Shikonin suppresses the migratory ability of hepatocellular carcinoma cells.
Shikonin is a traditional Oriental medical herb extracted from Lithospermum erythrorhizon. Many studies have shown that shikonin possesses anticancer ability against many different cancers, including hepatocellular carcinoma (HCC). Recently, tumor metastasis has been become an important clinical obstacle. However, the effect of shikonin on metastasis by HCC is unknown. The 50% inhibitory concentration (IC50) of shikonin on HCC cells was determined by an MTT assay and the xCELLigence biosensor system. The migratory ability of HCC cells was detected by a transwell migration assay and the xCELLigence biosensor system. Matrix metalloproteinase-2 and -9 (MMP-2 and -9) expression levels were determined by Western blotting, and the activities of MMP-2 and -9 were determined by gelatin zymography. We found that IC50 values of HepJ5 and Mahlavu cells to shikonin treatment were around 2 μM. Exposure to a low dose of shikonin (0-0.4 μM) did not influence the survival of HCC cells. Interestingly, exposure to a low dose of shikonin inhibited the migratory ability on HepJ5 and Mahlavu cells. To further dissect the mechanism, we found that treatment with a low dose of shikonin reduced the activities and expression levels of MMP-2 and -9, which were correlated with the decreased cell migratory ability of HCC cells. In addition, we found a decrease of vimnetin expression, but no influence on the expression levels of N-cadherin, TWIST, or GRP78. In mechanism dissecting, we found that shikonin treatment may suppress the phosphorylation of AKT and then reduce the NF-κB (NF = nuclear factor) levels, but has no influence on the levels of c-Fos and c-Jun. Furthermore, we also found that shikonin may also reduce the phosphorylation of IκB. We concluded that a low dose of shikonin can suppress the migratory ability of HCC cells through downregulation of expression levels of vimentin and MMP-2 and -9. Our findings suggest that shikonin may be a new compound to prevent the migration of HCC cells.


Prevalence of atopic dermatitis in Leicester: a study of methodology and examination of possible ethnic variation.
A study was undertaken to investigate and compare various methods of estimating the prevalence of atopic dermatitis (AD), and to investigate a possible ethnic difference in our local community. Preschool children attending routine child health surveillance clinics and Social Services day nurseries were examined by a trained observer, and their parents were interviewed. In addition, general practice records from a health centre were scrutinized. Three hundred and twenty-two children aged 1-4 years were examined, and the point prevalence of AD was 14% [95% confidence interval (CI) 10-18]. There was no apparent ethnic difference in prevalence. Twenty-seven per cent (95% CI 22-32) of parents reported that their children had suffered from 'eczema' at some time. General practitioners' records contained a diagnosis of 'eczema' in 32% (95% CI 28-36) of 446 children aged 1-4 years. It is clear that methodology must be carefully standardized if comparisons are to be made between different studies. Accurate estimations of the prevalence of AD can probably only be obtained by examination of a population sample by a trained observer. However, the estimates obtained in this study are high, and would tend to support existing evidence that the prevalence of AD is rising.


Cold physiology: postprandial blood flow dynamics and metabolism in the Antarctic fish Pagothenia borchgrevinki.
Previous studies on metabolic responses to feeding (i.e. the specific dynamic action, SDA) in Antarctic fishes living at temperatures below zero have reported long-lasting increases and small peak responses. We therefore hypothesized that the postprandial hyperemia also would be limited in the Antarctic fish Pagothenia borchgrevinki. The proportion of cardiac output directed to the splanchnic circulation in unfed fish was 18%, which is similar to temperate fish species. Contrary to our prediction, however, gastrointestinal blood flow had increased by 88% at twenty four hours after feeding due to a significant increase in cardiac output and a significant decrease in gastrointestinal vascular resistance. While gastric evacuation time appeared to be longer than in comparable temperate species, digestion had clearly commenced twenty four hours after feeding as judged by a reduction in mass of the administered feed. Even so, oxygen consumption did not increase suggesting an unusually slowly developing SDA. Adrenaline and angiotensin II was injected into unfed fish to investigate neuro-humoral control mechanisms of gastrointestinal blood flow. Both agonists increased gastrointestinal vascular resistance and arterial blood pressure, while systemic vascular resistance was largely unaffected. The hypertension was mainly due to increased cardiac output revealing that the heart and the gastrointestinal vasculature, but not the somatic vasculature, are important targets for these agonists. It is suggested that the apparently reduced SDA in P. borchgrevinki is due to a depressant effect of the low temperature on protein assimilation processes occurring outside of the gastrointestinal tract, while the gastrointestinal blood flow responses to feeding and vasoactive substances resemble those previously observed in temperate species.


Heart Failure in Africa, Asia, the Middle East and South America: The INTER-CHF study.
There are few data on heart failure (HF) patients from Africa, Asia, the Middle East and South America. INTER-CHF is a prospective study that enrolled HF patients in 108 centers in 16 countries from 2012 to 2014. Consecutive ambulatory or hospitalized adult patients with HF were enrolled. Baseline data were recorded on sociodemographics, clinical characteristics, HF etiology and treatments. Age- and sex-adjusted results are reported. We recruited 5813 HF patients: mean(SE) age=59(0.2)years, 39% female, 65% outpatients, 31% from rural areas, 26% with HF with preserved ejection fraction, with 1294 from Africa, 2661 from Asia, 1000 from the Middle-East, and 858 from South America. Participants from Africa-closely followed by Asians-were younger, had lower literacy levels, and were less likely to have health or medication insurance or be on beta-blockers compared with participants from other regions, but were most likely to be in NYHA class IV. Participants from South America were older, had higher insurance and literacy levels, and, along with Middle Eastern participants, were more likely to be on beta-blockers, but had the lowest proportion in NYHA IV. Ischemic heart disease was the most common HF etiology in all regions except Africa where hypertensive heart disease was most common. INTER-CHF describes significant regional variability in socioeconomic and clinical factors, etiologies and treatments in HF patients from Africa, Asia, the Middle East and South America. Opportunities exist for improvement in health/medication insurance rates and proportions of patients on beta blockers, particularly in Africa and Asia.


Prevalence & factors associated with chronic obstetric morbidities in Nashik district, Maharashtra.
In India, community based data on chronic obstetric morbidities (COM) are scanty and largely derived from hospital records. The main aim of the study was to assess the community based prevalence and the factors associated with the defined COM--obstetric fistula, genital prolapse, chronic pelvic inflammatory disease (PID) and secondary infertility among women in Nashik district of Maharashtra State, India. The study was cross-sectional with self-reports followed by clinical and gynaecological examination. Six primary health centre areas in Nashik district were selected by systematic random sampling. Six months were spent on rapport development with the community following which household interviews were conducted among 1560 women and they were mobilized to attend health facility for clinical examination. Of the 1560 women interviewed at household level, 1167 women volunteered to undergo clinical examination giving a response rate of 75 per cent. The prevalence of defined COM among 1167 women was genital prolapse (7.1%), chronic PID (2.5%), secondary infertility (1.7%) and fistula (0.08%). Advancing age, illiteracy, high parity, conduction of deliveries by traditional birth attendants (TBAs) and obesity were significantly associated with the occurrence of genital prolapse. History of at least one abortion was significantly associated with secondary infertility. Chronic PID had no significant association with any of the socio-demographic or obstetric factors. The study findings provided an insight in the magnitude of community-based prevalence of COM and the factors associated with it. The results showed that COM were prevalent among women which could be addressed by interventions at personal, social and health services delivery level.


Hospital care or home care after allogeneic hematopoietic stem cell transplantation--patients' experiences of care and support.
Treatment at home during the pancytopenic phase after allogeneic hematopoietic stem cell transplantation (HSCT) has been an option for patients at our center since 1998. Earlier studies have shown that home care is safe and has medical advantages. In this study, we present patients' experiences of care and support while being treated in hospital or at home during the acute post-transplantation phase. Patients (n = 41, 22 in hospital care and 19 in home care) answered the SAUC questionnaire at discharge (when home, or from hospital). Both statistical analysis and deductive content analysis were used. The patients were highly satisfied with the care and support during the acute post-transplantation phase. Patients in home care were found to be more satisfied with care in general than patients in hospital care. The importance of safety, empathy, and encouragement from healthcare staff were expressed regardless of where care was given. Patients also felt that receipt of continuous, updated information during treatment was important and they had a strong belief in HSCT but were uncertain of the future regarding recovery. The main findings of this study were that in comparison to hospital care, home care does not appear to have a significant negative effect on patients' experiences of care and support during the acute post-transplantation phase. In addition patients in home care felt safe, seen as a person and encouragement seem to empower the patients at home. Thus, this study may encourage other transplantation centers to provide home care if the patients want it.


Teacher-child relationships in the context of poverty: the role of frequent school mobility.
Children's relationships with their teachers are critical for classroom-based learning, but children growing up in poverty may be at risk for lower-quality relationships with teachers. Little is known about how changing schools, one poverty-related risk, affects teacher-child relationships. Using growth curve models that control for a host of other poverty-related risks, this study explores the association between children changing schools frequently (defined as three or more school moves) between preschool and third grade and the quality of their relationships with their teachers over these five years in a low-income, ethnic-minority sample. Children who frequently moved schools were reported to be less close to their teachers in third grade and experienced steeper declines in closeness than children who did not change schools frequently. Moreover, the effects of frequent school mobility at third grade were robust to other poverty-related risks, including residential mobility, parental education risk, family income, and single-parent households. Changing schools was unrelated to children's conflict with teachers. We discuss these findings in the context of policies that support students' transitions when changing schools.


