Proteolytic processing of the Alzheimer's disease amyloid precursor protein in brain and platelets.
Proteolytic processing of the amyloid precursor protein by beta -and gamma-secretases results in the production of Alzheimer's disease (AD) Abeta amyloid peptides. Modulation of secretase activity is being investigated as a potential therapeutic approach. Recent studies with human brain have revealed that the beta-secretase protein, BACE, is increased in cortex of AD patients. Analysis of betaCTF (or C99), the amyloid precursor protein (APP) product of BACE cleavage that is the direct precursor to Abeta, shows it is also elevated in AD, underlying the importance of beta-secretase cleavage in AD pathogenesis. The C-terminal product of gamma-secretase cleavage of APP, epsilonCTF (or AICD), is enriched in human brain cortical nuclear fractions, a subcellular distribution appropriate for a putative involvement of APP cytosolic domain in signal transduction. Analysis of AD cortex samples, particularly that of a carrier of a familial APP mutation, suggests that processing of APP transmembrane domain generates an alternative CTF product. All these particularities observed in the AD brain demonstrate that APP processing is altered in AD. The transgenic mouse model Tg2576 seems to be a promising laboratory tool to test potential modulators of Abeta formation. Indeed, C-terminal products of alpha-, beta-, and gamma-secretase cleavage are readily detectable in the brain of these transgenic mice. Finally, the finding of the same secretase products in platelets and neurons make platelets a potentially useful and easily accessible clinical tool to monitor effects of novel therapies based on inhibition of beta- or gamma-secretase.


Health services research: why and how?
It is useful to divide medical research into three areas: biomedical, clinical, and health services research. The areas partly overlap, and health services research is also related to social services research. Research is carried out to solve problems and is an instrument for change. Health services research has developed over the last ten years in response to increasing problems in many health services. Superficially, these problems are caused by insufficient resources, but no service can hope to pay its way out of them. Some may be fairly accurately investigated, like need, demand, and utilization of care. Others are more complicated, e.g. evaluation of care, defining standards, and cost--benefit analyses. A few deal with fundamental values, like quality of life and responsibility of individuals and societies. So far, health services research has led to greater emphasis on primary care, but it is fair to say that it has not managed to infiltrate the service and influence people's attitudes and ambitions. In the future, one must bring health services research inside the service and involve the professionals more deeply. One must support prevention studies, attack the ethical and clinical problems related to quality of life, study the potential of non-professional support in the community, and promote rational attitudes among professions, patients, people and politicians. The task is never-ending and health services research, therefore, must be part of the programme of all medical schools.


ACPA-negative RA consists of two genetically distinct subsets based on RF positivity in Japanese.
HLA-DRB1, especially the shared epitope (SE), is strongly associated with rheumatoid arthritis (RA). However, recent studies have shown that SE is at most weakly associated with RA without anti-citrullinated peptide/protein antibody (ACPA). We have recently reported that ACPA-negative RA is associated with specific HLA-DRB1 alleles and diplotypes. Here, we attempted to detect genetically different subsets of ACPA-negative RA by classifying ACPA-negative RA patients into two groups based on their positivity for rheumatoid factor (RF). HLA-DRB1 genotyping data for totally 954 ACPA-negative RA patients and 2,008 healthy individuals in two independent sets were used. HLA-DRB1 allele and diplotype frequencies were compared among the ACPA-negative RF-positive RA patients, ACPA-negative RF-negative RA patients, and controls in each set. Combined results were also analyzed. A similar analysis was performed in 685 ACPA-positive RA patients classified according to their RF positivity. As a result, HLA-DRB1*04:05 and *09:01 showed strong associations with ACPA-negative RF-positive RA in the combined analysis (p = 8.8×10(-6) and 0.0011, OR: 1.57 (1.28-1.91) and 1.37 (1.13-1.65), respectively). We also found that HLA-DR14 and the HLA-DR8 homozygote were associated with ACPA-negative RF-negative RA (p = 0.00022 and 0.00013, OR: 1.52 (1.21-1.89) and 3.08 (1.68-5.64), respectively). These association tendencies were found in each set. On the contrary, we could not detect any significant differences between ACPA-positive RA subsets. As a conclusion, ACPA-negative RA includes two genetically distinct subsets according to RF positivity in Japan, which display different associations with HLA-DRB1. ACPA-negative RF-positive RA is strongly associated with HLA-DRB1*04:05 and *09:01. ACPA-negative RF-negative RA is associated with DR14 and the HLA-DR8 homozygote.


Parental bonds in children at high and low familial risk for panic disorder.
A rejecting and overprotective parenting style is considered to be an important risk factor for the development of anxiety disorders. This study examined the role of perceived parental bonding as a potential environmental risk factor for panic disorder (PD) in unaffected offspring with parental PD. Children with a biological parent with PD (n = 71) and children of parents with no psychiatric history (n = 80) participated in the study. Results indicate that high risk children do not perceive their parents as being more protective and less caring than low risk controls. The optimal bonding type (high care, low protection) was the most frequently reported parenting style across groups. The constraining type of maternal bonding (high care, high protection) was less frequently reported by high risk children (p < 0.05). Overall, these data suggest that parental PD does not compromise the parent-child bonds in never-ill offspring.


Carboplatin and vinblastine in advanced non-small-cell lung cancer: a phase II study.
Between July 2, 1987, and August 21, 1987, Cancer and Leukemia Group B (CALGB) conducted a phase II evaluation of carboplatin (CBDCA) and vinblastine (VBL) in advanced non-small-cell lung cancer. Of the 58 patients who entered the study, 55 were eligible and produced follow-up data. Chemotherapy, which was carried out in 28-day cycles, consisted of 4 mg/m2 VBL given on days 1 and 3 and 125 mg/m2 CBDCA given on days 1-3. Partial responses were observed in 10 cases (18%), and 1 patient (2%) exhibited regression of evaluable disease. No complete responses were achieved. The overall objective response rate was 20%. The median survival was 6.1 months, and the median time to treatment failure was 3.3 months. Life-threatening (grade 4) toxicity was mainly leukopenia (20%), followed by anemia (7%), infection (4%), thrombocytopenia (2%), fever (2%), nausea and vomiting (2%), and weight loss (2%). There were two deaths due to infection. The results of this study demonstrate that the combination CBDCA/VBL is active in advanced NSCLC; however, whether this combination is more active than either CBDCA or VBL alone is unknown.


The influence of learning style in understanding analogies and 2D animations in embryology course.
Undergraduate students struggle to comprehend embryology because of its dynamic nature. Studies have recommended using a combination of teaching methods to match the student's learning style. But there has been no study to describe the effect of such teaching strategy over the different types of learners. In the present study, an attempt has been made to teach embryology using the combination of analogies and simple 2D animations made with Microsoft powerpoint software. The objective of the study is to estimate the difference in academic improvement and perception scale between the different types of learners after introducing analogies and 2D animation in a lecture environment. Based on Visual, Aural, Read/Write, and Kinesthetic (VARK) scoring system the learners were grouped into unimodal and multimodal learners. There was significant improvement in post-test score among the unimodal (P<0.001) and multimodal learners (P<0.001). When the post-test score was compared between the two groups, the multimodal learners performed better the unimodal learners (P=0.018). But there was no difference in the perception of animations and analogies and long-term assessment between the groups. The multimodal learners performed better than unimodal learners in short term recollection, but in long term retention of knowledge the varied learning style didn't influence its outcome.


ERS International Congress, Madrid, 2019: highlights from the Allied Respiratory Professionals' Assembly.
This article provides an overview of outstanding sessions that were (co)organised by the Allied Respiratory Professionals' Assembly during the European Respiratory Society International Congress 2019 in Madrid, Spain. Session content was mainly targeted at allied respiratory professionals such as respiratory physiologists, respiratory physiotherapists and respiratory nurses, and is summarised in this document. Short take-home messages related to pulmonary function testing highlight the importance of quality control. Furthermore, novel findings regarding the assessment of functional status call attention to bodily factors that can affect functional status. Regarding pulmonary rehabilitation, data were presented about the use of equipment and type of exercise training in COPD and lung cancer. Recent developments in physical activity-related research give insight in enablers of physical activity after hospital admission. The importance of integrated respiratory care was also highlighted, with the occupational therapist, nurse, and nutritional and psychological counsellor playing a pivotal role, which relates directly to research in the field of respiratory nursing that formulates the need for more nursing led-interventions in the future. To conclude, this review provides readers with valuable insight into some of the emerging and future areas affecting clinical practice of allied healthcare professionals.


Disparities in surgical care for children across Brazil: Use of geospatial analysis.
Health systems for surgical care for children in low- and middle-income countries remain poorly understood. Our goal was to characterize the delivery of surgical care for children across Brazil and to identify associations between surgical resources and childhood mortality. We performed a cross-sectional, ecological study to analyze surgical care for children in the public health system (Sistema Único de Saúde) across Brazil from 2010 to 2015. We collected data from several national databases, and used geospatial analysis (two-step floating catchment, Getis-Ord-Gi analysis, and geographically weighted regression) to explore relationships between infrastructure, workforce, access, procedure rate, under-5 mortality rate (U5MR), and perioperative mortality rate (POMR). A total of 246,769 surgical procedures were performed in 6,007 first level/ district hospitals and 491 referral hospitals across Brazil over the study period. The surgical workforce is distributed unevenly across the country, with 0.13-0.26 pediatric surgeons per 100,000 children in the poorer North, Northeast and Midwest regions, and 0.6-0.68 pediatric surgeons per 100,000 children in the wealthier South and Southeast regions. Hospital infrastructure, procedure rate, and access to care is also unequally distributed across the country, with increased resources in the South and Southeast compared to the Northeast, North, and Midwest. The U5MR varies widely across the country, although procedure-specific POMR is consistent across regions. Increased access to care is associated with lower U5MR across Brazil, and access to surgical care differs by geographic region independent of socioeconomic status. There are wide disparities in surgical care for children across Brazil, with infrastructure, manpower, and resources distributed unevenly across the country. Access to surgical care is associated with improved U5MR independent of socioeconomic status. To address these disparities, policy should direct the allocation of surgical resources commensurate with local population needs.


