Behavioral bone-conduction thresholds for infants with normal hearing.
Bone-conduction thresholds have been used in audiologic assessments of both infants and adults to differentiate between conductive and sensorineural hearing losses. However, air- and bone-conduction thresholds estimated for infants with normal hearing using physiological measures have identified an "air-bone gap" in the low frequencies that does not result from conductive hearing impairment but, rather, from maturational differences in sensitivity. This maturational air-bone gap appears to be present up to at least 2 yr of age. Because most infants older than 6 mo of age are clinically assessed behaviorally, rather than physiologically, it is necessary to determine whether a similar maturational air-bone gap is present for behavioral air- and bone-conduction thresholds. The purpose of this study was to estimate behavioral bone-conduction thresholds for infants using a standard clinical visual reinforcement audiometry (VRA) protocol to determine whether frequency-dependent maturational patterns exist as previously reported for physiological bone-conduction thresholds. Behavioral bone-conduction minimum response levels were estimated at 500, 1000, 2000, and 4000 Hz using VRA for each participant. Young (7-15 mo; N = 17) and older (18-30 mo; N = 20) groups of infants were assessed. All infants were screened and considered to be at low risk for hearing loss. Preliminary "normal levels" were determined by calculating the 90th percentile for responses present as a cumulative percentage. Mean bone-conduction thresholds were compared and analyzed using a mixed-model analysis of variance across frequency and age group. Linear regression analysis was also performed to assess the effect of age on bone-conduction thresholds. Results of this study indicate that, when measured behaviorally, infants under 30 mo of age show frequency-dependent bone-conduction thresholds whereby their responses at 500 and 1000 Hz are significantly better than those at 2000 and 4000 Hz. However, thresholds obtained from the younger group of infants (mean age of 10.6 mo) were not significantly different from those obtained from the older group of infants (mean age of 23.0 mo) at any frequency. The findings of the present study are similar to the results obtained from previous physiological studies. Compared to previously documented air-conduction thresholds of infants using similar VRA techniques, a maturational air-bone gap is observed in the low frequencies. Therefore, differences between infant and adult bone-conduction thresholds persist until at least 30 mo of age. As a result, different "normal levels" should be used when assessing bone-conduction hearing sensitivity of infants using behavioral methods.


Robust and discriminating method for face recognition based on correlation technique and independent component analysis model.
We demonstrate a novel technique for face recognition. Our approach relies on the performances of a strongly discriminating optical correlation method along with the robustness of the independent component analysis (ICA) model. Simulations were performed to illustrate how this algorithm can identify a face with images from the Pointing Head Pose Image Database. While maintaining algorithmic simplicity, this approach based on ICA representation significantly increases the true recognition rate compared to that obtained using our previously developed all-numerical ICA identity recognition method and another method based on optical correlation and a standard composite filter.


Mig12, a novel Opitz syndrome gene product partner, is expressed in the embryonic ventral midline and co-operates with Mid1 to bundle and stabilize microtubules.
Opitz G/BBB syndrome is a genetic disorder characterized by developmental midline abnormalities, such as hypertelorism, cleft palate, and hypospadias. The gene responsible for the X-linked form of this disease, MID1, encodes a TRIM/RBCC protein that is anchored to the microtubules. The association of Mid1 with the cytoskeleton is regulated by dynamic phosphorylation, through the interaction with the alpha4 subunit of phosphatase 2A (PP2A). Mid1 acts as an E3 ubiquitin ligase, regulating PP2A degradation on microtubules. In spite of these findings, the biological role exerted by the Opitz syndrome gene product is still unclear and the presence of other potential interacting moieties in the Mid1 structure prompted us to search for additional cellular partners. Through a yeast two-hybrid screening approach, we identified a novel gene, MIG12, whose protein product interacts with Mid1. We confirmed by immunoprecipitation that this interaction occurs in vivo and that it is mediated by the Mid1 coiled-coil domain. We found that Mig12 is mainly expressed in the neuroepithelial midline, urogenital apparatus, and digits during embryonic development. Transiently expressed Mig12 is found diffusely in both nucleus and cytoplasm, although it is enriched in the microtubule-organizing center region. Consistently with this, endogenous Mig12 protein is partially detected in the polymerized tubulin fraction after microtubule stabilization. When co-transfected with Mid1, Mig12 is massively recruited to thick filamentous structures composed of tubulin. These microtubule bundles are resistant to high doses of depolymerizing agents and are composed of acetylated tubulin, thus representing stabilized microtubule arrays. Our findings suggest that Mig12 co-operates with Mid1 to stabilize microtubules. Mid1-Mig12 complexes might be implicated in cellular processes that require microtubule stabilization, such as cell division and migration. Impairment in Mig12/Mid1-mediated microtubule dynamic regulation, during the development of embryonic midline, may cause the pathological signs observed in Opitz syndrome patients.


Novel agents for the treatment of multiple myeloma: proteasome inhibitors and immunomodulatory agents.
The integration of novel agents into the treatment of multiple myeloma (MM) has shifted the focus from an incurable disease to one that is chronic, with a realistic hope of someday achieving a cure. Proteasome inhibitors and immunomodulatory agents are the backbone of novel therapies for MM. These agents are particularly important for patients with relapsed or refractory disease, a fate faced by the majority of myeloma patients over the course of their disease. Review of recent clinical trial data for the proteasome inhibitors and immunomodulatory agents, including clinical efficacy and safety information, will assist the advanced practitioner in oncology with integrating these data into the current treatment guidelines for MM.


Pericardial patch graft repair of severe localized scleral thinning encountered during strabismus surgery.
This article presents a surgical technique using a pericardial patch for the permanent repair of severe scleral thinning encountered during strabismus surgery. In the present case scleral thinning resulted from buckle removal. Familiarity with this technique may prove important for the strabismus surgeon treating patients with a history of surface ocular hardware or disease-induced scleral thinning. This video article may be viewed atjaapos.org.


Effect of increased body mass index (BMI) on time to tumour progression (TTP) in unresectable metastatic colorectal cancer (mCRC) patients treated with bevacizumab-based therapy.
High BMI is a well-known risk factor for the development and recurrence of several solid tumours, including CRC. Obesity is associated with increased levels of vascular endothelial growth factor (VEGF). Bevacizumab is the main targeted therapy for inhibiting tumour angiogenesis by blocking the VEGF/VEGF receptor pathway. Elevated VEGF in obese patients might provoke resistance to anti-VEGF therapy. We evaluated the efficacy of bevacizumab on TTP among mCRC patients through stratifying them according to their BMI. Patients with mCRC who had been treated with fluoropyrimidine-based combination chemotherapy with bevacizumab were included in the study. Patients were assigned according to their BMI before initiation of therapy (group A: BMI < 25 kg/m(2), group B: BMI ≥ 25 kg/m(2)). Multivariate analysis was performed to evaluate the risk of tumour progression. Between April 2007 and June 2011, 80 patients were treated with chemotherapy and bevacizumab as first-line therapy (n = 37 for group A, n = 43 for group B). Tumours in 56.3 % of the patients in group A (n = 21) and 76.3 % of the patients in group B (n = 33) progressed during a median 10-months (3-57 months) follow-up. The median TTP was 11.7 months in the group A and 6 months in the group B (p = 0.004). In a multivariate analysis, high BMI (≥25 kg/m(2)) was associated with significantly shorter TTP (p = 0.01; HR: 4.37). High BMI among mCRC patients treated with bevacizumab is associated with shorter TTP. Further study in larger databases is warranted for confirming the negative prognostic effect of obesity during treatment with anti-VEGF agents.


Southern Ocean Biogeochemical Float Deployment Strategy, With Example From the Greenwich Meridian Line (GO-SHIP A12).
Biogeochemical Argo floats, profiling to 2,000-m depth, are being deployed throughout the Southern Ocean by the Southern Ocean Carbon and Climate Observations and Modeling program (SOCCOM). The goal is 200 floats by 2020, to provide the first full set of annual cycles of carbon, oxygen, nitrate, and optical properties across multiple oceanographic regimes. Building from no prior coverage to a sparse array, deployments are based on prior knowledge of water mass properties, mean frontal locations, mean circulation and eddy variability, winds, air-sea heat/freshwater/carbon exchange, prior Argo trajectories, and float simulations in the Southern Ocean State Estimate and Hybrid Coordinate Ocean Model (HYCOM). Twelve floats deployed from the 2014-2015 Polarstern cruise from South Africa to Antarctica are used as a test case to evaluate the deployment strategy adopted for SOCCOM's 20 deployment cruises and 126 floats to date. After several years, these floats continue to represent the deployment zones targeted in advance: (1) Weddell Gyre sea ice zone, observing the Antarctic Slope Front, and a decadally-rare polynya over Maud Rise; (2) Antarctic Circumpolar Current (ACC) including the topographically steered Southern Zone chimney where upwelling carbon/nutrient-rich deep waters produce surprisingly large carbon dioxide outgassing; (3) Subantarctic and Subtropical zones between the ACC and Africa; and (4) Cape Basin. Argo floats and eddy-resolving HYCOM simulations were the best predictors of individual SOCCOM float pathways, with uncertainty after 2 years of order 1,000 km in the sea ice zone and more than double that in and north of the ACC.


Molecular cloning and enzymatic characterization of sheep CYP2J.
Cytochrome P450 (CYP) 2Js have been studied in various mammals, but not in sheep, as an animal model used to test veterinary drug metabolism. Sheep CYP2J was cloned from liver messenger RNA (mRNA) by RACE. The cDNA, after modification at its N- and C-terminals, was expressed in Escherichia coli and the sheep CYP2J protein, purified by chromatography, was 80% homologous to human and monkey CYP2J2. Reverse transcriptase-polymerase chain reaction (RT-PCR) experiments showed that CYP2J mRNA was expressed in liver, cortex, respiratory and olfactory mucosa, heart, bronchi, lung, spleen, small intestine and kidney. The purified enzyme was catalytically active towards aminopyrine, all-trans-retinoic acid, and particularly arachidonic acid forming 20-HETE, 19-HETE, and 18-HETE (about 86% of the total) and 14,15-, 11,12-, 8,9-, and 5,6-EETs (cis-epoxyeicosatrienoic acids; about 14% of total), with a regioselectivity similar to that shown by the mammalian CYP2J2s.


