Association Between NRAS and BRAF Mutational Status and Melanoma-Specific Survival Among Patients With Higher-Risk Primary Melanoma.
NRAS and BRAF mutations in melanoma inform current treatment paradigms, but their role in survival from primary melanoma has not been established. Identification of patients at high risk of melanoma-related death based on their primary melanoma characteristics before evidence of recurrence could inform recommendations for patient follow-up and eligibility for adjuvant trials. To determine tumor characteristics and survival from primary melanoma by somatic NRAS and BRAF status. A population-based study with a median follow-up of 7.6 years (through 2007), including 912 patients from the United States and Australia in the Genes, Environment, and Melanoma (GEM) Study, with first primary cutaneous melanoma diagnosed in the year 2000 and analyzed for NRAS and BRAF mutations. Tumor characteristics and melanoma-specific survival of primary melanoma by NRAS and BRAF mutational status. The melanomas were 13% NRAS+, 30% BRAF+, and 57% with neither NRAS nor BRAF mutation (wildtype [WT]). In a multivariable model including clinicopathologic characteristics, relative to WT melanoma (with results reported as odds ratios [95% CIs]), NRAS+ melanoma was associated with presence of mitoses (1.8 [1.0-3.3]), lower tumor-infiltrating lymphocyte (TIL) grade (nonbrisk, 0.5 [0.3-0.8]; and brisk, 0.3 [0.5-0.7] [vs absent TILs]), and anatomic site other than scalp/neck (0.1 [0.01-0.6] for scalp/neck vs trunk/pelvis), and BRAF+ melanoma was associated with younger age (ages 50-69 years, 0.7 [0.5-1.0]; and ages >70 years, 0.5 [0.3-0.8] [vs <50 years]), superficial spreading subtype (nodular, 0.5 [0.2-1.0]; lentigo maligna, 0.4 [0.2-0.7]; and unclassified/other, 0.2 [0.1-0.5] [vs superficial spreading]), and presence of mitoses (1.7 [1.1-2.6]) (P < .05 for all). There was no significant difference in melanoma-specific survival (reported as hazard ratios [95% CIs]) for melanoma harboring mutations in NRAS (1.7 [0.8-3.4]) or BRAF (1.5 [0.8-2.9]) compared with WT melanoma, as adjusted for age, sex, site, American Joint Committee on Cancer (AJCC) tumor stage, TIL grade, and study center. However, melanoma-specific survival was significantly poorer for higher-risk (T2b or higher stage) tumors with NRAS (2.9 [1.1-7.7]) or BRAF (3.1 [1.2-8.5]) mutations (P = .04) but not for lower-risk (T2a or lower) tumors with NRAS (0.9 [0.3-3.0]) or BRAF (0.6 [0.2-1.7]) (P = .65), as adjusted for age, sex, site, AJCC tumor stage, TIL grade, and study center. Lower TIL grade for NRAS+ melanoma suggests it has a more immunosuppressed microenvironment, which may affect its response to immunotherapies. The approximate 3-fold increased risk of death for higher-risk tumors harboring NRAS or BRAF mutations after adjusting for other prognostic factors compared with WT melanomas indicates that the prognostic implication of these mutations deserves further investigation, particularly in higher–AJCC stage primary melanomas.


Rituximab treatment for chronic steroid-dependent Henoch-Schonlein purpura: 8 cases and a review of the literature.
Henoch-Schonlein purpura (HSP) is a small vessel vasculitis that is characterized by non-thrombocytopenic purpura, abdominal pain, arthritis, and glomerulonephritis. Typically, HSP is self-limited requiring only supportive care, but more severe cases may require corticosteroid (CS) treatment. Rarely, a subset of these patients has persistent rash, arthritis, abdominal involvement, or renal disease despite treatment with CS, or has disease recurrence on CS tapering. Refractory HSP has been effectively treated with a variety of CS sparing therapies. For life-threatening refractory HSP, the B cell depleting agent, rituximab (RTX), has been reported as beneficial for children with substantial renal or central nervous system involvement. However, RTX use for children with less severe HSP, but chronic CS dependent disease refractory to CS sparing immunomodulatory agents, has been less well explored. Herein, we describe 8 children treated with RTX for chronic refractory HSP and report a reduction in recurrent hospitalizations and eventual CS discontinuation. This is a retrospective analysis of eight children who were treated with RTX for chronic CS dependent HSP during the years 2006-2014 at a single institution. A chart review of the electronic medical record was performed to determine the presenting symptoms, the type and duration of treatment received, and the number of hospitalizations prior to and after RTX. The number of hospitalizations and oral corticosteroid burden were analyzed using the Wilcoxon signed rank test. Prior to receiving RTX, seven patients had at least one hospitalization for HSP (median 1.5, range 0-3). Following RTX, only two patients were hospitalized, each a single time for recurrent abdominal pain. The median oral CS burden was 0.345 mg/kg/day before RTX and 0 mg/kg/day at 6 months (p = 0.078), 1 year (p = 0.0625), and 2 years (p = 0.03) following RTX infusion. Seven out of eight children met remission criteria, defined as no active rash, arthritis, nephritis (hematuria and proteinuria), or gastrointestinal distress following RTX. No serious adverse events were noted. Overall, RTX effectively reduced the number of hospital admissions and oral CS burden. RTX also helped most all children achieve clinical remission. RTX appears to be an effective and safe alternative for chronic CS dependent and immunomodulatory refractory childhood HSP.


Radiation therapy for the whole breast: Executive summary of an American Society for Radiation Oncology (ASTRO) evidence-based guideline.
The purpose of this guideline is to offer recommendations on fractionation for whole breast irradiation (WBI) with or without a tumor bed boost and guidance on treatment planning and delivery. The American Society for Radiation Oncology (ASTRO) convened a task force to address 5 key questions focused on dose-fractionation for WBI, indications and dose-fractionation for tumor bed boost, and treatment planning techniques for WBI and tumor bed boost. Guideline recommendations were based on a systematic literature review and created using a predefined consensus-building methodology supported by ASTRO-approved tools for grading evidence quality and recommendation strength. For women with invasive breast cancer receiving WBI with or without inclusion of the low axilla, the preferred dose-fractionation scheme is hypofractionated WBI to a dose of 4000 cGy in 15 fractions or 4250 cGy in 16 fractions. The guideline discusses factors that might or should affect fractionation decisions. Use of boost should be based on shared decision-making that considers patient, tumor, and treatment factors, and the task force delineates specific subgroups in which it recommends or suggests use or omission of boost, along with dose recommendations. When planning, the volume of breast tissue receiving >105% of the prescription dose should be minimized and the tumor bed contoured with a goal of coverage with at least 95% of the prescription dose. Dose to the heart, contralateral breast, lung, and other normal tissues should be minimized. WBI represents a significant portion of radiation oncology practice, and these recommendations are intended to offer the groundwork for defining evidence-based practice for this common and important modality. This guideline also seeks to promote appropriately individualized, shared decision-making regarding WBI between physicians and patients.


Rural-urban differences in the long-term care of the disabled elderly in China.
In China, the rapid rate of population aging and changes in the prevalence of disability among elderly people could have significant effects on the demand for long-term care. This study aims to describe the urban-rural differences in use and cost of long-term care of the disabled elderly and to explore potential influencing factors. This study uses data from a cross-sectional survey and a qualitative investigation conducted in Zhejiang province in 2012. The participants were 826 individuals over 60 years of age, who had been bedridden or suffered from dementia for more than 6 months. A generalized linear model and two-part regression model were applied to estimate costs, with adjustment of covariates. Pensions provide the main source of income for urban elderly, while the principal income source for rural elderly is their family. Urban residents spend more on all services than do rural residents. Those who are married spend less on daily supplies and formal care than the unmarried do. Age, incapacitation time, comorbidity number, level of income, and bedridden status influence spending on medical care (β=-0.0316, -0.0206, 0.1882, 0.3444, and -0.4281, respectively), but the cost does not increase as the elderly grow older. Urban residents, the married, and those with a higher income level tend to spend more on medical equipment. Urban residence and living status are the two significant factors that affect spending on personal hygiene products. The use of long-term care services varies by living area. Long-term care of the disabled elderly imposes a substantial burden on families. Our study revealed that informal care involves huge opportunity costs to the caregivers. Chinese policy makers need to promote community care and long-term care insurance to relieve the burden of families of disabled elderly, and particular attention should be given to the rural elderly.


Injectable ammonium chloride used enterally for the treatment of persistent metabolic alkalosis in three pediatric patients.
Enteral administration of injectable ammonium chloride may offer an effective method for the treatment of persistent metabolic alkalosis, without the adverse effects associated with the intravenous route. This case series describes 3 pediatric patients who received ammonium chloride enterally for the treatment of persistent metabolic alkalosis. The patients were a 2-month-old female infant, a 6-week-old male infant, and a 3-year-old male toddler. Four to 18 doses of ammonium chloride were administered enterally (range, 3-144 mEq/dose). Two of the 3 patients achieved resolution of metabolic alkalosis with ammonium chloride, while 1 patient's condition was refractory to treatment. Resolution of metabolic alkalosis occurred at 4 and 8 days, which required a total weight-based dose of 10.7 mEq/kg and 18 mEq/kg, respectively. No adverse effects were recorded. The use of ammonium chloride injection administered enterally was a safe and effective option in 2 of the 3 pediatric patients with persistent metabolic alkalosis.


TP53 gene mutation analysis in chronic lymphocytic leukemia by nanopore MinION sequencing.
The assessment of TP53 mutational status is becoming a routine clinical practice for chronic lymphocytic leukemia patients (CLL). A broad spectrum of molecular techniques has been employed so far, including both direct Sanger sequencing and next generation sequencing. Oxford Nanopore Technologies recently released the MinION an USB-interfaced sequencer. In this paper we report our experience, with the MinION technology for the detection of the TP53 gene mutation in CLL patients. Twelve CLL patients at diagnosis were included in this study. All except one patient showed the TP53 gene deletion in Fluorescence in situ hybridization experiments. Patients were investigated for TP53 mutation by Sanger and by MinION sequencing. Analysis by Sanger was performed according with the IARC protocol. Analysis by MinION was performed adopting a strategy based on long template PCR, read error correction, and post variant calling filtering. Due to the high error rate of nanopore technology, sequence data were both used directly and before correction with two different in silico methods: ALEC and nanocorrect. A mean error rate of 15 % was detected before correction that was reduced to 4-5 % after correction. Analysis by Sanger sequencing was able to detect four patients mutated for TP53. MinION analysis detected one more mutated patient previously not detected from Sanger. In our hands, the Nanopore technology shows correlation with Sanger sequencing but more sensitive, manageable and less expensive, and therefore has proven to be a useful tool for TP53 gene mutation detection.