Comparative metabolism and disposition of 1-chloro- and 3-chloro-2-methylpropene in rats and mice.
A recent 2-year carcinogenicity study found that gavage administration of 3-chloro-2-methylpropene (CMP), containing 5% 1-chloro-2-methylpropene (dimethylvinyl chloride, DMVC), caused forestomach neoplasms in rats and mice. Similar chronic studies revealed that DMVC caused forestomach neoplasms in both rats and mice; neoplasms of the nasal and oral cavities were observed in rats but not in mice. In the current studies we have investigated the metabolic basis of these differences. Daily doses of 150 mg/kg of 2-[14C]DMVC or 2-[14C]CMP were administered to rats for 1, 2, or 4 consecutive days. One daily dose of 150 mg/kg of DMVC was administered to mice. Both DMVC and CMP were rapidly metabolized; however, CMP was cleared at a slightly lower rate. Rats exhaled approximately 25 and 10% of the DMVC and CMP as CO2, respectively. Mice exhaled 25% of the DMVC as CO2. Rats expired 30% of the administered DMVC unchanged in the 24 hr after dosing compared to only 7% of the administered CMP. Mice expired 5% of the administered DMVC in the same time period. This observation may explain the occurrence of tumors in the nasal and oral cavities of rats treated with DMVC but not in rats treated with CMP or in mice treated with DMVC in 2-year carcinogenicity studies. The 24-hr urinary excretion in rats was 35% of the administered DMVC compared to 58% of CMP. Mice excreted 47% of the administered DMVC in 24 hr in the urine. An unusual urinary metabolite of DMVC, 2-amino-6-methyl-4-thia-5-heptene-1,7-dioic acid, was identified.(ABSTRACT TRUNCATED AT 250 WORDS)


Development and Validation of a 28-gene Hypoxia-related Prognostic Signature for Localized Prostate Cancer.
Hypoxia is associated with a poor prognosis in prostate cancer. This work aimed to derive and validate a hypoxia-related mRNA signature for localized prostate cancer. Hypoxia genes were identified in vitro via RNA-sequencing and combined with in vivo gene co-expression analysis to generate a signature. The signature was independently validated in eleven prostate cancer cohorts and a bladder cancer phase III randomized trial of radiotherapy alone or with carbogen and nicotinamide (CON). A 28-gene signature was derived. Patients with high signature scores had poorer biochemical recurrence free survivals in six of eight independent cohorts of prostatectomy-treated patients (Log rank test P < .05), with borderline significances achieved in the other two (P < .1). The signature also predicted biochemical recurrence in patients receiving post-prostatectomy radiotherapy (n = 130, P = .007) or definitive radiotherapy alone (n = 248, P = .035). Lastly, the signature predicted metastasis events in a pooled cohort (n = 631, P = .002). Prognostic significance remained after adjusting for clinic-pathological factors and commercially available prognostic signatures. The signature predicted benefit from hypoxia-modifying therapy in bladder cancer patients (intervention-by-signature interaction test P = .0026), where carbogen and nicotinamide was associated with improved survival only in hypoxic tumours. A 28-gene hypoxia signature has strong and independent prognostic value for prostate cancer patients.


Urine steroid metabolomics as a biomarker tool for detecting malignancy in adrenal tumors.
Adrenal tumors have a prevalence of around 2% in the general population. Adrenocortical carcinoma (ACC) is rare but accounts for 2-11% of incidentally discovered adrenal masses. Differentiating ACC from adrenocortical adenoma (ACA) represents a diagnostic challenge in patients with adrenal incidentalomas, with tumor size, imaging, and even histology all providing unsatisfactory predictive values. Here we developed a novel steroid metabolomic approach, mass spectrometry-based steroid profiling followed by machine learning analysis, and examined its diagnostic value for the detection of adrenal malignancy. Quantification of 32 distinct adrenal derived steroids was carried out by gas chromatography/mass spectrometry in 24-h urine samples from 102 ACA patients (age range 19-84 yr) and 45 ACC patients (20-80 yr). Underlying diagnosis was ascertained by histology and metastasis in ACC and by clinical follow-up [median duration 52 (range 26-201) months] without evidence of metastasis in ACA. Steroid excretion data were subjected to generalized matrix learning vector quantization (GMLVQ) to identify the most discriminative steroids. Steroid profiling revealed a pattern of predominantly immature, early-stage steroidogenesis in ACC. GMLVQ analysis identified a subset of nine steroids that performed best in differentiating ACA from ACC. Receiver-operating characteristics analysis of GMLVQ results demonstrated sensitivity = specificity = 90% (area under the curve = 0.97) employing all 32 steroids and sensitivity = specificity = 88% (area under the curve = 0.96) when using only the nine most differentiating markers. Urine steroid metabolomics is a novel, highly sensitive, and specific biomarker tool for discriminating benign from malignant adrenal tumors, with obvious promise for the diagnostic work-up of patients with adrenal incidentalomas.


Predicting Primary and Secondary Abstinence Among Adolescent Boys and Girls in the Western Cape, South Africa.
Two of the most effective health behaviours with regard to HIV prevention are condom use and sexual abstinence. While determinants of condom use among sub-Saharan African adolescents have been studied extensively, factors related to abstinence have received far less attention. This study identified socio-cognitive determinants of primary and secondary abstinence intentions and of early sexual activity. This study also assessed whether these factors had a direct or indirect association with intentions to abstain from sex. A longitudinal design was used in which 1670 students (age 12-16) of non-private South African high schools filled in a questionnaire, with a follow-up after 6 months, concerning sexual abstinence, attitudes, social norms, self-efficacy, risk perception and knowledge. Logistic and linear regression analysis with latent factors was used to assess determinants of intentions and abstinence, and structural equation modelling was used to assess indirect effects. Results showed that among sexually inactive students, social norms predicted the intention to abstain from sex in the next 6 months. Among sexually active students, reporting less disadvantages of abstinence predicted the intention to abstain. Sexual activity at follow-up was predicted by attitudes and intention among sexually inactive girls, and by knowledge among sexually inactive boys. No predictors were found for sexually active adolescents. Structural equation modelling further showed that risk perception was indirectly related to intentions to abstain from sexual intercourse. We conclude that addressing socio-cognitive factors in order to motivate adolescents to delay sex is more likely to be successful before they experience sexual debut. In addition, this study shows that the effect of increasing risk perceptions, a strategy often applied by parents and HIV prevention programmes, is to a large extent mediated by more proximal cognitive factors such as attitude. Research is needed to identify factors that influence the execution of intentions to abstain from sex.


A comprehensive investigation of memory impairment in attention deficit hyperactivity disorder and oppositional defiant disorder.
We conducted a comprehensive and systematic assessment of memory functioning in drug-naïve boys with attention deficit hyperactivity disorder (ADHD) and oppositional defiant disorder (ODD). Boys performed verbal and spatial working memory (WM) component (storage and central executive) and verbal and spatial storage load tasks, and the spatial span, spatial executive WM, spatial recognition memory and verbal recognition memory tasks from the Cambridge Neuropsychological Test Automated Battery. Groups comprised: (a) ADHD only (N = 21); (b) ADHD+ODD (N = 27); (c) ODD only (N = 21); and (d) typically developing (TYP) boys (N = 26). Groups were matched for age (M = 9.7 years) and sex (all boys). Confirmatory factor analyses confirmed the presence of five factors: verbal functioning, spatial functioning, WM storage, WM central executive and long-term memory (LTM). All three clinical groups demonstrated impaired memory performance. Boys with ODD and ODD+ADHD but not ADHD alone performed poorly on verbal memory tasks, whilst all three clinical groups showed impaired performance on spatial memory tasks. All three clinical groups performed poorly on the storage and central executive WM factors and the LTM factor. ADHD and ODD are characterised by impaired performance storage and central executive WM tasks and LTM tasks. This is, we believe, the first report of impaired WM and LTM performance in ODD. This study suggests that verbal memory difficulties are more closely associated with ODD than ADHD symptoms and that combined ADHD+ODD represents a true comorbidity. The data also support a small but growing number of suggestions in the literature of impaired LTM in ADHD.


Microbial counts, fermentation products, and aerobic stability of whole crop corn and a total mixed ration ensiled with and without inoculation of Lactobacillus casei or Lactobacillus buchneri.
Whole crop corn (DM 29.2%) and a total mixed ration (TMR, DM 56.8%) containing wet brewers grains, alfalfa hay, dried beet pulp, cracked corn, soybean meal, and molasses at a ratio of 5:1:1:1:1:1 on fresh weight basis, were ensiled with and without Lactobacillus casei or Lactobacillus buchneri in laboratory silos. The effects of inoculation on microbial counts, fermentation products, and aerobic stability were determined after 10 and 60 d. Untreated corn silage was well preserved with high lactic acid content, whereas large numbers of remaining yeasts resulted in low stability on exposure to air. Inoculation with L. casei suppressed heterolactic fermentation, but no improvements were found in aerobic stability. The addition of L. buchneri markedly enhanced the aerobic stability, while not affecting the DM loss and NH3-N production. Large amounts of ethanol were found when the TMR was ensiled, and the content of ethanol overwhelmed that of lactic acid in untreated silage. This fermentation was related to high yeast populations and accounted for a large loss of DM found in the initial 10 d. The ethanol production decreased when inoculated with L. casei and L. buchneri, but the effects diminished at 60 d of ensiling. Inoculation with L. buchneri lowered the yeasts in TMR silage from the beginning of storage; however, the populations decreased to undetectable levels when stored for 60 d, regardless of inoculation. No heating was observed in TMR silage during aerobic deterioration test for 7 d. This stability was achieved even when a high population of yeasts remained and was not affected by either inoculation or ensiling period. The results indicate that inoculation with L. buchneri can inhibit yeast growth and improve aerobic stability of corn and TMR silage; however, high stability of TMR silage can be obtained even when no treatments were made and high population (>10(5) cfu/g) of yeasts were detected.


Anticoagulation in patients with atrial fibrillation undergoing coronary stent implantation.
In patients with atrial fibrillation (AF) undergoing coronary stent implantation, the optimal antithrombotic strategy is unclear. We evaluated whether use of oral anticoagulation (OAC) was associated with any benefit in morbidity or mortality in patients with AF, high risk of thromboembolism (TE) (CHA2DS2-VASC score ≥ 2) and coronary stent implantation. Among 8,962 unselected patients with AF seen between 2000 and 2010, a total of 2,709 (30%) had coronary artery disease and 417/2,709 (15%) underwent stent implantation while having CHA2DS2-VASC score ≥ 2. During follow-up (median=650 days), all TE, bleeding episodes, and major adverse cardiac events (i.e. death, acute myocardial infarction, target lesion revascularisation) were recorded. At discharge, 97/417 patients (23%) received OAC, which was more likely to be prescribed in patients with permanent AF and in those treated for elective stent implantation. The incidence of outcome event rates was not significantly different in patients treated and those not treated with OAC. However, in multivariate analysis, the lack of OAC at discharge was independently associated with increased risk of death/stroke/systemic TE (relative risk [RR] =2.18, 95% confidence interval [CI] 1.02-4.67, p=0.04), with older age (RR =1.12, 1.04-1.20, p=0.003), heart failure (RR =3.26, 1.18-9.01, p=0.02), and history of stroke (RR =18.87, 3.11-111.11, p=0.001). In conclusion, in patients with AF and high thromboembolic risk after stent implantation, use of OAC was independently associated with decreased risk of subsequent death/stroke/systemic TE, suggesting that OAC should be systematically used in this patient population.