Primary multiple cerebral hydatid disease: still symptomatic despite pathologically confirmed death of the cyst.
Hydatid disease is a life-threatening parasitic infestation caused by Echinococcus granulosus. Infection with E. granulosus typically results in the formation of hydatid cysts in liver, lungs, kidney and spleen. Majority of the intracranial cysts are secondary and solitary. Multiple primary cerebral cysts are uncommon. Surgical and medical management of a 14-year-old boy with multiple primary hydatid cysts are presented. 14 cysts, which were symptomatic due to their mass effect, were surgically removed, whereas a deep-seated asymptomatic cyst was followed-up with medical treatment. Despite proper antibiotic regimen the patient was admitted with epileptic seizures six months later. The deep-seated lesion was also surgically removed. Intraoperative observations and pathological examination demonstrated different characteristics, with pericystic gliosis, gel-like cyst content and death scolices within the cavity. In addition to the fact, that the presented case is an additional example for the rare primary multiple cerebral hydatid cysts, to our knowledge it is the first case of a dead cerebral hydatid cyst, causing symptoms despite effective medical treatment.


Strategies for metabolic flux analysis in plants using isotope labelling.
Flux measurements through metabolic pathways generate insights into the integration of metabolism, and there is increasing interest in using such measurements to quantify the metabolic effects of mutation and genetic manipulation. Isotope labelling provides a powerful approach for measuring metabolic fluxes, and it gives rise to several distinct methods based on either dynamic or steady-state experiments. We discuss the application of these methods to photosynthetic and non-photosynthetic plant tissues, and we illustrate the different approaches with an analysis of the pathways interconverting hexose phosphates and triose phosphates. The complicating effects of the pentose phosphate pathway and the problems arising from the extensive compartmentation of plant cell metabolism are considered. The non-trivial nature of the analysis is emphasised by reference to invalid deductions in earlier work. It is concluded that steady-state isotopic labelling experiments can provide important information on the fluxes through primary metabolism in plants, and that the combination of stable isotope labelling with detection by nuclear magnetic resonance is particularly informative.


FOXE1 polymorphism rs965513 predisposes to thyroid cancer in a European cohort.
FOXE1 is an intronless gene on chromosome 9 which plays a significant role in thyroid morphogenesis. Mutations in FOXE1 are associated with thyroid phenotypes including congenital hypothyroidism, thyroid dysgenesis and thyroid cancer. This study aims to investigate the frequency and impact of a SNP (rs965513, G>A) at 9q22.23 in a Western European cohort of patients with differentiated thyroid cancer(DTC), compared to controls. This is a candidate gene case control study. 277 patients with histologically confirmed DTC were recruited from tertiary referral centres in Ireland and France. 309 cancer-free controls were recruited from the community. DNA was extracted from buccal swabs or whole blood of control subjects and patients with DTC. Allelic and genotypic frequencies among patients were compared with controls, to assess the risk for disease conferred by homozygous and heterozygous carriers compared to WT genotypes. Genotyping was performed using Taqman-based PCR. 277 patients with confirmed DTC and 309 non-cancer controls were genotyped for the variant (rs965513). The frequency of the minor allele among cases was 0.45 compared to 0.34 among controls. The genotypic odds ratio for heterozygotes was 1.66 (CI 1.16-2.39, P =0.00555), increasing to 2.93 (CI 1.70-5.05, P =0.00007) for rare homozygotes. All subjects were in Hardy-Weinberg equilibrium (±χ2, P =0.09, P =0.07 respectively). This FOXE1 polymorphism is a low penetrance variant associated with DTC susceptibility in this cohort. The minor allele was identified among patients with thyroid cancer significantly more frequently than controls. An allele dosage effect was observed, with rare homozygous genotypes conferring greater risk than heterozygotes.


Surgical management of sacral chordoma.
Treatment results of 17 patients who were diagnosed with sacral chordoma between 1993 and 2007, were analyzed retrospectively. The mean duration of symptoms was 30.2 months. The mean tumour size was 10.7 cm; the location was S2 or more proximal in ten-patients. A wide resection was achieved in 14 patients, a marginal resection in one patient and two patients had intralesional excision. Seven patients had a recurrence after a mean time interval of 36.3 months. Wound dehiscence and infection were the major problems. Four patients required continuous urinary catheterization because of incontinence, and ten patients had colostomy or ileostomy simultaneously with tumour resection. This study showed that tumour size and a wide surgical margin are importance factors for survival in chordoma patients. Use of a combined anterior and posterior approach could increase the chance of achieving a wide margin. A multidisciplinary approach is required to achieve this goal.


Cryopreservation of schistosomules of Schistosoma mansoni in quantity.
Large quantities of Schistosoma mansoni schistosomules were cryopreserved in 35% ethanediol, and attempts were made to improve their morphology and infectivity after thawing. Several variables affected the appearance of the thawed organisms. These included: 1) the particular in vitro method used to transform cercariae to schistosomules; 2) addition of serotonin to the thawing medium; 3) changing the thawing temperature; and 4) culturing schistosomules for extended post-thaw times before assessing their condition. Varying either the postemergence age of the cercariae before transformation or the concentration of organisms in the freezing suspension had no effect on their survival. Although damage to many of the thawed schistosomules became apparent only upon extended in vitro cultivation, the addition of fetal calf serum to the thawing medium usually retarded this deterioration. When injected into mice, approximately 5% of cryopreserved schistosomules matured to adult worms, representing 71% of the value for maturation of unfrozen schistosomules. These studies define conditions of importance for the successful preservation of very large quantities of schistosomules (up to 500,000 in 1 ml volume) in liquid nitrogen.


Spumaretroviruses: Updated taxonomy and nomenclature.
Spumaretroviruses, commonly referred to as foamy viruses, are complex retroviruses belonging to the subfamily Spumaretrovirinae, family Retroviridae, which naturally infect a variety of animals including nonhuman primates (NHPs). Additionally, cross-species transmissions of simian foamy viruses (SFVs) to humans have occurred following exposure to tissues of infected NHPs. Recent research has led to the identification of previously unknown exogenous foamy viruses, and to the discovery of endogenous spumaretrovirus sequences in a variety of host genomes. Here, we describe an updated spumaretrovirus taxonomy that has been recently accepted by the International Committee on Taxonomy of Viruses (ICTV) Executive Committee, and describe a virus nomenclature that is generally consistent with that used for other retroviruses, such as lentiviruses and deltaretroviruses. This taxonomy can be applied to distinguish different, but closely related, primate (e.g., human, ape, simian) foamy viruses as well as those from other hosts. This proposal accounts for host-virus co-speciation and cross-species transmission.


The Fat1 cadherin is overexpressed and an independent prognostic factor for survival in paired diagnosis-relapse samples of precursor B-cell acute lymphoblastic leukemia.
Improved survival of patients with acute lymphoblastic leukemia (ALL) has emerged from identifying new prognostic markers; however, 20% of children still suffer recurrence. Previously, the altered expression of Fat1 cadherin has been implicated in a number of solid tumors. In this report, in vitro analysis shows that Fat1 protein is expressed by a range of leukemia cell lines, but not by normal peripheral blood (PB) and bone marrow (BM) cells from healthy donors. In silico analysis of expression of array data from clinical leukemias found significant levels of Fat1 transcript in 11% of acute myeloid leukemia, 29% and 63% of ALL of B and T lineages, respectively, and little or no transcript present in normal PB or BM. Furthermore, in two independent studies of matched diagnosis-relapse of precursor B-cell (preB) ALL pediatric samples (n=32 and n=27), the level of Fat1 mRNA expression was prognostic at the time of diagnosis. High Fat1 mRNA expression was predictive of shorter relapse-free and overall survival, independent of other traditional prognostic markers, including white blood cell count, sex and age. The data presented demonstrate that Fat1 expression in preB-ALL has a role in the emergence of relapse and could provide a suitable therapeutic target in high-risk preB-ALL.


Determination of degree of ionization of poly(allylamine hydrochloride) (PAH) and poly[1-[4-(3-carboxy-4 hydroxyphenylazo)benzene sulfonamido]-1,2-ethanediyl, sodium salt] (PAZO) in layer-by-layer films using vacuum photoabsorption spectroscopy.
Electrostatic and hydrophobic interactions govern most of the properties of supramolecular systems, which is the reason determining the degree of ionization of macromolecules has become crucial for many applications. In this paper, we show that high-resolution ultraviolet spectroscopy (VUV) can be used to determine the degree of ionization and its effect on the electronic excitation energies of layer-by-layer (LbL) films of poly(allylamine hydrochloride) (PAH) and poly[1-[4-(3-carboxy-4 hydroxyphenylazo)benzene sulfonamido]-1,2-ethanediyl, sodium salt] (PAZO). A full assignment of the VUV peaks of these polyelectrolytes in solution and in cast or LbL films could be made, with their pH dependence allowing us to determine the pK(a) using the Henderson-Hasselbach equation. The pK(a) for PAZO increased from ca. 6 in solution to ca. 7.3 in LbL films owing to the charge transfer from PAH. Significantly, even using solutions at a fixed pH for PAH, the amount adsorbed on the LbL films still varied with the pH of the PAZO solutions due to these molecular-level interactions. Therefore, the procedure based on a comparison of VUV spectra from solutions and films obtained under distinct conditions is useful to determine the degree of dissociation of macromolecules, in addition to permitting interrogation of interface effects in multilayer films.


The influence of experimentally induced copper deficiency on the fertility of rams. I. Semen parameters and peripheral plasma androgen concentration.
The influence of molybdenum and molybdenum plus sulphate-induced copper deficiencies on semen quality and peripheral plasma testosterone concentrations in South African Mutton Merino rams was investigated. All animals received an identical ration, supplemented with molybdenum in one group (M) and molybdenum plus sulphate in another (MS) while the control group (C) received additional copper supplementation. After almost a year on these rations, rams in Group MS produced ejaculates of lower volume, lower sperm concentration, poorer sperm motility and morphology than rams of the other 2 groups. The fructose concentration in the ejaculates of group MS rams was also lower than that of rams in the other groups. Peripheral plasma testosterone concentrations in Group MS were lower than in Groups C or M. Liver copper concentrations and clinical signs were consistent with a severe copper deficiency in Group MS and a mild deficiency in Group M. After the copper deficiency was reversed, the above parameters reverted to normal. It was concluded that an experimentally induced copper deficiency produced reversible impairment of testicular function in rams.