Laminar flow effects during laser-induced oxidative labeling for protein structural studies by mass spectrometry.
Laser-induced oxidative labeling of proteins provides insights into biomolecular structures and interactions. In these experiments, the hydroxyl radical ((*)OH) formed by photolysis of H(2)O(2) generates covalent modifications that are detectable by mass spectrometry. Under conditions where individual protein molecules are irradiated only once, the short (*)OH lifetime (approximately 1 micros) ensures that covalent modifications are formed before any oxidation-induced conformational changes take place. This feature implies that the method should be free of structural artifacts. It has been proposed that single-exposure conditions can be achieved by passing the solution through a capillary where successive laser pulses generate a string of irradiated flow segments that are well separated from one another. The current work explores the convection phenomena within the labeling capillary in more detail. The experiments are conducted at Reynolds numbers <<2000, resulting in laminar flow. The associated parabolic velocity profile causes a portion of each irradiated segment to remain in the labeling window during the subsequent laser pulse. Achieving a genuine single-exposure regime is, therefore, not possible. We estimate the fraction of labeled protein formed under laminar flow conditions, as well as the occurrence of multiple exposure events for any combination of experimental parameters (laser spot width, pulse frequency, and solution flow rate). A proper choice of these parameters provides extensive labeling, while keeping multiple exposure events at an acceptably low level. The theoretical framework developed here is supported by experimental data. Overall, this study reaffirms the feasibility of the use of flow devices for meaningful laser-induced oxidative labeling studies. At the same time, we provide a theoretical underpinning of this technique that goes beyond previously suggested plug flow models.


The effect of Arthrospira platensis capsules on CD4 T-cells and antioxidative capacity in a randomized pilot study of adult women infected with human immunodeficiency virus not under HAART in Yaoundé, Cameroon.
Dietary supplements are often used to improve the nutritional status of people living with HIV/AIDS (PLHIV). Arthrospira platensis (Asp), also known as Spirulina, is a cyanobacterium rich in proteins and micronutrients. Cell and animal trials described immune-modulating, antiretroviral and antioxidant activities. This pilot study describes the effects of the supplementation of 5 g/day of Asp on a pre-highly-active antiretroviral therapy (pre-HAART), HIV-infected, adult female population. It was conducted as a three-month randomized controlled trial (RCT) that compared a cup supplementation of five grams/day of Asp with a placebo of equal protein content and energy. The study included 73 HIV-infected women. The immediate outcome variables were CD4 T-cells, viral load and immune activation by CD8 T-cells expressing CD38. The antioxidant status was assessed by way of the total antioxidant capacity of the serum (TAOS). The renal function was documented by way of creatinine, urea and the calculated glomerular filtration rate. Statistical analyses were carried out with non-parametric tests, and the effect size of each interaction was calculated. No differences in the immunological and virological markers between the Asp and the placebo group could be observed. In the placebo group, 21 of 30 patients (70%) developed concomitant events, while in the Asp group, only 12 of 28 patients (43%) did. Both groups registered a significant weight increase; 0.5 kg (p < 0.05) in the Asp group and 0.65 kg (p < 0.05) in the placebo group. The antioxidant capacity increase of 56 (1-98) µM for Asp was significantly different from the decrease observed in the placebo group (p < 0.001). A slight increase in the creatinine level of 0.1 g/dL (p < 0.001) was observed in the Asp group, and no effect was observed in the urea levels. The improvement of the antioxidant capacity under Asp, shown for the first time on PLHIV, could become a focus for future research on the nutritional and health effects of Spirulina. The observed slight, but significant increase of serum creatinine needs further evaluation, especially with varying doses of Asp.


Lower Vocal Tract Morphologic Adjustments Are Relevant for Voice Timbre in Singing.
The vocal tract shape is crucial to voice production. Its lower part seems particularly relevant for voice timbre. This study analyzes the detailed morphology of parts of the epilaryngeal tube and the hypopharynx for the sustained German vowels /a/, /e/, /i/, /o/, and /u/ by thirteen male singer subjects who were at the beginning of their academic singing studies. Analysis was based on two different phonatory conditions: a natural, speech-like phonation and a singing phonation, like in classical singing. 3D models of the vocal tract were derived from magnetic resonance imaging and compared with long-term average spectrum analysis of audio recordings from the same subjects. Comparison of singing to the speech-like phonation, which served as reference, showed significant adjustments of the lower vocal tract: an average lowering of the larynx by 8 mm and an increase of the hypopharyngeal cross-sectional area (+ 21:9%) and volume (+ 16:8%). Changes in the analyzed epilaryngeal portion of the vocal tract were not significant. Consequently, lower larynx-to-hypopharynx area and volume ratios were found in singing compared to the speech-like phonation. All evaluated measures of the lower vocal tract varied significantly with vowel quality. Acoustically, an increase of high frequency energy in singing correlated with a wider hypopharyngeal area. The findings offer an explanation how classical male singers might succeed in producing a voice timbre with increased high frequency energy, creating a singer`s formant cluster.


The Epidemiology of Injuries in Australian Professional Rugby Union 2014 Super Rugby Competition.
Rugby union is a collision-based ball sport played at the professional level internationally. Rugby union has one of the highest reported incidences of injury of all team sports. To identify the characteristics, incidence, and severity of injuries occurring in Australian professional Super Rugby Union. Descriptive epidemiology study. The present study was a prospective epidemiology study on a cohort of 180 professional players from 5 Australian Super Rugby teams during the 2014 Super Rugby Union Tournament. Team medical staff collected and submitted daily training and match-play injury data through a secure, web-based electronic platform. The injury data included the main anatomic location of the injury, specific anatomic structure of the injury, injury diagnosis, training or match injury occurrence, main player position, mechanism of injury, and the severity of the injury quantified based on the number of days lost from training and/or competition due to injury. The total combined incidence rate for injury during training and match-play across all Australian Super Rugby Union teams was 6.96 per 1000 hours, with a mean injury severity of 37.45 days lost from training and competition. The match-play injury incidence rate was 66.07 per 1000 hours, with a mean severity of 39.80 days lost from training and competition. No significant differences were observed between forward- and back-playing positions for match or training injury incidence rate or severity. The incidence of injury for the present study was lower during match-play than has previously been reported in professional rugby union; however, the overall time loss was higher compared with previous studies in professional rugby union. The high overall time loss was due fundamentally to a high incidence of injuries with greater than 28 days' severity.


Cdk1-phosphorylated CUEDC2 promotes spindle checkpoint inactivation and chromosomal instability.
Aneuploidy and chromosomal instability are major characteristics of human cancer. These abnormalities can result from defects in the spindle assembly checkpoint (SAC), which is a surveillance mechanism for accurate chromosome segregation through restraint of the activity of the anaphase-promoting complex/cyclosome (APC/C). Here, we show that a CUE-domain-containing protein, CUEDC2, is a cell-cycle regulator that promotes spindle checkpoint inactivation and releases APC/C from checkpoint inhibition. CUEDC2 is phosphorylated by Cdk1 during mitosis. Depletion of CUEDC2 causes a checkpoint-dependent delay of the metaphase-anaphase transition. Phosphorylated CUEDC2 binds to Cdc20, an activator of APC/C, and promotes the release of Mad2 from APC/C-Cdc20 and subsequent APC/C activation. CUEDC2 overexpression causes earlier activation of APC/C, leading to chromosome missegregation and aneuploidy. Interestingly, CUEDC2 is highly expressed in many types of tumours. These results suggest that CUEDC2 is a key regulator of mitosis progression, and that CUEDC2 dysregulation might contribute to tumour development by causing chromosomal instability.


Estrogen and progestin receptor immunocytochemistry in lactotropes versus gonadotropes of monkey pituitary cell cultures.
The distribution of estrogen receptors (ER), progesterone receptors (PR), PRL, and gonadotropins in different cell types of the monkey pituitary was examined by immunocytochemical (ICC) labeling of pituitary cell cultures. Dispersed monkey pituitary cells were cultured on extracellular matrix and in serum-free medium for 6-14 days. Individual cultures were singly stained for ER, PR, PRL, LH, and FSH or double labeled for PR and one of the protein hormones. ICC reaction product localizes over the nuclei of cells that are positive for the steroid receptors, whereas reaction product localizes over the cytoplasm of cells that are positive for the protein hormones. Sixty-one percent of the parenchymal cells were positive for PRL, while 1-3% were positive for LH or FSH. Sixty-two percent of the parenchymal cells were ER positive. ER staining was localized over the nuclei of two morphologically distinct cell types. One cell type is smaller and more prevalent than the second cell type. Based on single staining for each of the protein hormones, we propose that the smaller cells are lactotropes, and the larger cells are gonadotropes. PR-positive cells averaged 7.7% of the parenchymal cells. Double ICC staining for PR and the protein hormones demonstrated that PR localize in the nuclei of gonadotropes, but not lactotropes, of monkey pituitary cell cultures. The absence of PR in lactotropes is consistent with our observation that progesterone has no direct effect on PRL secretion in monkey pituitary cell cultures. In contrast, the presence of PR in gonadotropes suggests that progesterone may act directly at the pituitary to modulate gonadotropin secretion in the primate. In conclusion, ER are present in both lactotropes and gonadotropes. PR are present in gonadotropes, but not lactotropes, of the primate pituitary.


Wearable technologies to measure clinical outcomes in multiple sclerosis: A scoping review.
Wearable technology refers to any sensor worn on the person, making continuous and remote monitoring available to many people with chronic disease, including multiple sclerosis (MS). Daily monitoring seems an ideal solution either as an outcome measure or as an adjunct to support rater-based monitoring in both clinical and research settings. There has been an increase in solutions that are available, yet there is little consensus on the most appropriate solution to use in either MS research or clinical practice. We completed a scoping review (using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Reviews (PRISMA-ScR) guidelines) to summarise the wearable solutions available in MS, to identify those approaches that could potentially be utilised in clinical trials, by evaluating the following: scalability, cost, patient adaptability and accuracy. We identified 35 unique products that measure gait, cognition, upper limb function, activity, mood and fatigue, with most of these solutions being phone applications.


