will enable their use in high-, middle-, and low-income countries. So far, digital biomarkers have been focused primarily on accelerometer data and heart rate due to well-established sensors originating from the consumer market. Novel emerging smart biosensors will detect biomarkers (or compounds) independent of a lab and noninvasively in sweat, saliva, and exhaled breath. These molecular digital biomarkers are a promising novel approach to reduce the burden from 2 major infectious diseases with urgent unmet needs: tuberculosis and infections with multidrug resistant pathogens. Active tuberculosis (aTbc) is one of the deadliest diseases from an infectious agent. However, a simple and reliable test for its detection is still missing. Furthermore, inappropriate antimicrobial use leads to the development of antimicrobial resistance, which is associated with high mortality and health care costs. From this perspective, we discuss the innovative approach of a noninvasive and lab-independent collection of novel biomarkers to detect aTbc, which at the same time may additionally serve as a scalable therapeutic drug monitoring approach for antibiotics. These molecular digital biomarkers are next-generation digital biomarkers and have the potential to shape the future of infectious diseases.


An unusually dry story.
We present a middle-aged woman with a prior history of central nervous system (CNS) demyelinating disorder who presented with an acute onset quadriparesis and respiratory failure. The evaluation revealed distal renal tubular acidosis with hypokalemia and medullary nephrocalcinosis. Weakness persisted despite potassium correction, and ongoing evaluation confirmed recurrent CNS and long-segment spinal cord demyelination with anti-aquaporin-4 antibodies. There was no history of dry eyes or dry mouth. Anti-Sjogren's syndrome A antigen antibodies were elevated, and there was reduced salivary flow on scintigraphy. Coexistent antiphospholipid antibody syndrome with inferior vena cava thrombosis was also found on evaluation. The index patient highlights several rare manifestations of primary Sjogren's syndrome (pSS) as the presenting features and highlights the differential diagnosis of the clinical syndromes in which pSS should be considered in the Intensive Care Unit.


Isolation and characterization of rat complement factor B and its interaction with cell-bound human C3.
Factor B was isolated from fresh rat plasma by sequential chromatography on QAE-A50, Biorex-70, gel filtration on Sephadex G-200 superfine and rechromatography on QAE-A50. That Brat was isolated in its native form was indicated by its migration during immunoelectrophoresis and by its capacity to react with cobra venom factor (CoVF) in the presence of human D to form a C3 convertase capable of cleaving purified rat C3 and human C3. The recovery of Brat was between 8 and 15%; the final material was homogeneous according to SDS-PAGE analysis. Reduction of Brat with DTT in the presence of urea and SDS did not produce detectable peptides of lower molecular weight. Both reduced and unreduced Brat had an apparent molecular weight of 100,000. An antiserum against Brat induced in rabbits recognized only one protein in fresh rat plasma as indicated by immunodiffusion and immunoelectrophoretic analysis. Zymosan treatment of rat serum resulted in the cleavage of Brat into two fragments with alpha and gamma mobility. Native Brat has a beta electrophoretic mobility. The plasma concentration of Brat in Wistar rats was 215 +/- 38 microgram/ml (mean +/- SD).


The relationship between dietary intake, growth and body composition in Phenylketonuria.
Phenylketonuria (PKU) is an inborn error of protein metabolism that results from perturbation in phenylalanine hydroxylase activity leading to elevated blood levels of phenylalanine (phe). We aimed to explore the relationships between dietary patterns (total-protein, natural-protein, amino-acid formula), and the ratio of protein to energy intake with growth and body composition. Longitudinal prospective data (1-6 measurements) of growth, dietary intake and body composition in patients treated with phe-restricted diet only (D-PKU; n=32), and tetrahydrobiopterin (BH4)±phe-restricted diet (BH4-PKU; n=5) were collected over a two-year period. Healthy siblings provided control data (n=21). There were no significant differences in weight-, height-, BMI z-score or percent body fat mass (%fatmass) between the D-PKU, BH4-PKU and control groups or between the all-types of PKU combined and controls, which confirmed 'normal' growth in the PKU cohort. Total-protein intake in the all-types of PKU group met or exceeded WHO safe protein recommendations. There were no significant relationships between anthropometric and dietary variables. Significant negative correlations were found in body composition: %fatmass and total-protein intake (rs=-0.690, p≤0.001), natural-protein intake (rs=-0.534, p=0.001), and AAF intake (rs=-0.510, p=0.001). Age was significantly correlated with %fatmass (rs=0.493, p=0.002) A total-protein intake of 1.5-2.6g/kg/day and natural-protein intake >0.5g/kg/day were associated with improved body composition. An apparent safe P:E ratio of 3.0-4.5g protein/100kcal was strongly associated with appropriate growth outcomes. Clinical decision-making needs to consider both the enhancement of natural-protein tolerance and the application of an apparent 'safe' protein to energy ratio to support optimal growth and body composition in PKU.


MicroRNA-184 is a downstream effector of albuminuria driving renal fibrosis in rats with diabetic nephropathy.
Renal fibrosis is a common complication of diabetic nephropathy and is a major cause of end-stage renal disease. Despite the suggested link between renal fibrosis and microRNA (miRNA) dysregulation in diabetic nephropathy, the identification of the specific miRNAs involved is still incomplete. The aim of this study was to investigate miRNA profiles in the diabetic kidney and to identify potential downstream targets implicated in renal fibrosis. miRNA expression profiling was investigated in the kidneys of 8-month-old Zucker diabetic fatty (ZDF) rats during overt nephropathy. Localisation of the most upregulated miRNA was established by in situ hybridisation. The candidate miRNA target was identified by in silico analysis and its expression documented in the diabetic kidney associated with fibrotic markers. Cultured tubule cells served to assess which of the profibrogenic stimuli acted as a trigger for the overexpressed miRNA, and to investigate underlying epigenetic mechanisms. In ZDF rats, miR-184 showed the strongest differential upregulation compared with lean rats (18-fold). Tubular localisation of miR-184 was associated with reduced expression of lipid phosphate phosphatase 3 (LPP3) and collagen accumulation. Transfection of NRK-52E cells with miR-184 mimic reduced LPP3, promoting a profibrotic phenotype. Albumin was a major trigger of miR-184 expression. Anti-miR-184 counteracted albumin-induced LPP3 downregulation and overexpression of plasminogen activator inhibitor-1. In ZDF rats, ACE-inhibitor treatment limited albuminuria and reduced miR-184, with tubular LPP3 preservation and tubulointerstitial fibrosis amelioration. Albumin-induced miR-184 expression in tubule cells was epigenetically regulated through DNA demethylation and histone lysine acetylation and was accompanied by binding of NF-κB p65 subunit to miR-184 promoter. These results suggest that miR-184 may act as a downstream effector of albuminuria through LPP3 to promote tubulointerstitial fibrosis, and offer the rationale to investigate whether targeting miR-184 in association with albuminuria-lowering drugs may be a new strategy to achieve fully anti-fibrotic effects in diabetic nephropathy.