Do the stars align? Distribution of high-quality ratings of healthcare sectors across US markets.
The US government created five-star rating systems to evaluate hospital, nursing homes, home health agency and dialysis centre quality. The degree to which quality is a property of organisations versus geographical markets is unclear. To determine whether high-quality healthcare service sectors are clustered within US healthcare markets. Using data from the Centers for Medicare and Medicaid Services' Hospital, Dialysis, Nursing Home and Home Health Compare databases, we calculated the mean star ratings of four healthcare sectors in 304 US hospital referral regions (HRRs). For each sector, we ranked HRRs into terciles by mean star rating. Within each HRR, we assessed concordance of tercile rank across sectors using a multirater kappa. Using t-tests, we compared characteristics of HRRs with three to four top-ranked sectors, one to two top-ranked sectors and zero top-ranked sectors. Six HRRs (2.0% of HRRs) had four top-ranked healthcare sectors, 38 (12.5%) had three top-ranked health sectors, 71 (23.4%) had two top-ranked sectors, 111 (36.5%) had one top-ranked sector and 78 (25.7%) HRRs had no top-ranked sectors. A multirater kappa across all sectors showed poor to slight agreement (K=0.055). Compared with HRRs with zero top-ranked sectors, those with three to four top-ranked sectors had higher median incomes, fewer black residents, lower mortality rates and were less impoverished. Results were similar for HRRs with one to two top-ranked sectors. Few US healthcare markets exhibit high-quality performance across four distinct healthcare service sectors, suggesting that high-quality care in one sector may not be dependent on or improve care quality in other sectors. Policies that promote accountability for quality across sectors (eg, bundled payments and shared quality metrics) may be needed to systematically improve quality across sectors.


Induction of transmission-blocking immunity in Aotus monkeys by vaccination with a Plasmodium vivax clinical grade PVS25 recombinant protein.
Aotus monkeys were used to determine the immunogenicity of Pvs25 protein expressed in the zygote/ookinete surface. Animals were immunized in three times with 100 microg of Pvs25 formulated in Montanide ISA-720. Antibodies to Pvs25 detected by an enzyme-linked immunosorbent assay appeared by day 30 after the first immunization, with a peak of antibodies levels on day 150. These antibodies were still detectable on day 300. Plasma samples on day 150 from experimental group were able to completely block the development of the parasite in Anopheles albimanus mosquitoes artificially fed with human isolates of Plasmodium vivax. Immunized Aotus monkeys were infected with blood forms of the P. vivax Salvador I strain and no boosting effect of blood infection on titers of antibodies to Pvs25 was observed despite the presence of infective gametocytes. In conclusion, Pvs25 protein formulated in Montanide ISA-720 induces efficient and long-lasting transmission-blocking antibodies that cannot be boosted by parasite infection.


Urban Water Services in Fragile States: An Analysis of Drinking Water Sources and Quality in Port Harcourt, Nigeria, and Monrovia, Liberia.
Establishing and maintaining public water services in fragile states is a significant development challenge. In anticipation of water infrastructure investments, this study compares drinking water sources and quality between Port Harcourt, Nigeria, and Monrovia, Liberia, two cities recovering from political and economic instability. In both cities, access to piped water is low, and residents rely on a range of other private and public water sources. In Port Harcourt, geographic points for sampling were randomly selected and stratified by population density, whereas in Monrovia, locations for sampling were selected from a current inventory of public water sources. In Port Harcourt, the sampling frame demonstrated extensive reliance on private boreholes and a preference, in both planned and unplanned settlements, for drinking bottled and sachet water. In Monrovia, sample collection focused on public sources (predominantly shallow dug wells). In Port Harcourt, fecal indicator bacteria (FIB) were detected in 25% of sources (N = 566), though concentrations were low. In Monrovia, 57% of sources contained FIB and 22% of sources had nitrate levels that exceeded standards (N = 204). In Monrovia, the convenience of piped water may promote acceptance of the associated water tariffs. However, in Port Harcourt, the high prevalence of self-supply and bottled and sachet drinking water suggests that the consumer's willingness to pay for ongoing municipal water supply improvements may be determined by service reliability and perceptions of water quality.


Thymus dictates major histocompatibility complex (MHC) specificity and immune response gene phenotype of class II MHC-restricted T cells but not of class I MHC-restricted T cells.
Athymic H-2b nude mice received grafts from C57BL/6 (Sendai virus and H-Y antigen cytotoxic T lymphocyte [CTL] responder type), bm1 (H-2Kb mutant, Sendai CTL nonresponder type), or bm12 (H-21-A mutant, H-Y CTL nonresponder type) neonates. In observations of the CTL response to H-Y, both recipients and thymus donors were female. All types of thymus engraftment resulted in mature H-2b splenic T lymphocyte surface phenotype in nude hosts. T cell immunocompetence (as measured by major histocompatibility complex [MHC] CTL responses to allogeneic cells) was restored, and induced nonresponsiveness to the MHC determinants of the engrafted thymus in the nude host. The CTL reaction to Sendai virus in both responder type C57BL/6 and nonresponder type bm1 neonatal thymuses allowed maturation of Sendai-specific, H-2Kb-restricted CTL. For the CTL reaction to H-Y, only responder type C57BL/6 thymuses restored the CTL response, whereas this was not achieved with thymuses from nonresponder type bm12 neonatal females. Results of double thymus (B6 and bm12) engraftment excluded the possibility that this latter effect was caused by suppression. In addition, athymic bm1 mice were engrafted with thymuses from either B6 (Sendai CTL responder type) or syngeneic bm1 neonates (Sendai CTL nonresponder type). Again, both types of neonate thymuses restored T cell competence as measured by MHC/CTL responses to allogeneic cells. However, neither responder B6 nor nonresponder bm1 neonate thymus grafts allowed maturation of Sendai-specific CTL. In conclusion, the thymus dictates MHC specificity and immune response gene phenotype of T cells restricted to class II MHC molecules but not of T cells restricted to class I MHC molecules.


Intracranial plaque enhancement from high resolution vessel wall magnetic resonance imaging predicts stroke recurrence.
Intracranial atherosclerosis is associated with frequent stroke recurrence. High resolution vessel wall magnetic resonance imaging (HRMRI) can provide atheroma information related to its vulnerability. We performed HRMRI in stroke patients with intracranial atherosclerosis to determine whether plaque characteristics from vessel wall imaging can predict future stroke recurrence. Between July 2011 and June 2013, acute stroke patients with symptomatic intracranial atherosclerosis were prospectively enrolled and 3-tesla HRMRI was performed on the relevant artery. The plaque enhancement was visually determined from T1 post-gadolinium enhancement image. Stroke recurrence was monitored after index event and multivariate Cox proportional hazards model was constructed to identify factors related to future stroke recurrence. A total of 138 patients were included with a median follow-up of 18 months. There were 39 stroke recurrences. Plaque enhancement was detected in 108 patients (78.3%), and 37 of them experienced stroke recurrence. Among 30 stroke patients without plaque enhancement, two patients experienced stroke recurrence. Kaplan-Meier curves demonstrated a significant difference in event free survival between the patients with plaque enhancement and those patients without plaque enhancement (event rates at year 1: 30.3% vs. 6.8%, log-rank test, p = 0.004). Multivariate Cox-regression analysis showed that the plaque enhancement from HRMRI was independently associated with stroke recurrence (hazard ratio: 7.42, 95% confidence interval: 1.74-31.75, p = 0.007). Intracranial plaque enhancement from HRMRI is associated with stroke recurrence among the patients with symptomatic intracranial atherosclerosis.


Ghrelin Axis Reveals the Interacting Influence of Central Obesity and Hypertension.
Objective: This study aimed to investigate how central obesity and hypertension modulate unacylated ghrelin (UnAG), acylated ghrelin (AG), obestatin, growth hormone (GH), and the ratios of UnAG/obestatin, AG/obestatin, and total ghrelin/obestatin. Methods: Circulatory abundances of UnAG, AG, obestatin and GH were determined in 387 Hong Kong Chinese female adults with age between 24 to 86 years based on a 2 × 2 factorial design of hypertension (blood pressure ≥140/90 mmHg) and central obesity (waist circumference or WC ≥80 cm). Participants were categorized as neither hypertensive nor centrally obese (NHNO; n = 105), hypertensive but not centrally obese (HNO; n = 102), centrally obese but not hypertensive (NHO; n = 74) and hypertensive and centrally obese (NO; n = 106). Pearson's correlation analyses were performed to detect the association between the peptides examined with WC and blood pressure. The main and interaction effects of hypertension and central obesity were examined by generalized estimating equations analyses. Results: Correlation analyses revealed that systolic blood pressure was negatively correlated with AG/obestatin, UnAG/obestatin and total ghrelin/obestatin ratios, AG, total ghrelin, and GH, while diastolic blood pressure was negatively correlated with UnAG/obestatin, total ghrelin/obestatin ratios, and GH. WC was negatively correlated with AG/obestatin, UnAG/obestatin, and total ghrelin/obestatin ratios, UnAG, AG, total ghrelin, GH, and obestatin. Interaction effects of hypertension and central obesity were observed on UnAG/obestatin, AG/obestatin and total ghrelin/obestatin ratios, and obestatin. Obestatin in NHO group was significantly higher compared to NHNO and HO groups. UnAG/obestatin, AG/obestatin, and total ghrelin/obestatin ratios were higher in NHNO group compared to HNO and HO groups. Main effects of central obesity and hypertension were observed in UnAG, total ghrelin and GH. The HO group manifested the lowest level of UnAG, total ghrelin and GH among all the groups studied. Main effect of hypertension was observed on AG, suggesting that hypertensive individuals exhibited lower levels of AG regardless of central obesity. Conclusion: Circulatory ghrelin gene products and GH exhibit different modes of modulation in response to the co-manifestation of multiple cardiovascular risk factors compared with a single risk factor alone.


The Novel Ribonucleotide Reductase Inhibitor COH29 Inhibits DNA Repair In Vitro.
COH29 [N-(4-(3,4-dihydroxyphenyl)-5-phenylthiazol-2-yl)-3,4-dihydroxybenzamide], a novel antimetabolite drug developed at City of Hope Cancer Center, has anticancer activity that stems primarily from the inhibition of human ribonucleotide reductase (RNR). This key enzyme in deoxyribonucleotide biosynthesis is the target of established clinical agents such as hydroxyurea and gemcitabine because of its critical role in DNA replication and repair. Herein we report that BRCA-1-defective human breast cancer cells are more sensitive than wild-type BRCA-1 counterparts to COH29 in vitro and in vivo. Microarray gene expression profiling showed that COH29 reduces the expression of DNA repair pathway genes, suggesting that COH29 interferes with these pathways. It is well established that BRCA1 plays a role in DNA damage repair, especially homologous recombination (HR) repair, to maintain genome integrity. In BRCA1-defective HCC1937 breast cancer cells, COH29 induced more double-strand breaks (DSBs) and DNA-damage response than in HCC1937 + BRCA1 cells. By EJ5- and DR-green fluorescent protein (GFP) reporter assay, we found that COH29 could inhibit nonhomologous end joining (NHEJ) efficiency and that no HR activity was detected in HCC1937 cells, suggesting that repression of the NHEJ repair pathway may be involved in COH29-induced DSBs in BRCA1-deficient HCC1937 cells. Furthermore, we observed an accumulation of nuclear Rad51 foci in COH29-treated HCC1937 + BRCA1 cells, suggesting that BRCA1 plays a crucial role in repairing and recovering drug-induced DNA damage by recruiting Rad51 to damage sites. In summary, we describe here additional biologic effects of the RNR inhibitor COH29 that potentially strengthen its use as an anticancer agent.