Molecular diagnosis and translymphatic chemotherapy targeting sentinel lymph nodes of patients with early gastrointestinal cancers.
The sentinel lymph node (SLN) is defined as the lymph node(s) that first receives lymphatic drainage from the site of the primary tumor. Recent progress in molecular techniques has demonstrated the presence of micrometastatic tumor cells in SLNs. Quantitative real-time RT-PCR assay, which enables rapid analysis, is currently being undertaken for intraoperative molecular diagnosis of SLNs. We developed an intraoperative real-time RT-PCR assay to detect micrometastasis in SLNs for early gastric cancer. All SLNs and randomly selected non-SLNs in 96 cT1 or cT2 gastric cancer patients were biopsied intraoperatively and examined by routine hematoxylin and eosin staining, immunohistochemistry with anticytokeratin antibody (AE1/AE3), and multimarker real-time RT-PCR assay including cytokeratin (CK) 19, CK20, and carcinoembryonic antigen. All patients with histopathologically verified metastasis in their SLNs demonstrated positive results by RT-PCR assay. Forty percent of patients with histopathologically negative SLNs showed positive SLNs by RT-PCR assay. RT-PCR assay revealed that 4 patients (4%) with negative SLNs had positive non-SLNs; however, these positive non-SLNs were identified within each SLN basin. We recently developed a new drug delivery system targeting SLNs with a phospholipid polymer, using 2-methacryloyloxyethyl phosphorylcholine conjugated with paclitaxel. Our preliminary data suggest that this novel drug delivery system may be feasible for translymphatic chemotherapy targeting SLNs of patients with cN0 early gastrointestinal cancer, who have the potential for occult metastasis in SLNs. Endoscopic resection of the primary tumor followed by translymphatic chemotherapy targeting SLNs may become a promising minimally invasive multidisciplinary therapy.


Gene pathways associated with prognosis and chemotherapy sensitivity in molecular subtypes of breast cancer.
We hypothesized that distinct biological processes might be associated with prognosis and chemotherapy sensitivity in the different types of breast cancers. We performed gene set analyses with BRB-ArrayTools statistical software including 2331 functionally annotated gene sets (ie, lists of genes that correspond to a particular biological pathway or biochemical function) assembled from Ingenuity Pathway Analysis and Gene Ontology databases corresponding to almost all known biological processes. Gene set analysis was performed on gene expression data from three cohorts of 234, 170, and 175 patients with HER2-normal lymph node-negative breast cancer who received no systemic adjuvant therapy to identify gene sets associated prognosis and three additional cohorts of 198, 85, and 62 patients with HER2-normal stage I-III breast cancer who received preoperative chemotherapy to identify gene sets associated with pathological complete response to therapy. These analyses were performed separately for estrogen receptor (ER)-positive and ER-negative breast cancers. Interaction between gene sets and survival and treatment response by breast cancer subtype was assessed in individual datasets and also in pooled datasets. Statistical significance was estimated with permutation test. All statistical tests were two-sided. For ER-positive cancers, from 370 to 434 gene sets were associated with prognosis (P ≤ .05) and from 209 to 267 gene sets were associated with chemotherapy response in analysis by individual dataset. For ER-positive cancers, 131 gene sets were associated with prognosis and 69 were associated with pathological complete response (P ≤.001) in pooled analysis. Increased expression of cell cycle-related gene sets was associated with poor prognosis, and B-cell immunity-related gene sets were associated with good prognosis. For ER-negative cancers, from 175 to 288 gene sets were associated with prognosis and from 212 to 285 gene sets were associated with chemotherapy response. In pooled analyses of ER-negative cancers, 14 gene sets were associated with prognosis and 23 were associated with response. Gene sets involved in sphingolipid and glycolipid metabolism were associated with better prognosis and those involved in base excision repair, cell aging, and spindle microtubule regulation were associated with chemotherapy response. Different biological processes were associated with prognosis and chemotherapy response in ER-positive and ER-negative breast cancers.


Azido and diazarinyl analogues of bis-tyrphostin as asymmetrical inhibitors of dynamin GTPase.
Probing the dynamin binding site: Bis-tyrphostin (1, Bis-T), is a potent inhibitor of the phospholipid-stimulated GTPase activity of dynamin I. Analogues of Bis-T have significant potential as a biological probes for the dissection of endocytic pathways. Bis-T-derived compounds were synthesised and evaluated for their ability to inhibit the GTPase activity of dynamin I. Two analogues (23 and 24) represent the first asymmetrically substituted Bis-T analogues to retain dynamin inhibition.Two azidobenzyl amide (4 and 23) and one 3-trifluoromethyl-3H-diazirin-3-ylphenyl (24) analogues of bis-tyrphostin (1, Bis-T) were synthesised as potential photoaffinity labels for the elucidation of the binding site of compound 1 in dynamin I. Of the two azidobenzyl amide analogues (4 and 23), the terminally substituted 23 retained dynamin I GTPase inhibition (IC(50)=6.4+/-2.8 microM) whilst 4, which was substituted on the central carbon of the amide linker, displayed no activity. Analogue 24 also retained inhibitory activity (IC(50)=36+/-9 microM). Photoaffinity labelling experiments did not unequivocally elucidate the binding pocket of compound 1. However, compounds 23 and 24 represent the first asymmetrically substituted Bis-T analogues to retain dynamin inhibitory activity, providing a new direction for analogue synthesis.


Enhanced immune presentation of a single-chain major histocompatibility complex class I molecule engineered to optimize linkage of a C-terminally extended peptide.
Major histocompatibility complex class I molecules can be expressed as single polypeptides wherein the antigenic peptide, beta2-microglobulin, and heavy chain are attached by flexible linkers. These molecules, single-chain trimers (SCTs), are remarkably stable at the cell surface compared with native (noncovalently attached) class I molecules. In this study, we used a structure-based approach to engineer an F pocket variant SCT of the murine class I molecule Kb that presents the SIINFEKL epitope of ovalbumin. Mutation of heavy chain residue Tyr84 (Y84A) in the SCT resulted in enhanced serological and cytolytic CD8 T cell recognition of the covalently linked peptide due to better accommodation of the linker extending from the C terminus of the peptide. These SCTs exhibit significant cell-surface stability, which we hypothesize is rendered by their ability to continuously and efficiently rebind the covalently attached peptide. In addition, we demonstrate that SCT technology can be applied to tetramer construction using recombinant SCTs expressed in Escherichia coli. SCT-based tetramers could have applications for the enumeration of T and natural killer cells that recognize peptide.class I complexes prone to dissociation.


The double burden of diabetes and global infection in low and middle-income countries.
Four out of five people in the world with diabetes now live in low- and middle-income countries (LMIC), and the incidence of diabetes is accelerating in poorer communities. Diabetes increases susceptibility to infection and worsens outcomes for some of the world's major infectious diseases such as tuberculosis, melioidosis and dengue, but the relationship between diabetes and many neglected tropical diseases is yet to be accurately characterised. There is some evidence that chronic viral infections such as hepatitis B and HIV may predispose to the development of type 2 diabetes by chronic inflammatory and immunometabolic mechanisms. Helminth infections such as schistosomiasis may be protective against the development of diabetes, and this finding opens up new territory for discovery of novel therapeutics for the prevention and treatment of diabetes. A greater understanding of the impact of diabetes on risks and outcomes for infections causing significant diseases in LMIC is essential in order to develop vaccines and therapies for the growing number of people with diabetes at risk of infection, and to prioritise research agendas, public health interventions and policy. This review seeks to give an overview of the current international diabetes burden, the evidence for interactions between diabetes and infection, immune mechanisms for the interaction, and potential interventions to tackle the dual burden of diabetes and infection.


Amide I Raman optical activity of polypeptides: fragment approximation.
Vibrational optical activity (VOA) is an important property used to determine the absolute configuration of a chiral molecule in condensed phases. In particular, vibrational circular dichroism and Raman optical activity (ROA) are two representative VOA measurement techniques that have been extensively used to study structures and dynamics of biomolecules. Recently, the amide I vibrational circular dichroism of polypeptides was theoretically described by using fragment approximation methods, which are based on the assumption that amide I VOA can be described as a linear combination of those of constituent fragment peptide units. Here, we develop a fragment approximation theory applicable to numerical simulations of Raman and Raman optical activity spectra for the amide I vibrations in polypeptides. For an alanine dipeptide and pentapeptide analogs, we carried out density functional theory calculations of polarizability, magnetic dipole-, and electric quadrupole-ROA tensors. Numerically simulated spectra using the fragment approximation are directly compared to density functional theory results. Furthermore, the simulated ROA spectra of alanine-based right-handed alpha-helix and polyproline II polypeptides are directly compared to the previously reported experimental results. The agreements were found to be excellent, which suggests that the fragment approximation method developed for the numerical simulation of ROA spectrum of polypeptide in solution is valid and useful.


Insig-dependent ubiquitination and degradation of mammalian 3-hydroxy-3-methylglutaryl-CoA reductase stimulated by sterols and geranylgeraniol.
The endoplasmic reticulum enzyme 3-hydroxy-3-methylglutaryl-CoA reductase produces mevalonate, which is converted to sterols and to other products, including geranylgeraniol groups attached to proteins. The enzyme is known to be ubiquitinated and rapidly degraded when sterols and nonsterol end products of mevalonate metabolism accumulate in cells. Here, we use RNA interference to show that sterol-accelerated ubiquitination of reductase requires Insig-1 and Insig-2, membrane-bound proteins of the endoplasmic reticulum that were shown previously to accelerate degradation of reductase when overexpressed by transfection. Alanine substitution experiments reveal that binding of reductase to Insigs and subsequent ubiquitination require the tetrapeptide sequence YIYF in the second membrane-spanning helix of reductase. The YIYF peptide is also found in the sterol-sensing domain of SCAP, another protein that binds to Insigs in a sterol-stimulated fashion. When lysine 248 of reductase is substituted with arginine, Insig binding persists, but the reductase is no longer ubiquitinated and degradation is markedly slowed. Lysine 248 is predicted to lie immediately adjacent to a membrane-spanning helix, suggesting that a membrane-bound ubiquitin transferase is responsible. Finally, we show that Insig-dependent, sterol-stimulated degradation of reductase is further accelerated when cells are also supplied with the 20-carbon isoprenoid geranylgeraniol, but not the 15-carbon farnesol, raising the possibility that the nonsterol potentiator of reductase regulation is a geranylgeranylated protein.