Effects of initial cell density and hydrodynamic culture on osteogenic activity of tissue-engineered bone grafts.
This study aimed to study the effects of initial cell density and in vitro culture method on the construction of tissue-engineered bone grafts and osteogenic activities. Human mesenchymal stem cells (hMSCs) were seeded onto cubic scaffolds prepared from demineralized bone matrix (DBM) by three methods - static, hydrodynamic, or fibrin hydrogel-assisted seeding. The resulting cell-scaffold constructs were cultured in vitro by static flask culture or hydrodynamic culture. The initial cell density and the subsequent in vitro proliferation and alkaline phosphate activities of the constructs were analyzed. The constructs were also subcutaneously implanted in nude mice to examine their in vivo osteogenic activities. Hydrogel-assisted seeding gave the highest seeding efficiency, followed by hydrodynamic and conventional static seeding. During in vitro culture, hydrodynamic culture produced higher plateau cell densities, alkaline phosphatase (ALP) activities, and extracellular matrix production than static culture. After subcutaneous implantation in nude mice, the implants prepared by the combination of hydrogel-assisted seeding and hydrodynamic culture produced higher wet weight and bone mineral density than implants prepared by other methods. The results suggest that the hydrogel-assisted seeding can substantially increase the initial seed cell density in scaffolds. Subsequent hydrodynamic culture can promote the proliferation and osteoblastic differentiation of the seeded cells. Correspondingly, bone grafts produced by the combination of these two methods achieved the highest osteogenic activity among the three methods employed.


Chemotyping of diverse Eucalyptus species grown in Egypt and antioxidant and antibacterial activities of its respective essential oils.
The chemical composition of the essential oil from the leaves of Eucalyptus camaldulensis, Eucalyptus camaldulensis var. obtusa and Eucalyptus gomphocephala grown in northern Egypt was analysed by using GC-FID and GC-MS techniques. The antibacterial (agar disc diffusion and minimum inhibitory concentration methods) and antioxidant activities (2,2'-diphenypicrylhydrazyl) were examined. The main oils constituents were 1,8-cineole (21.75%), β-pinene (20.51%) and methyleugenol (6.10%) in E. camaldulensis; spathulenol (37.46%), p-cymene (17.20%) and crypton (8.88%) in E. gomphocephala; spathulenol (18.37%), p-cymene (19.38%) and crypton (16.91%) in E. camaldulensis var. obtusa. The essential oils from the leaves of Eucalyptus spp. exhibited considerable antibacterial activity against Gram-positive and Gram-negative bacteria. The values of total antioxidant activity were 70 ± 3.13%, 50 ± 3.34% and 84 ± 4.64% for E. camaldulensis, E. camaldulensis var. obtusa and E. gomphocephala, respectively. The highest antioxidant activity value of 84 ± 4.64% could be attributed to the high amount of spathulenol (37.46%).


Effects of job-related stress and burnout on asthenopia among high-tech workers.
Eye- and vision-related symptoms are the most frequent health problems among computer users. The findings of eye strain, tired eyes, eye irritation, burning sensation, redness, blurred vision and double vision, when appearing together, have recently been termed 'computer vision syndrome', or asthenopia. To examine the frequency and intensity of asthenopia among individuals employed in research and development departments of high-tech firms and the effects of job stress and burnout on ocular complaints, this study included 106 subjects, 42 high-tech workers (study group) and 64 bank employees (control group). All participants completed self-report questionnaires covering demographics, asthenopia, satisfaction with work environmental conditions, job-related stress and burnout. There was a significant between-group difference in the intensity of asthenopia, but not in its frequency. Burnout appeared to be a significant contributing factor to the intensity and frequency of asthenopia. This study shows that burnout is a significant factor in asthenopic complaints in high-tech workers. This manuscript analyses the effects of psychological environmental factors, such as job stress and burnout, on ocular complaints at the workplace of computer users. The findings may have an ergonomic impact on how to improve health, safety and comfort of the working environment among computer users, for better perception of the job environment, efficacy and production.


Risk factors for prostate cancer.
The incidence of prostate cancer in the United States has recently undergone dramatic and unprecedented changes. Exposure to prostate-specific antigen testing has led to a steep increase in reported incidence. The data indicate that the strongest risk factors for prostate cancer are age and African American race/ethnicity. Family history is also an important risk factor for prostate cancer, although only a small proportion of cases will be due to high-penetrance genes such as those at the putative susceptibility loci (eg, ELAC2) recently identified through linkage analysis. International variation in the risk of prostate cancer is profound, as is the frequency with which migrants from low- to high-risk areas adopt the risk pattern of the host country, possibly within a single generation. This article reviews the current state of knowledge regarding risk factors for prostate cancer, including factors related to diet, anthropometrics, hormone profiles, and concomitant medical conditions. The need for unifying, overarching hypotheses is emphasized, along with identification of some of the barriers to future progress in the field.


Interleukin 2 restores CD3-zeta chain expression but fails to generate tumour-specific lytic activity in tumour-infiltrating lymphocytes derived from human colorectal hepatic metastases.
Metastatic colorectal cancer is usually progressive despite infiltration of the tumours by T lymphocytes, suggesting that these tumour-infiltrating lymphocytes (TILs) are functionally deficient. Recently, TILs from other tumours have been shown to express reduced levels of the T-cell receptor signal-transducing CD3-zeta chain. We were interested to determine whether a similar abnormality existed in TILs from human colorectal hepatic metastasis (CHM) and, if so, whether correcting the abnormality in vitro would restore anti-tumour activity and provide support for the development of immunotherapy for colorectal hepatic metastases. Twelve of 19 TILs from colorectal hepatic metastases were successfully expanded in vitro in high-dose recombinant interleukin 2 (rlL-2) and their specific anti-tumour cytolytic activity was determined. CD3-positive (CD3+) TILs were HLA-Drhigh and CD69high, suggesting that they had been activated by exposure to antigen but expressed low levels of CD25, CD71 and the nuclear proliferation antigen Ki-67. Furthermore, they showed reduced expression of CD3-zeta compared with autologous peripheral blood T cells (PBTs) and failed to proliferate in the absence of high-dose rIL-2. Expansion of TILs in rIL-2 resulted in restoration of CD3-zeta expression and the ability to lyse K562 and Daudi cells but not autologous tumour cells. The absence of autologous tumour-specific cytolytic T-cell (CTL) activity may be due to the poor immunogenicity of colorectal tumour cells, which we found expressed only low levels of MHC I antigens and CD54 and failed to express MHC II antigens or the co-stimulatory molecules CD80, CD86 or CD106. The inability of rIL-2 to generate tumour-specific CTLs despite restoration of CD3-zeta expression and the presence of an intact lytic mechanism suggests that successful immunotherapy may require the development of strategies to increase the immunogenicity of this tumour.


Evaluation of intermittent long-term negative-pressure ventilation in patients with severe chronic obstructive pulmonary disease.
We tested the hypothesis that intermittent ventilatory assistance in patients with severe chronic obstructive pulmonary disease (COPD) improves pulmonary function and exercise capacity. Twenty stable patients with severe COPD were recruited from outpatient pulmonary clinics and were randomized to use a poncho wrap, negative-pressure ventilator or to receive standard care. After 6 months, the patients receiving standard care were switched over to the ventilator and vice versa, and follow-up was continued for an additional 6 months. After 3 to 6 months of ventilator use, we observed no clinically significant improvements in FEV1, FVC, blood gas determinations, maximal inspiratory and expiratory pressures, and exercise duration. However, 11 of our patients dropped out of the study because of an inability to tolerate the ventilator, and all but one of the nine who completed the study expressed dissatisfaction with it, using it for less time (4.1 h/day) than we recommended. Musculoskeletal pain and inconvenience were the most frequently voiced complaints. Because we did not document that ventilator use actually rested the respiratory muscles in our patients and because duration of ventilator use may have been too brief, we cannot conclude that intermittent rest of respiratory muscles in patients with severe COPD fails to bring about improvement. On the other hand, our results demonstrate that the poncho wrap ventilator is poorly tolerated by patients with severe COPD in a typical outpatient setting. We suggest that future trials seek to utilize better tolerated ventilatory assist devices.