In vivo delivery of cytoplasmic RNA virus-derived miRNAs.
The discovery of microRNAs (miRNAs) revealed an unappreciated level of post-transcriptional control used by the cell to maintain optimal protein levels. This process has represented an attractive strategy for therapeutics that is currently limited by in vivo delivery constraints. Here, we describe the generation of a single-stranded, cytoplasmic virus of negative polarity capable of producing functional miRNAs. Cytoplasmic RNA virus-derived miRNAs accumulated to high levels in vitro, generated significant amounts of miRNA star strand, associated with the RNA-induced silencing complex (RISC), and conferred post transcriptional gene silencing in a sequence-specific manner. Furthermore, we demonstrate that these vectors could deliver miRNAs to a wide range of tissues, and sustain prolonged expression capable of achieving measurable knockdown of physiological targets in vivo. Taken together, these results validate noncanonical processing of cytoplasmic-derived miRNAs and provide a novel platform for small RNA delivery.


A minimal model for G protein-mediated synaptic facilitation and depression.
G protein-coupled receptors are ubiquitous in neurons, as well as other cell types. Activation of receptors by hormones or neurotransmitters splits the G protein heterotrimer into Galpha and Gbetagamma subunits. It is now clear that Gbetagamma directly inhibits Ca2+ channels, putting them into a reluctant state. The effects of Gbetagamma depend on the specific beta and gamma subunits present, as well as the beta subunit isoform of the N-type Ca2+ channel. We describe a minimal mathematical model for the effects of G protein action on the dynamics of synaptic transmission. The model is calibrated by data obtained by transfecting G protein and Ca2+ channel subunits into tsA-201 cells. We demonstrate with numerical simulations that G protein action can provide a mechanism for either short-term synaptic facilitation or depression, depending on the manner in which G protein-coupled receptors are activated. The G protein action performs high-pass filtering of the presynaptic signal, with a filter cutoff that depends on the combination of G protein and Ca2+ channel subunits present. At stimulus frequencies above the cutoff, trains of single spikes are transmitted, while only doublets are transmitted at frequencies below the cutoff. Finally, we demonstrate that relief of G protein inhibition can contribute to paired-pulse facilitation.


The cost-effectiveness of fluorodeoxyglucose 18-F positron emission tomography in the N0 neck.
Although surgery and radiation are effective treatments of regional lymphatics for classification N0 head and neck squamous cell carcinoma (HNSCC) patients, both have morbidities that could be avoided in approximately 70% of patients without lymph node disease with better diagnostic information. 18-F fluoro-2-deoxyglucose positron emission tomography (FDG-PET) has shown promise in detecting subclinical lymph node disease, but its cost and availability have limited its use. Here, we sought to determine whether the use of FDG-PET was cost-effective as part of a treatment strategy for classification N0 HNSCC patients. The cost-effectiveness of proceeding from classification of N0 by computed tomography to a PET scan was estimated using standard methods of economic evaluation. Costs were for a large, Midwestern university medical center. Probabilities were computed from a review of the literature. Utilities were obtained by a time-tradeoff method, and life expectancy was estimated using the Surveillance, Epidemiology, and End Results database. Outcomes measures were cost per year of life saved and cost per quality-adjusted life-year. Modified radical neck dissection was associated with the lowest morbidity (utility [u] = 0.93), and radical neck dissection plus radiation was associated with the highest (u = 0.68). Life expectancy was estimated to be 5.9 and 11.5 years for patients with and without lymph node disease, respectively. The incremental cost-effectiveness ratio for the PET strategy was $8718 per year of life saved, or $2505 per quality-adjusted life-year. A diagnostic and treatment strategy that proceeds from classification of N0 to a PET scan is cost-effective. Prospective studies that evaluate this strategy are important to assure that these simulation results are realized in clinical practice.


Clinical efficacy of icatibant in the treatment of acute hereditary angioedema during the FAST-3 trial.
Bradykinin is the key mediator of symptoms of hereditary angioedema (HAE), a rare genetic disorder characterized by recurrent episodes of edema of the skin, mucosa and muscle. Icatibant, a bradykinin B(2) receptor antagonist, is an effective and generally well-tolerated treatment option for acute attacks of type I and II HAE. A Phase III randomized, double-blind, placebo-controlled study, FAST-3 (NCT00912093), was designed to further evaluate the efficacy and safety of icatibant in patients presenting with moderate to very severe cutaneous and/or abdominal or mild-to-moderate laryngeal symptoms. Severe laryngeal attacks were treated with open-label icatibant. The controlled phase of FAST-3, completed in October 2010 with results published in December 2011, demonstrated that compared with placebo, icatibant evoked clinically meaningful and statistically significant efficacy across multiple end points in the treatment of type I and II HAE attacks. In addition, icatibant was generally well tolerated and no drug-related serious adverse events were experienced.


Specialist involvement and referral patterns in ambulatory medical care for patients with dementia in Germany: results of a claims data based case-control study.
To analyze the referral processes from general practitioners to specialists and among specialists for dementia patients in the time periods before, during and after the diagnosis in Germany. In this case-control study claims data from 1,848 insurants with incident dementia aged 65 years and more and 7,392 matched controls were compared over a two-year period covering the pre-incidence, incidence and post-incidence time periods. We found an increase in referrals of 30% in the incidence quarter, mainly from general practice to neuropsychiatry and from there to radiology. Referrals to clinical chemistry and other disciplines for dementia-specific reasons were negligible in amount. 34% of incident cases had at least one contact with a neuropsychiatrist during the year of incidence, and the majority of them visited this specialist repeatedly during that year. Only a minority (13.5%) of patients was referred to radiology for imaging. Referrals to other specialists declined whereas self-referrals did not increase. The referral rates to relevant specialists (neuropsychiatry, radiology and clinical chemistry) are far less frequent than proposed in German guidelines. More research is needed to explain the gape between guidelines and daily care and to find ways for a better implementation of guidelines in ambulatory care. Guidelines should not only deal with diagnostic procedures and therapeutic options but also consider questions of applicability in daily clinical practice and propose effective organizational models of care provision.


A System for Multi-Domain Contextualization of Personal Health Data.
Current telehealth systems are used to improve the treatment of chronic diseases by collecting medical data at the patient and transferring them to a remote medical institution. Research shows that such medical practice can be substantially improved if the measured parameters are greater in number and of more diverse nature. Emerging consumer solutions for monitoring personal health and wellness, as well as various resources from domains like internet, telecommunications and smart living, can be used as possible sources for exploration of an individual's wider health context. It was our aim to design an internet of things solutions, which would combine these sources of information into context information, complementary to health data. An internet of things platform was designed and implemented and integration with an established e-health system was provided to enrich telehealth data with context information by aggregating and processing cross-domain inputs from various sources. The approach was validated on a use case scenario. The concept was tried in a scenario related to prevention and management of heart disease. The system's advanced graphic correlation features are expected to help physicians and patients identify true roots of health problems. Medical researchers are also expected to benefit from a deeper insight into complex cross-domain parameter dependencies that determine an individual's health.


A bilayered hybrid microfibrous PLGA--acellular matrix scaffold for hollow organ tissue engineering.
Various synthetic and natural biomaterials have been used for regeneration of tissues and hollow organs. However, clinical outcome of reconstructive procedures remained challenging due to the lack of appropriate scaffold materials, supporting the needs of various cell types and providing a barrier function required in hollow organs. To address these problems, we have developed a bilayered hybrid scaffold comprising unique traits of polymeric microfibers and naturally derived acellular matrices and tested its potential for hollow organ regeneration in a rat bladder model. Hybrid scaffolds were fabricated by electrospinning of PLGA microfibers directly onto the abluminal surface of a bladder acellular matrix. Stability of this bilayered construct was established using modified spinning technique. The resulting 3-dimensional framework provided good support for growth, attachment and proliferation of primary bladder smooth muscle cells. Histological analysis in vivo at 4 and 8 weeks post implantation, revealed regeneration of bladder tissue structures consisting of urothelium, smooth muscle and collagen rich layers infiltrated with host cells and micro vessels. Furthermore, hybrid scaffolds maintained normal bladder capacity, whereas BAM recipients showed a significant distension of the bladder. These results demonstrate that this adaptable hybrid scaffold supports bladder regeneration and holds potential for engineering of bladder and other hollow organs.


Preferences for vertical position of the maxillary lateral incisors.
To evaluate the preferences for vertical position of the maxillary lateral incisors among orthodontists, general dentists, and laypeople. The study group consisted of 120 judges, 40 in each of 3 study groups. Judges were asked to give their preference for maxillary lateral incisor position while viewing an animation of an idealized smile. Perceptometrics software animated the photograph and collated responses. Each judge participated in 2 similar experiments, 1 to determine what he/she found to be the most pleasing position and another to define that position's range of acceptability. The processed data were analyzed with ANOVA (analysis of variance) and post-hoc Scheffé tests to determine mean differences among the 3 study groups. The most pleasing protocol revealed no significant difference (P >.05) in the mean preferences of the 3 groups. The average preference was 0.6 mm above the incisal plane. No subjects chose a level incisal plane as being most pleasing. Orthodontists and general dentists had a smaller range of acceptability than laypeople. The means for lower and upper limits of acceptability for all subjects were 0.3 mm and 1.0 mm, respectively, above the incisal plane. The judges in this study preferred that the maxillary lateral incisor to be set about 0.5 mm above the incisal plane, slightly offset from the incisal plane, and not level. The lateral incisors were best perceived when they were offset slightly from the incisal plane, and there was an acceptable range within which the lateral incisors may be placed.


Genetic transformation of putative cariogenic properties in Streptococcus mutans.
Rough colonial morphology and bacteriocin production, two properties which may be associated with the cariogenicity of Streptococcus mutans, were transformed into several strain GS-5 mutants defective in each respective property. Transformation was determined by observing the frequency of cotransfer of these properties with different reference markers. The rough colonial transformants were identical to the parental GS-5 strain with respect to ability to synthesize water-insoluble glucans and undergo in vitro sucrose-dependent colonization of glass surfaces. Alterations in the growth medium and the concentration of the initial cell inoculum resulted in an approximate 10-fold increase in the frequency of transformation of strain GS-5 compared to previous observations.