Potential benefits of weight loss in coronary heart disease.
The prevalence of overweight, obesity and insulin resistance in patients with coronary heart disease (CHD) exceeds that of the general population. Obesity is associated with a constellation of coronary risk factors that predispose to the development and progression of CHD. Intentional weight loss, accomplished through behavioral weight loss and exercise, improves insulin sensitivity and associated cardio-metabolic risk factors such as lipid measures, blood pressure, measures of inflammation and vascular function both in healthy individuals and patients with CHD. Additionally, physical fitness, physical function and quality of life all improve. There is evidence that intentional weight loss prevents the onset of CHD in high risk overweight individuals. While weight loss associated improvements in insulin resistance, fitness and related risk factors strongly supports favorable prognostic effects in individuals with established CHD, further study is needed to determine if long-term clinical outcomes are improved.


HIV risk behaviors among Dominican brothel and street prostitutes in New York City.
Latina women are overrepresented among AIDS cases in the United States. To assist in developing appropriate prevention and intervention programs, information regarding HIV risk behaviors is needed on the many diverse Latina subgroups. This study examined sociodemographic characteristics and HIV risk behaviors of Dominican female prostitutes, comparing those who worked primarily in brothels with those who were street workers. A total of 77 Dominican prostitutes (54 brothel; 23 street) were recruited in New York City to participate in a structured interview and were offered HIV testing. Ethnographic interviews were conducted with a subscale. Results indicated that there were many significant differences in demographics and risk behaviors between the two groups, and those working in brothels engaged in lower levels of risk behaviors. In addition, those working in brothels had closer ties to the Dominican and Spanish cultures. Implications for AIDS prevention efforts are discussed.


The use of nontreated mosquito-net mesh cloth for a tension free inguinal hernia repair: our experience.
The prohibitive costs and scarcity of the imported prosthetic mesh for hernioplasty, has prevented its widespread use in most developing countries. We then set out to ascertain the outcome, complications (undue pain, wound infection, recurrence, and mesh extrusion) and cost implications in the use of a nontreated mosquito-net for inguinal hernioplasty. A prospective study of all consecutive adult patients with uncomplicated inguinal hernia who were admitted for open herniorrhaphy between January 2012 and December, 2013 at the Federal Medical Centre, Ido - Ekiti, South West, Nigeria. A sheet of the nontreated mosquito-net mesh 10 cm Χ 8 cm, autoclaved, a day prior to surgery was used for each patient's hernia repair. The operation sites were exposed and examined 3(rd) and 6(th) postoperative days. Findings were documented for analysis. A total of 130 adult patients were recruited for this study of which 115 of the patients were males and 15 were females. Forty-four (41.53%) had inguinal hernia and 76 (58.46%) of them had inguinoscrotal hernia. They all had successful repair and were followed-up for complications for a period of 6 weeks to 6 months at the surgical out-patient department. Locally-sourced and autoclaved mosquito-net mesh is an effective alternative for hernioplasty especially in situations where commercial mesh is not readily available or affordable.


Reversal of cellular phenotypes in neural cells derived from Huntington's disease monkey-induced pluripotent stem cells.
Huntington's disease (HD) is a dominant neurodegenerative disorder caused by the expansion of glutamine residues in the N-terminal region of the huntingtin (HTT) protein. The disease results in progressive neuronal loss, leading to motor, cognitive, and psychiatric impairment. Here, we report the establishment of neural progenitor cell (NPC) lines derived from induced pluripotent stem cells (iPSCs) of transgenic HD monkeys. Upon differentiation to neurons, HD neural cells develop cellular features of HD, including the formation of nuclear inclusions and oligomeric mutant HTT (mHTT) aggregates, as well as increased apoptosis. These phenotypes are rescued by genetic suppression of HTT and pharmacological treatment, demonstrating the ability of our HD cell model to respond to therapeutic treatment. The development and reversal of HD-associated phenotypes in neural cells from HD monkeys provides a unique nonhuman primate (NHP) model for exploring HD pathogenesis and evaluating therapeutics that could be assessed further in HD monkeys.


An evaluation of epidemiological and reporting characteristics of complementary and alternative medicine (CAM) systematic reviews (SRs).
Systematic reviews (SRs) are abundant. The optimal reporting of SRs is critical to enable clinicians to use their findings to make informed treatment decisions. Complementary and alternative medicine (CAM) therapies are widely used therefore it is critical that conduct and reporting of systematic research in this field be of high quality. Here, methodological and reporting characteristics of a sample of CAM-related SRs and a sample of control SRs are evaluated and compared. MEDLINE(®) was searched to identify non-Cochrane SRs indexed from January 2010 to May 2011. Control SRs were retrieved and a search filter was used to identify CAM SRs. Citations were screened and publications that met a pre-specified definition of a SR were included. Pre-designed, standardized data extraction forms were developed to capture reporting and methodological characteristics of the included reviews. Where appropriate, samples were compared descriptively. A total of 349 SRs were identified, of which 174 were CAM-related SRs and 175 were conventional SRs. We compared 131 CAM-related non-Cochrane SRs to the 175 conventional non-Cochrane reviews. Fifty-seven percent (75/131) of CAM SRs specified a primary outcome compared to 21% (37/175) of conventional sample reviews. Reporting of publication bias occurred in less than 5% (6/131) of the CAM sample versus 46% (80/175) of the conventional sample of SRs. Source of funding was frequently and consistently under-reported. Less than 5% (11/306) of all SRs reported public availability of a review protocol. The two samples of reviews exhibited different strengths and weaknesses. In some cases there were consistencies across items which indicate the need for continued improvements in reporting for all SR reports. We advise authors to utilise the PRISMA Statement or other SR guidance when reporting SRs.


Heterogeneity in association of remote herpesvirus infections and pediatric MS.
While prior Epstein-Barr virus (EBV) infection has been consistently associated with subsequent risk of developing multiple sclerosis (MS), the association with other common herpesviruses has been more controversial. Our objectives were to determine whether remote infection with EBV and other common herpesviruses affect the susceptibility to pediatric MS and if there are interactions between genetic and demographic factors and viral infections. Cases with pediatric-onset MS or clinically isolated syndrome within 4 years of disease onset, and controls were recruited from 16 American pediatric MS centers. Logistic regression models adjusted for potential confounders assessed the association between case status and serological evidence for past infection with EBV, cytomegalovirus (CMV), Herpes Simplex viruses-1 (HSV-1) and -2. We determined the heterogeneity of the effect of viral infection on the risk of having MS according to race, ethnicity and HLA-DRB1:1501 status. A total of 356 pediatric cases and 493 controls were recruited. In multivariable models, EBV-viral capsid antigen (VCA) seropositivity was associated with increased odds of having MS by 7.4 times (95% CI: 4.5-12.0, P < 0.001). Seropositivity for HSV-1 was also associated with increased odds of having MS (OR 1.54, 95% CI: 1.06-2.25, P = 0.025) but this increase was seen only in Whites (OR = 2.18, 95% CI 1.35-3.52, P < 0.001) and those negative for HLA-DRB1*1501 (OR = 1.89, 95% CI 1.17-3.03, P = 0.009). The effect of remote EBV infection on the risk of pediatric MS depended on race and HLA-DRB1*15:01 status. EBV seropositivity is strongly associated with pediatric MS, as is HSV-1 seropositivity in subjects negative for HLA-DRB1*15:01. Our report of interactions between select viral exposures, and age, race and DRB1 status suggests a complex effect of environmental and genetic risk factors on MS development.


Multipurpose prevention technologies: the future of HIV and STI protection.
Every day, more than 1 million people are newly infected with sexually transmitted infections (STIs) that can lead to morbidity, mortality, and an increased risk of human immunodeficiency virus (HIV) acquisition. Existing prevention and management strategies, including behavior change, condom promotion, and therapy have not reduced the global incidence and prevalence, pointing to the need for novel innovative strategies. This review summarizes important issues raised during a satellite session at the first HIV Research for Prevention (R4P) conference, held in Cape Town, on October 31, 2014. We explore key STIs that are challenging public health today, new biomedical prevention approaches including multipurpose prevention technologies (MPTs), and the scientific and regulatory hurdles that must be overcome to make combination prevention tools a reality.


Stable isotope analysis of production and trophic relationships in a tropical marine hard-bottom community.
Seagrasses produce much of the organic carbon in the shallow waters of the Caribbean and it has long been assumed that a substantial portion of this carbon is exported to nearby habitats, contributing substantially to their food webs. In the shallow coastal waters of the Florida Keys (USA), seagrass intersperses with hard-bottom habitat where bushy, red macroalgae are the most prominent primary producers. However, the relative importance of seagrass-derived carbon versus autochthonous algal production or phytoplankton in supporting higher trophic levels within hard-bottom communities has never been investigated. We compared the carbon and nitrogen isotopic values of potential primary producers and representative higher trophic level taxa from hard-bottom sites on the bay-side and ocean-side of the Florida Keys. We also included in our study a set of bay-side sites that experienced significant ecological disturbances over the past decade (e.g., cyanobacteria blooms, seagrass die-off, and sponge die-offs) that may have altered trophic relationships in those regions. We did not detect any differences among regions in the trophic status of hard-bottom taxa that might be associated with ecosystem disturbance. However, our results suggest that autochthonous production of algal detritus is an important source of secondary production in these hard-bottom communities, with seagrass and phytoplankton contributing smaller fractions.