Solid-phase extraction and nanoflow liquid chromatography-nanoelectrospray ionization mass spectrometry for improved global urine metabolomics.
Global urine metabolomics is a rapidly expanding field with the potential to discover biomarkers of disease and exposure. To date, most methods focus on rapid sample preparation, using neat or diluted urine, together with high-throughput analyses, and are poorly suited for detection of low abundance metabolites present in urine samples. In this study, novel methods have been developed to analyze urine by splitless nanoflowUHPLC-nanoESI-TOFMS (nUHPLC-nESI-TOFMS) after preconcentration by solid-phase extraction (SPE), thus enabling significant improvements in analytical sensitivity and coverage of the urinary metabolome. In initial work, urine samples were extracted by both anion and cation exchange mixed-mode polymeric SPE cartridges and qualitatively compared with those using conventional sample preparations using UHPLC-ESI-TOFMS analyses. Compared with neat or diluted urine samples, SPE concentration of urine resulted in detection of additional metabolites including bile acids, lipids, pharmaceuticals, and markers of lifestyle, with little loss of other components of the metabolome. Analyses of SPE preparations by nUHPLC-nESI-TOFMS revealed excellent retention time repeatability with <1% coefficient of variation (CV) for 96% of analyzed peaks. The repeatability of the MS response was <30% CV for >79% of MS features in both negative and positive nESI modes. Compared with UHPLC-ESI-TOFMS, analysis by the nanoplatform enabled detection of signaling molecules important in disease processes including sex steroids, glucocorticoids, eicosanoids, and neurotransmitter metabolites. The significant improvement in sensitivity arising from use of splitless nUHPLC-nESI-TOFMS analyses of SPE-concentrated samples represents a step change in coverage of the urinary metabolome, thereby increasing the potential for biomarker discovery.


Preparation of a hybrids APT@MIL by one-step solvent-thermal method for effectively degrading organics.
The attapulgite (APT), a typical nano-rod structured clay was introduced to MIL-101(Fe), a typical eco-friendly iron-based Metal-Organic Framework material (MOF), during the preparation by a one-step solvothermal method, which afforded a novel APT and MOF hybrid (APT@MIL). Based on the characterization of SEM, FT-IR and XRD, it was found that the rod-like crystals of APT determined the size of MIL-101(Fe) while maintaining its regular octahedral crystal form, and the crystal size of MIL-101(Fe) in APT@MIL enlarged 4 times. It was also discovered that the rod-like APT were evenly distributed in MIL-101(Fe) crystals. Using APT@MIL as the photocatalyst, some organic dyes were photodegraded in simulated sunlight. The analysis indicated that APT@MIL has high adsorption and photodegradation activity, the removal rate of methylene blue was up to 99.5%. Finally, the photocatalytic activity of APT@MIL was verified by UV-Vis DRS, photoluminescence spectra. The thermodynamic adsorption, kinetic characteristics adsorption, and removal mechanism of APT@MIL are also discussed. In summary, a novel hybrid material APT@MIL was successfully prepared with good adsorption and photocatalytic performance. It is expected to be used in photocatalytic degradation of dye wastewater.


[Gene transfer efficiencies and cell tropism of recombinant adeno-associated virus in the hippocampus of adult rat].
To explore the gene transfer efficiencies and different cell tropism of recombinant adeno-associated virus 1 (rAAV1), rAAV 2, and rAAV 5 in the hippocampus of adult rats, and select more suitable gene vectors for central nervous system (CNS) gene therapy. Eighteen SD male adult rat were divided into 3 groups randomly (n = 6), with every group being injected with the titre and volume matched rAAV1, rAAV2, and rAAV5 vectors. All these vectors contained enhanced green fluorescent protein (EGFP) sequences as a reporter gene. Animals were killed after 8 weeks. The coronal cryosections of brains were processed, and the EGFP gene expression was observed with fluorescence microscopy; the expression area and the number of EGFP positive cells were automatically measured using Image-Pro Plus 4.5 software. To identify their cell tropism in the CNS, the sections were counterstained with neuronal marker neuron-specific nuclear protein (NeuN) and the astrocyte marker glial fibrillary acidic protein (GFAP) and were examined with a confocal laser scanning microscope. Eight weeks after adeno-associated virus gene transfer, the expression profile of EGFP demonstrated significant difference. Most of the pyramidal cell layers of CA1 to CA3 area and granular cell of dent gyrus were strongly transduced by rAAV1; whereas rAAV2 primarily transduced the cells of multiform layer in hilar region of the dentate gyrus; only a few pyramidal cells were transduced by rAAV5. Moreover, rAAV1 showed significantly wider distribution throughout the hippocampus, and the quantity of EGFP positive cells and the EGFP positive area were significantly more than those of rAAV2 and rAAV5 (P < 0.01). The counterstaining for NeuN and GFAP showed that rAAV1 was able to transduce both neurons and glia cells, whereas rAAV2 and rAAV5 transduced the neurons only. rAAV1 is an excellent transgene vector with higher efficiency and broader cell tropism in the CNS.


Effects of local anesthesia with bupivacaine plus epinephrine on blepharoptosis and levator palpebrae muscle function.
To evaluate the effect of local anesthesia with bupivacaine plus epinephrine on the extent of blepharoptosis and levator palpebrae muscle function. : In this prospective interventional case series, patients with blepharoptosis who were candidates for aponeurotic surgery were included. After initial preparations in the operating room, a total of 1 ml of a mixture of bupivacaine 0.5% plus epinephrine 1:100,000 were injected into the upper eyelid. The margin reflex distance 1 (MRD1) and the extent of levator muscle function were measured before and 2, 5, 10, and 15 minutes after injection. A total of 36 eyes including 21 men (58.3%) and 15 women (41.7%) with an average age of 41.81 ± 23.09 (17-83 years) were studied. There were 21 eyes with myogenic and 15 eyes with aponeurotic blepharoptosis. The mean MRD1 was 1.18 ± 1.06 mm before injection and -0.02 ± 0.85, 0.52 ± 0.98, 0.98 ± 1.05, and 1.02 ± 1.06 mm at 2, 5, 10, and 15 minutes after injections, respectively. The changes in the MRD1 measurements were statistically significant at all time points. The MRD1 values decreased during the first 2 minutes after injection in 88.8% of eyes, but returned to initial value after 15 minutes in 84.3%. The change in the levator muscle function measurements was statistically significant at 2 and 5 minutes after injections; however, the differences were clinically negligible. : The local anesthesia of the eyelid with 1 ml bupivacaine plus epinephrine causes a temporary increase of blepharoptosis within the first few minutes with minimal effect on levator muscle function measurements.


Improved survival after pulmonary metastasectomy for soft tissue sarcoma.
Survival after pulmonary metastasectomy for soft tissue sarcoma (STS) has been lower than in osteosarcoma (14-40% versus 40-50%). With improved patient selection criteria and advanced chemotherapy agents, we hypothesized that survival after metastasectomy for STS has improved in recent years. Retrospective study of 48 patients undergoing pulmonary metastasectomy for STS between 1995 and 2007. Potential predictors of overall survival and disease-free survival (DFS) were examined using the log-rank test or Cox regression. Multivariate analysis was conducted using Cox regression. Overall survival after initial metastasectomy was 67% and 52% at 3 and 5 years, respectively; DFS was 17% and 10% at 3 and 5 years. Univariate analysis indicated that ≤2 pulmonary metastases (p = 0.03), diameter of largest metastasis ≤2 cm (p = 0.09), and the absence of extrapulmonary metastases (p = 0.10) were associated with longer overall survival. Absence of extrapulmonary metastases (p = 0.07) and smaller size of the largest pulmonary metastasis (p = 0.06) were associated with longer DFS. Before 2001, 46.7% of patients received adjuvant chemotherapy versus 72.7% after (p = 0.10). Neither use of chemotherapy nor chemotherapy type was related to overall survival or DFS. Five-year overall survival is substantially higher after pulmonary metastasectomy for STS in our study relative to previously published results (52% versus 14-40%). This improvement does not seem to be the result of greater use of, or newer, chemotherapeutic regimens. Among potential explanations, improved patient selection is the most likely factor.