Spontaneous simultaneous bilateral malignant glaucoma of a patient with no antecedent history of medical or surgical eye diseases.
Malignant glaucoma, or aqueous misdirection syndrome, is a condition characterized by sudden intraocular pressure (IOP) elevation, and it is usually unilateral and induced by ocular surgical intervention or by medical therapy. Here, we report a case of simultaneous bilateral malignant glaucoma in a young patient with no history of any ocular diseases. A case of a 24-year-old female with no apparent previous history of ocular medical or surgical conditions was referred to our hospital because of recent bilateral IOP elevation associated with a severe drop in vision and shallow anterior chamber with no posterior segment anomalies detected by ocular ultrasound in both eyes. Yttrium aluminum garnet (YAG) laser iridotomy dropped the IOP only temporarily and the patient received topical atropine treatment with combined trabeculectomy and anterior vitrectomy. In this case, the patient had a typical presentation of bilateral malignant glaucoma and her IOP dropped only temporarily following laser iridotomy to rise again shortly thereafter. Also, deepening of the anterior chamber and IOP decrease after topical atropine was very supportive of the diagnosis of malignant glaucoma. Successful management with trabeculectomy and limited vitrectomy also affirmed the diagnosis of malignant glaucoma. This is a very rare case of bilateral malignant glaucoma in a young adult without any prior eye conditions; only one similar case has been reported in the literature. We propose our own theory regarding this simultaneous occurrence of the pathology based on previously published studies about the presence of communication between the two eyes along the cerebrospinal fluid pathways.


Bone marrow scintigraphy as a useful method for estimating the physiological status of bone marrow and spleen in polycythaemia vera.
Bone marrow scintigraphy is a simple and noninvasive examination useful to define the status of the bone marrow and spleen in polycythaemia vera (P.V.). Despite the absence of specificity of Indium 111 labelled transferrin (In-Tf) for myelopoietic tissue, there is a close correlation between bone marrow In-Tf uptake and bone marrow cellularity and between splenic In-Tf uptake and splenic metaplasia. The results of scintigraphy are compared to clinical data, radioactive iron kinetics, bone marrow and spleen histology and the course of the disease. The diagnostic and prognostic value of bone marrow scintigraphy is discussed, particularly at the stage of transformation of P.V. into postpolycythaemia myeloid metaplasia (Post-P.V.M.M.).


Making fast cylindrical gradient-index lenses diffraction limited by using a wave-front-correction element.
A wave-front-correction element (WFCE) is produced to make a cylindrical Ag-ion-exchanged gradient-index (GRIN) lens with a high numerical aperture (0.53) diffraction limited (wave-front error, 0.02lambda rms). The wave-front aberrations of the cylindrical GRIN lens are measured by a phase-shifting shearing interferometer, with a conventional microscope objective used as a compensation lens. The continuous surface relief of the WFCE is produced by a lithographic process. The wave-front-corrected GRIN lens is applied to collimate the strongly divergent light (57 degrees full diverging angle measured at 1/e(2) of maximum intensity) emitted by a high-power diode laser. The power irradiated into a full angle of 2 mrad can be enhanced by a factor of 1.8 with the WFCE.


A tale of two models: mouse and zebrafish as complementary models for lymphatic studies.
Lymphatic vessels provide essential roles in maintaining fluid homeostasis and lipid absorption. Dysfunctions of the lymphatic vessels lead to debilitating pathological conditions, collectively known as lymphedema. In addition, lymphatic vessels are a critical moderator for the onset and progression of diverse human diseases including metastatic cancer and obesity. Despite their clinical importance, there is no currently effective pharmacological therapy to regulate functions of lymphatic vessels. Recent efforts to manipulate the Vascular Endothelial Growth Factor-C (VEGFC) pathway, which is arguably the most important signaling pathway regulating lymphatic endothelial cells, to alleviate lymphedema yielded largely mixed results, necessitating identification of new targetable signaling pathways for therapeutic intervention for lymphedema. Zebrafish, a relatively new model system to investigate lymphatic biology, appears to be an ideal model to identify novel therapeutic targets for lymphatic biology. In this review, we will provide an overview of our current understanding of the lymphatic vessels in vertebrates, and discuss zebrafish as a promising in vivo model to study lymphatic vessels.


Cytotoxic effects of loperamide hydrochloride on canine cancer cells.
Loperamide is a peripheral opiate agonist that can cause apoptosis and G2/M arrest in human cancer cell lines and may sensitize cells to chemotherapy. The objectives of this study were to investigate the effects of loperamide on viability, apoptosis and cell cycle kinetics in canine cancer cells and to establish whether the drug sensitizes cells to doxorubicin. Cell viability was assessed using Alamar Blue. Cell death and cell cycle were studied using flow cytometry with 7-Aminoactinomycin-D (7-AAD) and propidium iodide (PI), respectively. Loperamide decreased cell viability in a dose-dependent fashion and was most effective against canine osteosarcoma cells. In all cell lines, it induced a dose and time dependent apoptosis and resulted in accumulation in G0/G1. When co-incubated with doxorubicin, loperamide induced a synergistic cell kill in canine carcinoma cells. Investigation is warranted into the role of loperamide in the treatment of canine cancer.


Multiple recurrences of unknown primary tumor in a meningioma: A case report.
Metastasis of a systemic tumor into a primary brain tumor has been rarely reported in the literature. These metastases could be seen mostly in meningiomas and in less frequently in the other intracranial tumors. Carcinoma of an unknown primary site (CUP) metastasizing into an intracranial meningioma is not a common occasion. According to the best of our knowledge, in this report, we present the first case with multiple recurrent CUP, which metastasized to intracranial meningioma, reported in the literature. The whole body was investigated with FDG PET/CT and biomarkers of the most common carcinomas for primary cancer. However, these tests showed no primary affected region. Despite all adjuvant therapies, the tumor had multiple recurrences. Such cases are still a challenge to offer optimal management.


Glassy dynamics and kinetic vitrification of isotropic suspensions of hard rods.
A nonlinear Langevin equation (NLE) theory for the translational center-of-mass dynamics of hard nonspherical objects has been applied to isotropic fluids of rigid rods. The ideal kinetic glass transition volume fraction is predicted to be a monotonically decreasing function beyond an aspect ratio of two. The functional form of the decrease is weaker than the inverse aspect ratio. Vitrification occurs at lower volume fractions for corrugated tangent bead rods compared to their smooth spherocylinder analogs. The ideal glass transition signals a crossover to activated dynamics, which is estimated to be observable before the nematic phase boundary is encountered if the aspect ratio is less than roughly 25. Calculations of the glassy elastic shear modulus and absolute yield stress reveal a roughly exponential growth with volume fraction. The dependence of entropic barriers and mean barrier hopping times on concentration for rods of variable aspect ratios can be collapsed quite well based on a difference volume fraction variable that quantifies the distance from the ideal glass boundary. Full numerical solution of the NLE theory via stochastic trajectory simulation was performed for tangent bead rods, and the results were compared to their hard sphere analogs. With increasing shape anisotropy the characteristic length scales of the nonequilibrium free energy increase and the magnitude of the localization well and entropic barrier curvatures decreases. These changes result in a significant aspect ratio dependence of dynamical properties and time correlation functions including weaker intermediate time subdiffusive transport, stronger two-step decay of the incoherent dynamic structure factor, longer mean alpha relaxation time, and stronger wavevector-dependent decoupling of relaxation times and the self-diffusion constant. The theoretical results are potentially testable via computer simulation, confocal microscopy, and dynamic light scattering.


Microorganisms in the Placenta: Links to Early-Life Inflammation and Neurodevelopment in Children.
Prenatal exposure to various stressors can influence both early and later life childhood health. Microbial infection of the intrauterine environment, specifically within the placenta, has been associated with deleterious birth outcomes, such as preterm birth, as well as adverse neurological outcomes later in life. The relationships among microorganisms in the placenta, placental function, and fetal development are not well understood. Microorganisms have been associated with perinatal inflammatory responses that have the potential for disrupting fetal brain development. Microbial presence has also been associated with epigenetic modifications in the placenta, as well other tissues. Here we review research detailing the presence of microorganisms in the placenta and associations among such microorganisms, placental DNA methylation, perinatal inflammation, and neurodevelopmental outcomes.


Mitochondrial Dok-4 recruits Src kinase and regulates NF-kappaB activation in endothelial cells.
The downstream of kinase (Dok) family of adapter proteins consists of at least five members structurally characterized by an NH2-terminal tandem of conserved pleckstrin homology and phosphotyrosine binding domains linked to a unique COOH-terminal region. To determine the role of the novel adapter protein Dok-4 in endothelial cells, we first investigated the cell localization of Dok-4. Most surprisingly, immunofluorescence microscopy, cell fractionation studies, and studies with enhanced green fluorescent protein chimeras showed that wild type Dok-4 (Dok-4-WT) specifically localized in mitochondria. An NH2-terminal deletion mutant of Dok-4 (Dok-4-(deltaN11-29)), which lacks the mitochondrial targeting sequence, could not accumulate in mitochondria. Co-immunoprecipitation revealed an interaction of c-Src with Dok-4-WT in endothelial cells. Most interestingly, overexpression of Dok-4-WT, but not Dok-4-(deltaN1-99), increased mitochondrial c-Src expression, whereas knock-down of endogenous Dok-4 with a small interfering RNA vector greatly inhibited mitochondrial localization of c-Src, suggesting a unique function for Dok-4 as an anchoring protein for c-Src in mitochondria. Dok-4-WT significantly decreased 39-kDa subunit complex I expression. PP2, a specific Src kinase inhibitor, prevented the Dok-4-mediated complex I decrease, suggesting the involvement of Src kinase in regulation of complex I expression. Dok-4-WT enhanced tumor necrosis factor-alpha (TNF-alpha)-mediated reactive oxygen species (ROS) production, supporting the functional relevance of a Dok-4-Src-complex I/ROS signaling pathway in mitochondria. Finally, Dok-4 enhanced TNF-alpha-mediated NF-kappaB activation, whereas this was inhibited by transfection with Dok-4 small interfering RNA. In addition, Dok-4-induced NF-kappaB activation was also inhibited by transfection of a dominant negative form of c-Src. These data suggest a role for mitochondrial Dok-4 as an anchoring molecule for the tyrosine kinase c-Src, and in turn as a regulator of TNF-alpha-mediated ROS production and NF-kappaB activation.