Spin-flip methods in quantum chemistry.
This Perspective discusses salient features of the spin-flip approach to strong correlation and describes different methods that sprung from this idea. The spin-flip treatment exploits the different physics of low-spin and high-spin states and is based on the observation that correlation is small for same-spin electrons. By using a well-behaved high-spin state as a reference, one can access problematic low-spin states by deploying the same formal tools as in the excited-state treatments (i.e., linear response, propagator, or equation-of-motion theories). The Perspective reviews applications of this strategy within wave function and density functional theory frameworks as well as the extensions for molecular properties and spectroscopy. The utility of spin-flip methods is illustrated by examples. Limitations and proposed future directions are also discussed.


Combined effects of interleukin-7 and stem cell factor administration on lymphopoiesis after murine bone marrow transplantation.
The decreased ability of the thymus to generate T cells after bone marrow transplantation (BMT) is a clinically significant problem. Interleukin (IL)-7 and stem cell factor (SCF) induce proliferation, differentiation, and survival of thymocytes. Although previous studies have shown that administration of recombinant human IL-7 (rhIL-7) after murine and human BMT improves thymopoiesis and immune function, whether administration of SCF exerts similar effects is unclear. To evaluate independent or combinatorial effects of IL-7 and SCF in post-BMT thymopoiesis, bone marrow (BM)-derived mesenchymal stem cells transduced ex vivo with the rhIL-7 or murine SCF (mSCF) genes were cotransplanted with T cell-depleted BM cells into lethally irradiated mice. Although rhIL-7 and mSCF each improved immune reconstitution, the combination treatment had a significantly greater effect than either cytokine alone. Moreover, the combination treatment significantly increased donor-derived common lymphoid progenitors (CLPs) in BM, suggesting that transplanted CLPs expand more rapidly in response to IL-7 and SCF and may promote immune reconstitution. Our findings demonstrate that IL-7 and SCF might be therapeutically useful for enhancing de novo T cell development. Furthermore, combination therapy may allow the administration of lower doses of IL-7, thereby decreasing the likelihood of IL-7-mediated expansion of mature T cells.


Cell surface heparan sulfate participates in CXCL1-induced signaling.
The CXC subfamily of chemokines plays an important role in diverse processes, including inflammation, wound healing, growth regulation, angiogenesis, and tumorigenesis. The ELR-CXC chemokine, CXCL1 or MGSA/GROalpha, is traditionally considered to attract neutrophils to sites of inflammation. The non-ELR-CXC chemokine, CXCL10 or IP-10, is chemotactic for monocytes, B cells, and activated T lymphocytes. In addition to its role in leukocyte migration, CXCL10 inhibits the angiogenic functions of the ELR-CXC chemokines as well as bFGF and VEGF. Heparan sulfate proteoglycans (HSPGs) are required for the interaction of bFGF and vEGF ligands and their receptors. However, the role of HSPGs in regulating the ELR-chemokines signaling and biological functions is poorly understood. We show here that the CXCL1 maximal binding to CXCR2 expressed on HEK293 and CHO-K1 cells is dependent on the presence of cell surface HSPGs. The cell surface HSPGs on cells are required for CXCL1-induced PAK1 activation. Moreover, CXCL10 can inhibit CXCL1-induced PAK1 and ERK activation as well as the CXCL1-induced chemotaxis through decreasing CXCL1 binding to cell surface heparan sulfate. These data indicate that HSPGs are involved in modulating CXCL1-induced PAK1 activation and chemotaxis through regulating CXCL1 binding activity to CXCR2 receptor. CXCL10 inhibits CXCL1-induced PAK1 activation and chemotaxis by interfering with appropriate binding of CXCL1 to CXCR2 receptor.


Selection of polymers for application in scaffolds applicable for human pancreatic islet transplantation.
The liver is currently the site for transplantation of islets in humans. This is not optimal for islets, but alternative sites in humans are not available. Polymeric scaffolds in surgically accessible areas are a solution. As human donors are rare, the polymers should not interfere with functional survival of human-islets. We applied a novel platform to test the adequacy of polymers for application in scaffolds for human-islet transplantation. Viability, functionality, and immune parameters were included to test poly(D,L-lactide-co-ε-caprolactone) (PDLLCL), poly(ethylene oxide terephthalate)/polybutylene terephthalate (PEOT/PBT) block copolymer, and polysulfone. The type of polymer influenced the functional survival of human islets. In islets cultured on PDLLCL the glucagon-producing α-cells and insulin-producing β-cells contained more hormone granules than in islets in contact with PEOT/PBT or polysulfone. This was studied with ultrastructural analysis by electron microscopy (nanotomy) during 7 d of culture. PDLLCL was also associated with statistically significant lower release of double-stranded DNA (dsDNA, a so called danger-associate molecular pattern (DAMP)) from islets on PDLLCL when compared to the other polymers. DAMPs support undesired immune responses. Hydrophilicity of the polymers did not influence dsDNA release. Islets on PDLLCL also showed less cellular outgrowth. These outgrowing cells were mainly fibroblast and some β-cells undergoing epithelial to mesenchymal cell transition. None of the polymers influenced the glucose-stimulated insulin secretion. As PDLLCL was associated with less release of DAMPs, it is a promising candidate for creating a scaffold for human islets. Our study demonstrates that for sensitive, rare cadaveric donor tissue such as pancreatic islets it might be necessary to first select materials that do not influence functionality before proposing the biomaterial for in vivo application. Our presented platform may facilitate this selection of biomaterials.


Transesophageal two-dimensional echocardiography in young patients with cerebral ischemic events.
Using transesophageal echocardiography, cardiac structures can be imaged with high resolution. The aim of our study was to evaluate whether transesophageal echocardiography is superior in detecting mitral valve prolapse and other cardiac abnormalities compared with transthoracic echocardiography in an age-matched control group and in young patients with cerebral ischemic events (patient group). Forty patients with cerebral ischemic events (mean age 35.2 years) and 29 controls (mean age 30.4 years) were examined using both methods. Transthoracic and transesophageal echocardiography showed a significantly higher incidence of mitral valve prolapse in the patient group compared with the control group (p less than 0.001). By means of transesophageal echocardiography, it was possible to measure highly significant bulging in both the anterior and the posterior mitral leaflet in the patient group compared with the control group (p less than 0.001), and the thickness of the mitral leaflets was significantly higher in the patient group. In 9 of 20 (45%) patients with normal transthoracic echocardiograms, transesophageal echocardiography showed pathologic findings. We found transesophageal echocardiography to be a sensitive method for detecting mitral valve prolapse as well as valve changes and other cardiac abnormalities not detectable by conventional echocardiography. Our study underlines the role of mitral valve prolapse in young stroke patients as a relevant risk factor and emphasizes the importance of changed mitral valve morphology.


Phosphorylation of peptides derived from isolated soybean protein: effects on calcium binding, solubility and influx into Caco-2 cells.
Calcium homeostasis in the human body is maintained primarily via the absorption of calcium through the intestine. In order to maintain an efficient absorption of calcium with minimal calcium loss due to the formation of calcium phosphate precipitates in the small intestinal lumen, we developed a calcium-binding mediator using peptides derived from isolated soybean protein (ISP). ISP was modified via tryptic digestion and chemical phosphorylation using sodium trimetaphosphate, thereby generating soybean phosphopeptides (SPP), and this was followed by conducting a binding reaction with calcium chloride. We have established an optimized procedure and reaction conditions for maximal phosphorylation and calcium binding. Consequently, the phosphorylation of soybean peptides resulted in considerable improvement in their calcium binding activities. Next, we demonstrated that SPP was able to render calcium ions resistant to precipitate formation with inorganic phosphates, which suggested the enhancement of calcium bioavailability. Finally, we noted that the addition of calcium-bound SPP induced an increase in cytosolic calcium concentration in the intestinal Caco-2 cells, due to an influx of calcium. These findings provide a new basis by which we may assess the possibility that SPP, as a potent calcium carrier, can be utilized in the prevention of poor absorption of dietary calcium in animals.


Substructure and accessory proteins in scallop myosin filaments.
Native myosin filaments from scallop striated muscle fray into subfilaments of approximately 100-A diameter when exposed to solutions of low ionic strength. The number of subfilaments appears to be five to seven (close to the sevenfold rotational symmetry of the native filament), and the subfilaments probably coil around one another. Synthetic filaments assembled from purified scallop myosin at roughly physiological ionic strength have diameters similar to those of native filaments, but are much longer. They too can be frayed into subfilaments at low ionic strength. Synthetic filaments share what may be an important regulatory property with native filaments: an order-disorder transition in the helical arrangement of myosin cross-bridges that is induced on activation by calcium, removal of nucleotide, or modification of a myosin head sulfhydryl. Some native filaments from scallop striated muscle carry short "end filaments" protruding from their tips, comparable to the structures associated with vertebrate striated muscle myosin filaments. Gell electrophoresis of scallop muscle homogenates reveals the presence of high molecular weight proteins that may include the invertebrate counterpart of titin, a component of the vertebrate end filament. Although the myosin molecule itself may contain much of the information required to direct its assembly, other factors acting in vivo, including interactions with accessory proteins, probably contribute to the assembly of a precisely defined thick filament during myofibrillogenesis.


Common Data Elements for Unruptured Intracranial Aneurysms and Subarachnoid Hemorrhage Clinical Research: A National Institute for Neurological Disorders and Stroke and National Library of Medicine Project.
The goal for this project was to develop a comprehensive set of common data elements (CDEs), data definitions, case report forms and guidelines for use in unruptured intracranial aneurysm (UIA) and subarachnoid hemorrhage (SAH) clinical research, as part of a new joint effort between the National Institute of Neurological Disorders and Stroke (NINDS) and the National Library of Medicine of the US National Institutes of Health. These UIA and SAH CDEs will join several other neurological disease-specific CDEs that have already been developed and are available for use by research investigators. A Working Group (WG) divided into eight sub-groups and a Steering Committee comprised of international UIA and SAH experts reviewed existing NINDS CDEs and instruments, created new elements when needed and provided recommendations for UIA and SAH clinical research. The recommendations were compiled, internally reviewed by the entire UIA and SAH WG and posted online for 6 weeks for external public comments. The UIA and SAH WG and the NINDS CDE team reviewed the final version before posting the SAH Version 1.0 CDE recommendations. The NINDS UIA and SAH CDEs and supporting documents are publicly available on the NINDS CDE ( https://www.commondataelements.ninds.nih.gov/#page=Default ) and NIH Repository ( https://cde.nlm.nih.gov/home ) websites. The recommendations are organized into domains including Participant Characteristics and Outcomes and Endpoints. Dissemination and widespread use of CDEs can facilitate UIA and SAH clinical research and clinical trial design, data sharing, and analyses of observational retrospective and prospective data. It is vital to maintain an international and multidisciplinary collaboration to ensure that these CDEs are implemented and updated when new information becomes available.