Creating Extraordinary From Ordinary: High Resource Efficiency of Underdog Entrepreneurs and Its Mechanism.
Existing theory has not documented the potential benefits of facing the challenges of underdog entrepreneurs, who may succeed unexpectedly. This research explains why, and under what circumstances, the underdog status of entrepreneurs can promote entrepreneurial success rather than just hinder it. We predict that the underdog effect has the potential to boost entrepreneurial resource efficiency when entrepreneurs hold an incremental (vs. entity) theory, enter a low-barrier (vs. high-barrier) industry, and are in a favorable (vs. unfavorable) business environment. Study 1 provides support for the positive relationship between underdog status and resource efficiency through an ordinary least squares (OLS) regression analysis, which is accompanied by a moderating effect of the implicit theory, industry context, and business environment. The data was obtained from two nationwide surveys. By extending a qualitative comparative analysis (QCA) of multiple case studies, Study 2 reveals support for a synergistic effect of the above factors. Our research results examine the assumption that perceiving underdog status is detrimental and offer meaningful insights into why and when underdog entrepreneurs have good performance in entrepreneurial resource efficiency. We provide a psychological and behavioral explanation for the underdog effect, extending the underdog effect theory to the field of entrepreneurship for the first time from the perspective of the actors. Finally, theoretical contributions and practical implications are discussed by indicating the limitations of the research.


The NBS1-ATM connection revisited.
Nijmegen Breakage syndrome (NBS) is a rare autosomal recessive disorder characterized by microcephaly, immunodeficiency, and increased predisposition to the development of malignancy. Due to the overlap of clinical and cellular features of patients with ataxia telangiectasia (AT), NBS was described as an AT variant syndrome until the underlying gene product mutation was identified. Cells from both AT and NBS patients show increased sensitivity to ionizing radiation (IR), genomic instability and cell cycle checkpoint defects following DNA damage, suggesting that both gene products participate in the same DNA damage response pathway. Here we highlight recent developments and refinements in our understanding of the interplay between NBS1 and ATM in vivo.


Design, execution, and interpretation of plant RNA-seq analyses.
Genomics has transformed our understanding of the genetic architecture of traits and the genetic variation present in plants. Here, we present a review of how RNA-seq can be performed to tackle research challenges addressed by plant sciences. We discuss the importance of experimental design in RNA-seq, including considerations for sampling and replication, to avoid pitfalls and wasted resources. Approaches for processing RNA-seq data include quality control and counting features, and we describe common approaches and variations. Though differential gene expression analysis is the most common analysis of RNA-seq data, we review multiple methods for assessing gene expression, including detecting allele-specific gene expression and building co-expression networks. With the production of more RNA-seq data, strategies for integrating these data into genetic mapping pipelines is of increased interest. Finally, special considerations for RNA-seq analysis and interpretation in plants are needed, due to the high genome complexity common across plants. By incorporating informed decisions throughout an RNA-seq experiment, we can increase the knowledge gained.


Diagnosis Using CCTA and Management of Anomalous Right Coronary Artery from the Opposite Sinus.
Coronary anomalies can be observed in 1-1.2% of all angiograms performed. Majority of coronary anomalies are benign and do not lead to cardiac ischemia; however anomalous coronary arteries from the opposite sinus (ACAOS) are often associated with sudden cardiac deaths, typically in 0.11-0.35% of individuals who participate in vigorous physical activity (Peñalver et al., 2012). Left and right ACAOS have an incidence of 0.15% and 0.92%, respectively. Left ACAOS are often associated with higher incidence of sudden cardiac death; this could be secondary to greater territory of myocardial perfusion by the left coronary artery. ACAOS are often asymptomatic and initially present as sudden death following exertion in young athletes. The management of left ACAOS is clear and surgery is usually indicated. However there is a lack of consensus on the management of certain cases of right ACAOS. In this paper a case of 20 yo M with right coronary artery from left sinus is going to be presented with a discussion on pathophysiology, diagnosis, and management.


Diabetes and atherosclerosis. Metabolic links.
Atherosclerosis is the most frequent complication of diabetes. Two of the many factors that may contribute to this are dyslipoproteinaemias and hyperinsulinaemia. The most frequent dyslipoproteinaemia in diabetics is hypertriglyceridaemia. This probably reflects an increase in intermediate density lipoprotein (IDL) concentration. Plasma levels of triglyceride and insulin are often correlated. This partly reflects a vicious circle in which hypertriglyceridaemia itself can cause insulin resistance and compensatory hyperinsulinaemia. The chronic hyperinsulinaemic state in vivo is, in turn, associated with an increase in the rate of production of very low density lipoprotein (VLDL) triglyceride. Such a vicious circle is of potential importance in atherogenesis in view of the association of both hypertriglyceridaemia and hyperinsulinaemia with early macrovascular disease.


Ion beam doping of silicon nanowires.
We demonstrate n- and p-type field-effect transistors based on Si nanowires (SiNWs) implanted with P and B at fluences as high as 10(15) cm (-2). Contrary to what would happen in bulk Si for similar fluences, in SiNWs this only induces a limited amount of amorphization and structural disorder, as shown by electrical transport and Raman measurements. We demonstrate that a fully crystalline structure can be recovered by thermal annealing at 800 degrees C. For not-annealed, as-implanted NWs, we correlate the onset of amorphization with an increase of phonon confinement in the NW core. This is ion-dependent and detectable for P-implantation only. Hysteresis is observed following both P and B implantation.


Characterization and proliferation capacity of potentially pathogenic fungi in marine and freshwater fish commercial feeds.
In the aquaculture industry, the selection and quality of feed are highly relevant because their integrity and management have an impact on the health and development of organisms. In general, feeds contamination depends on storage conditions and formulation. Furthermore, it has been recognized that filamentous fungi are among the most important contaminating agent in formulated feeds. Therefore, the purpose of this research was to identify saprophytic fungi capable of proliferating in commercial feeds, as well as determining their prevalence, extracellular enzymes profile, ability to assimilate carbon sources, and finally their ability to produce aflatoxins. In order to do that, twenty-two fungi were isolated from commercial fish feeds. After, the species Aspergillus chevalieri, A. cristatus, A. sydowii, A. versicolor, A. flavus, A. creber, and Lichtheimia ramosa were identified. These fungi were able to produce extracellular enzymes, such as phosphatases, esterases, proteases, β-glucosidase, and N-acetyl-β-glucosaminidase. The isolated fungi showed no selective behavior in the assimilation of the different carbon sources, showing a strong metabolic diversity. Prevalence percentages above 85% were recorded. Among all fungi studied, A. flavus M3-C1 had the highest production of aflatoxins when this strain was inoculated directly in the feeds (295 ppb). The aflatoxin production by this strain under the experimental setting is above the permitted levels, and it has been established that high levels of aflatoxins in feeds can cause alterations in fish growth as well as the development of cancerous tumors in the liver, in addition to enhancing mortality.