Dorsal-stream motion processing deficits persist into adulthood in Williams syndrome.
Previous studies of children with Williams syndrome (WS) have found a specific deficit in dorsal cortical stream function, indicated by poor performance in coherence thresholds for motion compared to form. Here we investigated whether this is a transient developmental feature or a persisting aspect of cerebral organization in WS. Motion and form coherence thresholds were tested in a group of 45 WS individuals aged 16-42 years, and 19 normal adult controls. Although there was considerable variation in the coherence thresholds across individuals with WS, the WS group showed overall worse performance than controls. A significant group x threshold condition interaction showed a substantially greater performance deficit for motion than for form coherence in the WS group relative to controls. This result suggests that the motion deficit is an enduring feature in WS and is a marker for one aspect of dorsal-stream vulnerability.


Trends in Natural Decannulation in Patients with Robin Sequence: A Twenty-five Year Retrospective Review.
Robin sequence (RS) consists of micrognathia and glossoptosis that result in upper airway obstruction (UAO). In RS patients who undergo tracheostomy, long-term goals include natural decannulation (ND) without further surgical airway intervention. The objective of this study was to identify long-term trends in the rate and length of time to ND. Retrospective chart review on 144 patients with RS treated from 1995 to 2020 at a pediatric tertiary care center. Patients were grouped by year of tracheostomy. Demographic data, UAO management, postoperative care, complications, and time to decannulation were recorded. Thirty-six patients met the inclusion criteria. Tracheostomy was performed at a median age of 45.5 days. 19 (53%) patients experienced ND at a median time of 66.1 months. ND rate was higher in non-syndromic patients (93% non-syndromic vs 27% syndromic; P < .0001) and during the first study period (1995-2006: 78%, 2007-2020: 28%; P = .003). Cox proportional-hazard regression demonstrated that white race [aHR 0.15 (0.03-0.8); P = .023] and higher birthweight [aHR 0.9 (0.8-0.98); P = .018] were associated with a higher likelihood of ND while a syndromic diagnosis had a negative association with ND [aHR 12.5 (3.3-50.0); P < .001]. Our study documented that ND in patients with RS who underwent tracheostomy was significantly associated with ethnicity, birthweight, and syndromic status. The negative impact on successful ND was most observed in patients with syndromic associations. Additionally, ND rates are lower in the 2007 to 2020 subgroup. We suspect this is because alternative management techniques such as tongue lip adhesion and mandibular distraction osteogenesis became primary surgical treatment in severe RS upper airway obstruction at our institution starting in 2007.


The application of root mean square electrocardiography (RMS ECG) for the detection of acquired and congenital long QT syndrome.
Precise measurement of the QT interval is often hampered by difficulty determining the end of the low amplitude T wave. Root mean square electrocardiography (RMS ECG) provides a novel alternative measure of ventricular repolarization. Experimental data have shown that the interval between the RMS ECG QRS and T wave peaks (RTPK) closely reflects the mean ventricular action potential duration while the RMS T wave width (TW) tracks the dispersion of repolarization timing. Here, we tested the precision of RMS ECG to assess ventricular repolarization in humans in the setting of drug-induced and congenital Long QT Syndrome (LQTS). RMS ECG signals were derived from high-resolution 24 hour Holter monitor recordings from 68 subjects after receiving placebo and moxifloxacin and from standard 12 lead ECGs obtained in 97 subjects with LQTS and 97 age- and sex-matched controls. RTPK, QTRMS and RMS TW intervals were automatically measured using custom software and compared to traditional QT measures using lead II. All measures of repolarization were prolonged during moxifloxacin administration and in LQTS subjects, but the variance of RMS intervals was significantly smaller than traditional lead II measurements. TW was prolonged during moxifloxacin and in subjects with LQT-2, but not LQT-1 or LQT-3. These data validate the application of RMS ECG for the detection of drug-induced and congenital LQTS. RMS ECG measurements are more precise than the current standard of care lead II measurements.


Online cognitive-behavioural therapy for traumatically bereaved people: study protocol for a randomised waitlist-controlled trial.
The traumatic death of a loved one, such as death due to a traffic accident, can precipitate persistent complex bereavement disorder (PCBD) and comorbid post-traumatic stress disorder (PTSD) and depression. Waitlist-controlled trials have shown that grief-specific cognitive-behavioural therapy (CBT) is an effective treatment for such mental health problems. This is the first study that will examine the effectiveness of online CBT (vs waitlist controls) in a sample exclusively comprised of people bereaved by a traumatic death. Our primary hypothesis is that people allocated to the online CBT condition will show larger reductions in PCBD, PTSD and depression symptom levels at post-treatment than people allocated to a waitlist. We further expect that reductions in symptom levels during treatment are associated with reductions of negative cognitions and avoidance behaviours and the experience of fewer accident-related stressors. Moreover, the effect of the quality of the therapeutic alliance on treatment effects and drop-out rates will be explored. A two-arm (online CBT vs waiting list) open-label parallel randomised controlled trial will be conducted. Participants will complete questionnaires at pretreatment and 12 and 20 weeks after study enrolment. Eligible for participation are Dutch adults who lost a loved one at least 1 year earlier due to a traffic accident and report clinically relevant levels of PCBD, PTSD and/or depression. Multilevel modelling will be used. Ethics approval has been received by the Medical Ethics Review Board of the University Medical Center Groningen (METc UMCG: M20.252121). This study will provide new insights in the effectiveness of online CBT for traumatically bereaved people. If the treatment is demonstrated to be effective, it will be made publicly accessible. Findings will be disseminated among lay people (eg, through newsletters and media performances), our collaborators (eg, through presentations at support organisations), and clinicians and researchers (eg, through conference presentations and scientific journal articles). NL7497.


Estimation of Survival Probabilities for Use in Cost-effectiveness Analyses: A Comparison of a Multi-state Modeling Survival Analysis Approach with Partitioned Survival and Markov Decision-Analytic Modeling.
Modeling of clinical-effectiveness in a cost-effectiveness analysis typically involves some form of partitioned survival or Markov decision-analytic modeling. The health states progression-free, progression and death and the transitions between them are frequently of interest. With partitioned survival, progression is not modeled directly as a state; instead, time in that state is derived from the difference in area between the overall survival and the progression-free survival curves. With Markov decision-analytic modeling, a priori assumptions are often made with regard to the transitions rather than using the individual patient data directly to model them. This article compares a multi-state modeling survival regression approach to these two common methods. As a case study, we use a trial comparing rituximab in combination with fludarabine and cyclophosphamide v. fludarabine and cyclophosphamide alone for the first-line treatment of chronic lymphocytic leukemia. We calculated mean Life Years and QALYs that involved extrapolation of survival outcomes in the trial. We adapted an existing multi-state modeling approach to incorporate parametric distributions for transition hazards, to allow extrapolation. The comparison showed that, due to the different assumptions used in the different approaches, a discrepancy in results was evident. The partitioned survival and Markov decision-analytic modeling deemed the treatment cost-effective with ICERs of just over £16,000 and £13,000, respectively. However, the results with the multi-state modeling were less conclusive, with an ICER of just over £29,000. This work has illustrated that it is imperative to check whether assumptions are realistic, as different model choices can influence clinical and cost-effectiveness results.


Evolutionarily conserved and non-conserved retrovirus restriction activities of artiodactyl APOBEC3F proteins.
The APOBEC3 proteins are unique to mammals. Many inhibit retrovirus infection through a cDNA cytosine deamination mechanism. HIV-1 neutralizes this host defense through Vif, which triggers APOBEC3 ubiquitination and degradation. Here, we report an APOBEC3F-like, double deaminase domain protein from three artiodactyls: cattle, pigs and sheep. Like their human counterparts, APOBEC3F and APOBEC3G, the artiodactyl APOBEC3F proteins are DNA cytosine deaminases that locate predominantly to the cytosol and can inhibit the replication of HIV-1 and MLV. Retrovirus restriction is attributable to deaminase-dependent and -independent mechanisms, as deaminase-defective mutants retain significant anti-retroviral activity. However, unlike human APOBEC3F and APOBEC3G, the artiodactyl APOBEC3F proteins have an active N-terminal DNA cytosine deaminase domain, which elicits a broader dinucleotide deamination preference, and they are resistant to HIV-1 Vif. These data indicate that DNA cytosine deamination; sub-cellular localization and retrovirus restriction activities are conserved in mammals, whereas active site location, local mutational preferences and Vif susceptibility are not. Together, these studies indicate that some properties of the mammal-specific, APOBEC3-dependent retroelement restriction system are necessary and conserved, but others are simultaneously modular and highly adaptable.


Races of Phytophthora sojae in Arkansas Soybean Fields and Their Effects on Commonly Grown Soybean Cultivars.
Isolates of Phytophthora sojae were collected during 1995 to 1998 from soil samples collected in 23 Arkansas soybean fields in 14 counties, and characterized by race. A total of seven races (races 2, 10, 14, 15, 24, 26, and 38) were found. Races 10, 24, and 15 were the most common and comprised 47, 22, and 9% of the 32 isolates, respectively. A single isolate each of races 2, 14, 26, and 38 also was found. Three of the isolates collected could not be characterized to race due to inconsistent results. In 1997 and 1998, a portion of a single soybean field at the University of Arkansas Southeast Research and Experiment Center near Rohwer, AR was surveyed intensively for P. sojae. The area was planted each year to the P. sojae-susceptible cv. Williams and both plants and soil were collected to assay for P. sojae from 16 and 28 plots (4.9 by 7.6 m) in 1997 and 1998, respectively. A total of 83 isolates were collected (11 from plants and 72 from soil), and found to represent 13 pathotypes, including 6 with virulence formulae that have not been described previously. Nine commercial soybean cultivars representing a range of reported resistance and tolerance to Phytophthora root and stem rot were screened for resistance to races 10, 15, and 26 of P. sojae using both hypocotyl injection and inoculum layer techniques. Cvs. Manokin, Hartz Variety 5545, and Riverside 499 were consistently resistant to all of the races using both inoculation methods. These results indicate that, although considerable pathogenic variability in P. sojae exists in soybean fields in Arkansas, cultivars with effective resistance are available to help growers manage Phytophthora root and stem rot.