Assessing atrophy of the major white matter fiber bundles of the brain from diffusion tensor MRI data.
Brain atrophy is a typical feature of many neurological conditions. Therefore, quantitative evaluation and spatial characterization of atrophy are potentially useful for monitoring the evolution of central nervous system (CNS) disorders. In this study, a method for measuring atrophy of the major white matter (WM) fiber bundles in the brain using diffusion tensor (DT) MRI data is developed. To this end, an atlas was created from sets of diffusion anisotropy images from normal subjects, and the deformations necessary to match single subject anisotropy images to this atlas were then computed. Because diffusion anisotropy images were used, this approach should be sensitive to fiber bundle volume changes in the same way that using T1-weighted images allows gray matter volume changes to be measured. The Jacobian determinant of the deformation field for each subject was then used as a measure of contraction or expansion of the tissue at each image voxel. An overview of the nonlinear registration problem is given; then an optimization of the parameters for the chosen algorithm is performed and the method for producing the atlas is described. The effectiveness of the method was then tested on data from five patients with multiple sclerosis (MS) and two patients with amyotrophic lateral sclerosis (ALS).


Development of respiratory muscle contractile fatigue in the course of hyperpnoea.
To assess the development of inspiratory and expiratory muscle fatigue during normocapnic hyperpnoea, we studied fourteen healthy men performing 8min hyperpnoea, 6min pause, 8min hyperpnoea, etc., until task failure. Twitch transdiaphragmatic (P(di,tw)) and gastric (P(ga,tw)) pressures were measured during cervical and thoracic magnetic nerve stimulation, before hyperpnoea, after every 8min of hyperpnoea, and at task failure (i.e., at 25.3+/-4.7min). P(di,tw) decreased during the first 16min (-28+/-7%, p<0.001) and P(ga,tw) during the first 8min (-20+/-7%, p<0.001) of hyperpnoea without further change until task failure. During inspiration, the pressure-time-product of oesophageal pressure (PTP(oes)) increased relative to PTP(di) during the first 16min (+11+/-21%, p<0.05). Similarly, during expiration, PTP(oes) increased relative to PTP(ga) during the first 8min (+10+/-16%, p<0.05). Also, blood lactate concentration and respiratory sensations significantly increased during the first 8min (+1.0+/-0.5mmoll(-1), p<0.001) and 16min (breathlessness +1.6+/-1.8 points, respiratory effort +5.9+/-2.2points, p<0.001), respectively. We conclude that, during hyperpnoea, contractile fatigue of the diaphragm and abdominal muscles develops long before task failure and may trigger an increased recruitment of rib cage muscles.


Incidence and Outcome of CPAP Failure in Preterm Infants.
Data from clinical trials support the use of continuous positive airway pressure (CPAP) for initial respiratory management in preterm infants, but there is concern regarding the potential failure of CPAP support. We aimed to examine the incidence and explore the outcomes of CPAP failure in Australian and New Zealand Neonatal Network data from 2007 to 2013. Data from inborn preterm infants managed on CPAP from the outset were analyzed in 2 gestational age ranges (25-28 and 29-32 completed weeks). Outcomes after CPAP failure (need for intubation <72 hours) were compared with those succeeding on CPAP using adjusted odds ratios (AORs). Within the cohort of 19 103 infants, 11 684 were initially managed on CPAP. Failure of CPAP occurred in 863 (43%) of 1989 infants commencing on CPAP at 25-28 weeks' gestation and 2061 (21%) of 9695 at 29-32 weeks. CPAP failure was associated with a substantially higher rate of pneumothorax, and a heightened risk of death, bronchopulmonary dysplasia (BPD) and other morbidities compared with those managed successfully on CPAP. The incidence of death or BPD was also increased: (25-28 weeks: 39% vs 20%, AOR 2.30, 99% confidence interval 1.71-3.10; 29-32 weeks: 12% vs 3.1%, AOR 3.62 [2.76-4.74]). The CPAP failure group had longer durations of respiratory support and hospitalization. CPAP failure in preterm infants is associated with increased risk of mortality and major morbidities, including BPD. Strategies to promote successful CPAP application should be pursued vigorously.


A bacterial-type ABC transporter is involved in aluminum tolerance in rice.
Aluminum (Al) toxicity is a major factor limiting crop production in acidic soil, but the molecular mechanisms of Al tolerance are poorly understood. Here, we report that two genes, STAR1 (for sensitive to Al rhizotoxicity1) and STAR2, are responsible for Al tolerance in rice. STAR1 encodes a nucleotide binding domain, while STAR2 encodes a transmembrane domain, of a bacterial-type ATP binding cassette (ABC) transporter. Disruption of either gene resulted in hypersensitivity to aluminum toxicity. Both STAR1 and STAR2 are expressed mainly in the roots and are specifically induced by Al exposure. Expression in onion epidermal cells, rice protoplasts, and yeast showed that STAR1 interacts with STAR2 to form a complex that localizes to the vesicle membranes of all root cells, except for those in the epidermal layer of the mature zone. When expressed together in Xenopus laevis oocytes, STAR1/2 shows efflux transport activity specific for UDP-glucose. Furthermore, addition of exogenous UDP-glucose rescued root growth in the star1 mutant exposed to Al. These results indicate that STAR1 and STAR2 form a complex that functions as an ABC transporter, which is required for detoxification of Al in rice. The ABC transporter transports UDP-glucose, which may be used to modify the cell wall.


Induction of the Swallowing Reflex by Electrical Stimulation of the Posterior Oropharyngeal Region in Awake Humans.
We designed an electrical stimulation system to safely and reliably evoke the swallowing reflex in awake humans, and then examined the neural control of reflex swallowing initiated by oropharyngeal stimulation. A custom-made electrode connected to a flexible stainless-steel coil spring tube was introduced into the pharyngeal region through the nasal cavity and placed against the posterior wall of the oropharynx. Surface electrodes placed over the suprahyoid muscles recorded the electromyogram during swallowing. Swallowing reflexes were induced several times by 30 s of repetitive electrical pulse stimulation (intensity: 0.2-1.2 mA, frequency: 10-70 Hz, pulse duration: 1.0 ms). The onset latency of the swallowing reflex was measured over the 10-70 Hz frequency range. In addition, the two time intervals between the first three swallows were measured. The onset latency of the swallowing reflex became shorter as the stimulus frequency increased up to ≤30 Hz. Once the frequency exceeded 30 Hz, there was no further reduction in the latency. This finding was consistent with those of previous studies in anesthetized animals. The time intervals between successive swallowing reflexes did not change with increased stimulus frequencies. Furthermore, prolonged stimulation often failed to elicit multiple swallowing reflexes. The frequency dependence of onset latency suggests that temporal summation of pharyngeal afferents is required to activate the medullary swallowing center. This reliable stimulation method may help in rehabilitation of dysphagic patients without causing aspiration.


Association between type 2 diabetes loci and measures of fatness.
Type 2 diabetes (T2D) is a metabolic disorder characterized by disturbances of carbohydrate, fat and protein metabolism and insulin resistance. The majority of T2D patients are obese and obesity by itself may be a cause of insulin resistance. Our aim was to evaluate whether the recently identified T2D risk alleles are associated with human measures of fatness as characterized with Dual Energy X-ray Absorptiometry (DEXA). Genotypes and phenotypes of approximately 3,000 participants from cross-sectional ERF study were analyzed. Nine single nucleotide polymorphisms (SNPs) in CDKN2AB, CDKAL1, FTO, HHEX, IGF2BP2, KCNJ11, PPARG, SLC30A8 and TCF7L2 were genotyped. We used linear regression to study association between individual SNPs and the combined allelic risk score with body mass index (BMI), fat mass index (FMI), fat percentage (FAT), waist circumference (WC) and waist to hip ratio (WHR). Significant association was observed between rs8050136 (FTO) and BMI (p = 0.003), FMI (p = 0.007) and WC (p = 0.03); fat percentage was borderline significant (p = 0.053). No other SNPs alone or combined in a risk score demonstrated significant association to the measures of fatness. From the recently identified T2D risk variants only the risk variant of the FTO gene (rs8050136) showed statistically significant association with BMI, FMI, and WC.


Impact of calcium binding and thionylation of S100A1 protein on its nuclear magnetic resonance-derived structure and backbone dynamics.
S100 proteins play a crucial role in multiple important biological processes in vertebrate organisms acting predominantly as calcium signal transmitters. S100A1 is a typical representative of this family of proteins. After four Ca(2+) ions bind, it undergoes a dramatic conformational change, resulting in exposure, in each of its two identical subunits, a large hydrophobic cleft that binds to target proteins. It has been shown that abnormal expression of S100A1 is strongly correlated with a number of severe human diseases: cardiomyopathy and neurodegenerative disorders. A few years ago, we found that thionylation of Cys 85, the unique cysteine in two identical S100A1 subunits, leads to a drastic increase of the affinity of the protein for calcium. We postulated that the protein activated by thionylation becomes a more efficient calcium signal transmitter. Therefore, we decided to undertake, using nuclear magnetic resonance methods, a comparative study of the structure and dynamics of native and thionylated human S100A1 in its apo and holo states. In this paper, we present the results obtained for both forms of this protein in its holo state and compare them with the previously published structure of native apo-S100. The main conclusion that we draw from these results is that the increased calcium binding affinity of S100A1 upon thionylation arises, most probably, from rearrangement of the hydrophobic core in its apo form.