Soybean isoflavone exposure does not have feminizing effects on men: a critical examination of the clinical evidence.
To critically evaluate the clinical evidence, and when not available, the animal data, most relevant to concerns that isoflavone exposure in the form of supplements or soy foods has feminizing effects on men. Medline literature review and cross-reference of published data. In contrast to the results of some rodent studies, findings from a recently published metaanalysis and subsequently published studies show that neither isoflavone supplements nor isoflavone-rich soy affect total or free testosterone (T) levels. Similarly, there is essentially no evidence from the nine identified clinical studies that isoflavone exposure affects circulating estrogen levels in men. Clinical evidence also indicates that isoflavones have no effect on sperm or semen parameters, although only three intervention studies were identified and none were longer than 3 months in duration. Finally, findings from animal studies suggesting that isoflavones increase the risk of erectile dysfunction are not applicable to men, because of differences in isoflavone metabolism between rodents and humans and the excessively high amount of isoflavones to which the animals were exposed. The intervention data indicate that isoflavones do not exert feminizing effects on men at intake levels equal to and even considerably higher than are typical for Asian males.


The theory of planned behavior applied to continuing education for mental health professionals.
This study evaluated the effects of a continuing education class that applied the theory of planned behavior to the intentions and behavior of mental health practitioners. In 2006 a total of 94 mental health practitioners were randomly assigned to either a standard continuing education class or one that applied principles of the theory of planned behavior. The class topic was a brief, self-report tool that assesses felt need for employment among people with serious mental illnesses. Participants' intentions to apply the tool were evaluated before and after each class. Participants' implementation of the tool was measured three months after the class. The class guided by the theory of planned behavior significantly and substantially increased the participants' intentions to use the tool in comparison with the standard class. Significantly more participants in the theory-guided class than in the standard class (74% versus 42%) had applied the tool by the three-month follow-up. Among those who implemented the assessment tool, the participants in the theory of planned behavior class also assessed significantly more of their caseload. The theory of planned behavior can improve and may be well suited to continuing education in psychiatry.


[A phase II study of vindesine for pretreated non-small cell lung cancer].
A phase II evaluation of vindesine (VDS) was performed in 16 patients with non-small cell lung cancer (ten patients with adenocarcinoma, six patients with squamous cell carcinoma, and one patient with large cell carcinoma). All except one of the patients had had prior chemotherapy. VDS at a dose of 3 mg/m2 was given intravenously every week for more than three weeks. Among 16 evaluable patients, two patients with pretreated adenocarcinoma of the lung showed partial response. The response rate for VDS was 12.5%. Toxic effects included leukopenia (94%), anemia (44%), thrombopenia (13%), alopecia (38%), peripheral neurotoxicity (38%), liver injury (19%), constipation (13%), anorexia (13%), nausea (13%), stomatitis (6%) and fever (6%).


Renal cell cancers: unveiling the hereditary ones and saving lives-a tailored diagnostic approach.
The prevalence of RCC in Europe is 2-3% and increasing every year. Hereditary predisposition is found in 5-8% of all RCC cases. Hereditary syndromes associated with RCC include: Von Hippel-Lindau, hereditary papillary renal cell carcinoma, Birt-Hogg-Dube', hereditary leiomyomatosis, succinate dehydrogenase's deficiency, tuberous sclerosis complex and Cowden's syndrome. These syndromes are related to specific genetic mutations. So far the European Association of Urology and American Urological Association have not established guidelines for referral of patients with RCC for germline mutation screening. The scope of this article is to review which clinical manifestations should direct clinicians' thinking towards hereditary kidney carcinomas and therefore suggest which patients could benefit from genetic testing.


Inorganic-organic hybrid coatings on stainless steel by layer-by-layer deposition and surface-initiated atom-transfer-radical polymerization for combating biocorrosion.
To improve the biocorrosion resistance of stainless steel (SS) and to confer the bactericidal function on its surface for inhibiting bacterial adhesion and biofilm formation, well-defined inorganic-organic hybrid coatings, consisting of the inner compact titanium oxide multilayers and outer dense poly(vinyl-N-hexylpyridinium) brushes, were successfully developed. Nanostructured titanium oxide multilayer coatings were first built up on the SS substrates via the layer-by-layer sol-gel deposition process. The trichlorosilane coupling agent, containing the alkyl halide atom-transfer-radical polymerization (ATRP) initiator, was subsequently immobilized on the titanium oxide coatings for surface-initiated ATRP of 4-vinylpyridine (4VP). The pyridium nitrogen moieties of the covalently immobilized 4VP polymer, or P(4VP), brushes were quaternized with hexyl bromide to produce a high concentration of quaternary ammonium salt on the SS surfaces. The excellent antibacterial efficiency of the grafted polycations, poly(vinyl-N-pyridinium bromide), was revealed by viable cell counts and atomic force microscopy images of the surface. The effectiveness of the hybrid coatings in corrosion protection was verified by the Tafel plot and electrochemical impedance spectroscopy measurements.


Mucoepidermoid carcinoma of the tongue.
We report a 27-year-old Congolese man with mucoepidermoid carcinoma (MEC) of the tongue base, which presented as spontaneous intraoral bleeding. Optimal treatment of tongue base MEC is unknown. To our knowledge, this is the first reported case treated with transoral excision with carbon dioxide laser and selective neck dissection. Although immunohistochemical studies have revolutionised understanding of the disease, little else is known of the natural history of MEC. The majority of MEC is considered low-grade, with an indolent course without recurrence or metastasis. Nonetheless, MEC requires surgical management, postoperative radiotherapy and close long-term follow-up.


Polystyrene nanoplastic exposure induces immobilization, reproduction, and stress defense in the freshwater cladoceran Daphnia pulex.
The widespread occurrence and accumulation of plastic waste have been globally recognized as a critical issue. However, there is limited information on the adverse effects of nanoplastics on freshwater invertebrates. In this study, the effects of a polystyrene nanoplastic on physiological changes (e.g., survival, growth, and reproduction) and expression levels of stress defense genes (oxidative stress-mediated and heat shock proteins) in the freshwater flea Daphnia pulex were measured. The results showed that the digestive organs of D. pulex were strongly fluorescent after exposure to the nanoplastic particles, and the 48-h median lethal concentration (LC 50) of the nanoplastic was determined to be 76.69 mg/L. In the 21-day chronic toxicity test, dose- and time-dependent relationships were observed for body length, and the time to first eggs was significantly prolonged in the 0.5 and 1 mg/L groups. The time to clutch was delayed, and total offspring per female and number of clutches were decreased in all the treatment groups. In addition, the offspring per clutch were significantly decreased in the 0.1 mg/L group. As the nanoplastic concentration increased, expression of stress defense genes (SOD, GST, GPx, and CAT) was first induced and then inhibited. The gene expressions of heat shock proteins (HSP70 and HSP90) were induced in all the treatment groups. Our results suggest that nanoplastics can be ingested by the freshwater cladoceran D. pulex and affect its growth and reproduction as well as induce stress defense.


Novel in vitro screening system for monoclonal antibodies using hypermutating chicken B cell library.
Here, we report an in vitro screening system for monoclonal antibodies using a hypermutating chicken B cell line, DT40-SW. When switching on hypermutation, cultured DT40-SW cells constituted an antibody library, from which clones secreting antibodies to a test antigen were successfully isolated, and genetically stabilized by switching off the mutation machinery.


"He is suitable for her, of course he is our relative": a qualitative exploration of the drivers and implications of child marriage in Gezira State, Sudan.
Child marriage is a fundamental development challenge for women and girls, with significant negative health and social outcomes. Sudan has a high rate of child marriage, with 34% of women aged 20-24 married before their 18th birthday. Since limited preventive interventions exist, we aimed to inform the evidence base to strengthen strategic action, using mixed qualitative methods to enhance study credibility. This study is the first to conduct a rigorous qualitative examination of the drivers of child marriage from the perspective of key stakeholders involved in marriage decision making within Sudan, and makes a significant contribution towards global knowledge by developing an evidence-based conceptual framework. Initially, we completed 14 focus group discussions separated by gender with mothers, fathers, and girls married as adolescents, and 23 key informant interviews. We then used a critical incident case study approach to explore 11 'cases' of child marriage (46 interviews). Findings indicate that gendered social norms and values, underpinned by religious beliefs and educational accessibility, interconnect to shape marriage decisions. In this context, many child marriages are triggered by an intrakinship proposal and further enabled by the relative lack of autonomy and influence of girls and women in marriage decision-making processes. Interconnected drivers demand context-specific holistic and multisectoral approaches, which should include simultaneous strategies to expand access to education, health services and livelihood opportunities, and evoke legal change, and participatory social and attitudinal processes that include the engagement of religious leaders and men.