Particle size matters: diagnostics and treatment of small airways involvement in asthma.
Small airways are an important site of inflammation and obstruction in asthma, which contributes to the severity of airway hyperresponsiveness (AHR) that is usually measured by nebulisation of large-particle stimuli. We investigated whether small and large particle sizes of aerosolised adenosine monophospate (AMP) provoke similar severity of AHR. Additionally, effects of the small-particle inhaled corticosteroid (ICS) ciclesonide and large-particle ICS fluticasone on AHR to large- and small-particle size AMP were assessed. After a 4-week run-in period using open-label fluticasone (100 μg b.i.d.), 37 mild-to-moderate asthmatics underwent provocations with standard-size (3.7 μm), large-particle (9.9 μm) and small-particle (1.06 μm) AMP. Subjects received 4-week ciclesonide (160 μg s.i.d.) or fluticasone (100 μg b.i.d.) treatment (double-blind and double-dummy) followed by large- and small-particle AMP provocation. Small-particle AMP induced a 20% decrease in forced expiratory volume in 1 s (FEV(1)) at a significantly higher dose than large-particle AMP. Ciclesonide and fluticasone had comparable effects on AMP provocations. Not all subjects reached the provocative concentration causing a 20% fall in FEV(1) (PC(20)) at the highest AMP dose. In those who did, ciclesonide improved small-particle AMP PC(20) by 1.74 doubling doses (DD) (p = 0.03), whereas fluticasone did not. Conversely, fluticasone improved large-particle AMP PC(20) significantly (1.32 DD; p = 0.03), whereas ciclesonide did not. Small-particle AMP provocation appears to be a promising tool to assess changes in small airway inflammation. Future adjustments are necessary taking into account the very small particle size used, with large exhaled fractions. In asthmatics reaching a PC(20) with small- and large-particle AMP provocations, ciclesonide improves hyperresponsiveness to small particle size AMP, and fluticasone to large particle size. This warrants further research to target provocations and treatment to specific airway sizes.


Loss of perivascular aquaporin 4 may underlie deficient water and K+ homeostasis in the human epileptogenic hippocampus.
An abnormal accumulation of extracellular K+ in the brain has been implicated in the generation of seizures in patients with mesial temporal lobe epilepsy (MTLE) and hippocampal sclerosis. Experimental studies have shown that clearance of extracellular K+ is compromised by removal of the perivascular pool of the water channel aquaporin 4 (AQP4), suggesting that an efficient clearance of K+ depends on a concomitant water flux through astrocyte membranes. Therefore, we hypothesized that loss of perivascular AQP4 might be involved in the pathogenesis of MTLE. Whereas Western blot analysis showed an overall increase in AQP4 levels in MTLE compared with non-MTLE hippocampi, quantitative ImmunoGold electron microscopy revealed that the density of AQP4 along the perivascular membrane domain of astrocytes was reduced by 44% in area CA1 of MTLE vs. non-MTLE hippocampi. There was no difference in the density of AQP4 on the astrocyte membrane facing the neuropil. Because anchoring of AQP4 to the perivascular astrocyte endfoot membrane depends on the dystrophin complex, the localization of the 71-kDa brain-specific isoform of dystrophin was assessed by immunohistochemistry. In non-MTLE hippocampus, dystrophin was preferentially localized near blood vessels. However, in the MTLE hippocampus, the perivascular dystrophin was absent in sclerotic areas, suggesting that the loss of perivascular AQP4 is secondary to a disruption of the dystrophin complex. We postulate that the loss of perivascular AQP4 in MTLE is likely to result in a perturbed flux of water through astrocytes leading to an impaired buffering of extracellular K+ and an increased propensity for seizures.


Flexible expressed region analysis for RNA-seq with derfinder.
Differential expression analysis of RNA sequencing (RNA-seq) data typically relies on reconstructing transcripts or counting reads that overlap known gene structures. We previously introduced an intermediate statistical approach called differentially expressed region (DER) finder that seeks to identify contiguous regions of the genome showing differential expression signal at single base resolution without relying on existing annotation or potentially inaccurate transcript assembly.We present the derfinder software that improves our annotation-agnostic approach to RNA-seq analysis by: (i) implementing a computationally efficient bump-hunting approach to identify DERs that permits genome-scale analyses in a large number of samples, (ii) introducing a flexible statistical modeling framework, including multi-group and time-course analyses and (iii) introducing a new set of data visualizations for expressed region analysis. We apply this approach to public RNA-seq data from the Genotype-Tissue Expression (GTEx) project and BrainSpan project to show that derfinder permits the analysis of hundreds of samples at base resolution in R, identifies expression outside of known gene boundaries and can be used to visualize expressed regions at base-resolution. In simulations, our base resolution approaches enable discovery in the presence of incomplete annotation and is nearly as powerful as feature-level methods when the annotation is complete.derfinder analysis using expressed region-level and single base-level approaches provides a compromise between full transcript reconstruction and feature-level analysis. The package is available from Bioconductor at www.bioconductor.org/packages/derfinder.


Sensory attenuation of self-produced feedback: the Lombard effect revisited.
The Lombard effect describes the automatic and involuntary increase in vocal intensity that speakers exhibit in a noisy environment. Previous studies of the Lombard effect have typically focused on the relationship between speaking and hearing. Automatic and involuntary increases in motor output have also been noted in studies of finger force production, an effect attributed to mechanisms of sensory attenuation. The present study tested the hypothesis that sensory attenuation mechanisms also underlie expression of the Lombard effect. Participants vocalized phonemes in time with a metronome, while auditory and visual feedback of their performance were manipulated or removed during the course of the trial. We demonstrate that providing a visual reference to calibrate somatosensory-based judgments of current vocal intensity resulted in reduced expression of the Lombard effect. Our results suggest that sensory attenuation effects typically seen in fingertip force production play an important role in the control of speech volume.


Allometric Analysis Detects Brain Size-Independent Effects of Sex and Sex Chromosome Complement on Human Cerebellar Organization.
The cerebellum is a large hindbrain structure that is increasingly recognized for its contribution to diverse domains of cognitive and affective processing in human health and disease. Although several of these domains are sex biased, our fundamental understanding of cerebellar sex differences-including their spatial distribution, potential biological determinants, and independence from brain volume variation-lags far behind that for the cerebrum. Here, we harness automated neuroimaging methods for cerebellar morphometrics in 417 individuals to (1) localize normative male-female differences in raw cerebellar volume, (2) compare these to sex chromosome effects estimated across five rare sex (X/Y) chromosome aneuploidy (SCA) syndromes, and (3) clarify brain size-independent effects of sex and SCA on cerebellar anatomy using a generalizable allometric approach that considers scaling relationships between regional cerebellar volume and brain volume in health. The integration of these approaches shows that (1) sex and SCA effects on raw cerebellar volume are large and distributed, but regionally heterogeneous, (2) human cerebellar volume scales with brain volume in a highly nonlinear and regionally heterogeneous fashion that departs from documented patterns of cerebellar scaling in phylogeny, and (3) cerebellar organization is modified in a brain size-independent manner by sex (relative expansion of total cerebellum, flocculus, and Crus II-lobule VIIIB volumes in males) and SCA (contraction of total cerebellar, lobule IV, and Crus I volumes with additional X- or Y-chromosomes; X-specific contraction of Crus II-lobule VIIIB). Our methods and results clarify the shifts in human cerebellar organization that accompany interwoven variations in sex, sex chromosome complement, and brain size.SIGNIFICANCE STATEMENT Cerebellar systems are implicated in diverse domains of sex-biased behavior and pathology, but we lack a basic understanding of how sex differences in the human cerebellum are distributed and determined. We leverage a rare neuroimaging dataset to deconvolve the interwoven effects of sex, sex chromosome complement, and brain size on human cerebellar organization. We reveal topographically variegated scaling relationships between regional cerebellar volume and brain size in humans, which (1) are distinct from those observed in phylogeny, (2) invalidate a traditional neuroimaging method for brain volume correction, and (3) allow more valid and accurate resolution of which cerebellar subcomponents are sensitive to sex and sex chromosome complement. These findings advance understanding of cerebellar organization in health and sex chromosome aneuploidy.


Matrix-assisted laser desorption/ionization tandem mass spectrometry for identification of organic tattoo pigments in inks and tissue samples.
With regard to the increasing number of tattooed people, legal regulations for tattoo inks were implemented across Europe-aiming for higher consumer safety. To control the laws' abidance, analytical methods are needed to identify banned ingredients from the given negative lists. Since specific organic pigments are often associated with tattoo side effects, their identification in tattoo inks as well as in biological samples is of great importance. Particularly, poorly soluble organic pigments are challenging to detect. In the past, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was reported as a promising tool for organic pigment identification. Here, we present a MALDI tandem mass spectrometry (MS/MS) approach to increase identification specificity and sensitivity in the process based on pigment fragment ions which is of special importance in tissue samples. For further verification of pigment identities, alkali metal cation attachment was used. Sample preparation was optimized and included mechanical disruption followed by the application as dried droplets with the matrices α-cyano-4-phenylcinnamic acid and sinapinic acid as well as ethanol. Pigments were identified by spectral comparison to reference libraries containing 40 pigments and following a decision tree. Additionally, successful pigment identification in biological samples was carried out. The implemented automated MALDI-MS and -MS/MS acquisitions make the hereby proposed pigment identification suitable for routine application.


Radiation exposure to personnel performing endoscopic retrograde cholangiopancreatography.
Endoscopic retrograde cholangiopancreatography (ERCP) relies on the use of ionising radiation but risks to operator and patient associated with radiation exposure are unclear. The aim of this prospective study was to estimate the radiation dose received by personnel performing fluoroscopic endoscopic procedures, mainly ERCP. Consecutive procedures over a two month period were included. The use of thermoluminescent dosimeters to measure radiation exposure to the abdomen, thyroid gland, and hands of the operator permitted an estimation of the annual whole body effective dose equivalent. During the study period 66 procedures (61 ERCP) were performed and the estimated annual whole body effective dose equivalent received by consultant operators ranged between 3.35 and 5.87 mSv. These values are similar to those received by patients undergoing barium studies and equate to an estimated additional lifetime fatal cancer risk between 1 in 7000 and 1 in 3500. While within legal safety limits for radiation exposure to personnel, these doses are higher than values deemed acceptable for the general public. It is suggested that personnel as well as patients may be exposed to significant values of radiation during ERCP. The study emphasises the need to carefully assess the indication for, and to use measures that minimise radiation exposure during any fluoroscopic procedure.