Anti-Inflammatory Action of Blueberry Polyphenols in HIG-82 Rabbit Synoviocytes.
Rheumatoid arthritis (RA) is an autoimmune and chronic inflammatory disease resulting in joint destruction and disability in the adult population. The etiology of RA is not well understood and presently there is no known cure for this disease. The accumulation and proliferation of fibroblast-like synoviocytes may be involved in cartilage destruction. Both in vitro and in vivo studies support an anti-inflammatory role of dietary polyphenols, the bioactive constituents found in fruits and vegetables. The objective of this study was to investigate the anti-inflammatory role of blueberry polyphenols (BBPs) using rabbit synoviocytes stimulated with tumor necrosis factor alpha (TNFα). Rabbit synoviocytes (HIG-82) were treated with varying doses of BBPs and stimulated with TNFα. Stimulation of rabbit synoviocytes with the proinflammatory cytokine TNFα increased cell proliferation by ∼19% compared with the nonstimulated control. Cell proliferation was significantly decreased in a dose-dependent manner by the treatment with BBPs. Post-TNFα stimulation, cells treated with BBPs resulted in decreases in interleukin 1 beta and nuclear factor kappa B (NFκB) concentration. Reverse transcription-polymerase chain reaction analysis showed that matrix metalloproteinase 3 increased fivefold in the control TNFα-stimulated group, but was decreased by threefold in the blueberry treatment group. These results suggest that downregulation of proinflammatory cytokines and transcription factor NFκB by naturally occurring bioactives such as BBPs may be a potential therapeutic strategy for reducing inflammation associated with RA.


Brain changes associated with sleep disruption in cognitively unimpaired older adults: A short review of neuroimaging studies.
Ageing is characterized by a progressive decline of sleep quality. Sleep difficulties are increasingly recognized as a risk factor for Alzheimer's disease (AD), and have been associated with cognitive decline. However, the brain substrates underlying this association remain unclear. In this review, our objective was to provide a comprehensive overview of the relationships between sleep changes and brain structural, functional and molecular integrity, including amyloid and tau pathologies in cognitively unimpaired older adults. We especially discuss the topography and causality of these associations, as well as the potential underlying mechanisms. Taken together, current findings converge to a link between several sleep parameters, amyloid and tau levels in the CSF, and neurodegeneration in diffuse frontal, temporal and parietal areas. However, the existing literature remains heterogeneous, and the specific sleep changes associated with early AD pathological changes, in terms of topography and neuroimaging modality, is not clearly established yet. Notably, if slow wave sleep disruption seems to be related to frontal amyloid deposition, the brain correlates of sleep-disordered breathing and REM sleep disruption remain unclear. Moreover, sleep parameters associated with tau- and FDG-PET imaging are largely unexplored. Lastly, whether sleep disruption is a cause or a consequence of brain alterations remains an open question.


The vaginal epithelium of immature rats metabolizes androgens through an aromatase-like reaction: changes during the time of puberty.
Testosterone (T) at physiological levels can induce precocious vaginal opening without advancing the time of first ovulation. The present experiments were undertaken to test the hypothesis that the vaginal epithelium has the ability to aromatize androgens to estrogens. Using standardized conditions, we estimated aromatase activity using both 3H2O-release from [1 beta-3H]T and thin-layer chromatographic (TLC) characterization of estrogen formed after incubations with [1,2,6,7-3H] testosterone. Vaginal aromatase-like activity, as measured by the 3H2O-release assay, increased between the juvenile and peripubertal phases of development and remained elevated throughout puberty. In contrast, ovarian aromatase increased markedly during the early proestrus (EP) and late (first) proestrus (LP) phases of puberty but declined after the first ovulation. Vaginal aromatase-like activity was induced in vivo by either stimulation of ovarian steroidogenesis with pregnant mare's serum gonadotropin (PMSG), or by producing EP levels of serum T via testosterone-containing Silastic capsules. 4-Hydroxy androstenedione, a potent aromatase inhibitor, decreased both vaginal and ovarian aromatase activity in vitro in a concentration-dependent manner. Although the principal product of ovarian aromatase derived from [1,2,6,7-3H] T was identified as estradiol (E2), the identity of the vaginal estrogen product could not be firmly established. The vaginal metabolite comigrated with 16-keto-E2 in two TLC systems before and one TLC system after acetylation but failed to recrystallize as 16-keto-E2 diacetate and failed to co-elute with 16-keto-E2 diacetate on high performance liquid chromatography. This vaginal metabolite does not correspond to any of 13 steroids tested, including 2-hydroxy-E2, and it does not represent a 5 alpha-reduced metabolite of T.(ABSTRACT TRUNCATED AT 250 WORDS)


Leasing physician office space.
When leasing office space, physicians should determine the effective lease rate (ELR) for each building they are considering before making a selection. The ELR is based on a number of factors, including building quality, building location, basic form of lease agreement, rent escalators and add-on factors in the lease, tenant improvement allowance, method of square footage measurement, quality of building management, and other variables. The ELR enables prospective physician tenants to accurately compare lease rates being quoted by building owners and to make leasing decisions based on objective criteria.


[The effectiveness of continuing care models in patients with chronic diseases: a systematic review].
Population aging has caused significant rises in the prevalence of chronic diseases and the utilization of healthcare services in Taiwan. The current healthcare delivery system is fragmented. Integrating medical services may increase the quality of healthcare, enhance patient and patient family satisfaction with healthcare services, and better contain healthcare costs. This article introduces two continuing care models: discharge planning and case management. Further, the effectiveness and essential components of these two models are analyzed using a systematic review method. Articles included in this systematic review were all original articles on discharge-planning or case-management interventions published between February 1999 and March 2013 in any of 6 electronic databases (Medline, PubMed, Cinahl Plus with full Text, ProQuest, Cochrane Library, CEPS and Center for Chinese Studies electronic databases). Of the 70 articles retrieved, only 7 were randomized controlled trial studies. Three types of continuity-of-care models were identified: discharge planning, case management, and a hybrid of these two. All three models used logical and systematic processes to conduct assessment, planning, implementation, coordination, follow-up, and evaluation activities. Both the discharge planning model and the case management model were positively associated with improved self-care knowledge, reduced length of stay, decreased medical costs, and better quality of life. This study cross-referenced all reviewed articles in terms of target clients, content, intervention schedules, measurements, and outcome indicators. Study results may be referenced in future implementations of continuity-care models and may provide a reference for future research.


Burden of hypertension in China: A nationally representative survey of 174,621 adults.
Hypertension is a major cause of cardiovascular disease. Periodic nationwide surveys are essential for monitoring secular trend of hypertension and its control in population. We assessed prevalence of hypertension and related awareness, treatment and control rates in Chinese adults in 2013-14. A nationally representative survey recruited 174,621 adults aged>18years from 31 provinces in mainland China between 2013 and 2014. Population-weighted prevalence of hypertension and related rates of awareness, treatment and control were calculated and compared by age, sex, region and other factors of interest. Overall, 27.8% of Chinese adults were hypertensive, with the adjusted prevalence higher in men than in women and increasing steeply with rising age. Of those with hypertension, 31.9% were previously diagnosed, of those diagnosed, 82.9% were treated, and of those treated, 34.6% had their blood pressure properly controlled, resulting in an overall control rates of 9.7% among those with hypertension. Despite similar prevalence, the awareness, treatment and control were much better in urban areas than in rural areas. Among hypertensive individuals, older age, higher levels of education or household income tended to be associated with better awareness, treatment and control rates. During 2013-14, 292 million adults in China had hypertension, representing an absolute increase of 139 million individuals since year 2002. Among Chinese adults, more than one forth had hypertension and the prevalence has increased significantly during recent decades. Despite huge efforts, the levels of awareness, treatment and control rates of hypertension remain extremely low, foreshadowing substantial unnecessary disease burden.


Highly sensitive fiber pressure sensor based on off-center diaphragm reflection.
A fiber pressure sensor with a collimator at the off-center position of a diaphragm is demonstrated. The detection mechanism is incident-angle sensitive rather than traditional working-distance sensitive. Due to the small beam divergence of the collimator, the control on working distance is less stringent, and high sensitivity can be realized because the coupling efficiency of the collimator is very sensitive to the incident angle decided by the off-center diaphragm reflection. Sensitivity of 1.11 and 0.16 dB/KPa can be achieved with silicon diaphragm thicknesses of 100 and 150 μm, respectively. Moreover, the detection range can be continually shifted by changing the pressure in the sealed diaphragm cavity.


Quantifying and Mapping the Supply of and Demand for Carbon Storage and Sequestration Service from Urban Trees.
Studies that assess the distribution of benefits provided by ecosystem services across urban areas are increasingly common. Nevertheless, current knowledge of both the supply and demand sides of ecosystem services remains limited, leaving a gap in our understanding of balance between ecosystem service supply and demand that restricts our ability to assess and manage these services. The present study seeks to fill this gap by developing and applying an integrated approach to quantifying the supply and demand of a key ecosystem service, carbon storage and sequestration, at the local level. This approach follows three basic steps: (1) quantifying and mapping service supply based upon Light Detection and Ranging (LiDAR) processing and allometric models, (2) quantifying and mapping demand for carbon sequestration using an indicator based on local anthropogenic CO2 emissions, and (3) mapping a supply-to-demand ratio. We illustrate this approach using a portion of the Twin Cities Metropolitan Area of Minnesota, USA. Our results indicate that 1735.69 million kg carbon are stored by urban trees in our study area. Annually, 33.43 million kg carbon are sequestered by trees, whereas 3087.60 million kg carbon are emitted by human sources. Thus, carbon sequestration service provided by urban trees in the study location play a minor role in combating climate change, offsetting approximately 1% of local anthropogenic carbon emissions per year, although avoided emissions via storage in trees are substantial. Our supply-to-demand ratio map provides insight into the balance between carbon sequestration supply in urban trees and demand for such sequestration at the local level, pinpointing critical locations where higher levels of supply and demand exist. Such a ratio map could help planners and policy makers to assess and manage the supply of and demand for carbon sequestration.