Long-term effects of inadequate and excessive dietary ascorbate on bile acid metabolism in the guinea pig.
The effects of long-term chronic ascorbic acid deficiency and excessive ascorbic acid consumption on bile acid metabolism and biliary lipid composition were studied in guinea pigs. Male, weanling guinea pigs were fed a cereal-based scorbutigenic diet for 19 or 21 weeks. Ascorbic acid was administered either orally at 0.15 (group A) or 2.0 (group B) mg/100 g body weight, or it was mixed in the diet at levels of 500 (group C), 16-22 (group D), or 20,000 mg/kg (group E). Chronic ascorbic acid deficiency (groups A and D) caused depression of hepatic cytochrome P-450 levels and elevation of plasma cholesterol. Excessive ascorbate consumption did not alter these parameters relative to control levels. In contrast to results obtained in guinea pigs fed low or high amounts of ascorbate for 7-9 weeks, prolonged consumption of inadequate or excessive ascorbate resulted in little or no change in bile acid metabolism and biliary lipid composition except that bile acid pool size was increased 12% as a result of excessive ascorbate ingestion. Results of the present study suggest that there may be important differences in the guinea pig's metabolic response to ascorbic acid deficiency and ascorbic acid excess, depending on the length of the experimental period.


DRGs in Transfusion Medicine and Hemotherapy in Germany.
Patients requiring transfusion medicine and hemotherapy in an inpatient setting are incorporated into the German Diagnosis Related Groups (G-DRG) system in multiple ways. Different DRGs exist in Major Diagnostic Category 16 for patients that have been admitted for the treatment of a condition from the field of transfusion medicine. However, the reimbursement might be not cost covering for many cases, and efforts have to be intensified to find adequate definitions and prices. We believe that this can only be successful if health service research is intensified in this field. For patients requiring hemotherapy and transfusion medicine concomitant to the treatment of an underlying disease such as cancer, multiple systems exist to increase remuneration, among them the Patient Clinical Complexity Level (PCCL) and complex constellations to induce DRG splits. For direct reimbursement of high cost products, additional remuneration fees (Zusatzentgelte, ZE) are the most important. In addition, expensive innovations not reflected within the DRGs can be reimbursed after application and negotiation of the New Diagnostic and Treatment Methods (Neue Untersuchungs-und Behandlungsmethoden, NUB) system. The NUB system guarantees that medical progress is put rapidly into clinical practice and prevents financial issues from becoming a stumbling block for the use of innovative drugs and methods.


Structure-Metabolism Relationships in the Glucuronidation of d-Amino Acid Oxidase Inhibitors.
Representative d-amino acid oxidase (DAAO) inhibitors were subjected to in vitro liver microsomal stability tests in the absence or presence of uridine diphosphate glucuronic acid (UDPGA). While carboxylate-based DAAO inhibitors displayed little glucuronidation, most DAAO inhibitors containing α-hydroxycarbonyl moiety exhibited nearly complete glucuronidation within 30 min. The one exception was 6-[2-(3,5-difluorophenyl)ethyl]-4-hydroxypyridazin-3(2H)-one 10, which exhibited some degree of resistance to glucuronidation by liver microsomes from mice, rats, and humans.


Male siblings with dyserythropoiesis, microcephaly and intrauterine growth retardation.
Male siblings with intrauterine growth retardation, hydrops, mild liver dysfunction, chronic diarrhoea, failure to thrive and microcephaly are reported. In both patients, the intrauterine growth retardation was detected in the second trimester of pregnancy. Relatively severe early onset neonatal jaundice, microcytosis, anisocytosis and abnormal iron metabolism were also seen. Bone marrow examination in the second sibling showed marked ringed sideroblasts and multilobulated erythroblasts in late developmental stages. The brain was very small with enlarged cerebrospinal fluid space, a reduced number of gyri and a thin cortex. The clinical and laboratory findings in these patients appear to be unique.


Therapy of the murine plasmacytoma MOPC 104E: role of the immune response.
The murine plasmacytoma MOPC 104E was susceptible to cytotoxic therapy in female inbred BALB/c mice. Palpable subcutaneous tumors (0.6--1.2 X 10(8) cells) could be cured with a single administration of cyclophosphamide (5--250 mg/kg) or localized irradiation (800--2,400 R). Clonogenic assay showed that, following minimal curative doses of cyclophosphamide or radiation, 0.5--1.5 X 10(6) tumor cells should remain viable. Control animals succumbed to progressive, invariably lethal tumor growth after they were given sc injections of 2--3 X 10(3) tumor cells. Minimal doses of cyclophosphamide, which were curative in control tumor-bearing animals, were ineffective in treating tumor-bearing animals immunosuppressed by 450 R whole-body irradiation. Subsequent experiments measured the ability of animals cured of the murine plasmacytoma to reject secondary challenge with the same tumor. These experiments demonstrated and partially quantitated the substantial role of the immune response in effecting tumor cure following radiotherapy or chemotherapy.


[Types and prognosis of human papillomavirus infection in the male urethra and skin lesions].
To determine the types and investigate the distribution and prognosis of human papillomavirus (HPV) infection in different parts of the urethra and skin lesions in men. Using real-time fluorescence quantitative (RT-qPCR), we determined and analyzed the genotypes of HPV in the urethra and skin lesions of 620 male patients. The results of HPV examination were comparable in the urethra and skin lesions of 53.39% of the patients (331/620) and different in 46.61% (289/620). HPV positive was detected in 36.61% (227/620) in the urethra and 44.84% (278/620) in skin lesions. Various HPV subtypes were identified in both the urethra and skin lesions, mostly of the low-risk type. High-risk types in the urethra included types 16, 52, 18 and 56, and those in the skin lesions included types 16, 51, 52 and 58 types. Analysis of HPV infection in different parts and its prognosis helps to predict the risk of morbidity. Attention should be paid to the early detection of high-risk HPV infection in males so as to give positive and effective intervention and take targeted preventive measures for the high-risk population.


Valvular heart disease associated with taking low-dose pergolide for restless legs syndrome.
A 49-year-old female took low-dose pergolide (625 microg daily) for approx. 5 years (approximately cumulative dose 1.140 g/5 years) for the treatment of restless legs syndrome. She developed moderate to severe mitral and aortic valve insufficiency, requiring semi-urgent double-valve replacement. The initial diagnosis of rheumatic valve disease was refuted on histological examination of the valves due to the lack of typical calcification and neovascularization. Valvular heart disease is associated with the use of dopamine agonists for the treatment of Parkinson's disease and obesity, typically at much higher doses.