Heart Failure Telemonitoring in Japan and Sweden: A Cross-Sectional Survey.
Telemonitoring of heart failure (HF) patients is increasingly discussed at conferences and addressed in research. However, little is known about actual use in specific countries. We aimed to (1) describe the use of non-invasive HF telemonitoring, (2) clarify expectations of telemonitoring among cardiologists and nurses, and (3) describe barriers to the implementation of telemonitoring in Japan and Sweden. This study used a cross-sectional survey of non-invasive HF telemonitoring. A total of 378 Japanese (120 cardiologists, 258 nurses) and 120 Swedish (39 cardiologists, 81 nurses) health care professionals from 165 Japanese and 61 Swedish hospitals/clinics nationwide participated in the study (210 in Japan and 98 in Sweden were approached). Data were collected between November 2013 and May 2014 with a questionnaire that was adapted from a previous Dutch study on telemonitoring. The mean age of the cardiologists and nurses was 47 years and 41 years, respectively. Experience at the current position caring for HF patients was 19 years among the physicians and 15 years among the nurses. In total, 7 Japanese (4.2%) and none of the Swedish health care institutions used telemonitoring. One fourth (24.0%, 118/498) of the health care professionals were familiar with the technology (in Japan: 21.6%, 82/378; in Sweden: 30.0%, 36/120). The highest expectations of telemonitoring (rated on a scale from 0-10) were reduced hospitalizations (8.3 in Japan and 7.5 in Sweden), increased patient self-care (7.8 and 7.4), and offering high-quality care (7.8 and 7.0). The major goal for introducing telemonitoring was to monitor physical condition and recognize signs of worsening HF in Japan (94.1%, 352/374) and Sweden (88.7%, 102/115). The following reasons were also high in Sweden: to monitor effects of treatment and adjust it remotely (86.9%, 100/115) and to do remote drug titration (79.1%, 91/115). Just under a quarter of Japanese (22.4%, 85/378) and over a third of Swedish (38.1%, 45/118) health care professionals thought that telemonitoring was a good way to follow up stable HF patients. Three domains of barriers were identified by content analysis: organizational barriers "how are we going to do it?" (categories include structure and resource), health care professionals themselves "what do we need to know and do" (reservation), and barriers related to patients "not everybody would benefit" (internal and external shortcomings). Telemonitoring for HF patients has not been implemented in Japan or Sweden. However, health care professionals have expectations of telemonitoring to reduce patients' hospitalizations and increase patient self-care. There are still a wide range of barriers to the implementation of HF telemonitoring.


N-methyl-D-aspartate receptors in human erythroid precursor cells and in circulating red blood cells contribute to the intracellular calcium regulation.
The presence of N-methyl-d-aspartate receptor (NMDAR) was previously shown in rat red blood cells (RBCs) and in a UT-7/Epo human myeloid cell line differentiating into erythroid lineage. Here we have characterized the subunit composition of the NMDAR and monitored its function during human erythropoiesis and in circulating RBCs. Expression of the NMDARs subunits was assessed in erythroid progenitors during ex vivo erythropoiesis and in circulating human RBCs using quantitative PCR and flow cytometry. Receptor activity was monitored using a radiolabeled antagonist binding assay, live imaging of Ca(2+) uptake, patch clamp, and monitoring of cell volume changes. The receptor tetramers in erythroid precursor cells are composed of the NR1, NR2A, 2C, 2D, NR3A, and 3B subunits of which the glycine-binding NR3A and 3B and glutamate-binding NR2C and 2D subunits prevailed. Functional receptor is required for survival of erythroid precursors. Circulating RBCs retain a low number of the receptor copies that is higher in young cells compared with mature and senescent RBC populations. In circulating RBCs the receptor activity is controlled by plasma glutamate and glycine. Modulation of the NMDAR activity in RBCs by agonists or antagonists is associated with the alterations in whole cell ion currents. Activation of the receptor results in the transient Ca(2+) accumulation, cell shrinkage, and alteration in the intracellular pH, which is associated with the change in hemoglobin oxygen affinity. Thus functional NMDARs are present in erythroid precursor cells and in circulating RBCs. These receptors contribute to intracellular Ca(2+) homeostasis and modulate oxygen delivery to peripheral tissues.


Educating Future Environmental Health Professionals.
Future environmental health problems will require a new generation of educated and trained professionals. Efforts to enhance the environmental public health workforce have been promoted by several organizations. While progress has been measured by these organizations, many environmental health academic programs are experiencing budget reductions and lower enrollments. One of the reasons for this trend is the so-called higher education crisis. We argue that training is not equivalent to education in the environmental health sciences, albeit the two terms are often used interchangeably. Organizations involved with the education, training, and credentialing of environmental health professionals must work together to ensure the viability and effectiveness of environmental health academic programs.


Changes in thoracic kyphosis negatively impact sagittal alignment after lumbar pedicle subtraction osteotomy: a comprehensive radiographic analysis.
Consecutive, multicenter retrospective review. To evaluate if change in thoracic kyphosis (TK) has a positive or negative impact on spinopelvic alignment after lumbar pedicle subtraction osteotomy (PSO) with short fusions. In the setting of sagittal malalignment, the effect of large vertebral resections can now be anticipated in long fusions, but their impact on unfused segments (reciprocal changes [RC]) remains poorly understood. A total of 34 adult patients (mean age = 54 years; SD = 12) who underwent lumbar PSO with upper instrumented vertebra below T10 were included. Radiographic analysis included pre- and postassessment of TK, lumbar lordosis (LL), sagittal vertical axis (SVA), T1 spinopelvic inclination (T1SPI), pelvic tilt (PT), and pelvic incidence (PI). Final SVA and PT were analyzed to determine successful realignment. RC in the thoracic spine was designated favorable or unfavorable on the basis of impact on final SVA and PT. Mean PSO resection was 26°. LL increased from 20° to 49° (P < 0.001). SVA improved from 14 to 4 cm (P < 0.001), and PT improved from 33° to 25° (P < 0.001). Mean increase in TK was 13° (P = 0.002) but was unchanged in 11 patients. Five patients had a favorable RC, and 18 patients had an unfavorable RC. Unfavorable RC was attributed to junctional failure in 6 of 18 patients. Significant differences in the unfavorable RC group included age and greater preoperative PT, PI, SVA, and T1SPI. Significant postoperative alignment changes can occur through unfused thoracic spinal segments after lumbar PSO. Unfavorable RC may limit optimal correction and lead to clinical failures. Risk factors for unfavorable thoracic RC include older patients, larger preoperative PI and PT, and worse preoperative T1SPI and are not simply due to junctional failure. Care should be taken with selective lumbar fusion and PSO in older patients and in those with severe preoperative spinopelvic parameters.


Differential effect of ethanol on PC12 cell death.
Our goal was to examine the effects of ethanol on cell death using rat pheochromocytoma (PC12) cells as a neuronal model. Withdrawal of serum for 24 hr increased the number of PC12 cells labeled with ethidium homodimer indicating an increase in cell death. Inclusion of 50 mM ethanol during the serum deprivation further increased the amount of ethidium fluorescence by 39%. Although reducing the serum concentration from the usual 15 to 4% did not alter cellular viability, a significant increase in the amount of ethidium fluorescence was observed in PC12 cells incubated for 24 hr in the presence of 4% serum and 150 mM ethanol. No change in viability was observed in cells exposed to either 150 mM ethanol in the presence of 15% serum or 50 mM ethanol in the presence of 4% serum. Inclusion of ethanol during serum deprivation increased the amount of soluble DNA found in the 15,000 x g supernatant. Similarly, using the terminal deoxynucleotidyl transferase dUTP nick-end labeling method to visualize DNA fragmentation in situ, ethanol caused a 213% increase in the number of PC12 cells labeled during serum withdrawal. Agarose gel electrophoresis of the DNA isolated from cells maintained in the absence of serum yielded the classical DNA laddering pattern of 180 to 200 bp fragments suggestive of apoptosis. Ethanol caused a concentration-dependent increase in the amount of DNA laddering in cells deprived of serum. Furthermore, ethanol significantly potentiated the DNA laddering of cells maintained in 4% serum. In contrast to the ethanol-induced increase in cell death when serum factors were reduced or withdrawn, 150 mM ethanol lowered by 34% the number of ethidium-labeled PC12 cells observed after a 30-min exposure to 2 mM H2O2. Similarly, agarose gel electrophoresis of the DNA from H2O2-treated cells did not display DNA laddering. This study demonstrates that: 1) ethanol antagonizes the trophic action of serum factors; 2) pharmacologically relevant ethanol concentrations significantly potentiate apoptosis after serum withdrawal and 3) this enhancement appears specific for apoptosis.


Pseudo-Online BMI Based on EEG to Detect the Appearance of Sudden Obstacles during Walking.
The aim of this paper is to describe new methods for detecting the appearance of unexpected obstacles during normal gait from EEG signals, improving the accuracy and reducing the false positive rate obtained in previous studies. This way, an exoskeleton for rehabilitation or assistance of people with motor limitations commanded by a Brain-Machine Interface (BMI) could be stopped in case that an obstacle suddenly appears during walking. The EEG data of nine healthy subjects were collected during their normal gait while an obstacle appearance was simulated by the projection of a laser line in a random pattern. Different approaches were considered for selecting the parameters of the BMI: subsets of electrodes, time windows and classifier probabilities, which were based on a linear discriminant analysis (LDA). The pseudo-online results of the BMI for detecting the appearance of obstacles, with an average percentage of 63.9% of accuracy and 2.6 false positives per minute, showed a significant improvement over previous studies.