Testis-specific lactate dehydrogenase (LDH-C4; Ldh3) in murine oocytes and preimplantation embryos.
LDH-C(4) (Ldh3) is a member of the lactate dehydrogenase family of isozymes that catalyze the terminal reaction in the glycolytic pathway. In mammals, 3 genes, ldha, ldhb, and Ldhc, encode the subunits that assemble into catalytically active homo- and heterotetramers. Differential expression of these genes determines the lactate dehydrogenase (LDH) isozyme composition of tissues, and, as is well known, A subunits predominate in skeletal muscle and B subunits are abundantly produced in brain and heart, with the Ldh2 isozyme the most abundant form in oocytes. The C peptide can be detected first in pachytene spermatocytes and constitutes the primary LDH of spermatozoa. Originally the Ldhc gene (Ldh3 in terminology applied to murine cells) was considered to be testis specific on the basis of immunochemical, enzymatic, and molecular analyses. Here we report the detection of this isozyme in the murine oocyte and early embryo. Our results indicate that Ldh3 mRNA is transcribed in oocytes and cannot be detected in fertilized eggs. Ldh3 protein, however, persists to the blastocyst stage of embryonic development localizing mainly to the cortex region of oocytes, eggs, zygotes, and embryonic blastomeres.


Expanding tuberculosis case detection by screening household contacts.
Tuberculosis (TB) case detection remains low in many countries, compromising the efficacy of TB control efforts. Current global TB control policy emphasizes case finding through sputum smear microscopy for patients who self-report to primary health centers. Our objective was to assess the feasibility and yield of a simple active case finding strategy in a high incidence population in northern Lima, Peru. We implemented this pilot strategy in one health center's catchment area. Health workers visited household contacts of new TB case subjects to identify symptomatic individuals and collect sputum for screening. Neighboring households were screened in the same manner. Secondary analyses measured risk of TB by (1) sputum smear status of the index case subject, (2) compliance with testing, and (3) risk factors for disease detected through active contact tracing in contrast to self-report. The TB prevalence detected through combined active and passive case finding among 1,094 household contacts was 0.91% (914 per 100,000), much higher than with passive case finding alone (0.18%; 183 per 100,000; p=0.02). Among 2,258 neighbors, the combined strategy detected a TB prevalence of 0.22% (221 per 100,000) in contrast to 0.08% (80 per 100,000) detected through passive case finding alone (p=0.25). Risk factors for being diagnosed through active case finding in contrast with self-report included age >55 years (odds ratio [OR]=5.5; 95% confidence interval [CI] 1.2, 22.8) and female gender (OR=3.9; 95% CI 0.99, 22.3). Risk of active TB among symptomatic household contacts of active case subjects in this community is very high. Results suggest that contact tracing in such settings may be a powerful means of improving case detection rates for active TB disease.


Bootstrap estimation of diagnostic accuracy with patient-clustered data.
The purpose of this study was to describe a simple bootstrap approach for estimating sensitivity, specificity, and the area under the receiver operating characteristic curve for multisite test outcome data. The performance of bootstrap estimates was evaluated and compared with that of analytic estimates by using a simulation study. Bootstrapping was demonstrated by using data from a previous study comparing two angiographic methods. Analytic and bootstrap estimates had similar coverage rates for 95% confidence intervals. With many sites per patient, bootstrap estimates had slightly better coverage than analytic estimates. Bootstrap percentile intervals had better coverage than asymptotic normal bootstrap intervals. Bootstrapping is a useful method of estimating confidence intervals for the area under the receiver operating characteristic curve, sensitivity, and specificity when data are correlated.


Nortriptyline Compared to Amitriptyline for the Treatment of Persistent Masticatory Myofascial Pain.
To evaluate and compare the pharmacotherapeutic efficacies of two tricyclic antidepressant (TCA) drugs for masticatory myofascial pain (MFP): nortriptyline (NOR) and amitriptyline (AMI). Fifty patients with chronic MFP were included in the study; 30 were medicated with AMI only, and 20 took NOR after discontinuing AMI due to adverse effects. Pain diaries recording verbal pain scores (VPS) were utilized to compare posttreatment scores to baseline scores. Chi-square and t tests were used to analyze the data. Across both groups, the mean ± standard deviation VPS score at the end of treatment (2.92 ± 3.2) was significantly lower compared to baseline (6.4 ± 1.75; P < .0001) and was a clinically meaningful (≥ 50%) difference. Initial VPS scores were similar in the AMI and NOR groups (6.27 ± 1.92 and 6.78 ± 1.98). At the end of the study, NOR patients reported a lower final VPS compared to AMI patients (2.83 ± 3.06 vs 4.55 ± 2.92; P = .039). The 50% improvement rate with NOR treatment was better than with AMI treatment (P = .036). The same maximal dosages were used by the patients who achieved a ≥ 50% success rate (20.96 ± 5.036 mg) than those who did not (21.667 ± 5.036 mg). TCAs are effective in reducing pain in patients with chronic MFP. NOR seems more effective and better tolerated than AMI, but due to study limitations, more data are needed to confirm these results.


Effect of amorphous content on dissolution characteristics of rifampicin.
Rifampicin, one of the main first line anti-TB drugs, shows variable bioavailability in different marketed preparations and reasons cited include physiological, degradation, manufacturing/ processing, solid state, and bioavailability assessment procedure. Although the amorphous form of a drug is expected to exhibit higher solubility, the amorphous rifampicin has been reported to have a solubility disadvantage as compared to crystalline form II, which is used in marketed preparations. Amorphous form was generated and characterized by solid-state characterization techniques. Physical powder mixtures of form II with varying amounts of amorphous form were prepared, which were then subjected to solid-state characterization techniques and further evaluated for their dissolution behavior. Differential scanning calorimetry (DSC) scans show that area enclosed by integral of melting endotherm can be used for quantification of crystalline component, which can then be used to estimate amorphous content. No definite trend was evident in powder dissolution of mixtures that could implicate solubility difference of amorphous form. Intrinsic dissolution rate (IDR) results indicate that amorphous content has no effect on dissolution profiles of crystalline rifampicin.


Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17).
Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) is an autosomal dominant neurodegenerative disorder, which has three cardinal features: behavioral and personality changes, cognitive impairment, and motor symptoms. FTDP-17 was defined during the International Consensus Conference in Ann Arbor, Michigan, in 1996. The prevalence and incidence remain unknown but FTDP-17 is an extremely rare condition. It is caused by mutations in the tau gene, which encodes a microtubule-binding protein. Over 100 families with 38 different mutations in the tau gene have been identified worldwide. The phenotype of FTDP-17 varies not only between families carrying different mutations but also between and within families carrying the same mutations. The pathogenetic mechanisms underlying the disorder are thought to be related to the altered proportion of tau isoforms or to the ability of tau to bind microtubules and to promote microtubule assembly. Definitive diagnosis of FTDP-17 requires a combination of characteristic clinical and pathological features and molecular genetic analysis. Genetic counseling should be offered to affected and at-risk individuals; for most subtypes, penetrance is incomplete. Currently, treatment for FTDP-17 is only symptomatic and supportive. The prognosis and rate of the disease's progression vary considerably among individual patients and genetic kindreds, ranging from life expectancies of several months to several years, and, in exceptional cases, as long as two decades.


Aligned chitosan-polycaprolactone polyblend nanofibers promote the migration of glioblastoma cells.
In vitro models that accurately mimic the microenvironment of invading glioblastoma multiform (GBM) cells will provide a high-throughput system for testing potential anti-invasion therapies. Here, the ability of chitosan-polycaprolactone polyblend nanofibers to promote a migratory phenotype in human GBM cells by altering the nanotopography of the nanofiber membranes is investigated. Fibers are prepared with diameters of 200 nm, 400 nm, and 1.1 μm, and are either randomly oriented or aligned to produce six distinct nanotopographies. Human U-87 MG GBM cells, a model cell line commonly used for invasion assays, are cultured on the various nanofibrous substrates. Cells show elongation and alignment along the orientation of aligned fibers as early as 24 h and up to 120 h of culture. After 24 h of culture, human GBM cells cultured on aligned 200 nm and 400 nm fibers show marked upregulation of invasion-related genes including β-catenin, Snail, STAT3, TGF-β, and Twist, suggesting a mesenchymal change in these migrating cells. Additionally, cells cultured on 400 nm aligned fibers show similar migration profiles as those reported in vivo, and thus these nanofibers should provide a unique high-throughput in vitro culture substrate for developing anti-migration therapies for the treatment of GBM.


Occult bleeding from a perforated gastric duplication in an infant.
We report what we believe is the first case of a perforated gastric duplication with occult intraperitoneal hemorrhage and massive abdominal distention in the neonatal period. At operation, a perforated ulcer on the free anterior surface of the duplication was identified and the entire cyst excised by submucosal dissection.


The Effects of Ethnically Congruent Music on Eye Movements and Food Choice-A Cross-Cultural Comparison between Danish and Chinese Consumers.
Musical fit refers to the congruence between music and attributes of a food or product in context, which can prime consumer behavior through semantic networks in memory. The vast majority of research on this topic dealing with musical fit in a cultural context has thus far been limited to monocultural groups in field studies, where uncontrolled confounds can potentially influence the study outcome. To overcome these limitations, and in order to explore the effects of ethnically congruent music on visual attention and food choice across cultures, the present study recruited 199 participants from China (n = 98) and Denmark (n = 101) for an in-laboratory food choice paradigm with eye-tracking data collection. For each culture group, the study used a between-subject design with half of the participants listening to only instrumental "Eastern" music and the other half only listening to instrumental "Western" music, while both groups engaged in a food choice task involving "Eastern" and "Western" food. Chi-square tests revealed a clear ethnic congruency effect between music and food choice across culture, whereby Eastern (vs. Western) food was chosen more during the Eastern music condition, and Western (vs. Eastern) food was chosen more in the Western music condition. Furthermore, results from a generalized linear mixed model suggested that Chinese participants fixated more on Western (vs. Eastern) food when Western music was played, whereas Danish participants fixated more on Eastern (vs. Western) food when Eastern music was played. Interestingly, no such priming effects were found when participants listened to music from their own culture, suggesting that music-evoked visual attention may be culturally dependent. Collectively, our findings demonstrate that ambient music can have a significant impact on consumers' explicit and implicit behaviors, while at the same time highlighting the importance of culture-specific sensory marketing applications in the global food industry.