Oral health awareness in Croatian and Italian urban adolescents.
Purpose of this study was to investigate and compare differences in oral health awareness between Croatian and Italian urban adolescents. The sample consisted of primary school last grade students aged between 13 and 15 years, 300 children from Zagreb (Croatia) and 298 children from Bari (Italy). Oral health awareness was evaluated using a self-administered standardized questionnaire. Self-perception of oral health proved to be different between the two groups (p < 0.001). The Croatians reported that their oral health was "excellent" or "very-good" more often than the Italians (68.6% vs. 50.2%). The reasons given for visiting a dentist were different (p < 0.001). The Italians cleaned their teeth more often than the Croatians ("two or more times a day", 83.1% vs. 72.2%, p < 0.003). Wooden toothpicks were preferred by the Croatians (p < 0.001), while floss was preferred by the Italians (p = 0.03). The awareness regarding the use of fluoridated toothpaste was higher in the Italian group (95.6% vs. 72.5%, p < 0.001). The Croatians were consuming sweetened foods more often than the Italians (p < 0.001). Croatian adolescents reported more indicators of a lower level of oral health awareness than the Italians, while on the contrary Croatians had higher esteem of their oral health. Defining national preventive strategies is essential for improving adolescents' attitudes toward oral health in both countries, particularly in Croatia.


Epithelial myoepithelial carcinoma of the parotid gland with malignant ductal and myoepithelial components arising in a pleomorphic adenoma: a case report with cytologic, histologic and immunohistochemical correlation.
To describe the cytologic, histologic and immunohistochemical findings of a case of epithelial myoepithelial carcinoma (EMC) arising from a pleomorphic adenoma (PA) of the parotid with both malignant epithelial and myoepithelial components. A 29-year-old female presented with a 1.5 x 1.5-cm, palpable mass of the left parotid of 7-8 months' duration with recent enlargement and pain. Fine needle aspiration biopsy (FNAB) revealed biphasic epithelial (small cell) and myoepithelial (large/clear cell) clusters arranged in a pseudopapillary and trabecular pattern with abundant hyaline material with many naked nuclei, together with areas typical of pleomorphic adenoma (PA) was noted. The cytology was reported as salivary gland neoplasm, "suggestive of adenoid cystic carcinoma, less likely pleomorphic adenoma." The mass was excised and histologically reported as "pleomorphic adenoma, with focal invasion of one resected margin." Four months later the tumor recurred, and FNAB showed almost the same cytologic features as did the previous aspirate. Due to early recurrence, previous histologic sections were reviewed, and typical areas of a biphasic pattern of EMC with atypicality and mitosis of both components was found. The final diagnosis was EMC ex PA. Although previous reports mention the difficulties in diagnosing EMC and differentiation from the more common salivary gland neoplasms such as PA, we like to emphasize the cytologic confusion that results when the tumors coexist.


Cloning and characterization of monacolin K biosynthetic gene cluster from Monascus pilosus.
Monacolin K is a secondary metabolite synthesized by polyketide synthases (PKS) from Monascus, and it has the same structure as lovastatin, which is mainly produced by Aspergillus terreus. In the present study, a bacterial artificial chromosome (BAC) clone, mps01, was screened from the BAC library constructed from Monascus pilosus BCRC38072 genomic DNA. The putative monacolin K biosynthetic gene cluster was found within a 42 kb region in the mps01 clone. The deduced amino acid sequences encoded by the nine genes designated as mokA- mokI, which share over 54% similarity with the lovastatin biosynthetic gene cluster in A. terreus, were assumed to be involved in monacolin K biosynthesis. A gene disruption construct designed to replace the central part of mokA, a polyketide synthase gene, in wild-type M. pilosus BCRC38072 with a hygromycin B resistance gene through homologous recombination, resulted in a mokA-disrupted strain. The disruptant did not produce monacolin K, indicating that mokA encoded the PKS responsible for monacolin K biosynthesis in M. pilosus BCRC38072.


Biological Pacemakers: Still a Dream?
A biological pacemaker is one or more types of cellular components that, when implanted into certain regions of the heart, produce electrical stimuli that mimic that of the body's natural pacemaker cells. Somatic gene transfer, cell fusion, or cell transplantation provide a way to realize it as somatic reprogramming strategies, which involve transfer of genes encoding transcription factors to transform working myocardium into a surrogate sinoatrial node, are furthest along in the possibilities. The idea, no doubt, is bright and appealing. The objective herein intends to dig into the subject trying to find out how realizable it really is.


Analysis of expression of the gene encoding for the nuclear autoantigen La/SS-B using reporter gene constructs.
In earlier studies mRNA isoforms encoding for the nuclear autoantigen La were identified. In an alternative La mRNA form the exon 1 was replaced with the exon 1'. Moreover, exon 1' La mRNAs were found to start at different 5'-regions. In dependence on the 5'-start the exon 1' La mRNAs encoded for up to three open reading frames upstream of the La frame, which starts in the exon 2. The exon 1' was located in the intron about 70 nts downstream of the exon 1. The exon 1' La mRNA was proposed to be the result of a promoter switch in combination with an alternative splicing mechanism. The commonly used technique to study the expression of a eucaryotic gene is to fuse a reportergene immediately downstream of the proposed regulatory elements. Due to (i) the short distance between exon 1 and exon 1', (ii) the varying 5'-starts of the exon 1' La mRNAs, and (iii) the upstream open reading frames in the exon 1' La mRNAs this technique appeared to be difficult to apply to the La gene. In order to overcome these problems a luciferase reportergene construct was cloned which started about 2500 nts upstream of the exon 1 and contained the exon 1, the intron including the exon 1', and a portion of the exon 2. Luciferase was fused into the exon 2. This construct was used to prepare 5'-deletion mutants. The constructs were transiently transfected into HeLa cells. RNAs were isolated from the transiently transfected cells and analyzed using the 5'-Rapid Amplification of cDNA End technique. The PCR products were subcloned and sequenced. This analysis showed that exon 1 and exon 1' transcripts were correctly transcribed and spliced from the La luciferase fusion construct. Moreover, the 5'-start of the respective transcript allowed to identify those genomic regions in the La gene that were most likely being involved in determining the respective transcription initiation site. In parallel to the estimation of the 5'-start of the transcripts, the luciferase activity was measured. Thereby we detected a cryptic promoter element in the intron between the exon 1 and exon 2.


MAG2 and MAL Regulate Vesicle Trafficking and Auxin Homeostasis With Functional Redundancy.
Auxin is a central phytohormone and controls almost all aspects of plant development and stress response. Auxin homeostasis is coordinately regulated by biosynthesis, catabolism, transport, conjugation, and deposition. Endoplasmic reticulum (ER)-localized MAIGO2 (MAG2) complex mediates tethering of arriving vesicles to the ER membrane, and it is crucial for ER export trafficking. Despite important regulatory roles of MAG2 in vesicle trafficking, the mag2 mutant had mild developmental abnormalities. MAG2 has one homolog protein, MAG2-Like (MAL), and the mal-1 mutant also had slight developmental phenotypes. In order to investigate MAG2 and MAL regulatory function in plant development, we generated the mag2-1 mal-1 double mutant. As expected, the double mutant exhibited serious developmental defects and more alteration in stress response compared with single mutants and wild type. Proteomic analysis revealed that signaling, metabolism, and stress response in mag2-1 mal-1 were affected, especially membrane trafficking and auxin biosynthesis, signaling, and transport. Biochemical and cell biological analysis indicated that the mag2-1 mal-1 double mutant had more serious defects in vesicle transport than the mag2-1 and mal-1 single mutants. The auxin distribution and abundance of auxin transporters were altered significantly in the mag2-1 and mal-1 single mutants and mag2-1 mal-1 double mutant. Our findings suggest that MAG2 and MAL regulate plant development and auxin homeostasis by controlling membrane trafficking, with functional redundancy.