FOLFOX or CAPOX in Stage II to III Colon Cancer: Efficacy Results of the Italian Three or Six Colon Adjuvant Trial.
Purpose Given the cumulative neurotoxicity associated with oxaliplatin, a shorter duration of adjuvant therapy, if equally efficacious, would be advantageous for patients and health-care systems. Methods The Three or Six Colon Adjuvant trial is an open-label, phase III, multicenter, noninferiority trial randomizing patients with high-risk stage II or stage III colon cancer to receive 3 months or 6 months of FOLFOX (fluorouracil, leucovorin, and oxaliplatin) or CAPOX (capecitabine plus oxaliplatin). Primary end-point is relapse-free survival. Results 3,759 patients were accrued from 130 Italian sites, 64% receiving FOLFOX and 36% CAPOX. Two-thirds were stage III. The median time of follow up was 62 months and 772 relapses or deaths have been observed. The hazard ratio (HR) of the 3 months versus 6 months for relapse/death was 1.14 (95% CI, 0.99 to 1.32; P [for noninferiority] = .514) and the CI crossed the noninferiority limit of 1.20. However, the absolute difference in 3-year RFS was 1.9% (95% CI, -0.7% to 4.4%). Counter-intuitively, while the RFS curves were similar for stage III (HR, 1.07; 95% CI, 0.91 to 1.26) and for CAPOX treated patients (HR, 0.98; 95% CI, 0.77 to 1.26), they were not for stage II and for FOLFOX treated patients, with HR of 1.41 (95% CI, 1.05 to 1.89) and 1.23 (95% CI, 1.03 to 1.46), respectively, favoring the 6 months of treatment. Conclusion The Three or Six Colon Adjuvant trial failed to formally show noninferiority of 3 versus 6 months of treatment to the predefined margin of 20% relative increase. The results depended on the adjuvant regimen and risk. For CAPOX, 3 months were as good as 6 months; for FOLFOX, 6 months added extra benefit. Counter-intuitively, the low-risk patients benefitted more than the high-risk population from the 6-month duration. The choice of regimen and duration should depend on patient characteristics and be balanced against the extra toxicity of longer therapy.


Brainstem auditory evoked potentials and cochlear microphonics in the HMSN family with auditory neuropathy.
The aim of this work was to assess the hearing impairment in patients with hereditary motor and sensory neuropathy (HMSN). Elevation of pure tone thresholds in the presence of preserved inner ear function as suggested by cochlear microphonics (CM), absent or markedly abnormal brainstem auditory evoked potentials (BAEP), and elevation of speech perception out of proportion to the pure tone loss were found in the patients. From 28 members of a Gypsy family, we examined two siblings aged 31 and 30 years and their nephew aged 20 years, all suffering from HMSN that was associated with auditory neuropathy. All three affected members with difficulty of understanding speech had following investigations: pure tone and speech audiograms, BAEP, cochlear microphonics, and nerve conduction studies (NCV). the older two siblings had a flat 80 dB audiogram, whereas the younger one has flat 20 dB audiogram on the Lt. ear and 30 dB audiogram on the Rt. ear. All had no speech comprehension and no BAEP. Two patients had preserved cochlear microphonics on one ear. Peripheral nerves were electrically not elicitable, however, at the beginning of the disease nerve conduction was slow. in all three affected members with distinct clinical picture of HMSN their hearing impairment was proved to be due to severe auditory neuropathy in the presence of preserved inner ear function.


Perfect imaging of hypersurfaces via transformation optics.
Conventional optical imaging systems suffer from the presence of many imperfections, such as spherical aberrations, astigmatism, or coma. If the imaging system is corrected for spherical aberrations and fulfills the Abbe sine condition, perfect imaging is guaranteed between two parallel planes but only in a small neighborhood of the optical axis. It is therefore worth asking for optical systems that would allow for perfect imaging between arbitrary smooth surfaces without restrictions in shape or extension. In this Letter, we describe the application of transformation optics to design refractive index distributions that allow perfect, aberration-free imaging for various imaging configurations in R(n). A special case is the imaging between two extended parallel lines in R(2), which leads to the well-known hyperbolic secant index distribution that is used for the fabrication of gradient index lenses.


Hemorheological Approach for Early Detection of Chronic Kidney Disease and Diabetic Nephropathy in Type 2 Diabetes.
Hemorheologic alterations or changes in blood viscosity have been suggested to play a role in the pathogenesis of microvascular complications in diabetes. We measured various hemorheologic parameters in type 2 diabetes patients at different stages of chronic kidney disease (CKD) and assessed their possible role as early markers of diabetic nephropathy and renal insufficiency. One hundred-five patients with type 2 diabetes were divided into four groups according to glomerular filtration rate (GFR), which represents the kidney function. Hemorheologic parameters, including erythrocyte deformability, fibrinogen/elongation index (EI), and aggregation index (AI) were measured using a microfluidic hemorheometer, and critical shear stress (CSS) was measured using a microfluidic technique. Various metabolic parameters were assessed from fasting blood samples, and the urine albumin-to-creatinine ratio (ACR) was calculated from first morning voided urine. There were significant differences in red blood cell (RBC) deformability, AI, CSS, fibrinogen/EI, and ACR among patients in different stages of CKD (all P<0.05). RBC deformability and fibrinogen/EI significantly differed between normal (GFR >90 mL/min/1.73 m(2)) and CKD stage 2 (GFR 60-90 mL/min/1.73 m(2)) patients, whereas there was no such difference in ACR. In multiple regression analysis, fibrinogen/EI under a moderate shear stress of 3 Pa was an independent predictor of GFR (β=-0.328, P<0.05). Also, AI, CSS, and fibrinogen/EI were significantly different among patients at different stages of diabetic nephropathy, with a significant difference in fibrinogen/EI between normal and microalbuminuric patients (all P<0.05). RBC deformability and fibrinogen/EI are sensitive parameters measured via point-of-care testing for detecting erythrocyte alterations in early CKD and nephropathy in patients with type 2 diabetes. Further studies are warranted to verify their use as screening tools for diabetic nephropathy and renal impairment.


State of science: occupational slips, trips and falls on the same level.
Occupational slips, trips and falls on the same level (STFL) result in substantial injuries worldwide. This paper summarises the state of science regarding STFL, outlining relevant aspects of epidemiology, biomechanics, psychophysics, tribology, organisational influences and injury prevention. This review reaffirms that STFL remain a major cause of workplace injury and STFL prevention is a complex problem, requiring multi-disciplinary, multi-faceted approaches. Despite progress in recent decades in understanding the mechanisms involved in STFL, especially slipping, research leading to evidence-based prevention practices remains insufficient, given the problem scale. It is concluded that there is a pressing need to develop better fall prevention strategies using systems approaches conceptualising and addressing the factors involved in STFL, with considerations of the full range of factors and their interactions. There is also an urgent need for field trials of various fall prevention strategies to assess the effectiveness of different intervention components and their interactions. Practitioner Summary: Work-related slipping, tripping and falls on the same level are a major source of occupational injury. The causes are broadly understood, although more attention is needed from a systems perspective. Research has shown preventative action to be effective, but further studies are required to understand which aspects are most beneficial.


The osteoclast-not always guilty.
Osteopetrosis is usually regarded as the disease of nonfunctioning osteoclasts, with a consequent accumulation of osseocartilaginous material in the bone marrow. A recent report shows that, in some patients with the rare autosomal recessive form of the disease, the osteoclast defect may be non-cell autonomous, in this case due to a mutation in RANKL, the protein that normally controls osteoclast formation and activity.


FedExosomes: Engineering Therapeutic Biological Nanoparticles that Truly Deliver.
Many aspects of intercellular communication are mediated through "sending" and "receiving" packets of information via the secretion and subsequent receptor-mediated detection of biomolecular species including cytokines, chemokines, and even metabolites. Recent evidence has now established a new modality of intercellular communication through which biomolecular species are exchanged between cells via extracellular lipid vesicles. A particularly important class of extracellular vesicles is exosomes, which is a term generally applied to biological nanovesicles ~30-200 nm in diameter. Exosomes form through invagination of endosomes to encapsulate cytoplasmic contents, and upon fusion of these multivesicular endosomes to the cell surface, exosomes are released to the extracellular space and transport mRNA, microRNA (miRNA) and proteins between cells. Importantly, exosome-mediated delivery of such cargo molecules results in functional modulation of the recipient cell, and such modulation is sufficiently potent to modulate disease processes in vivo. It is possible that such functional delivery of biomolecules indicates that exosomes utilize native mechanisms (e.g., for internalization and trafficking) that may be harnessed by using exosomes to deliver exogenous RNA for therapeutic applications. A complementary perspective is that understanding the mechanisms of exosome-mediated transport may provide opportunities for "reverse engineering" such mechanisms to improve the performance of synthetic delivery vehicles. In this review, we summarize recent progress in harnessing exosomes for therapeutic RNA delivery, discuss the potential for engineering exosomes to overcome delivery challenges and establish robust technology platforms, and describe both potential challenges and advantages of utilizing exosomes as RNA delivery vehicles.


Evaluation of Phytocannabinoids from High Potency Cannabis sativa using In Vitro Bioassays to Determine Structure-Activity Relationships for Cannabinoid Receptor 1 and Cannabinoid Receptor 2.
Cannabis has been around for thousands of years and has been used recreationally, medicinally, and for fiber. Over 500 compounds have been isolated from Cannabis sativa with approximately 105 being cannabinoids. Of those 105 compounds, Δ9-tetrahydrocannabinol has been determined as the primary constituent, which is also responsible for the psychoactivity associated with Cannabis. Cannabinoid receptors belong to the large superfamily of G protein-coupled receptors. Targeting the cannabinoid receptors has the potential to treat a variety of conditions such as pain, neurodegeneration, appetite, immune function, anxiety, cancer, and others. Developing in vitro bioassays to determine binding and functional activity of compounds has the ability to lead researchers to develop a safe and effective drug that may target the cannabinoid receptors. Using radioligand binding and functional bioassays, a structure-activity relationship for major and minor cannabinoids was developed.


Microalgae cultivation in agro-industrial effluents for biodiesel application: effects of the availability of nutrients.
The present study evaluated the cultivation of microalgae in a photobioreactor using effluents from the meat-processing industry, which had been previously treated at the primary and secondary levels. Scenedesmus sp. was the dominant genus in the phytoplankton community in both of the evaluated effluents. The different nutritional conditions affected the production of biomass, which reached 1,160 mg/L of volatile suspended solids (VSS) and 371 mg/L of VSS with cultivation in the primary (PE) and secondary effluents (SE), respectively. In both effluents, great removal efficiencies close to quantification limits were observed for ammoniacal nitrogen and soluble phosphorus. Regarding the accumulation of lipids, there were no considerable differences between the effluents. The highest lipid productivity that was observed in the PE, which reached 3.7 g/m²·d, was attributed to its larger production of biomass as a consequence of its better nutritional condition in relation to the SE.