Candida in burns: risk factors and outcomes.
Sepsis due to Candida is an uncommon but a significant cause of death in burns patients. Colonization is common, but consensus guidelines for prophylaxis and empirical therapy do not specifically include this cohort. Our aim was to define predictive factors for candidaemia in a burns unit, to guide protocols for prevention and early treatment. We conducted a 10-year review (July 1998-December 2007) of patients admitted to the Victorian Adult Burns Service, Melbourne, Australia. Of 1929 patients admitted with acute burn injury, 143 had Candida isolated at any site, most commonly Candida albicans. There were 12 episodes of candidaemia. Prior colonization was an important risk factor for candidaemia, and the risk increased substantially with the number of colonized sites; indeed 43% of patients colonized at more than three sites (and not on antifungals) developed candidaemia. Other risk factors were higher total burn surface area, higher full-thickness surface area, prolonged admission, number and duration of intensive care unit admissions, number of visits to the operating theatre, alcohol as a contributing factor to burn, prior treatment with total parenteral nutrition, or certain antibiotics (ceftriaxone, vancomycin, amikacin, co-trimoxazole). The attributable mortality of candidaemia was 15% (n = 2). Initiation of antifungal therapy was often delayed. Our results support early empirical antifungal therapy in septic burns patients who are colonized, before the results of cultures become known. The role of prophylactic antifungals is less clear, but should be strongly considered for patients colonized at multiple sites.


Autosomal dominant aplasia cutis in three generations and one case with preaxial polydactyly in the last generation.
Aplasia Cutis Congenita (ACC), characterized by the focal absence of the skin and skin adnexia resulting from a developmental failure, may occur as part of Adams-Oliver Syndrome (AOS) which can be defined as a congenital inherited disorder, consisting of terminal transverse limb defects and vascular anomalies in addition to ACC. Coexistence of isolated preaxial polydactyly without terminal extremity defect and ACC is extremely rare. Furthermore, ACC and preaxial polydactyly has not been reported previously. Here we report a three-generation family with autosomal dominant aplasia cutis congenita and preaxial polydactyly in the last generation and discuss whether it is a coincidence or not.


Global Gene Expression Profile of Acinetobacter baumannii During Bacteremia.
Acinetobacter baumannii is a pathogen of major importance in intensive care units worldwide, with the potential to cause problematic outbreaks and acquire high-level resistance to antibiotics. There is an urgent need to understand the mechanisms of A. baumannii pathogenesis for the future development of novel targeted therapies. In this study we performed an in vivo transcriptomic analysis of A. baumannii isolated from a mammalian host with bacteremia. Mice were infected with A. baumannii American Type Culture Collection 17978 using an intraperitoneal injection, and blood was extracted at 8 hours to purify bacterial RNA for RNA-Seq with an Illumina platform. Approximately one-quarter of A. baumannii protein coding genes were differentially expressed in vivo compared with in vitro (false discovery rate, ≤0.001; 2-fold change) with 557 showing decreased and 329 showing increased expression. Gene groups with functions relating to translation and RNA processing were overrepresented in genes with increased expression, and those relating to chaperone and protein turnover were overrepresented in the genes with decreased expression. The most strongly up-regulated genes corresponded to the 3 recognized siderophore iron uptake clusters, reflecting the iron-restrictive environment in vivo. Metabolic changes in vivo included reduced expression of genes involved in amino acid and fatty acid transport and catabolism, indicating metabolic adaptation to a different nutritional environment. Genes encoding types I and IV pili, quorum sensing components, and proteins involved in biofilm formation all showed reduced expression. Many genes that have been reported as essential for virulence showed reduced or unchanged expression in vivo. This study provides the first insight into A. baumannii gene expression profiles during a life-threatening mammalian infection. Analysis of differentially regulated genes highlights numerous potential targets for the design of novel therapeutics.


Sustained vertebral fracture risk reduction after withdrawal of teriparatide in postmenopausal women with osteoporosis.
Teriparatide (recombinant human parathyroid hormone [1-34]) reduces fracture risk in postmenopausal women with osteoporosis. We assessed the safety and incidence of new vertebral fractures after withdrawal of teriparatide. This study is a follow-up to the Fracture Prevention Trial (FPT), a randomized, placebo-controlled study of postmenopausal women with osteoporosis treated with teriparatide (20 or 40 microg) once daily for a mean of 18 months. More than 90% of the women remaining at the end of the FPT continued into the follow-up study (n = 1262). Patients and investigators were unblinded to original treatment group assignment. Women were treated according to standard clinical practice, including elective use of osteoporosis drugs. New vertebral fractures were determined by semiquantitative scoring of lateral thoracic lumbar spine radiographs 18 months after the end of the FPT. During the follow-up study, the reduction in fracture risk associated with previous treatment with teriparatide, 20 and 40 microg, was 41% (P = .004) and 45% (P = .001), respectively, vs placebo. The absolute reduction from the FPT baseline to the 18-month follow-up visit was 13% for both doses. Osteoporosis drugs were used by 47% of women during follow-up, with greater use in the former placebo group (P = .04); nevertheless, persistent fracture protection of previous teriparatide therapy was evident. Post hoc analysis also suggests that teriparatide treatment substantially reduced the increased risk of subsequent fracture in women who sustained a fracture during the FPT (P = .05). Vertebral fracture risk reduction by teriparatide administration persists for at least 18 months after discontinuation of therapy.


Review: Head and neck squamous cell carcinoma in sub-Saharan Africa.
Review the literature from 1990 to 2013 to determine known anatomic sites, risk factors, treatments, and outcomes of head and neck squamous cell carcinoma (HNSCC) in sub-Saharan Africa. Using a systematic search strategy, literature pertaining to HNSCC in sub-Saharan Africa was reviewed and patient demographics, anatomic sites, histology, stage, treatment, and outcomes were abstracted. The contributions of human immunodeficiency virus (HIV), human papillomavirus (HPV) and behavioural risk factors to HNSCC in the region were assessed. Of the 342 papers identified, 46 were utilized for review, including 8611 patients. In sub-Saharan Africa, the oropharyngeal/oral cavity was found to be the most common site, with 7750 cases (90% of all cases). Few papers distinguished oropharyngeal from oral cavity, making identification of possible HPV-associated oropharyngeal squamous cell carcinoma (SCC) difficult. SCC of the nasopharynx, nasal cavity, or paranasal sinuses was identified in 410 patients (4.8% of all cases). Laryngeal SCC was found in 385 patients (4.5% of all cases), and only 66 patients (0.8% of all cases) with hypopharyngeal SCC were identified. In 862 patients with data available, 43% used tobacco and 42% used alcohol, and reported use varied widely and was more common in laryngeal SCC than that of the oropharyngeal/oral cavity. Toombak and kola nut use was reported to be higher in patients with HNSCC. Several papers reported HIV-positive patients with HNSCC, but it was not possible to determine HNSCC prevalence in HIV-positive compared to negative patients. Reports of treatment and outcomes were rare. The oropharyngeal/oral cavity was by far the most commonly reported site of HNSCC reported in sub-Saharan Africa. The roles of risk factors in HNSCC incidence in sub-Saharan Africa were difficult to delineate from the available studies, but a majority of patients did not use tobacco and alcohol.


Utility of flow cytometry immunophenotyping and DNA ploidy studies for diagnosis and characterization of blood involvement in CD4+ Sézary's syndrome.
The exact immunophenotypic criteria for the identification of Sézary cells in the blood are still poorly defined. We analyzed the immunophenotype and DNA cell content of blood T cells in a series of 18 consecutive cases of Sézary's syndrome (SS), 21 normal individuals and 10 patients with reactive erythroderma, and correlated them with molecular and morphological findings. Phenotypically abnormal CD3+/TCRalphabeta+/CD4+ T cells were found in all SS patients but in none of the reactive erythroderma cases; small diploid, or less frequently hypodiploid Sézary's cells coexisted with large nearly tetraploid Sézary's cells in some cases. The most frequent phenotypic aberrations consisted in decreased expression of CD3/TCRalphabeta (94%), CD4 (94%), CD7 (100%) and/or CD2 (83%). In addition, Sézary's cells were constantly CD28+ and CD5+ and they did not express natural-killer associated (NKa) antigens. Phenotypic heterogeneity was a common finding and phenotypic changes over time were frequently observed. In contrast to what was found in patients with reactive erythroderma, flow cytometry analysis of the T-cell receptor (TCR) repertoire revealed a major TCR-Vbeta expansion in all SS cases. The presence of CD28+/CD5+/NKa-/CD4+ T cells expressing abnormally low levels of CD3, TCRalphabeta, CD4, CD7 and/or CD2 would support the diagnosis of SS in patients with erythroderma. Further analyses on larger series of patients are necessary in order to cover less frequent phenotypic patterns, establish the preferential usage of specific TCR-Vb families and investigate the specificity of these phenotypic abnormalities for diagnosing SS.


Pharmacogenomics of Methotrexate Membrane Transport Pathway: Can Clinical Response to Methotrexate in Rheumatoid Arthritis Be Predicted?
Methotrexate (MTX) is widely used for rheumatoid arthritis (RA) treatment. Single nucleotide polymorphisms (SNPs) could be used as predictors of patients' therapeutic outcome variability. Therefore, this study aims to evaluate the influence of SNPs in genes encoding for MTX membrane transport proteins in order to predict clinical response to MTX. Clinicopathological data from 233 RA patients treated with MTX were collected, clinical response defined, and patients genotyped for 23 SNPs. Genotype and haplotype analyses were performed using multivariate methods and a genetic risk index (GRI) for non-response was created. Increased risk for non-response was associated to SLC22A11 rs11231809 T carriers; ABCC1 rs246240 G carriers; ABCC1 rs3784864 G carriers; CGG haplotype for ABCC1 rs35592, rs2074087 and rs3784864; and CGG haplotype for ABCC1 rs35592, rs246240 and rs3784864. GRI demonstrated that patients with Index 3 were 16-fold more likely to be non-responders than those with Index 1. This study revealed that SLC22A11 and ABCC1 may be important to identify those patients who will not benefit from MTX treatment, highlighting the relevance in translating these results to clinical practice. However, further validation by independent studies is needed to develop the field of personalized medicine to predict clinical response to MTX treatment.