Use of the desmopressin test in the differential diagnosis of pseudo-Cushing state from Cushing's disease.
The desmopressin (DDAVP) test has been proposed to discriminate Cushing's disease (CD) from pseudo-Cushing states (PC); however, current information on its value is scarce and contradictory. The aim of the study was to assess the ability of the DDAVP test in distinguishing between these conditions, with emphasis on subjects with mild hypercortisolism. We conducted a retrospective/prospective study at the Division of Endocrinology, Polytechnic University of Marche, Ancona, Italy. The study included 52 subjects with CD, 28 with PC, and 31 control subjects (CT). We performed the DDAVP test and standard diagnostic procedures for the diagnosis of Cushing's syndrome. The diagnosis/exclusion of CD was measured. Interpretation of the DDAVP test based on percentage and absolute increment of cortisol and ACTH did not afford acceptable values of both sensitivity (SE) and specificity (SP). CD diagnosis based on simultaneous positivity for basal serum cortisol greater than 331 nmol/liter and absolute ACTH increment greater than 4 pmol/liter and its exclusion in subjects negative for one or both measures yielded an SE of 90.3% and an SP of 91.5%. The approach was also highly effective in distinguishing PC from: 1) CD with moderate values of urinary free cortisol (SE, 86.9%; SP, 92.8%); 2) CD with moderate values of serum cortisol after dexamethasone suppression (SE, 86.6%; SP, 92.8%); and 3) CD with moderate values of midnight serum cortisol (SE, 100%; SP, 92.8%). Interpretation of the DDAVP test through a combination of parameters allowed effective discrimination of CD from PC, even in subjects with mild hypercortisolism.


Hyperglycemia Impairs Neutrophil-Mediated Bacterial Clearance in Mice Infected with the Lyme Disease Pathogen.
Insulin-insufficient type 1 diabetes is associated with attenuated bactericidal function of neutrophils, which are key mediators of innate immune responses to microbes as well as pathological inflammatory processes. Neutrophils are central to immune responses to the Lyme pathogen Borrelia burgdorferi. The effect of hyperglycemia on host susceptibility to and outcomes of B. burgdorferi infection has not been examined. The present study investigated the impact of sustained obesity-independent hyperglycemia in mice on bacterial clearance, inflammatory pathology and neutrophil responses to B. burgdorferi. Hyperglycemia was associated with reduced arthritis incidence but more widespread tissue colonization and reduced clearance of bacterial DNA in multiple tissues including brain, heart, liver, lung and knee joint. B. burgdorferi uptake and killing were impaired in neutrophils isolated from hyperglycemic mice. Thus, attenuated neutrophil function in insulin-insufficient hyperglycemia was associated with reduced B. burgdorferi clearance in target organs. These data suggest that investigating the effects of comorbid conditions such as diabetes on outcomes of B. burgdorferi infections in humans may be warranted.


Staged Bilateral Nucleus Caudalis DREZ Lesioning for Persistent Idiopathic Facial Pain: Exceptional Case Report.
Persistent idiopathic facial pain (PIFP) is a rare but challenging type of facial pain. How to manipulate such a problem becomes more contentious in cases with bilateral complaints. In situations when conventional treatments fail to reduce the pain, some treatment options like nucleus caudalis dorsal root entry zone (DREZ) lesioning shine as valuable means of managing such intractable problems. Herein, we report an individual with bilateral PIFP in which the patient suffered from intractable pain and was the first to undergo bilateral nucleus caudalis DREZ operation for PIFP. The patient was followed up and the procedure resulted in immediate and complete pain relief.


Allergic Bronchopulmonary Aspergillosis in Children with Cystic Fibrosis: An Update on the Newest Diagnostic Tools and Therapeutic Approaches.
Cystic fibrosis (CF), the most common autosomal-recessive genetic disease in the Caucasian population, is characterized by frequent respiratory infections and progressive lung disease. Fungal species are commonly found in patients with CF, and among them, Aspergillus fumigatus is the most frequently isolated. While bacteria, particularly Pseudomonas aeruginosa, have a well-established negative effect on CF lung disease, the impact of fungal infections remains unclear. In patients with CF, inhalation of Aspergillus conidia can cause allergic bronchopulmonary aspergillosis (ABPA), a Th2-mediated lung disease that can contribute to disease progression. Clinical features, diagnostic criteria and treatment of ABPA are still a matter of debate. Given the consequences of a late ABPA diagnosis or the risk of ABPA overdiagnosis, it is imperative that the diagnostic criteria guidelines are reviewed and standardized. Along with traditional criteria, radiological features are emerging as tools for further classification as well as novel immunological tests. Corticosteroids, itraconazole and voriconazole continue to be the bedrock of ABPA therapy, but other molecules, such as posaconazole, vitamin D, recombinant INF-γ and Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) modulators, have been showing positive results. However, few studies have been conducted recruiting CF patients, and more research is needed to improve the prevention and the classification of clinical manifestations as well as to personalize treatment. Early recognition and early treatment of fungal infections may be fundamental to prevent progression of CF disease. The aim of this narrative review is to give an update on ABPA in children with CF.


Maastricht Delphi consensus on event definitions for classification of recurrence in breast cancer research.
In breast cancer studies, many different endpoints are used. Definitions are often not provided or vary between studies. For instance, "local recurrence" may include different components in similar studies. This limits transparency and comparability of results. This project aimed to reach consensus on the definitions of local event, second primary breast cancer, regional and distant event for breast cancer studies. The RAND-UCLA Appropriateness method (modified Delphi method) was used. A Consensus Group of international breast cancer experts was formed, including representatives of all involved clinical disciplines. Consensus was reached in two rounds of online questionnaires and one meeting. Twenty-four international breast cancer experts participated. Consensus was reached on 134 items in four categories. Local event is defined as any epithelial breast cancer or ductal carcinoma in situ (DCIS) in the ipsilateral breast, or skin and subcutaneous tissue on the ipsilateral thoracic wall. Second primary breast cancer is defined as epithelial breast cancer in the contralateral breast. Regional events are breast cancer in ipsilateral lymph nodes. A distant event is breast cancer in any other location. Therefore, this includes metastasis in contralateral lymph nodes and breast cancer involving the sternal bone. If feasible, tissue sampling of a first, solitary, lesion suspected for metastasis is highly recommended. This project resulted in consensus-based event definitions for classification of recurrence in breast cancer research. Future breast cancer research projects should adopt these definitions to increase transparency. This should facilitate comparison of results and conducting reviews as well as meta-analysis.


The efficacy and safety of glucose control algorithms in intensive care: a pilot study of the Survival Using Glucose Algorithm Regulation (SUGAR) trial.
The benefits, harms and feasibility of intensive insulin therapy in critically ill patients remain unclear. Several single center studies have attempted to demonstrate the benefit of intensive insulin therapy in critically ill patients with variable results. We conducted a pilot randomized trial to assess the feasibility, safety and clinical outcomes of preprinted glucose management algorithms before the initiation of a large multicenter trial. Within 48 hours of admission to the intensive care unit, we randomized mechanically ventilated patients to either the "high" group (target serum glucose concentration 9-11 mmol/l) or the "low" group (target serum glucose concentration 5-7 mmol/l). To assess feasibility we measured the time to reach target glucose range, time in target range, morning glucose concentrations, average daily glucose concentrations, and number of crossovers. To assess safety, we measured the number of hypoglycemic events (serum glucose <2.2 mmol/l), and other serious adverse events such as cardiac arrests and seizures. Sixty-eight patients were enrolled (35 in the high group and 33 in the low group). During the first week, the median proportions of time spent in the target range were 35.7% and 53.0% for the high and low groups, respectlively (p = 0.0001). Morning glucose concentrations were 8.3 +/-1.6 mmol/l and 6.2 -/+1.2 mmol/l. One (2.9%) and 8 (24.2%) episodes of hypoglycemia (<2.2 mmol/l) occurred in the high and low groups, reflecting 0.002 and 0.03 hypoglycemic events per patient-day, respectlively. This pilot trial of intensive insulin therapy identified numerous challenges that helped in the preparation of an international multicenter randomized trial of intensive insulin therapy to evaluate benefits and harms.


Recently activated naive CD4 T cells can help resting B cells, and can produce sufficient autocrine IL-4 to drive differentiation to secretion of T helper 2-type cytokines.
Development of T cells during primary responses was investigated using pigeon cytochrome C-specific naive Th from TCR transgenic mice. Naive CD4 cells did not activate and help resting B cells. This failure was found to be primarily because the resting B cells were incapable of stimulating the naive Th. Provision of a costimulatory signal such as anti-CD28, or addition of APCs that express costimulatory molecules, such as dendritic cells, activated B cells, and B7+ and B7+ICAM(+)-expressing fibroblasts, induced naive Th activation and promoted T cell-dependent help for IgM secretion. T cell activation for as little as 24 h promoted helper activity, and Ig secretion required production of small amounts of IL-4 by the activated naive Th. On initial stimulation, naive Th secrete only IL-2. By mRNA analysis, activated naive Th were also shown to produce IL-4, however induction of IL-4 message only occurred 24 h after initial activation and required additional stimulation with Ag. A single exposure of naive CD4 to Ag/APC followed by 4 to 12 days in culture led to generation of effector Th which secreted IL-2 and some IFN-gamma, and no detectable IL-4 or IL-5, and which could only help B cells to IgM secretion. In contrast, similar cultures that received Ag/APC one or more times during this period generated effector cells capable of secreting easily detectable titers of IL-4 and IL-5, as well as IL-2 and IFN-gamma, and able to now promote IgG1 and IgE responses. Generation of these Th0-like effectors was accompanied by increasing amounts of IL-4 secreted during the culture period after each restimulation, and addition of anti-IL-4 in culture inhibited development of the capacity to produce Th2 cytokines. These studies reinforce the notion that naive CD4 must interact with a costimulatory professional APC, rather than a resting B cell, for initiation of the primary response, but show that such an interaction can result in rapid development of the ability to interact with and provide cognate help to B cells. They also suggest that if activated naive CD4 cells receive multiple stimulations from Ag/APC, enough endogenous IL-4 can be produced to drive differentiation into effectors secreting type 2 cytokines. The existence of such an autocrine feedback mechanism suggests that the amount and availability of Ag could influence the nature and polarization of the Th response.


Acute axonal dystrophy caused by fluorocitrate: the role of mitochondrial swelling.
Fluorocitrate, a Krebs cycle inhibitor, induices neurons to rapidly expel multitudinous lysosomes, mitochondria, and other cytoplasmic constituents into their axons. After convulsive seizures commence, spectacular axonal balloons develop owing to obstruction of axonal flow by "log jams" of extruded organelles. A swelling of neuronal mitochondria is apparently responsible for the disgorgement of cytoplasmic material into axons.