Braincase simplification and the origin of lissamphibians.
Dissorophoidea, a group of temnospondyl tetrapods that first appear in the Late Carboniferous, is made up of two clades ⎼ Olsoniformes and Amphibamiformes (Branchiosauridae and Amphibamidae) ⎼ the latter of which is widely thought to have given rise to living amphibians (i.e., Lissamphibia). The lissamphibian braincase has a highly derived morphology with several secondarily lost elements; however, these losses have never been incorporated into phylogenetic analyses and thus the timing and nature of these evolutionary events remain unknown. Hindering research into this problem has been the lack of phylogenetic analyses of Dissorophoidea that includes both taxonomically dense sampling and specific characters to document changes in the braincase in the lineage leading to Lissamphibia. Here we build on a recent, broadly sampled dissorophoid phylogenetic analysis to visualize key events in the evolution of the lissamphibian braincase. Our ancestral character state reconstructions show a clear, step-wise trend towards reduction of braincase ossification leading to lissamphibians, including reduction of the sphenethmoid, loss of the basioccipital at the Amphibamiformes node, and further loss of both the basisphenoid and the hypoglossal nerve foramina at the Lissamphibia node. Our analysis confirms that the highly derived condition of the lissamphibian braincase is characterized by overall simplification in terms of the number and extent of chondrocranial ossifications.


The vitronectin receptor serves as an accessory molecule for the activation of a subset of gamma/delta T cells.
Constitutive production of cytokines was observed in 3 of 12 gamma/delta T cell lines derived from murine epidermis and correlated with the expression of the C gamma 4, V delta 6 T cell receptor (TCR). After adaptation of one of the lines (T195/BW) to serum-free culture conditions, cessation of the "spontaneous" production of interleukin 4 (IL-4) was observed and IL-4 production could then by induced by the addition of RGD-containing extracellular matrix (ECM) proteins to the culture. The response to the ECM proteins could be completely inhibited by a mAb to the murine vitronectin receptor (VNR). However, the induction of IL-4 production could also be inhibited by anti-CD3 and by an anti-clonotypic mAb to the TCR-gamma/delta of T195/BW. As TCR-gamma/delta loss mutants of T195/BW also failed to respond to ECM proteins, these data demonstrate that engagement of the VNR by its ligand is necessary, but not sufficient, for the induction of IL-4 production. Furthermore, the VNR is expressed by many other T cell clones (both gamma/delta and alpha/beta), none of which produce lymphokines constitutively. Taken together, these observations strongly favor the view that not only is coexpression of the VNR and TCR required for the induction of IL-4 production, but that the TCR must also be engaged by its ligand, most likely a cell surface antigen expressed by the hybridoma itself.


Antihepatoma activity of Artocarpus communis is higher in fractions with high artocarpin content.
Extracts from natural plants have been used in traditional medicine for many centuries worldwide. Artocarpus communis is one such plant that has been used to treat liver cirrhosis, hypertension, and diabetes. To our knowledge, this study is the first to investigate the antihepatoma activity of A. communis toward HepG2 and PLC/PRF/5 cells and the first to explore the relationship between antihepatoma activity and the active compound artocarpin content in different fractions of A. communis. A. communis methanol extract and fractions induced dose-dependent reduction of tumor cell viability. DNA laddering analysis revealed that A. communis extract and fractions did not induce apoptosis in HepG2 and PLC/PRF/5 cells. Instead, acridine orange staining revealed that A. communis triggered autophagic cell death in a dose-dependent manner. The antihepatoma activity of A. communis is attributable to artocarpin. The fractions with the highest artocarpin content were also the fractions with the highest antihepatoma activity in the following order: dichloromethane fraction > methanol extract > ethyl acetate fraction > n-butanol fraction > n-hexane fraction. Taken together, A. communis showed antihepatoma activity through autophagic cell death. The effect was related to artocarpin content. Artocarpin could be considered an indicator of the anticancer potential of A. communis extract.


Isolation, culture and immortalisation of hepatic oval cells from adult mice fed a choline-deficient, ethionine-supplemented diet.
Oval cells have great potential for use in cell therapy to treat liver disease, however this cannot be achieved until the factors which govern their proliferation and differentiation are better understood. We describe a method to establish primary cultures of murine oval cells, and the derivation of two novel lines from these. Primary cultures from the livers of wildtype or TAT-GRE lacZ transgenic mice subjected to a choline-deficient, ethionine-supplemented diet comprised up to 80% oval cells at day 7 based on A6 or CK19 staining. Cell lines were clonally derived, which underwent spontaneous immortalisation following prolonged maintenance in culture. Immunostaining and RT-PCR demonstrated they express hepatocytic and biliary markers and they were therefore termed "bipotential murine oval liver" (BMOL) cells. Under proliferating culture conditions, BMOL or BMOL-TAT cells abundantly expressed oval cell and biliary markers, whereas mature hepatocytic markers were upregulated when the growth conditions were changed to facilitate differentiation. Hepatic differentiation of BMOL-TAT cells could be traced by measuring the expression of their lacZ transgene, which is driven by a promoter element from tyrosine aminotransferase (TAT), a marker of adult hepatocytes. Interestingly, haematopoietic markers were upregulated in superconfluent cultures, indicating a possible multipotentiality. None of the cell lines grew in semi-solid agar, nor did they form tumours in nude mice, suggesting they are non-tumourigenic. These novel murine oval cell lines, together with a reliable method for isolation and culture of primary oval cells, will provide a useful tool for investigating the contribution of oval cells to liver regeneration.


Characterization of RAD9 of Saccharomyces cerevisiae and evidence that its function acts posttranslationally in cell cycle arrest after DNA damage.
In eucaryotic cells, incompletely replicated or damaged chromosomes induce cell cycle arrest in G2 before mitosis, and in the yeast Saccharomyces cerevisiae the RAD9 gene is essential for the cell cycle arrest (T.A. Weinert and L. H. Hartwell, Science 241:317-322, 1988). In this report, we extend the analysis of RAD9-dependent cell cycle control. We found that both induction of RAD9-dependent arrest in G2 and recovery from arrest could occur in the presence of the protein synthesis inhibitor cycloheximide, showing that the mechanism of RAD9-dependent control involves a posttranslational mechanism(s). We have isolated and determined the DNA sequence of the RAD9 gene, confirming the DNA sequence reported previously (R. H. Schiestl, P. Reynolds, S. Prakash, and L. Prakash, Mol. Cell. Biol. 9:1882-1886, 1989). The predicted protein sequence for the Rad9 protein bears no similarity to sequences of known proteins. We also found that synthesis of the RAD9 transcript in the cell cycle was constitutive and not induced by X-irradiation. We constructed yeast cells containing a complete deletion of the RAD9 gene; the rad9 null mutants were viable, sensitive to X- and UV irradiation, and defective for cell cycle arrest after DNA damage. Although Rad+ and rad9 delta cells had similar growth rates and cell cycle kinetics in unirradiated cells, the spontaneous rate of chromosome loss (in unirradiated cells) was elevated 7- to 21-fold in rad9 delta cells. These studies show that in the presence of induced or endogenous DNA damage, RAD9 is a negative regulator that inhibits progression from G2 in order to preserve cell viability and to maintain the fidelity of chromosome transmission.


Radioactive excretion in human milk following administration of technetium Tc 99m macroaggregated albumin.
Albumin-tagged sodium pertechnetate (technetium) is routinely used in nuclear medicine for scanning procedures of the lung. The rate of excretion of this radionuclide into breast milk and the resultant potential radiation hazard to the nursing infant have received little attention. Therefore the milk from a nursing mother who required a lung scan because of suspected pulmonary emboli using an intravenous injection of 4 mCi of technetium Tc 99 macroaggregated human serum albumin was monitored. Albumin tagging severely limited the entrance of technetium into her milk and the radioactivity of the milk returned to base line by 24 hours. A total of 2.02 muCi of technetium was measured in the 24-hour milk collection after technetium injection and 94% of this amount was excreted by 15.5 hours. This amount of technetium administered orally to a newborn would deliver a total body radiation dose of .3 mrad. Therefore, an infant would receive trivial doses of radiation if breast-feeding were resumed 15.5 hours after administration of the radionuclide to the mother and nursing can clearly be resumed safely 24 hours after injection.


Trifluoroacetic acid in 2,2,2-trifluoroethanol facilitates S(N)Ar reactions of heterocycles with arylamines.
Small-molecule drug discovery requires reliable synthetic methods for attaching amino compounds to heterocyclic scaffolds. Trifluoroacetic acid-2,2,2-trifluoroethanol (TFA-TFE) is as an effective combination for achieving SN Ar reactions between anilines and heterocycles (e.g., purines and pyrimidines) substituted with a leaving group (fluoro-, chloro-, bromo- or alkylsulfonyl). This method provides a variety of compounds containing a "kinase-privileged fragment" associated with potent inhibition of kinases. TFE is an advantageous solvent because of its low nucleophilicity, ease of removal and ability to solubilise polar substrates. Furthermore, TFE may assist the breakdown of the Meisenheimer-Jackson intermediate by solvating the leaving group. TFA is a necessary and effective acidic catalyst, which activates the heterocycle by N-protonation without deactivating the aniline by conversion into an anilinium species. The TFA-TFE methodology is compatible with a variety of functional groups and complements organometallic alternatives, which are often disadvantageous because of the expense of reagents, the frequent need to explore diverse sets of reaction conditions, and problems with product purification. In contrast, product isolation from TFA-TFE reactions is straightforward: evaporation of the reaction mixture, basification and chromatography affords analytically pure material. A total of 45 examples are described with seven discrete heterocyclic scaffolds and 2-, 3- and 4-substituted anilines giving product yields that are normally in the range 50-90 %. Reactions can be performed with either conventional heating or microwave irradiation, with the latter often giving improved yields.


Chronic application of NMDA decreases the NMDA sensitivity of the evoked tectal potential in the frog.
The activity-dependent mechanism that refines the topography of the retinotectal projection in frogs is mediated by the NMDA receptor. Earlier studies found that chronic treatment of the optic tectum with the NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (DL-AP5) desegregated eye-specific stripes in three-eyed frogs, while chronic treatment with NMDA sharpened stripe borders (Cline et al., 1987; Cline and Constantine-Paton, 1990). We now report that this same chronic treatment with NMDA decreases the electro-physiologically measured sensitivity of the optic tectum to applied NMDA: acute application of a given concentration of NMDA decreased the evoked tectal potential to a lesser extent in animals chronically treated with NMDA than it did in normal and sham-treated animals. This is observed as a shift to the right in the NMDA dose-response curves for both the positive and negative postsynaptic components of the evoked tectal response. We believe that this decreased NMDA receptor effectiveness further restricts the intermingling of axon branches from the two eyes by limiting synapse stabilization to areas where afferent activity is most correlated. This would account for the anatomical sharpening of stripe borders (i.e., increased afferent segregation). Quantitative autoradiographic analysis of 3H-glutamate binding to NMDA receptors indicated that binding densities within the tectum did not differ between control groups and NMDA chronically treated groups. We suggest that in the experimental animals the response to NMDA may be decreased by a change in the effectiveness of individual NMDA receptors rather than by decreases in receptor number. This experimentally induced change may be analogous to naturally occurring decreases in receptor function that correlate with the end of some periods of visual plasticity in mammals.