Incidence of FGFR2 Amplification and FGFR2 Fusion in Patients with Metastatic Cancer Using Clinical Sequencing.
Aberrations in the fibroblast growth factor receptor2 (FGFR2) gene, including genetic alterations and chromosomal rearrangements, lead to the development and progression of cancer with poor prognosis. However, the mechanisms underlying the FGFR2 signaling pathway to facilitate the development of FGFR2-targeted therapies have not been fully explored. Here, we examined the clinicopathological features of FGFR2 amplification and fusion in gastrointestinal tract/genitourinary tract cancers. FGFR2 amplification and fusion were identified in approximately 1.5% and 1.1% of all cancer types in 1,373 patients, respectively, with both FGFR2 amplification and fusion occurring together at a rate of approximately 0.6%. Of all cancer types screened, gastric cancer (GC) was the most common cancer type with FGFR2 amplification (87.5% of all FGFR2 amplification case) or fusion (46.7% of all cases). In addition, FGFR2 alteration had poorer overall survival (OS, 13.7 months vs. 50.2 months, P = 0.0001) and progression-free survival (PFS, 5.6 months vs. 11.4 months, P = 0.0005) than did those without FGFR2 alteration, respectively. Taken together, our data underscore to screen solid cancer patients for FGFR2 aberrations in oncology clinic.


Decreasing the immunogenicity of arginine deiminase enzyme via structure-based computational analysis.
The clinical applications of therapeutic enzymes are often limited due to their immunogenicity. B-cell epitope removal is an effective approach to solve this obstacle. The identification of hot spot epitopic residues is a critical step in the removal of protein B-cell epitope. Hereof, computational approaches are a suitable alternative to costly and labor-intensive experimental approaches. Arginine deiminase, a Mycoplasma arginine-catabolizing enzyme, is in the clinical trial for treating arginine auxotrophic cancers, especially hepatocellular carcinomas and melanomas through depleting plasma arginine and causing cell starvation. In this study, arginine deiminase from Mycoplasma hominis (MhADI) was computationally analyzed for recognizing and locating its immune-reactive regions. The 3D structure of the bioactive form of MhADI was modeled. The B-cell epitope mapping of protein was performed using various servers with different algorithms. Six segments: 31-40, 48-55, 131-140, 196-206, 294-314, and 331-344 were predicted to be the consensus immunogenic regions. The modification of epitopic hot spot residue was performed to reduce immune-reactiveness. The hot spot residue was selected considering a high B-cell epitope score, convexity index, surface accessibility, flexibility, and hydrophilicity. The structure stability of native and mutant proteins was evaluated through molecular dynamics simulation. The E304L mutein was suggested as a lower antigenic and stable enzyme derivative.


Effects of Plasmin on von Willebrand Factor and Platelets: A Narrative Review.
Plasmin is the major fibrinolytic protease responsible for dissolving thrombi by cleavage of its primary substrate fibrin. In addition, emerging evidence points to other roles of plasmin: (1) as a back-up for ADAMTS13 in proteolysis of ultra-large von Willebrand factor (VWF) multimers and (2) as an activator of platelets. Although the molecular mechanisms of fibrinolysis are well defined, insights on the effects of plasmin on VWF and platelets are relatively scarce and sometimes conflicting. Hence, this review provides an overview of the literature on the effects of plasmin on VWF multimeric structures, on VWF binding to platelets, and on platelet activation. This information is placed in the context of possible applications of thrombolytic therapy for the condition thrombotic thrombocytopenic purpura.


High cumulative risks of cancer in patients with PTEN hamartoma tumour syndrome.
PTEN hamartoma tumour syndrome (PHTS) encompasses several clinical syndromes with germline mutations in the PTEN tumour suppressor gene, including Cowden syndrome which is characterised by an increased risk of breast and thyroid cancers. Because PHTS is rare, data regarding cancer risks and genotype-phenotype correlations are limited. The objective of this study was to better define cancer risks in this syndrome with respect to the type and location of PTEN mutations. 154 PHTS individuals with a deleterious germline PTEN mutation were recruited from the activity of the Institut Bergonié genetic laboratory. Detailed phenotypic information was obtained for 146 of them. Age and sex adjusted standardised incidence ratio (SIR) calculations, cumulative cancer risk estimations, and genotype-phenotype analyses were performed. Elevated SIRs were found mainly for female breast cancer (39.1, 95% CI 24.8 to 58.6), thyroid cancer in women (43.2, 95% CI 19.7 to 82.1) and in men (199.5, 95% CI 106.39 to 342.03), melanoma in women (28.3, 95% CI 7.6 to 35.4) and in men (39.4, 95% CI 10.6 to 100.9), and endometrial cancer (48.7, 95% CI 9.8 to 142.3). Cumulative cancer risks at age 70 were 85% (95% CI 70% to 95%) for any cancer, 77% (95% CI 59% to 91%) for female breast cancer, and 38% (95% CI 25% to 56%) for thyroid cancer. The risk of cancer was two times greater in women with PHTS than in men with PHTS (p<0.05). This study shows a considerably high cumulative risk of cancer for patients with PHTS, mainly in women without clear genotype-phenotype correlation for this cancer risk. New recommendations for the management of PHTS patients are proposed.


Highly efficient single cell arraying by integrating acoustophoretic cell pre-concentration and dielectrophoretic cell trapping.
To array rare cells at the single-cell level, the volumetric throughput may become a bottleneck in the cell trapping and the subsequent single-cell analysis, since the target cells per definition commonly exist in a large sample volume after purification from the original sample. Here, we present a novel approach for high throughput single cell arraying by integrating two original microfluidic devices: an acoustofluidic chip and an electroactive microwell array. The velocity of the cells is geared down in the acoustofluidic chip while maintaining a high volume flow rate at the inlet of the microsystem, and the cells are subsequently trapped one by one into the microwell array using dielectrophoresis. The integrated system exhibited a 10 times improved sample throughput compared to trapping with the electroactive microwell array chip alone, while maintaining a highly efficient cell recovery above 90%. The results indicate that the serial integration of the acoustophoretic pre-concentration with the dielectrophoretic cell trapping drastically improves the performance of the electroactive microwell array for highly efficient single cell analysis. This simple and effective system for high throughput single cell arraying with further possible integration of additional functions, including cell sorting and downstream analysis after cell trapping, has potential for development to a highly integrated and automated platform for single-cell analysis of rare cells.


Cardiac myosin binding protein-C Ser302 phosphorylation regulates cardiac β-adrenergic reserve.
Phosphorylation of cardiac myosin binding protein-C (MyBP-C) modulates cardiac contractile function; however, the specific roles of individual serines (Ser) within the M-domain that are targets for β-adrenergic signaling are not known. Recently, we demonstrated that significant accelerations in in vivo pressure development following β-agonist infusion can occur in transgenic (TG) mouse hearts expressing phospho-ablated Ser282 (that is, TGS282A) but not in hearts expressing phospho-ablation of all three serines [that is, Ser273, Ser282, and Ser302 (TG3SA)], suggesting an important modulatory role for other Ser residues. In this regard, there is evidence that Ser302 phosphorylation may be a key contributor to the β-agonist-induced positive inotropic responses in the myocardium, but its precise functional role has not been established. Thus, to determine the in vivo and in vitro functional roles of Ser302 phosphorylation, we generated TG mice expressing nonphosphorylatable Ser302 (that is, TGS302A). Left ventricular pressure-volume measurements revealed that TGS302A mice displayed no accelerations in the rate of systolic pressure rise and an inability to maintain systolic pressure following dobutamine infusion similar to TG3SA mice, implicating Ser302 phosphorylation as a critical regulator of enhanced systolic performance during β-adrenergic stress. Dynamic strain-induced cross-bridge (XB) measurements in skinned myocardium isolated from TGS302A hearts showed that the molecular basis for impaired β-adrenergic-mediated enhancements in systolic function is due to the absence of protein kinase A-mediated accelerations in the rate of cooperative XB recruitment. These results demonstrate that Ser302 phosphorylation regulates cardiac contractile reserve by enhancing contractile responses during β-adrenergic stress.