An automated clinical alert system for newly-diagnosed atrial fibrillation.
Clinical decision support systems that notify providers of abnormal test results have produced mixed results. We sought to develop, implement, and evaluate the impact of a computer-based clinical alert system intended to improve atrial fibrillation stroke prophylaxis, and identify reasons providers do not implement a guideline-concordant response. We conducted a cohort study with historical controls among patients at a tertiary care hospital. We developed a decision rule to identify newly-diagnosed atrial fibrillation, automatically notify providers, and direct them to online evidence-based management guidelines. We tracked all notifications from December 2009 to February 2010 (notification period) and applied the same decision rule to all patients from December 2008 to February 2009 (control period). Primary outcomes were accuracy of notification (confirmed through chart review) and prescription of warfarin within 30 days. During the notification period 604 notifications were triggered, of which 268 (44%) were confirmed as newly-diagnosed atrial fibrillation. The notifications not confirmed as newly-diagnosed involved patients with no recent electrocardiogram at our institution. Thirty-four of 125 high-risk patients (27%) received warfarin in the notification period, compared with 34 of 94 (36%) in the control period (odds ratio, 0.66 [95% CI, 0.37-1.17]; p = 0.16). Common reasons to not prescribe warfarin (identified from chart review of 151 patients) included upcoming surgical procedure, choice to use aspirin, and discrepancy between clinical notes and the medication record. An automated system to identify newly-diagnosed atrial fibrillation, notify providers, and encourage access to management guidelines did not change provider behaviors.


Evidence of Hexadecapole Deformation in Uranium-238 at the Relativistic Heavy Ion Collider.
State-of-the-art hydrodynamic simulations of the quark-gluon plasma are unable to reproduce the elliptic flow of particles observed at the BNL Relativistic Heavy Ion Collider (RHIC) in relativistic ^{238}U+^{238}U collisions when they rely on information obtained from low-energy experiments for the implementation of deformation in the colliding ^{238}U ions. We show that this is due to an inappropriate treatment of well-deformed nuclei in the modeling of the initial conditions of the quark-gluon plasma. Past studies have identified the deformation of the nuclear surface with that of the nuclear volume, though these are different concepts. In particular, a volume quadrupole moment can be generated by both a surface hexadecapole and a surface quadrupole moment. This feature was so far neglected in the modeling of heavy-ion collisions, and is particularly relevant for nuclei like ^{238}U, which is both quadrupole deformed and hexadecapole deformed. With rigorous input from Skyrme density functional calculations, we show that correcting for such effects in the implementation of nuclear deformations in hydrodynamic simulations restores agreement with BNL RHIC data. This brings consistency to the results of nuclear experiments across energy scales, and demonstrates the impact of the hexadecapole deformation of ^{238}U on high-energy collisions.


An electrochemical enzyme immunoassay for chicken luteinizing hormone: extension of the detection limit by adequate control of the nonspecific adsorption.
A noncompetitive heterogeneous enzyme immunoassay for the determination of chicken luteinizing hormone (LH) was equipped with an electrochemical endpoint in order to further enhance its sensitivity. The immunological principle of the original ELISA remained essentially unchanged, except for the fact that the peroxidase label was replaced by alkaline phosphatase, since in the upgraded version of the assay, p-aminophenyl phosphate was to be used as the substrate of alkaline phosphatase. Enzyme-generated p-aminophenol was injected into a flow-injection system and detected amperometrically in a thin-layer flow cell with a glassy carbon electrode at 0.325 V vs Ag/AgCl. A classical problem associated with this type of solid-phase immunoassay is the adsorption of proteins other than the capture antibody to the solid phase. The detection sensitivity is therefore often limited by a large background signal observed in the absence of antigen. In the present study, an experiment was designed to examine in each step of the assay the contribution of each of the potential sources of background current. It was shown that the major contribution to the background current was caused by the nonspecific adsorption of biotinylated secondary antibody. Adsorption of the secondary antibody (biotinylated goat anti-rabbit IgG) to the capture antibody (mouse anti-chicken LHbeta) was clearly a case of specific aspecificity, whereas adsorption to the solid phase itself had to be treated as a nonspecific aspecificity. Addition of 0.25% mouse serum to the secondary antibody as a source of mouse immunoglobulin could overcome the cross-reaction and markedly reduced adsorption to capture antibody. The second part of nonspecific adsorption was eliminated by using combinations of Tween 20 and bovine serum albumin as blocking agents. Controlling the adsorption of the biotinylated secondary antibody in this way decreased the detection limit from 39 pg/ml in the original assay to 2.5 pg/ml in the electrochemical version. This way, the plasma volume of samples containing on the order of 1 ng/ml LH was reduced to less than 10 microl. The linear range was 2.5-625 pg/ml. The method allowed us to measure LH in buffer and in adult and juvenile chicken plasma.


An enquiry into the process of categorization of pictures and words.
This paper reports a series of experiments conducted to study the categorization of pictures and words. Whereas some studies reported in the past have found a picture advantage in categorization, other studies have yielded no differences between pictures and words. This paper used an experimental paradigm designed to overcome some methodological problems to examine picture-word categorization. The results of one experiment were consistent with an advantage for pictures in categorization. To identify the source of the picture advantage in categorization, two more experiments were conducted. Findings suggest that semantic relatedness may play an important role in the categorization of both pictures and words. We explain these findings by suggesting that pictures simultaneously access both their concept and visually salient features whereas words may initially access their concept and may subsequently activate features. Therefore, pictures have an advantage in categorization by offering multiple routes to semantic processing.


Lipid peroxidation: Its effects on the formulation and use of pharmaceutical emulsions.
Pharmaceutical delivery systems are developed to improve the physicochemical properties of therapeutic compounds. Emulsions are one of these drug delivering systems formulated using water, oils and lipids as main ingredients. Extensive data are usually generated on the physical and chemical characteristics of these oil-in-water and lipid emulsions. However, the oxidative tendency of emulsions is often overlooked. Oxidation impacts the overall quality and safety of these pharmaceutical emulsions. Additionally, introducing oxidatively unstable emulsions into biological systems further promotes oxidation in situ. Products of these reactions then continue to pose serious harm to cells and fuel other physiological oxidation reactions. Consequently, the increase of oxidation products leads to oxidative damage to biological systems. Thus, emulsions with lower lipid peroxidation are more stable and will reduce the negative effects of oxidation in situ. Preventive measures during the formulation of emulsions are important. Many naturally occurring and cost effective substances possess low oxidation tendencies and confer oxidative protection when used in emulsions. Additionally, certain preparatory methods should be employed to reduce or better control lipid peroxidation. Finally, emulsions must be evaluated for their oxidation susceptibility using the various techniques available. Careful attention to the preparation of emulsions and assessment of their oxidative stability will help produce safer emulsions without compromising efficacy.


Cooperative synergy between NFAT and MyoD regulates myogenin expression and myogenesis.
Calcineurin/NFAT signaling is involved in multiple aspects of skeletal muscle development and disease. The myogenic basic helix-loop-helix transcription factors, MyoD, myogenin, Myf5, and MRF4 specify the myogenic lineage. Here we show that calcineurin/NFAT (nuclear factor of activated T cells) signaling is required for primary myogenesis by transcriptional cooperation with the basic helix-loop-helix transcription factor MyoD. Calcineurin/NFAT signaling is involved in myogenin expression in differentiating myoblasts, where the myogenic regulatory factor MyoD synergistically cooperates with NFATc2/c3 at the myogenin promoter. Using gel shift and chromatin immunoprecipitation assays, we identified two conserved NFAT binding sites in the myogenin promoter that were occupied by NFATc3 upon skeletal muscle differentiation. The transcriptional integration between NFATc3 and MyoD is crucial for primary myogenesis in vivo, as myogenin expression is weak in myod:nfatc3 double null embryos, whereas myogenin expression is unaffected in embryos with null mutations for either factor alone. Thus, the combined findings provide a novel transcriptional paradigm for the first steps of myogenesis, where a calcineurin/NFATc3 pathway regulates myogenin induction in cooperation with MyoD during myogenesis.


The biologic properties of recombinant human thrombopoietin in the proliferation and megakaryocytic differentiation of acute myeloblastic leukemia cells.
Thrombopoietin (TPO) is implicated as a primary regulator of megakaryopoiesis and thrombopoiesis. However, the biologic effects of TPO on human acute myeloblastic leukemia (AML) cells are largely unknown. To determine if recombinant human (rh) TPO has proliferation-supporting and differentiation-inducing activities in AML cells, 15 cases of AML cells that were exclusively composed of undifferentiated leukemia cells and showed growth response to rhTPO in a short-term culture (72 hours) were subjected to long-term suspension culture with or without rhTPO. Of 15 cases, rhTPO supported proliferation of AML cells for 2 to 4 weeks in 4 cases whose French-American-British subtypes were M0, M2, M4, and M7, respectively. In addition to the proliferation-supporting activity, rhTPO was found to induce AML cells to progress to some degree of megakaryocytic differentiation at both morphologic and surface-phenotypic level in 2 AML cases with M0 and M7 subtypes. The treatment of AML cells with rhTPO resulted in rapid tyrosine phosphorylation of the TPO-receptor, c-mpl, and STAT3 in all of cases tested. By contrast, the expression of erythroid/megakaryocyte-specific transcription factors (GATA-1, GATA-2, and NF-E2) was markedly induced or enhanced in only 2 AML cases that showed megakaryocytic differentiation in response to rhTPO. These results suggested that, at least in a fraction of AML cases, TPO could not only support the proliferation of AML cells irrespective of AML subtypes, but could also induce megakaryocytic differentiation, possibly through activation of GATA-1, GATA-2, and NF-E2.


The early development of transient and permanent hypothyroidism following radioiodine therapy for hyperthyroid Graves' disease.
The outcome in 110 patients first treated with radioiodine (mean dose 6.56 mCi) for hyperthyroid Graves' disease in 1980 was reviewed. In 23% of the patients the disease had not been controlled by the initial dose after 3 months, and 17% were given one or two more doses. Within 2 years 65% of the patients required replacement thyroxine therapy. Although about half of the patients were biochemically hypothyroid 3 months after the last dose of iodine 131, this condition was transient in a third of them; five of these patients even became hyperthyroid again. Patients with transient, as opposed to permanent, hypothyroidism at 3 months tended to be clinically euthyroid but to have residual palpable thyroid tissue and only a modest reduction in the serum thyroxine level. It is therefore recommended that patients not overtly hypothyroid 3 months after treatment with 131I be observed still longer before thyroxine replacement therapy is instituted.