Moose-tree interactions: rebrowsing is common across tree species.
Plant strategies to resist herbivory include tolerance and avoidance. Tolerance strategies, such as rapid regrowth which increases the palatability of new shoots, can lead to positive feedback loops between plants and herbivores. An example of such a positive feedback occurs when moose (Alces alces) browse trees in boreal forests. We described the degree of change in tree morphology that accumulated over time in response to repeated browsing by moose, using an index of accumulated browsing. We evaluated whether accumulated browsing could predict the probability and extent of current browsing across woody species in a Norwegian boreal forest, and how our accumulated browsing index related to changes in tree height, shoot availability and shoot size. The probability and extent of current browsing increased with the degree of accumulated browsing in all tree species. Plants highly modified by previous browsing were the most attractive, with no indication of decreased preference with repeated browsing over time. The preference for previously browsed trees is most likely driven by increased relative availability of shoots within browsing height and maybe increased palatability. This response to previous browsing was general for both preferred and avoided forage species, in both conifers and deciduous trees. Our results suggest that the adaptation for rapid regrowth after browsing does not reduce herbivory on trees. Rather, our results indicate that plant responses to browsing increase the probability of future herbivory. This response could potentially lead to higher plant mortality where cervid populations are maintained at stable high densities and has implications for plant population dynamics and forestry practices.


Rapid and longer-term antidepressant effects of repeated ketamine infusions in treatment-resistant major depression.
Ketamine is reported to have rapid antidepressant effects; however, there is limited understanding of the time-course of ketamine effects beyond a single infusion. A previous report including 10 participants with treatment-resistant major depression (TRD) found that six ketamine infusions resulted in a sustained antidepressant effect. In the current report, we examined the pattern and durability of antidepressant effects of repeated ketamine infusions in a larger sample, inclusive of the original. Participants with TRD (n = 24) underwent a washout of antidepressant medication followed by a series of up to six IV infusions of ketamine (.5 mg/kg) administered open-label three times weekly over a 12-day period. Participants meeting response criteria were monitored for relapse for up to 83 days from the last infusion. The overall response rate at study end was 70.8%. There was a large mean decrease in Montgomery-Åsberg Depression Rating Scale score at 2 hours after the first ketamine infusion (18.9 ± 6.6, p < .001), and this decrease was largely sustained for the duration of the infusion period. Response at study end was strongly predicted by response at 4 hours (94% sensitive, 71% specific). Among responders, median time to relapse after the last ketamine infusion was 18 days. Ketamine was associated with a rapid antidepressant effect in TRD that was predictive of a sustained effect. Future controlled studies will be required to identify strategies to maintain an antidepressant response among patients who benefit from a course of ketamine.


Overview of Treatment Guidelines and Clinical Practical Guidelines That Recommend the Use of Acupuncture: A Bibliometric Analysis.
As positive evidence emerges for the use of an intervention to treat a health problem, the intervention gradually becomes incorporated into treatment guidelines (TGs) or clinical practice guidelines (CPGs) that are related to that health problem. To assess whether this general hypothesis can apply to acupuncture, 96 health problems were identified for which positive conclusions in systematic reviews and meta-analyses regarding the effectiveness of acupuncture have been made and then searched for TGs or CPGs that have recommended the use of acupuncture. Through August 31, 2017, searches were performed in relevant medical databases and Google using "treatment guideline," "clinical practice guideline," and the names of the 96 medical conditions as search terms. A "snow-balling" search approach was adopted. All positive recommendations were added into the registry. A total of 1311 publications were found that recommended using acupuncture published between 1991 and 2017. The number per year reached 50 in 2005 and 100 in 2009. In addition, 2189 positive recommendations were found for the use of acupuncture. Of these, 1486 were related to 107 pain indications and 703 were related to 97 nonpain indications. These recommendations were made by a wide range of groups, such as government health institutions, national guideline, and medical specialty groups. The recommendations came from around the world but were especially abundant in North America, Europe, and Australasia. Considerably more recommendations were found for the use of acupuncture than are known within the acupuncture or medical communities. A trend by year was also found; a rise in the number of positive statements about acupuncture was typically followed by a rise in the number of recommendations of acupuncture. Thus, the recommendations followed the emergent evidence for acupuncture. Better implementation plans need to be developed for the CPG/TG recommendations about acupuncture to be more effective/efficient.


Three- and four-element Windkessel models: assessment of their fitting performance in a large cohort of healthy middle-aged individuals.
Lumped-parameter models are used to estimate the global arterial properties by fitting the model to measured (aortic) pressure and flow. Different model configurations coexist, and it is still an open question as to which model optimally reflects the arterial tree and leads to correct estimates of arterial properties. An assessment was made of the performance of (a) the three-element Windkessel model (WK3) consisting of vascular resistance R, total arterial compliance C, and characteristic impedance Zc; (b) a four-element model with an inertance element L placed in parallel with Zc (WK4-p); and (c) a four-element model with L placed in series with Zc (WK4-s). Models were fitted to data measured non-invasively in 2404 healthy subjects, aged between 35 and 55 years. It was found that model performance segregated into two groups. In a group containing 20 per cent of the dataset (characterized by low blood pressure and wave reflection) the WK4-p model outperformed the other models, with model behaviour as envisioned by its promoters. In these cases, the WK3 and WK4-s models led to increased overestimation of total arterial compliance and underestimation of characteristic impedance. However, in about 80 per cent of the cases, the WK4-p model showed a behaviour that was very similar to that of the WK3 and WK4-s models. Here, the WK4-s model yielded the best quality of fit, although model parameters reached physically impossible values for L in about 12 per cent of all cases. The debate about which lumped-parameter model is the better approximation of the arterial tree is therefore still not fully resolved.


Both T cell receptor (TcR)-CD3 complex and CD2 increase the tyrosine kinase activity of p56lck. CD2 can mediate TcR-CD3-independent and CD45-dependent activation of p56lck.
T cell activation by triggering the T cell receptor (TcR)-CD3 complex leads to a dramatic increase in tyrosine phosphorylation of multiple cellular proteins. To date, there has been no direct evidence on the identity of the tyrosine kinase activity implicated in this signaling pathway. In this study, we demonstrate that activation of human T cells with anti-CD3 monoclonal antibody increases tyrosine kinase activity of p56lck. This extends our previous findings which demonstrated the involvement of p56lck kinase activity in the CD2 signal transduction pathway. The results from peripheral blood lymphocytes and Jurkat cell line showed in both cases an early and transient change in the specific activity of p56lck, followed by a shift to a higher apparent molecular mass. Therefore, to test directly the role of TcR-CD3 in CD2-induced activation of p56lck, we utilized mutant variants of the Jurkat cell line lacking in cell surface TcR-CD3. We found that cell surface expression of TcR-CD3 is not required for the activation of p56lck via CD2 but is necessary for the appearance of the reduced-electrophoretic-mobility form of p56lck observed after CD2 triggering. By isolating CD45- mutants from Jurkat cells, we observed that surface expression of the tyrosine phosphatase CD45 is required in order to increase p56lck activity following CD2 stimulation, while CD4-induced activation of the kinase remained unchanged. These data provide evidence for a specific functional linkage between CD2 and p56lck, in which CD45 may play an essential role.


First-line treatment of malignant glioma with carmustine implants followed by concomitant radiochemotherapy: a multicenter experience.
Randomized phase III trials have shown significant improvement of survival 1, 2, and 3 years after implantation of 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU) wafers for patients with newly diagnosed malignant glioma. But these studies and subsequent non-phase III studies have also shown risks associated with local chemotherapy within the central nervous system. The introduction of concomitant radiochemotherapy with temozolomide (TMZ) has later demonstrated a survival benefit in a phase III trial and has become the current treatment standard for newly diagnosed malignant glioma patients. Lately, this has resulted in clinical protocols combining local chemotherapy with BCNU wafers and concomitant radiochemotherapy with TMZ although this may carry the risk of increased toxicity. We have compiled the treatment experience of seven neurosurgical centers using implantation of carmustine wafers at primary surgery followed by 6 weeks of radiation therapy (59-60 Gy) and 75 mg/m(2)/day TMZ in patients with newly diagnosed glioblastoma followed by TMZ monochemotherapy. We have retrospectively analyzed the postoperative clinical course, occurrence and severity of adverse events, progression-free interval, and overall survival in 44 patients with newly diagnosed glioblastoma multiforme. All patients received multimodal treatment including tumor resection, BCNU wafer implantation, and concomitant radiochemotherapy. Of 44 patients (mean age 59 ± 10.8 years) with glioblastoma who received Gliadel wafer at primary surgery, 28 patients (64%) had died, 16 patients (36%) were alive, and 15 patients showed no evidence of clinical or radiographic progression after a median follow-up of 15.6 months. At time of analysis of adverse events in this patient population, the median overall survival was 12.7 months and median progression-free survival was 7.0 months. Surgical, neurological, and medical adverse events were analyzed. Twenty-three patients (52%) experienced adverse events of any kind including complications that did not require treatment. Nineteen patients (43%) experienced grade 3 or grade 4 adverse events. Surgical complications included cerebral edema, healing abnormalities, cerebral spinal fluid leakage, meningitis, intracranial abscess, and hydrocephalus. Neurological adverse events included newly diagnosed seizures, alteration of mental status, and new neurological deficits. Medical complications were thromboembolic events (thrombosis, pulmonary embolism) and hematotoxicity. Combination of both treatment strategies, local chemotherapy with BCNU wafer and concomitant radiochemotherapy, appears attractive in aggressive multimodal treatment schedules and may utilize the sensitizing effect of TMZ and carmustine on MGMT and AGT on their respective drug resistance genes. Our data demonstrate that combination of local chemotherapy and concomitant radiochemotherapy carries a significant risk of toxicity that currently appears underestimated. Adverse events observed in this study appear similar to complication rates published in the phase III trials for BCNU wafer implantation followed by radiation therapy alone, but further add the toxicity of concomitant radiochemotherapy with systemic TMZ. Save use of a combined approach will require specific prevention strategies for multimodal treatments.


Thermochemical remanent magnetization and thermal remanent magnetization: comparison in a basalt.
Recent studies have shown that the remanent magnetization carried by an extrusive igneous rock may not be entirely thermal remanent magnetization (TRM). Some may be thermochemical remanent magnetization (TCRM) acquired by the rock at temperatures at least as low as 300 degrees C during oxidation of the contained titanomagnetite grains. Results from a study of a set of basaltic samples from one locality indicate that the intensity of TCRM acquired by a sample in a known magnetic field is equal to that of TRM subsequently produced in the same sample in the same field. On the assumption that the samples we studied are not magnetically unique, we tentatively conclude that paleointensity studies are valid in spite of the presence of TCRM, as long as the rock acquired the magnetization during the initial cooling.