Radiology and endoscopy in acute upper gastrointestinal bleeding.
Of 112 patients admitted with acute upper gastrointestinal bleeding, the presumed bleeding site was detected in 61-5% of cases by radiology and in 57% of cases on endoscopy. Thirty-one patients who had barium-meal examination were operated on and the surgical and radiological findings agreed in 26 (84%). Twenty-three patients who had endoscopy were operated on and the surgical and endoscopic findings agreed in 15 (65%). In 10 cases radiology detected a lesion not identified on endoscopy and in nine endoscopy detected a lesion not seen at radiology. We suggest that when there are two potential sources of bleeding radiology as well as endoscopy can detect the actively bleeding lesion. The supplementary nature of radiology and endoscopy is emphasised and we conclude that both methods should be used if there is any doubt at the initial radiological or endoscopic examination about the source of the bleeding.


The predictability of dentoskeletal factors for soft-tissue chin strain during lip closure.
To investigate the dentoskeletal factors which may predict soft-tissue chin strain during lip closure. The pretreatment frontal and lateral facial photographs and lateral cephalograms of 209 women (aged 18-30 years) with Angle's Class I or II malocclusion were examined. The subjects were categorized by three examiners into the no-strain and strain groups according to the soft-tissue chin tension or deformation during lip closure. Relationships of the cephalometric measurements with the group classification were analyzed by logistic regression analysis, and a classification and regression tree (CART) model was used to define the predictive variables for the group classification. The lower the value of the overbite depth indicator (ODI) and the higher the values of upper incisor to Nasion-Pogonion (U1-NPog, mm), overjet, and upper incisor to upper lip (U1-upper lip, mm), the more likely was the subject to be classified into the strain group. The CART showed that U1-NPog was the most prominent predictor of soft-tissue chin strain (cut-off value of 14.2 mm), followed by overjet. To minimize strain of the soft-tissue chin, orthodontic treatment should be oriented toward increasing the ODI value while decreasing the U1-NPog, overjet, and U1 upper lip values.


Risk of gastrointestinal perforation in patients taking oral fluoroquinolone therapy: An analysis of nationally representative cohort.
Fluoroquinolone is a commonly prescribed antimicrobial agent, and up to 20% of its users registers adverse gastroenterological symptoms. We aimed to evaluate the association between use of fluoroquinolone and gastrointestinal tract perforation. We conducted a nested case-control study on a national health insurance claims database between 1998 and 2011. The use of fluoroquinolones was classified into current (< 60 days), past (61-365 days prior to the index date) and any prior year use of fluoroquinolones. We used the conditional logistic regression model to estimate rate ratios (RRs), adjusting or matching by a disease risk score (DRS). We identified a cohort of 17,510 individuals diagnosed with gastrointestinal perforation and matched them to 1,751,000 controls. Current use of fluoroquinolone was associated with the greatest increase in risk of gastrointestinal perforations after DRS score adjustment (RR, 1.90; 95% CI, 1.62-2.22). The risk of gastrointestinal perforation was attenuated for past (RR, 1.33; 95% CI, 1.20-1.47) and any prior year use (RR, 1.46; 95% CI, 1.34-1.59). To gain insights into whether the observed association can be explained by unmeasured confounder, we compared the risk of gastrointestinal perforation between fluoroquinolone and macrolide. Use of macrolide, an active comparator, was not associated with a significant increased risk of gastrointestinal perforation (RR, 1.11, 95%CI, 0.15-7.99). Sensitivity analysis focusing on perforation requiring in-hospital procedures also demonstrated an increased risk associated with current use. To mitigate selection bias, we have also excluded people who have never used fluoroquinolone before or people with infectious colitis, enteritis or gastroenteritis. In both of the analysis, a higher risk of gastrointestinal perforation was still associated with the use of fluoroquinolone. We found that use of fluoroquinolones was associated with a non-negligible increased risk of gastrointestinal perforation, and physicians should be aware of this possible association.


A novel feedback mechanism by Ephrin-B1/B2 in T-cell activation involves a concentration-dependent switch from costimulation to inhibition.
Bidirectional signals via Eph receptors/ephrins have been recognized as major forms of contact-dependent cell communications such as cell attraction and repulsion. T cells express EphBs, and their ligands, the ephrin-Bs, have been known as costimulatory molecules for T-cell proliferation. Recently, another remarkable feature of ephrin-As has emerged in the form of a concentration-dependent transition from promotion to inhibition in axon growth. Here we examined whether this modification plays a role in ephrin-B costimulation in murine primary T cells. Low doses of ephrin-B1 and ephrin-B2 costimulated T-cell proliferation induced by anti-CD3, but high concentrations strongly inhibited it. In contrast, ephrin-B3 showed a steadily increasing stimulatory effect. This modulation was virtually preserved in T cells from mice simultaneously lacking four genes, EphB1, EphB2, EphB3, and EphB6. High concentrations of ephrin-B1/B2, but not ephrin-B3, inhibited the anti-CD3-induced phosphorylation of Lck and its downstream signals such as Erk and Akt. Additionally, high doses of any ephrin-Bs could phosphorylate EphB4. However, only ephrin-B1/B2 but not ephrin-B3 recruited SHP1, a phosphatase to suppress the phosphorylation of Lck. These data suggest that EphB4 signaling could engage in negative feedback to TCR signals. T-cell activation may be finely adjusted by the combination and concentration of ephrin-Bs expressed in the immunological microenvironment.


Drug-induced reactivation of apoptosis abrogates HIV-1 infection.
HIV-1 blocks apoptosis, programmed cell death, an innate defense of cells against viral invasion. However, apoptosis can be selectively reactivated in HIV-infected cells by chemical agents that interfere with HIV-1 gene expression. We studied two globally used medicines, the topical antifungal ciclopirox and the iron chelator deferiprone, for their effect on apoptosis in HIV-infected H9 cells and in peripheral blood mononuclear cells infected with clinical HIV-1 isolates. Both medicines activated apoptosis preferentially in HIV-infected cells, suggesting that the drugs mediate escape from the viral suppression of defensive apoptosis. In infected H9 cells, ciclopirox and deferiprone enhanced mitochondrial membrane depolarization, initiating the intrinsic pathway of apoptosis to execution, as evidenced by caspase-3 activation, poly(ADP-ribose) polymerase proteolysis, DNA degradation, and apoptotic cell morphology. In isolate-infected peripheral blood mononuclear cells, ciclopirox collapsed HIV-1 production to the limit of viral protein and RNA detection. Despite prolonged monotherapy, ciclopirox did not elicit breakthrough. No viral re-emergence was observed even 12 weeks after drug cessation, suggesting elimination of the proviral reservoir. Tests in mice predictive for cytotoxicity to human epithelia did not detect tissue damage or activation of apoptosis at a ciclopirox concentration that exceeded by orders of magnitude the concentration causing death of infected cells. We infer that ciclopirox and deferiprone act via therapeutic reclamation of apoptotic proficiency (TRAP) in HIV-infected cells and trigger their preferential elimination. Perturbations in viral protein expression suggest that the antiretroviral activity of both drugs stems from their ability to inhibit hydroxylation of cellular proteins essential for apoptosis and for viral infection, exemplified by eIF5A. Our findings identify ciclopirox and deferiprone as prototypes of selectively cytocidal antivirals that eliminate viral infection by destroying infected cells. A drug-based drug discovery program, based on these compounds, is warranted to determine the potential of such agents in clinical trials of HIV-infected patients.


Changes in Clavicle Length and Maturation in Americans: 1840-1980.
Secular changes refer to short-term biological changes ostensibly due to environmental factors. Two well-documented secular trends in many populations are earlier age of menarche and increasing stature. This study synthesizes data on maximum clavicle length and fusion of the medial epiphysis in 1840-1980 American birth cohorts to provide a comprehensive assessment of developmental and morphological change in the clavicle. Clavicles from the Hamann-Todd Human Osteological Collection (n = 354), McKern and Stewart Korean War males (n = 341), Forensic Anthropology Data Bank (n = 1,239), and the McCormick Clavicle Collection (n = 1,137) were used in the analysis. Transition analysis was used to evaluate fusion of the medial epiphysis (scored as unfused, fusing, or fused). Several statistical treatments were used to assess fluctuations in maximum clavicle length. First, Durbin-Watson tests were used to evaluate autocorrelation, and a local regression (LOESS) was used to identify visual shifts in the regression slope. Next, piecewise regression was used to fit linear regression models before and after the estimated breakpoints. Multiple starting parameters were tested in the range determined to contain the breakpoint, and the model with the smallest mean squared error was chosen as the best fit. The parameters from the best-fit models were then used to derive the piecewise models, which were compared with the initial simple linear regression models to determine which model provided the best fit for the secular change data. The epiphyseal union data indicate a decline in the age at onset of fusion since the early twentieth century. Fusion commences approximately four years earlier in mid- to late twentieth-century birth cohorts than in late nineteenth- and early twentieth-century birth cohorts. However, fusion is completed at roughly the same age across cohorts. The most significant decline in age at onset of epiphyseal union appears to have occurred since the mid-twentieth century. LOESS plots show a breakpoint in the clavicle length data around the mid-twentieth century in both sexes, and piecewise regression models indicate a significant decrease in clavicle length in the American population after 1940. The piecewise model provides a slightly better fit than the simple linear model. Since the model standard error is not substantially different from the piecewise model, an argument could be made to select the less complex linear model. However, we chose the piecewise model to detect changes in clavicle length that are overfitted with a linear model. The decrease in maximum clavicle length is in line with a documented narrowing of the American skeletal form, as shown by analyses of cranial and facial breadth and bi-iliac breadth of the pelvis. Environmental influences on skeletal form include increases in body mass index, health improvements, improved socioeconomic status, and elimination of infectious diseases. Secular changes in bony dimensions and skeletal maturation stipulate that medical and forensic standards used to deduce information about growth, health, and biological traits must be derived from modern populations.