[Speech intelligibility and psychosocial adaptation in various voice rehabilitation methods following laryngectomy].
Intelligibility and psychosocial adjustment are measured by the PLTT (Post-Laryngectomy-Telephone-Test) and a newly developed questionnaire (FPAL). 110 male patients with total or partial laryngectomies are tested. Three different groups are compared with each other: laryngectomees with voice prostheses, laryngectomees with oesophageal voice or artificial larynx, and patients with partial laryngectomies. As was to be expected, intelligibility (single words and sentences) was best in patients with partial laryngectomies. The intelligibility of the laryngectomees with voice prostheses is significantly higher than the intelligibility of patients with oesophageal voice or artificial larynx. There was no difference between the groups with regard to the subjective assessment of intelligibility in relation to various communication partners. Partial laryngectomies have the lowest negative impact on the social and psychological situation of the patients. Laryngectomees, however, are more lonely, indicate more psychological stress in social interactions, and have a less positive evaluation of their life. Statistical analyses reveal no difference in psychosocial adjustment between laryngectomees with voice prostheses, oesophageal voice and artificial larynges.


Characterisation of a synergistic interaction between a thymidylate synthase inhibitor, ZD1694, and a novel lipophilic topoisomerase I inhibitor karenitecin, BNP1100: mechanisms and clinical implications.
We developed a combination protocol for inhibitors of thymidylate synthase (TS) and DNA topoisomerase I (Topo I) that can exert highly lethal effects in vitro against HCT-8 human colorectal cancer cells. The specific schedule was constructed so that a TS inhibitor could induce not only primary DNA damage but also cellular conditions optimal for the efficient action of a Topo I inhibitor. The initial drug treatment consisted of a brief exposure to a quinazoline-based antifolate, ZD1694. After an interval of approximately one cell-doubling time, cells were exposed for 8-24 h to BNP1100, a Karenitecin-class 7-thiomethyl-camptothecin, in the presence of 1-10 microM thymidine; the latter acted as a crucial factor to promote the collision of moving replication forks with the drug-stabilised DNA-Topo I cleavable complexes even under continuous TS inhibition. Clonogenic analyses confirmed that these mechanistically distinct drugs at clinically achievable concentrations worked in a highly synergistic manner, with a maximum effect abolishing the viability of virtually all cancer cells (> 99.9%). The pretreatment with ZD1694 increased the amount of DNA-bound Topo I by up to 4-fold and the DNA-damaging capability of BNP1100 by up to 15-fold. The possibility of at least four DNA-damaging pathways is proposed which might have resulted from the individual actions of TS and Topo I inhibitors as well as their concerted actions. Taken together, the present findings provided a logically permissible explanation as to why TS and Topo I inhibitors in concerted interactions induced a highly lethal effect which was more than a simple additive effect. Since these drugs are effective specifically on actively proliferating cancer cells, but not on non-cycling G0/G1 cells, this mechanism-based protocol may warrant consideration for clinical verification.


Smartphone assisted oral health data recording - an android based software application development.
Smartphone compared to the traditional pen-paper method could enhance oral health data recording procedure by reducing the cost of data collection, risk of data loss, early detection of errors and reducing data entry time. The present research developed a mobile/tablet-based software application to capture oral health data and test its adaptability and operations in oral health surveys. A comparative cross-sectional study was conducted among the general population of Sanwer town, Indore district. The initial testing of the application was done on 120 individuals. A random sampling (lottery method) followed by a systematic sampling strategy was employed to select 120 households. A "one per household" design was implemented for the survey. The initial oral health data collection was done using mobile-assisted software application followed by a second examination scheduled after 15 days on the same participants using the conventional Pen-paper method to collect oral health data. Six Investigator Recorder (IR) teams conducted the oral health data collection. Data collection through Smartphone-based application displayed less meantime (3.57 minutes) in comparison to pen-paper method (4.87 minutes) (p≤0.001). Survey team response showed the majority of investigators having strong agreement on user satisfaction and speed of data entry using software application. The initial testing of mobile-assisted recording system (MARS) efficiently captured oral health data among the general population with wide variations in oral disease level. The application facilitated minimal or no wastage of paper and had a high level of user-satisfaction, accuracy, speed of entry and low potential for any data loss.


Agminated pigmented matricoma: a case of a unique tumor with a multifocal appearance composed of neoplastic matrical cells with a significant component of melanocyte.
Matricoma is benign follicular neoplasm with the same constituent cells as pilomatricoma but with a different silhouette. Matricoma consists mostly of solid aggregations of matrical cells, as opposed to pilomatricoma, which is often a cystic lesion. We herein describe a case of agminated pigmented matricoma. A 27-year-old Japanese man visited our hospital complaining of three separate, pigmented, firm and sessile nodules on the back. Histopathologic examination revealed mostly solid aggregations of matrical and supramatrical cells located mainly within the dermis and the upper part of the subcutis. Shadow cells were present mostly within the aggregations. At higher magnifications, occasional indication of inner sheath differentiation, in addition to differentiation toward hair in the form of shadow cells, could be discerned in the center of the aggregations. The matrical and supramatrical cells within the aggregations were relatively uniform in size and shape, and contained several mitotic figures. Heavily pigmented melanocytes were present within the aggregations of matrical and supramatrical cells, and heavily pigmented melanophages were found in adjacent stroma. Pigmentation is a rare event in benign and malignant adnexal tumors, and to our knowledge, no similar case has been reported.


[Mood disorders in childhood and adolescence: continuities and discontinuities to adulthood].
Studies in child and adolescent psychiatry show that both internalizing and externalizing problems tend to persist in adulthood, e.g. it has been found that almost 40% of the adolescents who were classified as deviant initially were still deviant 4 years later. In a more recent study, over a large period of 24 years, continuity of psychopathology was found from childhood into adulthood; anxious/ depressed and other problems in childhood were core predictors for adult psychopathology. Other studies also show that there is substantial continuity, morbidity, and potential mortality from suicide into adulthood in patients with adolescent-onset major depressive disorder. Formerly depressed adolescents are at higher risk for recurrence of major depressive disorder during young adulthood; more specifically, depressed adolescents are at 2-7 times increased odds of being depressed as adults, compared with non-depressed controls. In addition, recent studies indicate that symptoms of depression in adolescence strongly predict an episode of major depression, suicidal ideation and rates for treatment for depression in adulthood, even among adolescents without major depression. The Maudsley long-term follow-up of child and adolescent depression showed that adolescent depression carries an elevated risk of adult depression irrespective of comorbidity, and that comorbid conduct disorder in childhood is associated with raised rates of other psychiatric outcomes. Personality disorders, i.e. of dependent, antisocial, passive-aggressive, and histrionic type, may represent alternative pathways of continuity for major depressive disorder and other Axis I disorders across the child-adult transition. Bipolar disorder in childhood is often found to be associated with long delays to first treatment. Studies have shown that both childhood onset and treatment delay are associated with a persistently more adverse course of illness in adulthood. Earlier and more effective treatment of bipolar disorder in children and adolescents would result in a more benign illness and a better prognosis in adulthood. Lastly, parental depression is a strong and consistent risk factor for offspring major depression and anxiety disorder. When offspring mature into young adulthood, effects of parental depression and family discord persist. Because child and especially adolescent mood disorders are likely to continue into adulthood, early identification, assessment and treatment are warranted; capacity for treatment of children at earlier stages of risk and disorder needs to be developed in other public sectors, such as primary health care and schools. In addition, continuity of care from childhood and adolescence to adulthood is needed.


Chitosan-alginate multilayered films with gradients of physicochemical cues.
Tissues presenting continuous variations of properties in one direction have inspired the development of functional graded materials. In this work, we developed a new facile method for the development of continuous gradients in chitosan (CHIT) and alginate (ALG) polyelectrolyte multilayers (PEMs) obtained layer-by-layer based on the gradual dipping of CHIT/ALG coated glass slides in genipin solution. Stiffness gradients were produced in the cm scale by varying the reaction time with genipin. Quartz crystal microbalance, colorimetric measurements, trypan blue assay, attenuated total reflection-Fourier transform infrared spectroscopy, swelling ability, water contact angle and dynamic mechanical analysis (DMA) were used to find suitable conditions for the stiffness gradient. The PEMs can be successfully built up and cross-linked with genipin to yield surfaces with uniform physicochemical properties or with gradients of different physicochemical properties. It was found that a large reduction in the hydrophobic nature of the CHIT/ALG PEMs could be produced with higher cross-linking reaction times, regardless of the decrease in their swelling ability. Moreover, the mechanical properties were evaluated using an innovative and non-conventional DMA to monitor the cross-linking reaction in situ. The results confirm an enhancement on the tensile storage modulus with increasing reaction times from 60 to 140 MPa. In another original DMA testing protocol the local compression storage modulus was also measured directly on the films along the stiffness gradient, with results consistent with the tensile tests obtained on the freestanding membranes with different cross-linking degrees. The in vitro biological performance demonstrates that L929 adhered and spread more in the stiffer regions. This work demonstrates the versatility and feasibility of the LbL methodology to generate functional biomimetic surfaces with tuned mechanical and physicochemical properties, which hold great promise for the study of cell-substrate interactions.


Nanotechnology: Newer Approach in Insulin Therapy.
Insulin is a peptide hormone released by pancreatic beta cells. An autoimmune reaction in diabetes mellitus type 1 causes the beta cells to die, preventing insulin from being produced or released into the bloodstream; that impacts 30 million people globally and is linked to shortened lifespan due to acute and chronic repercussions. Insulin therapy aims to replicate normal pancreatic insulin secretion, which includes low levels of insulin that are always present to support basic metabolism, as well as the two-phase secretion of additional insulin in response to high blood sugar - an initial spike in secreted insulin, followed by an extended period of continued insulin secretion. This is performed by combining various insulin formulations at varying rates and lengths of time. Since the beginning of human insulin use, several advances in insulin formulations have been made to help meet these aims as much as possible, resulting in improved glycaemic control while limiting hypoglycemia. In this review, we looked at devices used by patients with type 1 diabetes, such as insulin pumps, continuous glucose monitors, and, more recently, systems that combine a pump with a monitor for algorithm-driven insulin administration automation. We intend to provide insight into supplementary therapies and nanotechnology employed in insulin therapy as a result of our review.