In vivo quantification of demyelination and recovery using compartment-specific diffusion MRI metrics validated by electron microscopy.
There is a need for accurate quantitative non-invasive biomarkers to monitor myelin pathology in vivo and distinguish myelin changes from other pathological features including inflammation and axonal loss. Conventional MRI metrics such as T2, magnetization transfer ratio and radial diffusivity have proven sensitivity but not specificity. In highly coherent white matter bundles, compartment-specific white matter tract integrity (WMTI) metrics can be directly derived from the diffusion and kurtosis tensors: axonal water fraction, intra-axonal diffusivity, and extra-axonal radial and axial diffusivities. We evaluate the potential of WMTI to quantify demyelination by monitoring the effects of both acute (6weeks) and chronic (12weeks) cuprizone intoxication and subsequent recovery in the mouse corpus callosum, and compare its performance with that of conventional metrics (T2, magnetization transfer, and DTI parameters). The changes observed in vivo correlated with those obtained from quantitative electron microscopy image analysis. A 6-week intoxication produced a significant decrease in axonal water fraction (p<0.001), with only mild changes in extra-axonal radial diffusivity, consistent with patchy demyelination, while a 12-week intoxication caused a more marked decrease in extra-axonal radial diffusivity (p=0.0135), consistent with more severe demyelination and clearance of the extra-axonal space. Results thus revealed increased specificity of the axonal water fraction and extra-axonal radial diffusivity parameters to different degrees and patterns of demyelination. The specificities of these parameters were corroborated by their respective correlations with microstructural features: the axonal water fraction correlated significantly with the electron microscopy derived total axonal water fraction (ρ=0.66; p=0.0014) but not with the g-ratio, while the extra-axonal radial diffusivity correlated with the g-ratio (ρ=0.48; p=0.0342) but not with the electron microscopy derived axonal water fraction. These parameters represent promising candidates as clinically feasible biomarkers of demyelination and remyelination in the white matter.


Oxygen pathway modeling estimates high reactive oxygen species production above the highest permanent human habitation.
The production of reactive oxygen species (ROS) from the inner mitochondrial membrane is one of many fundamental processes governing the balance between health and disease. It is well known that ROS are necessary signaling molecules in gene expression, yet when expressed at high levels, ROS may cause oxidative stress and cell damage. Both hypoxia and hyperoxia may alter ROS production by changing mitochondrial Po2 (PmO2). Because PmO2 depends on the balance between O2 transport and utilization, we formulated an integrative mathematical model of O2 transport and utilization in skeletal muscle to predict conditions to cause abnormally high ROS generation. Simulations using data from healthy subjects during maximal exercise at sea level reveal little mitochondrial ROS production. However, altitude triggers high mitochondrial ROS production in muscle regions with high metabolic capacity but limited O2 delivery. This altitude roughly coincides with the highest location of permanent human habitation. Above 25,000 ft., more than 90% of exercising muscle is predicted to produce abnormally high levels of ROS, corresponding to the "death zone" in mountaineering.


Comparative analysis of the Sokal, Euro and European Treatment and Outcome Study score in prognostication of Indian chronic myeloid leukemia-chronic phase patients on imatinib.
The ultimate goal for CML management is risk stratification of the patients to design the appropriate treatment approach. The Sokal, Euro and EUTOS risk scores were established to prognosticate the patients on therapy. To perform a comparative assessment of the Sokal, Euro and EUTOS prognostic score in Indian CML-CP patients on imatinib. This is a retrospective study performed in 260 Ph+ CML-CP patients who were administered oral imatinib (400 mg/day). 166/260 were males and 94/260 were females (M: F::1.6:1) with median age 35 years (range 20-70). 92 (35.38%), 125 (48.07%) and 43 (16.5%) patients were divided into low, intermediate and high risk Sokal score respectively. 102 (39.23%), 106 (40.76%) and 52 (20%) patients were discriminated into low, intermediate and high risk Euro score respectively. 210 (80.7%) and 50 (19.2%) patients were divided into low and high risk EUTOS score. Cumulative incidence of MMR for low, intermediate and high-risk Sokal score was 87%, 76% and 84% respectively (P = 0.016). Incidence of MMR in low, intermediate and high-risk Euro score was 93%, 85% and 68% respectively (P = 0.001). Incidence of MMR was 80 % and 81% for low and high risk EUTOS score (P = 0.764). Both EFS and OS are significantly correlated with Sokal score (P = 0.004, P = 0.007) and Euro score (P = 0.009, P = 0.001) but not with EUTOS score (P = 0.581, P = 0.927). The present study highlights the significant prognostic role of Sokal and Euro score in predicting the treatment outcome of the CML- CP patients on imatinib.


PAX6 Expression and Retinal Cell Death in a Transgenic Mouse Model for Acute Angle-Closure Glaucoma.
PAX6 is a highly conserved protein essential for the control of eye development both in invertebrates and vertebrates. PAX6 expression persists in the adult inner retina, but little is known about its functions after completion of retinal differentiation. Therefore, we investigated PAX6 expression in wild-type and calcitonin receptor-like receptor transgenic (CLR(SMαA)) mice with angle-closure glaucoma. Intraocular pressure was measured by indentation tonometry in anesthetized mice. Eyes of mice of both genotypes were enucleated at various ages and retinas were processed for morphological analysis and PAX6 immunostaining. The content of PAX6 in retinal extracts was estimated by Western blot analysis. Retinal expression of glaucoma-related genes was analyzed by reverse transcription-polymerase chain reaction. Control mice showed normal retinal morphology between p22 and p428 with steady PAX6 expression in the ganglion cell layer (GCL) and the inner nuclear layer (INL). CLR(SMαA) mice examined between p22 and p82 exhibited increased intraocular pressure and a progressive decrease in cell number including PAX6-expressing cells in the GCL. The INL was not affected up to postnatal day 42. Later, a significant increase in PAX6-expressing cells concomitant with an overall loss of cells was observed in the INL of CLR(SMαA) as compared with control mice. Retinal up-regulation of glaucoma-related genes was furthermore observed. Distinctive changes of PAX6 expression in the inner retina of CLR(SMαA) mice suggest a role in regulatory mechanisms involved in glaucoma-related retinal cell death. The selective increase of PAX6 expression in the degenerating INL of CLR(SMαA) mice may represent an attempt to preserve retinal cytoarchitecture.


Prevalence of Depression and Depressive Symptoms Among Resident Physicians: A Systematic Review and Meta-analysis.
Physicians in training are at high risk for depression. However, the estimated prevalence of this disorder varies substantially between studies. To provide a summary estimate of depression or depressive symptom prevalence among resident physicians. Systematic search of EMBASE, ERIC, MEDLINE, and PsycINFO for studies with information on the prevalence of depression or depressive symptoms among resident physicians published between January 1963 and September 2015. Studies were eligible for inclusion if they were published in the peer-reviewed literature and used a validated method to assess for depression or depressive symptoms. Information on study characteristics and depression or depressive symptom prevalence was extracted independently by 2 trained investigators. Estimates were pooled using random-effects meta-analysis. Differences by study-level characteristics were estimated using meta-regression. Point or period prevalence of depression or depressive symptoms as assessed by structured interview or validated questionnaire. Data were extracted from 31 cross-sectional studies (9447 individuals) and 23 longitudinal studies (8113 individuals). Three studies used clinical interviews and 51 used self-report instruments. The overall pooled prevalence of depression or depressive symptoms was 28.8% (4969/17,560 individuals, 95% CI, 25.3%-32.5%), with high between-study heterogeneity (Q = 1247, τ2 = 0.39, I2 = 95.8%, P < .001). Prevalence estimates ranged from 20.9% for the 9-item Patient Health Questionnaire with a cutoff of 10 or more (741/3577 individuals, 95% CI, 17.5%-24.7%, Q = 14.4, τ2 = 0.04, I2 = 79.2%) to 43.2% for the 2-item PRIME-MD (1349/2891 individuals, 95% CI, 37.6%-49.0%, Q = 45.6, τ2 = 0.09, I2 = 84.6%). There was an increased prevalence with increasing calendar year (slope = 0.5% increase per year, adjusted for assessment modality; 95% CI, 0.03%-0.9%, P = .04). In a secondary analysis of 7 longitudinal studies, the median absolute increase in depressive symptoms with the onset of residency training was 15.8% (range, 0.3%-26.3%; relative risk, 4.5). No statistically significant differences were observed between cross-sectional vs longitudinal studies, studies of only interns vs only upper-level residents, or studies of nonsurgical vs both nonsurgical and surgical residents. In this systematic review, the summary estimate of the prevalence of depression or depressive symptoms among resident physicians was 28.8%, ranging from 20.9% to 43.2% depending on the instrument used, and increased with calendar year. Further research is needed to identify effective strategies for preventing and treating depression among physicians in training.


Emergency Department care of childhood epistaxis.
The aim of this review is to determine an efficient and safe primary strategy care for paediatric epistaxis. We searched PubMed and Cochrane databases for studies referenced with key words 'epistaxis AND childhood'. This search yielded 32 research articles about primary care in childhood epistaxis (from 1989 to 2015). Bibliographic references found in these articles were also examined to identify pertinent literature. We compared our results to the specific management of adult epistaxis classically described in the literature. Epistaxis is one of the most common reasons for referral of children to a hospital ENT outpatient department. The bleeding usually originates from the anterior septum, as opposed to adults. Crusting, digital trauma, foreign bodies and nasal colonisation with Staphylococcus aureus have been suggested as specific nosebleed factors in children. Rare aetiologies as juvenile nasopharyngeal angiofibroma appear later during adolescence. There are different modes of management of mild epistaxis, which begin with clearing out blood clots and bidigital compression. An intranasal topical local anaesthetic and decongestant can be used over 6 years of age. In case of active bleeding, chemical cauterisation is preferred to anterior packing and electric cauterisation but is only feasible if the bleeding site is clearly visible. In case of non-active bleeding in children, and in those with recurrent idiopathic epistaxis, antiseptic cream is easy to apply and can avoid 'acrobatic' cauterisation liable to cause further nasal cavity trauma. Aetiologies and treatment vary with patient age and the existence or not of active bleeding at the time of the examination. Local treatments are usually easy to perform, but physicians have to ponder their indications depending on the possible complications in order to inform parents and to know paediatric epistaxis specificities.