Tourniquet use in total knee arthroplasty: a meta-analysis.
The use of an intraoperative tourniquet for total knee arthroplasty (TKA) is a common practice. However, the effectiveness and safety are still questionable. A systematic review was conducted to examine that whether using a tourniquet in TKA was effective without increasing the risk of complications. A comprehensive literature search was done in PubMed Medicine, Embase, and other internet database. The review work and the following meta-analysis were processed to evaluate the role of tourniquet in TKA. Eight randomized controlled trials and three high-quality prospective studies involving 634 knees and comparing TKA with and without the use of a tourniquet were included in this analysis. The results demonstrated that using a tourniquet could decrease the measured blood loss but could not decrease the calculated blood loss, which indicated actual blood loss. Patients managed with a tourniquet might have higher risks of thromboembolic complications. Using the tourniquet with late release after wound closure could shorten the operation time; whereas early release did not show this benefit. The current evidence suggested that using tourniquet in TKA may save time but may not reduce the blood loss. Due to the higher risks of thromboembolic complications, we should use a tourniquet in TKA with caution.


Safety, immunogenicity, and efficacy of an inactivated avian polyomavirus vaccine.
To determine safety, immunogenicity, and efficacy of an inactivated avian polyomavirus vaccine in nonbudgerigar psittacine birds that varied in age, species, and immunologic status. Safety of the vaccine was evaluated in 1,823 psittacines representing more than 80 species. Immunogenicity was evaluated in 285 birds (260 of various Psittaciformes species, 25 chickens). Efficacy was evaluated in 104 birds (78 of various Psittaciformes species, 26 chickens). Safety was evaluated by vaccinating birds that were determined to be seronegative or seropositive (titer > 1:10) prior to vaccination. Birds were then evaluated for clinically detectable systemic or local reactions for 2 months to 2 years. Immunogenicity was evaluated by testing for virus-neutralizing antibodies, vaccinating each bird twice, and then testing for a significant change in antibody titer. Efficacy was evaluated by vaccinating birds, followed in 2 to 4 weeks by intramuscular or intravenous challenge exposure. After challenge exposure, protection was evaluated by attempting to recover virus from tissues or by observing birds for clinical signs of disease and testing for a significant change in titer. Avian polyomavirus vaccine is safe, immunogenic, and efficacious for use in multiple species of mature and immature psittacines. Until now, prevention of polyomavirus infection in psittacine birds could only be accomplished through strict isolation to reduce potential exposure to the virus. The USDA-registered inactivated avian polyomavirus vaccine can safely be used to protect vaccinates from infection and control spread of this virus in flocks.


The Space Between: Nature and Machine Heuristics in Evaluations of Organisms, Cyborgs, and Robots.
Cues delivered by agents are known to trigger mental shortcuts associated with ontological category or the kind of thing an agent is. Two such heuristics are key to considering organic and machine agents, and result in biased evaluations: the machine heuristic (MH) (if machine, then systematic/unbiased, therefore its products are good) and the nature heuristic (NH) (if natural, then pure/innate, therefore good). As machine agents such as robots are increasingly integrated into human spheres, it is yet unknown (a) if invocation of agent-cued heuristics is inherently tied to activities and (b) whether either/both heuristics are evoked when agents exhibit both organic and machinic properties (as with cyborgs). To investigate these open questions, a 3 × 2 experiment tasked individuals with considering a magazine article about an agent (organism, cyborg, robot) performing behaviors (natural, technical) to solve a widespread problem, and then evaluating the agent and its solution for markers of machine and NHs. Findings indicate that the NH may be dominant over the MH; however, this primacy may be driven by operational contexts. Post hoc analysis suggests that agent category grounds interpretations of agent behaviors that, in turn, drive biased evaluations of behavioral outcomes.


Statistical methods for mapping quantitative trait loci from a dense set of markers.
Lander and Botstein introduced statistical methods for searching an entire genome for quantitative trait loci (QTL) in experimental organisms, with emphasis on a backcross design and QTL having only additive effects. We extend their results to intercross and other designs, and we compare the power of the resulting test as a function of the magnitude of the additive and dominance effects, the sample size and intermarker distances. We also compare three methods for constructing confidence regions for a QTL: likelihood regions, Bayesian credible sets, and support regions. We show that with an appropriate evaluation of the coverage probability a support region is approximately a confidence region, and we provide a theroretical explanation of the empirical observation that the size of the support region is proportional to the sample size, not the square root of the sample size, as one might expect from standard statistical theory.


Homocysteine, folate, and vitamin B-12 in mild cognitive impairment, Alzheimer disease, and vascular dementia.
Evidence supports an independent association between plasma total homocysteine concentrations and the risk of vascular disease. Recent epidemiologic studies reappraised the possibility that vascular risk factors might play a role in the pathogenesis not only of vascular dementia (VaD) but also of Alzheimer disease (AD). The objective was to investigate the relations of mild cognitive impairment, AD, and VaD with blood homocysteine, folate, and vitamin B-12. The study population consisted of 314 consecutive subjects, 228 of whom were eligible for analyses. Plasma total homocysteine, serum folate, and serum vitamin B-12 concentrations were measured in 55 nondemented elderly control subjects, 81 mildly cognitively impaired subjects (Clinical Dementia Rating: 0.5), and 92 demented patients prevalently in a mild disease stage and with a clinical diagnosis of AD (n = 74) or VaD (n = 18). Subjects in the lowest folate tertile had significantly higher adjusted odds ratios (ORs) for mild cognitive impairment (OR: 3.1; 95% CI: 1.2, 8.1) and dementia (3.8; 1.3, 11.2). Hyperhomocysteinemia was significantly associated with dementia (adjusted OR: 4.3; 1.3, 14.7) and AD (adjusted OR: 3.7; 1.1, 13.1). In subjects with a Clinical Dementia Rating of 0.5, the mean (+/- SE) Mini-Mental State Examination score was significantly lower (P < 0.05) in the highest homocysteine tertile (24.5 +/- 0.5) than in the lowest tertile (26.6 +/- 0.5). No significant associations were found between minimum medial temporal lobe thickness or leukoaraiosis and any biochemical measure in the dementia and AD groups. These findings suggest that relative folate deficiency may precede AD and VaD onset. Hyperhomocysteinemia might also be an early risk factor for cognitive decline in the elderly, but its role in dementia development must be addressed in future longitudinal studies.


Comparison of 3% Hypertonic Saline and 20% Mannitol for Reducing Intracranial Pressure in Patients Undergoing Supratentorial Brain Tumor Surgery: A Randomized, Double-blind Clinical Trial.
In the present study, we hypothesized that 3% hypertonic saline (HS) is more effective than 20% mannitol to reduce intracranial pressure (ICP) and to modify brain bulk in patients undergoing an elective supratentorial craniotomy. After institutional review board approval, patients scheduled to undergo supratentorial craniotomy were enrolled into this prospective, randomized, double-blind study. The patients were monitored for routine hemodynamic parameters, depth of anesthesia, and ICP. They received 5 mL/kg 20% mannitol (n=20) or 3% HS (n=19) as infusion for 15 minutes. The patients' ICP values were monitored during hypertonic fluid infusion and throughout 30 minutes after infusion as a primary outcome. Secondary outcomes were hemodynamic variables, serum sodium value, blood gases, and surgeon brain relaxation assessment score (1=relaxed, 2=satisfactory, 3=firm, 4=bulging). In addition, the length of intensive care unit and hospital stay were recorded. Demographic and tumor characteristics were similar between groups. The basal (before hypertonic infusion, ICPT0) and last (30 min after hypertonic infusion finished, ICPT45) ICP values were 13.7±3.0 and 9.5±1.9 mm Hg, respectively, for the M group, which were comparable with the corresponding levels of 14.2±2.8 and 8.7±1.1 mm Hg in the HS group (P>0.05). The median amount of ICP reduction between T0 and T45 timepoints were 4 (1 to 7) and 5 (1 to 9) mm Hg for group M and group HS, respectively (P=0.035). Baseline central venous pressure, pulse pressure variation, and serum sodium and lactate values were similar between groups, but the last measured pulse pressure variation and lactate value were lower, and sodium value was higher in group HS than in group M (P<0.05). Duration of hospital and intensive care unit stay were similar between groups. Our results suggest that 3% HS provided more effective ICP reduction than 20% mannitol during supratentorial brain tumor surgery.


Preservation of episodic visual recognition memory in aging.
Visual episodic recognition memory was assessed in young (mean age 22.5 years) and older (mean age 74.1 years) adults. To isolate purely visual memory, recognition was tested with sets of briefly-presented compound sinusoidal gratings, which minimized age-related differences in visual processing, and resisted verbal encoding. Recognition was measured after delays of 1, 2 or 4 seconds. Overall, neither accuracy of recognition nor speed of response differed significantly between groups, or with probe delay, which strengthens recent claims that visual memory tends to be spared during the course of normal aging.


Purine nucleoside kinases in human T- and B-lymphoblasts.
Purine nucleoside kinases in human B- and T-lymphoblasts were fractionated by DEAE-cellulose chromatography. Human B-lymphoblast cell extracts showed three peaks of nucleoside kinase activities, adenosine kinase (EC 22.214.171.124), deoxyguanosine kinase and deoxycytidine kinase (EC 18.104.22.168). However, T-lymphoblast cell extracts showed a nucleoside kinase activity which phosphorylates deoxycytidine, deoxyadenosine and deoxyguanosine, similar to deoxycytidine kinase, in addition to the three nucleoside kinases. The Km values of T-lymphoblast-specific nucleoside kinase for deoxyadenosine and deoxyguanosine, 15 and 26 microM, respectively, were smaller than those of deoxycytidine kinase, 150 and 330 microM, respectively. Deoxyadenosine phosphorylation by deoxycytidine kinase was strongly inhibited by dCTP, but the phosphorylation by T-lymphoblast-specific nucleoside kinase was only weakly inhibited by dCTP. Deoxyadenosine phosphorylating activity in B-lymphoblast extracts was more distinctly inhibited by dCTP than that in T-lymphoblast extracts.