Design and mechanical properties of insect cuticle.
Since nearly all adult insects fly, the cuticle has to provide a very efficient and lightweight skeleton. Information is available about the mechanical properties of cuticle-Young's modulus of resilin is about 1 MPa, of soft cuticles about 1 kPa to 50 MPa, of sclerotised cuticles 1-20 GPa; Vicker's Hardness of sclerotised cuticle ranges between 25 and 80 kgf mm(-2); density is 1-1.3 kg m(-3)-and one of its components, chitin nanofibres, the Young's modulus of which is more than 150 GPa. Experiments based on fracture mechanics have not been performed although the layered structure probably provides some toughening. The structural performance of wings and legs has been measured, but our understanding of the importance of buckling is lacking: it can stiffen the structure (by elastic postbuckling in wings, for example) or be a failure mode. We know nothing of fatigue properties (yet, for instance, the insect wing must undergo millions of cycles, flexing or buckling on each cycle). The remarkable mechanical performance and efficiency of cuticle can be analysed and compared with those of other materials using material property charts and material indices. Presented in this paper are four: Young's modulus-density (stiffness per unit weight), specific Young's modulus-specific strength (elastic hinges, elastic energy storage per unit weight), toughness-Young's modulus (fracture resistance under various loading conditions), and hardness (wear resistance). In conjunction with a structural analysis of cuticle these charts help to understand the relevance of microstructure (fibre orientation effects in tendons, joints and sense organs, for example) and shape (including surface structure) of this fibrous composite for a given function. With modern techniques for analysis of structure and material, and emphasis on nanocomposites and self-assembly, insect cuticle should be the archetype for composites at all levels of scale.


Applying the Theory of Planned Behavior to reporting of forced sex by African-American college women.
Forced sex is a public health issue affecting many college women. Despite physical and mental health consequences, and multiple prevention programs on college campuses, most sexual violence goes unreported (Fisher, Daigle, Cullen, & Turner, 2003). The purpose of this research was to determine the significant attitudes and beliefs that are associated with reporting of forced sexual experiences. Guided by the Theory of Planned Behavior (TPB), the study used a predictive exploratory design to explore the association of intention to report forced sex with attitudes and beliefs (Ajzen, 1991). A convenience sample of 144 African-American women who were attending a private college in the south completed a survey. Women who expressed more favorable attitudes towards reporting, perceived reporting as being supported by important referents, and perceived more control over reporting, reported stronger intentions to report forced sex. The analysis supported the utility of TPB in predicting the intention to report forced sex by African-American college women. Theoretically significant and clinically relevant prevention strategies should incorporate important referents, address salient beliefs, and determine ways to increase perceived behavioral control.


CSF-biomarkers in Olympic boxing: diagnosis and effects of repetitive head trauma.
Sports-related head trauma is common but still there is no established laboratory test used in the diagnostics of minimal or mild traumatic brain injuries. Further the effects of recurrent head trauma on brain injury markers are unknown. The purpose of this study was to investigate the relationship between Olympic (amateur) boxing and cerebrospinal fluid (CSF) brain injury biomarkers. The study was designed as a prospective cohort study. Thirty Olympic boxers with a minimum of 45 bouts and 25 non-boxing matched controls were included in the study. CSF samples were collected by lumbar puncture 1-6 days after a bout and after a rest period for at least 14 days. The controls were tested once. Biomarkers for acute and chronic brain injury were analysed. NFL (mean ± SD, 532±553 vs 135±51 ng/L p = 0.001), GFAP (496±238 vs 247±147 ng/L p<0.001), T-tau (58±26 vs 49±21 ng/L p<0.025) and S-100B (0.76±0.29 vs 0.60±0.23 ng/L p = 0.03) concentrations were significantly increased after boxing compared to controls. NFL (402±434 ng/L p = 0.004) and GFAP (369±113 ng/L p = 0.001) concentrations remained elevated after the rest period. Increased CSF levels of T-tau, NFL, GFAP, and S-100B in >80% of the boxers demonstrate that both the acute and the cumulative effect of head trauma in Olympic boxing may induce CSF biomarker changes that suggest minor central nervous injuries. The lack of normalization of NFL and GFAP after the rest period in a subgroup of boxers may indicate ongoing degeneration. The recurrent head trauma in boxing may be associated with increased risk of chronic traumatic brain injury.


Glyphosate induces neurotoxicity in zebrafish.
Glyphosate based herbicides (GBH) like Roundup(®) are used extensively in agriculture as well as in urban and rural settings as a broad spectrum herbicide. Its mechanism of action was thought to be specific only to plants and thus considered safe and non-toxic. However, mounting evidence suggests that GBHs may not be as safe as once thought as initial studies in frogs suggest that GBHs may be teratogenic. Here we utilize the zebrafish vertebrate model system to study early effects of glyphosate exposure using technical grade glyphosate and the Roundup(®) Classic formulation. We find morphological abnormalities including cephalic and eye reductions and a loss of delineated brain ventricles. Concomitant with structural changes in the developing brain, using in situ hybridization analysis, we detect decreases in genes expressed in the eye, fore and midbrain regions of the brain including pax2, pax6, otx2 and ephA4. However, we do not detect changes in hindbrain expression domains of ephA4 nor exclusive hindbrain markers krox-20 and hoxb1a. Additionally, using a Retinoic Acid (RA) mediated reporter transgenic, we detect no alterations in the RA expression domains in the hindbrain and spinal cord, but do detect a loss of expression in the retina. We conclude that glyphosate and the Roundup(®) formulation is developmentally toxic to the forebrain and midbrain but does not affect the hindbrain after 24 h exposure.


[Critical events in the operating room among 1,440,776 patients with ASA PS 1 for elective surgery].
The Japanese Society of Anesthesiologists (JSA) survey of critical incidents in the operating room has shown that preoperative complications are the leading causes of critical incidents, and affect the occurrence, severity and outcome of critical incidents which are due to causes other than preoperative complications. Causes of critical events in the operating room were examind in patients for elective surgery with American Society of Anesthesiologists physical status (ASA PS) 1. JSA has conducted annual surveys of critical incidents in the operating room by sending and collecting confidential questionnaires to all JSA Certified Training Hospitals. From 1999 to 2002, 3,855,384 anesthesia patients were registered. Among these, 1,440,776 patients with ASA PS 1 for elective surgery were analyzed. The causes of critical incidents were classified as follows: totally attributable to anesthetic management (AM), mainly to intraoperative pathological events (IP), to preoperative complications (PC), and to surgical management (SM). IP consists of coronary ischemia mainly due to coronary vasospasm, arrhythmias, pulmonary embolism, and other conditions. The incidences of cardiac arrest, critical incidents other than cardiac arrest and subsequent death were 9.86, 59.41 and 3.12 per 100,000 anesthesia cases, respectively. IP and SM were responsible for 36.6% and 34.5% of cardiac arrest, respectively. AM and SM were responsible for 46.7% and 26.8% of critical incidents other than cardiac arrest, respectively. SM, IP and AM were responsible for 66.7%, 22.2% and 4.4% of subsequent deaths (within 7 postoperative days), respectively. Coronary ischemia and pulmonary embolism were the main causes of death due to IP. The incidences of cardiac arrest and death totally attributable to AM were 1.87 and 0.14 per 100,000 anesthesia cases, respectively. Medication problems were responsible for 48.1% of arrests, while airway/ventilation problems were for 57.2% of critical incidents other than arrest. Human factors (SM combined with AM) were responsible for 53.5%, 73.5%, and 71.1% of cardiac arrest, critical incidents other than arrest and death, respectively. Even in elective patients with good physical status, non-lethal incidents were not rare, and lethal incidents were also reported. We should pay significant attention to the following findings, and take some measures to overcome these problems especially related to human factors. Firstly, SM badly harmed some operative patients. Secondly, coronary vasospasm and pulmonary embolism were the main causes of death due to IP. Thirdly, drug administration and airway/ventilation management were the major causes of critical incidents totally attributable to AM. Human factors were responsible for 70.6% of critical incidents and 71.1% of deaths.


Hi-Jack: a novel computational framework for pathway-based inference of host-pathogen interactions.
Pathogens infect their host and hijack the host machinery to produce more progeny pathogens. Obligate intracellular pathogens, in particular, require resources of the host to replicate. Therefore, infections by these pathogens lead to alterations in the metabolism of the host, shifting in favor of pathogen protein production. Some computational identification of mechanisms of host-pathogen interactions have been proposed, but it seems the problem has yet to be approached from the metabolite-hijacking angle. We propose a novel computational framework, Hi-Jack, for inferring pathway-based interactions between a host and a pathogen that relies on the idea of metabolite hijacking. Hi-Jack searches metabolic network data from hosts and pathogens, and identifies candidate reactions where hijacking occurs. A novel scoring function ranks candidate hijacked reactions and identifies pathways in the host that interact with pathways in the pathogen, as well as the associated frequent hijacked metabolites. We also describe host-pathogen interaction principles that can be used in the future for subsequent studies. Our case study on Mycobacterium tuberculosis (Mtb) revealed pathways in human-e.g. carbohydrate metabolism, lipids metabolism and pathways related to amino acids metabolism-that are likely to be hijacked by the pathogen. In addition, we report interesting potential pathway interconnections between human and Mtb such as linkage of human fatty acid biosynthesis with Mtb biosynthesis of unsaturated fatty acids, or linkage of human pentose phosphate pathway with lipopolysaccharide biosynthesis in Mtb. Datasets and codes are available at http://cloud.kaust.edu.sa/Pages/Hi-Jack.aspx


Association Between Internet Addiction and the Risk of Musculoskeletal Pain in Chinese College Freshmen - A Cross-Sectional Study.
It is well established that increased internet use is related to an increased risk of musculoskeletal pain among adolescents. The relationship between internet addiction (IA), a unique condition involving severe internet overuse, and musculoskeletal pain has, however, not been reported. This study aimed to investigate the association between IA and the risk of musculoskeletal pain among Chinese college students. A cross-sectional study was conducted among 4211 Chinese college freshmen. IA status was evaluated using the 20-item Young's Internet Addiction Test (IAT). IA was defined as internet addiction score ≥50 points. Musculoskeletal pain was assessed using a self-reported questionnaire. Multiple logistic regression analysis was performed to determine association between IA categories (normal, mild, and moderate-to-severe) and musculoskeletal pain. Among all participants; neck, shoulder, elbow, wrist/hand, and low back and waist pain was reported by 29.2, 33.9, 3.8, 7.9, and 27.9%, respectively. The prevalence of IA was 17.4%. After adjusting for potential confounders, the results showed significant differences in the risk of musculoskeletal pain among different IA categories. The odds ratios (ORs) and 95% confidence intervals (CI) for neck pain with IA categories were 1.000 (reference), 1.451 (1.221, 1.725), and 1.994 (1.608, 2.473), respectively (P for trends: < 0.001). For shoulder pain, these were 1.000 (reference), 1.520 (1.287, 1.795), and 2.057 (1.664, 2.542), respectively (P for trends: < 0.001). For elbow pain, ORs (95% CIs) were 1.000 (reference), 1.627 (1.016, 2.605), and 2.341 (1.382, 3.968), respectively (P for trends: 0.001). Those for wrist/hand pain were 1.000 (reference), 1.508 (1.104, 2.060), and 2.236 (1.561, 3.202), respectively (P for trends: < 0.001). For low back and waist pain with severe IA categories, these were 1.000 (reference), 1.635 (1.368, 1.955), and 2.261 (1.813, 2.819), respectively (P for trends: < 0.001). This cross-sectional study showed that severe IA was associated with a higher risk of musculoskeletal pain in Chinese college freshmen. In future research, it will be necessary to explore causality regarding this relationship using interventional studies.


Convenient method to assess chemical modification of protein thiols by electrophilic metals.
Although covalent modification of protein thiols by electrophilic metals is implicated in disruption of protein functions associated with toxicity, there are limited methods available to detect such modifications. In the present study, we established a convenient method to assess modification of protein thiols by electrophiles, referred to as a biotin-PEAC₅-maleimide (BPM)-labeling assay. In this assay, protein S-modification by electrophiles can be estimated by a decrease in protein modification by BPM, a thiol reactive probe. Using methylmercury (MeHg) as a model electrophilic metal, thiol modification of cellular proteins was detected by the BPM-labeling assay in SH-SY5Y cell lysates and primary mouse hepatocytes. The sensitivity and reliability of the assay was confirmed by atomic absorption spectrometry with recombinant Keap1 as a model thiol protein. This assay was applied to not only MeHg but also to other metals such as cadmium and lead. We also established a BPM-precipitation assay with avidin-agarose beads to separate BPM-modified cellular proteins followed by detection with the individual antibodies. This assay was available for detecting MeHg-induced S-modification of cellular Keap1 in SH-SY5Y cells. Taken together, we have developed reliable simple methods to estimate protein S-modification by electrophilic metals.


Respiratory effects of occupational exposure to tobacco dust.
Few investigations of the respiratory effects of occupational exposure to tobacco dust have been carried out and the threshold limit value has not well been established. A cross sectional survey on a sample of 223 male and female workers at a cigar and cigarette factory in Lucca (Tuscany) showed a significantly higher prevalence of wheezing, attacks of shortness of breath with wheezing, dyspnoea, and rhinitis than in a reference population. A trend towards a decrease in forced end expiratory flows according to smoking habit and work duration was evident. Positive skin prick tests were observed in 26% of men and 23% of women and were positively associated with duration of work and negatively with cigarette smoking. Thin interstitial space involvement was observed on chest x ray examination in almost half the female workers with more than 35 years exposure. These findings suggest that prolonged exposure to tobacco dust may have negative health effects and that it is advisable to establish a threshold limit value for tobacco dust different from that of inert dust.


Antiviral effect of ranpirnase against Ebola virus.
The recent epidemic of Ebola has intensified the need for the development of novel antiviral therapeutics that prolong and improve survival against deadly viral diseases. We sought to determine whether ranpirnase, an endoribonuclease from Rana pipiens with a demonstrated human safety profile in phase III oncology trials, can reduce titers of Ebola virus (EBOV) in infected cells, protect mice against mouse-adapted EBOV challenge, and reduce virus levels in infected mice. Our results demonstrate that 0.50 μg/ml ranpirnase is potently effective at reducing EBOV Zaire Kikwit infection in cultured Vero E6 cells (Selectivity Index 47.8-70.2). In a prophylactic study, a single intravenous dose of 0.1 mg/kg ranpirnase protected 70% of mice from progressive infection. Additionally, in a post-exposure prophylactic study, 100% of female mice survived infection after intraperitoneal administration of 0.1 mg/kg ranpirnase for ten days beginning 1 h post challenge. Most of the male counterparts were sacrificed due to weight loss by Study Day 8 or 9; however, the Clinical Activity/Behavior scores of these mice remained low and no significant microscopic pathologies could be detected in the kidneys, livers or spleens. Furthermore, live virus could not be detected in the sera of ranpirnase-treated mice by Study Day 8 or in the kidneys, livers or spleens by Study Day 12, and viral RNA levels declined exponentially by Study Day 12. Because ranpirnase is exceptionally stable and has a long track record of safe intravenous administration to humans, this drug provides a promising new candidate for clinical consideration in the treatment of Ebola virus disease alone or in combination with other therapeutics.


Eight challenges faced by general practitioners caring for patients after an acute coronary syndrome.
The general practitioner is essential in the management of the patient who has recently been discharged from hospital following an acute coronary syndrome (ACS), particularly as duration of hospital stay is shorter than in previous decades. GPs caring for patients after an ACS face numerous challenges. Often, the first of these is insufficient or delayed documentation from the discharging hospital, although electronic discharge summaries are alleviating this problem. Post-ACS patients often have comorbidities, and GPs play a key role in managing these. Patients taking dual antiplatelet therapy who need surgery, and post-ACS patients with atrial fibrillation, require particular care from GPs. Patients will often approach their GP for advice on the safety of other drugs, such as smoking cessation medication, and phosphodiesterase type 5 inhibitors for erectile dysfunction. For patients complaining of persistent lethargy after an ACS, GPs must consider several differential diagnoses, including depression, hypotension, hypovolaemia, and side effects of β-blockers. GPs play an important ongoing role in ensuring that target cholesterol levels are reached with statin therapy; this includes ensuring long-term adherence. They may also need to advise patients who want to stop statin therapy, usually due to perceived side effects. Many of these challenges can be met with improved and respectful communication between the hospital, the treating cardiologist and the GP. The patient needs to be closely involved in the decision-making process, particularly when balancing the risks of bleeding versus thrombosis.


Effectiveness of surgical and non-surgical interventions for managing diabetic shoulder pain: a systematic review.
This systematic review evaluated and compared the effectiveness of non-surgical and surgical interventions for managing shoulder pain in patients with diabetes. PubMed, Scopus, CINAHL, EMBASE, Sport Discus, and Cochrane library were searched for studies published in the last 20 years. Randomized clinical trials (RCTs) and cohort studies that assessed shoulder pain in patients with diabetes and implemented one or a combination of non-surgical and surgical interventions were eligible for inclusion. The quality of the included studies was assessed using the Structured Effectiveness Quality Evaluation Scale (SEQES) tool. Data extracted from the eligible studies included study design, patient characteristics, duration of symptoms, type of interventions, outcome measures used to assess pain, follow-up intervals, and research findings. A narrative synthesis with effect sizes (ES) or between-group differences was conducted. A total of 25 (14 non-surgical and 11 surgical) studies met the inclusion criteria. Six studies addressed physiotherapeutic interventions (three RCTs and three cohorts - ES = 0.07-1.3), three studies assessed the effect of steroid injections (two RCTs and one cohort - ES = 0.2-0.4), two cohorts addressed arthrographic capsular distension (between-group difference of 1.1 on Visual Analogue Scale), two cohorts addressed MUA, and one RCT addressed suprascapular nerve block (ES = 1-6). For the surgical studies, six cohorts addressed arthroscopic capsular release (ES = 0.2), three cohorts addressed arthroscopic rotator cuff repair (ES = 0.05-0.5), and one cohort addressed arthroplasty (ES = 0.3). Moderate- to very-low-quality evidence suggests large effects of physiotherapy modalities plus exercise and suprascapular nerve block, and trivial to small effects for surgical interventions for improving shoulder pain in patients with diabetes. Future well-designed studies are needed to provide accurate estimates of the true effects of these interventions on improving shoulder pain in patients with diabetes.Implications For RehabilitationShoulder pain may lead to disability in patients with diabetes.We recommend the use of physiotherapy interventions to reduce shoulder pain.Corticosteroid injections are recommended for short-term shoulder pain relief.


Case Report: An Unusual Presentation of Bilateral Subclavian Stenosis in a Patient with Asymptomatic Hypotension.
BACKGROUND Subclavian stenosis is an uncommon clinical condition associated with severe cardiovascular complications, usually presenting with claudication and subclavian steal syndrome. Here we describe the rare case of bilateral subclavian artery stenosis in an asymptomatic patient. CASE REPORT Our patient was a 63-year-old chronic smoker with no prior medical history whose chief complaint was dyspnea from an exacerbation of his chronic obstructive pulmonary disease (COPD). He was hypotensive with blood pressure 74/56 mmHg at admission, which raised suspicion for sepsis, adrenal insufficiency but the workup (renal panel, full blood count and synacthen tests) were normal. He quickly recovered after we treated his COPD exacerbation, but his hypotension persisted despite repeated fluid challenges. To evaluate for structural causes of his hypotension, we performed a full cardiovascular examination with 4 limb blood pressure measurements and found upper limb hypotension and lower limb hypertension. Subsequent imaging with ultrasound and computed tomography confirmed the presence of bilateral subclavian artery stenosis. Our diagnosis was thus bilateral subclavian artery stenosis secondary to atherosclerosis from chronic smoking. The patient was subsequently referred to vascular surgery for consideration of surgical revascularization. CONCLUSIONS Bilateral subclavian stenosis is extremely rare and requires a high index of clinical suspicion. Early diagnosis is important in the primary prevention of associated cardiovascular diseases.


The first pulse of the extremely bright GRB 130427A: a test lab for synchrotron shocks.
Gamma-ray burst (GRB) 130427A is one of the most energetic GRBs ever observed. The initial pulse up to 2.5 seconds is possibly the brightest well-isolated pulse observed to date. A fine time resolution spectral analysis shows power-law decays of the peak energy from the onset of the pulse, consistent with models of internal synchrotron shock pulses. However, a strongly correlated power-law behavior is observed between the luminosity and the spectral peak energy that is inconsistent with curvature effects arising in the relativistic outflow. It is difficult for any of the existing models to account for all of the observed spectral and temporal behaviors simultaneously.


Predicting positive and negative treatment responses to stimulants with brain SPECT imaging.
The goal of this study was to test whether clinician-rated regional cerebral blood flow (rCBF) as rendered by SPECT imaging is a meaningful predictor of patient response to CNS stimulants. Chart reviews were used to identify patients who reported prior significant positive and negative responses to CNS stimulants. Each patient in the study had received resting and concentration SPECT scans using Tc99m exametazime. Differences in cerebral blood flow for frontal regions of interest were assessed in three conditions (resting, concentration, and their difference, or "delta") using ANCOVAs and age-matched ANOVAs. Prefrontal pole deltas were found to be highly sensitive and specific predictors of response to CNS stimulants, with pole activation predicting adverse responses and pole deactivation predicting good responses. Positive and negative predictive values were greater than .75 for both poles. We conclude that SPECT renderings of rCBF, particularly in the prefrontal cortex, are a potentially powerful clinical tool for anticipating response to stimulant medications, both positive and adverse.