Coping and coping assistance among children with sickle cell disease and their parents.
The ways in which a family copes with the physical and psychosocial burdens of sickle cell disease (SCD) can influence the child and family functioning. However, few studies have examined SCD-related stressors beyond pain or how children and parents cope with these stressors. This study aimed to describe child coping and parent attempts to help their children cope (ie, coping assistance) with a range of SCD stressors by using a triangulated mixed methods design. We also explored convergence between findings from qualitative interviews and quantitative coping inventories. Fifteen children (aged 6 to 14 years) with SCD and their parents (N=15) completed semistructured interviews and self-report measures to assess SCD-related stressors, coping, and coping assistance strategies. Findings indicate that children experience numerous stressors related to SCD and its treatment, including, but not limited to, pain. To manage these stressors, families employ a range of approach and avoidance-oriented coping strategies. Quantitative and qualitative assessments provided complementary and unique contributions to understanding coping processes among children with SCD and their parents. Examining a broad range of stressors and integrating multiple assessment methods helps improve our understanding of coping with pediatric SCD, which may inform clinical practice and family-focused intervention development.


TRPV1 and TRPV1-Expressing Nociceptors Mediate Orofacial Pain Behaviors in a Mouse Model of Orthodontic Tooth Movement.
Orthodontic force produces mechanical irritation and inflammation in the periodontium, which is inevitably accompanied by pain. Despite its prevalence, treatment of orthodontic pain is ineffective. Elucidating underlying neural mechanisms is critical to improving the management of orthodontic pain. We have assessed the contribution of transient receptor potential vanilloid subtype 1 (TRPV1) and the TRPV1-expressing subset of nociceptive afferents to pain behaviors induced by orthodontic force in mice. Microfocus X-ray computed tomography analysis showed that application of an orthodontic force of 10 g to the maxillary first molar produced reliable tooth movement in mice. Mouse grimace scale (MGS) was evaluated as an indication of non-evoked spontaneous pain and bite force (BF) was measured for assessing bite-evoked nocifensive behaviors. Orthodontic force increased MGS and decreased BF, both of which were interpreted as increased levels of pain. These behaviors peaked at 1d and returned near to the sham level at 7d. Retrograde labeling and immunohistochemical assays showed TRPV1-expressing peptidergic afferents are abundantly projected to the periodontium. Direct injection of resiniferatoxin into trigeminal ganglia (TG) decreased TRPV1-expressing afferents by half in the targeted region of TG. The chemical ablation of TRPV1-expressing afferents significantly attenuated orthodontic pain behaviors assessed by MGS and BF. Consistently, the knockout of TRPV1 also attenuated orthodontic force-induced changes in MGS and BF. These results suggest that TRPV1 and TRPV1-expressing trigeminal nociceptors constitute a primary pathway mediating orthodontic pain behaviors in mice. This model will be useful for mechanistic studies on orthodontic pain aimed at developing novel approaches for painless orthodontics.


Spontaneous intraperitoneal rupture of pyonephrosis in a patient with unknown kidney carcinosarcoma: a case report.
Seventeen cases of peritonitis due to rupture of a pyonephrosis have been reported. The majority of these cases occur secondary to renal stones. Only two cases of ruptured pyonephrosis with concurrent kidney neoplasm have been described and only one of these presented as an acute peritonitis. In this presentation we discuss an unusual case of a 68 year old man with a chronic history of bilateral nephrolithiasis and recent pyonephrosis. He presented acutely with peritonitis and was later found to have a carcinosarcoma of the kidney. The case highlights the importance of recognizing the possibility of underling renal carcinoma in patients presenting with a ruptured pyonephrosis and discuss steps to avoid this serious complication.


Escherichia coli aggravates endoplasmic reticulum stress and triggers CHOP-dependent apoptosis in weaned pigs.
Intestinal cells can sense the presence of pathogens and trigger many important signaling pathways to maintain tissue homeostasis and normal function. Escherichia coli and lipopolysaccharides (LPS) are the main pathogenic factors of intestinal disease in pigs. However, the roles of endoplasmic reticulum stress (ERS) and its mediated apoptosis in intestinal malfunction induced by E. coli or LPS remain unclear. In the present study, we aimed to evaluate whether ERS could be activated by E. coli fed to piglets and whether the underlying mechanisms of this disease process could be exploited. Eighteen weaned pigs (21 days old) were randomly assigned to one of two treatment groups (n = 9 per group). After pre-feeding for 1 week, the diets of the piglets in one group were supplemented with E. coli (W25 K, 109 cells kg-1 diet) for 7 days. At the end of the experiment, all piglets were slaughtered to collect jejunum and ileum samples. Western blotting and immunofluorescence experiments were used to determine the expression levels and histological locations of ERS and its downstream signaling proteins. The intestinal porcine epithelial cell line J2 (IPEC-J2) was used as in vitro model to investigate the possible mechanism. The results showed that E. coli supplementation in the diet increased the GRP78 expression in the jejunum and ileum, especially in the jejunal epithelium and ileac germinal center, and elevated the expression levels of CHOP (in both the jejunum and ileum) and caspase-11 (in the ileum), indicating that ERS and CHOP-caspase-11 dependent apoptosis were activated in the porcine small intestine. Moreover, as demonstrated by in vitro experiments, the CHOP inhibitor 4-phenylbutyrate alleviated the damage to IPEC-J2 cells induced by LPS derived from E. coli. Taken together, these data strongly suggest that ERS can be triggered in the small intestine by dietary supplementation with E. coli and that CHOP-caspase-11 dependent apoptosis may play a key role in maintaining normal homeostasis of the intestine in response to pathogenic factors.


The effects in vivo of mutationally modified uroporphyrinogen decarboxylase in different hem12 mutants of baker's yeast (Saccharomyces cerevisiae).
Nine new hem12 haploid mutants of baker's yeast (Saccharomyces cerevisiae), totally or partially deficient in uroporphyrinogen decarboxylase activity, were subjected to both genetic and biochemical analysis. The mutations sites studied are situated far apart within the HEM12 gene located on chromosome IV. Uroporphyrinogen decarboxylase activity in the cell-free extracts of the mutants was decreased by 50-100%. This correlated well with the decrease of haem formation and the increased accumulation and excretion of porphyrins observed in vivo. The pattern of porphyrins (uroporphyrin and its decarboxylation products) accumulated in the cells of mutants partially deficient in uroporphyrinogen decarboxylase activity did not differ significantly, although differences in vitro were found in the relative activity of the mutant enzyme at the four decarboxylation steps. The excreted porphyrins comprised mainly dehydroisocoproporphyrin or pentacarboxyporphyrin. In heterozygous hem12-1/HEM12 diploid cells, a 50% decrease in decarboxylase activity led to an increased accumulation of porphyrins as compared with the wild-type HEM12/HEM12 diploid, which points to the semi-dominant character of the hem12-1 mutation. The biochemical phenotypes of both the haploid and the heterozygous diploid resembles closely the situation encountered in porphyria cutanea tarda, the most common human form of porphyria.


Beyond the GIST: mesenchymal tumors of the stomach.
Intramural gastric masses arise in the wall of the stomach (generally within the submucosa or muscularis propria), often with intact overlying mucosa. These tumors are typically mesenchymal in origin and have overlapping radiologic appearances. A combination of features such as location, attenuation, enhancement, and growth pattern may suggest one diagnosis over another. Gastrointestinal stromal tumors (GISTs) account for the majority of intramural tumors and can vary widely in appearance, from small intraluminal lesions to exophytic masses that protrude into the peritoneal cavity, commonly with areas of hemorrhage or necrosis. A well-circumscribed mass measuring -70 to -120 HU is a lipoma. Leiomyomas usually manifest as low-attenuation masses at the gastric cardia. Homogeneous attenuation is a noteworthy characteristic of schwannomas, particularly for larger lesions that might otherwise be mistaken for GISTs. A hypervascular mass in the antrum is a common manifestation of glomus tumors. Hemangiomas are also hypervascular but often manifest in childhood. Inflammatory fibroid polyps usually arise as a polypoid mass in the antrum. Inflammatory myofibroblastic tumors are infiltrative neoplasms with a propensity for local recurrence. Plexiform fibromyxomas are rare, usually antral tumors. Carcinoid tumors are epithelial in origin, but often submucosal in location, and therefore should be distinguished from other intramural lesions. Multiple carcinoid tumors are associated with hypergastrinemia, either in the setting of chronic atrophic gastritis or Zollinger-Ellison syndrome. Sporadic solitary carcinoid tumors not associated with hypergastrinemia have a higher rate of metastasis. Histopathologic analysis, including immunohistochemistry, is usually required for diagnosis of intramural masses.


The oligomerization of the coiled coil-domain of occludin is redox sensitive.
The transmembrane tight junction protein occludin is sensitive to oxidative stress. Occludin oligomerizes; however, its function in the tight junction is unknown. The cytosolic C-terminal tail contains a coiled coil-domain and forms dimers contributing to the oligomerization. The regulation of the oligomerization remains unclear. As the domain area contains sulfhydryl residues, we tested the hypothesis that the dimerization of the coiled coil-domain depends on these residues. We showed that the dimerization is modulated by the thiol concentration in the low-millimolar range, which is relevant both for physiological and pathophysiological conditions. Masking the sulfhydryl residues in the fragment by covalent binding of 4-vinyl pyridine prevented the dimerization but did not affect its helical structure and cylindric shape. The data demonstrate, for the first time, that disulfide bridge formation of murine cystein 408 is involved in the dimerization. This process is redox-sensitive but the secondary structure of the domain is not. It is concluded that the dimerization of occludin may play a regulatory role in the tight junction assembly under physiological and pathological conditions.


Involvement of BcYak1 in the Regulation of Vegetative Differentiation and Adaptation to Oxidative Stress of Botrytis cinerea.
Yak1, a member of the dual-specificity tyrosine phosphorylation-regulated protein kinases, plays an important role in diverse cellular processes in fungi. However, to date, the role of BcYak1 in Botrytis cinerea, the causal agent of gray mold diseases in various plant species, remains uncharacterized. Our previous study identified one lysine acetylation site (Lys252) in BcYak1, which is the first report of such a site in Yak1. In this study, the function of BcYak1 and its lysine acetylation site were investigated using gene disruption and site-directed mutagenesis. The gene deletion mutant ΔBcYak1 not only exhibits much lower pathogenicity, conidiation and sclerotium formation, but was also much more sensitive to H2O2 and the ergosterol biosynthesis inhibitor (EBI) triadimefon. The Lys252 site-directed mutagenesis mutant strain ΔBcYak1-K252Q (mimicking the acetylation of the site), however, only showed lower sclerotium formation and higher sensitivity to H2O2. These results indicate that BcYAK1 is involved in the vegetative differentiation, adaptation to oxidative stress and triadimefon, and virulence of B. cinerea.