Human Papillomavirus Vaccination Among Young Men Who Have Sex With Men and Transgender Women in 2 US Cities, 2012-2014.
Since 2011, in the United States, quadrivalent human papillomavirus (HPV) vaccine has been recommended for boys aged 11 to 12 years, men through age 21, and men who have sex with men (MSM) through age 26. We assessed HPV vaccination coverage and factors associated with vaccination among young MSM (YMSM) and transgender women (TGW) in 2 cities. During 2012-2014, 808 YMSM and TGW aged 18 to 26 years reported vaccination status in a self-administered computerized questionnaire at 3 sexually transmitted disease (STD) clinics in Los Angeles and Chicago. Associations with HPV vaccination were assessed using bivariate and multivariable models to calculate adjusted odds ratios (aORs) and 95% confidence intervals (CIs). Few of the diverse participants (Hispanic/Latino, 38.0%; white, 27.0%; and black/African American, 17.9%) reported receiving 1 or more HPV vaccine doses (n = 111 [13.7%]) and even fewer reported 3 doses (n = 37 [4.6%]). A multivariable model found associations between vaccination and having a 4-year college degree or higher (aOR, 2.83; CI, 1.55-5.17) and self-reported STDs (aOR, 1.21; CI, 1.03-1.42). In a model including recommendation variables, the strongest predictor of vaccination was a health care provider recommendation (aOR, 11.85; CI, 6.70-20.98). Human papillomavirus vaccination coverage was low among YMSM and TGW in this 2-US city study. Our findings suggest further efforts are needed to reach YMSM seeking care in STD clinics, increase strong recommendations from health care providers, and integrate HPV vaccination with other clinical services such as STD testing.


Pylorus preservation pancreatectomy or not.
Pancreaticoduodenectomy (PD) is the treatment of choice for various benign and malignant tumors of the pancreatic head or the periampullary region, and the only hope for cure in patients with cancer at this side. While it has been associated with high morbidity and mortality rates in the last century, its centralization in specialized institutions together with refinements in the operative technique and better management of postoperative complications have made PD a standardized, safe procedure. Besides the classic Whipple procedure including distal gastrectomy, two variations of PD with or without pylorus resection, but preservation of the entire stomach in either procedure exist today. Pylorus-preserving PD has gained wide acceptance as standard procedure and is being performed by an increasing number of pancreatic surgeons. After its oncological adequacy was questioned initially, pylorus-preserving PD was shown to be equivalent to the classic Whipple procedure regarding tumor recurrence and long-term survival. Moreover, operation time and blood loss were shown to be reduced in the pylorus-preserving procedure and benefits in nutritional status and quality of life were observed. However, preservation of the pylorus has been suggested to result in an increased incidence of postoperative delayed gastric emptying (DGE). In this context, pylorus-resecting PD has become popular especially in Japan with the aim to prevent DGE by removal of the pylorus but preservation of the stomach. In contrast to positive results from early studies, latest high-quality randomized controlled trial (RCT) data show that pylorus resection does not reduce DGE compared to the pylorus-preserving operation. Non-superiority of pylorus resection was also confirmed in current meta-analysis on this topic. This article summarizes the existing evidence on PD with or without pylorus preservation and derives recommendations for daily practice.


Chk1 promotes replication fork progression by controlling replication initiation.
DNA replication starts at initiation sites termed replication origins. Metazoan cells contain many more potential origins than are activated (fired) during each S phase. Origin activation is controlled by the ATR checkpoint kinase and its downstream effector kinase Chk1, which suppresses origin firing in response to replication blocks and during normal S phase by inhibiting the cyclin-dependent kinase Cdk2. In addition to increased origin activation, cells deficient in Chk1 activity display reduced rates of replication fork progression. Here we investigate the causal relationship between increased origin firing and reduced replication fork progression. We use the Cdk inhibitor roscovitine or RNAi depletion of Cdc7 to inhibit origin firing in Chk1-inhibited or RNAi-depleted cells. We report that Cdk inhibition and depletion of Cdc7 can alleviate the slow replication fork speeds in Chk1-deficient cells. Our data suggest that increased replication initiation leads to slow replication fork progression and that Chk1 promotes replication fork progression during normal S phase by controlling replication origin activity.


Quantifying cortical EEG responses to TMS in (un)consciousness.
We normally assess another individual's level of consciousness based on her or his ability to interact with the surrounding environment and communicate. Usually, if we observe purposeful behavior, appropriate responses to sensory inputs, and, above all, appropriate answers to questions, we can be reasonably sure that the person is conscious. However, we know that consciousness can be entirely within the brain, even in the absence of any interaction with the external world; this happens almost every night, while we dream. Yet, to this day, we lack an objective, dependable measure of the level of consciousness that is independent of processing sensory inputs and producing appropriate motor outputs. Theoretically, consciousness is thought to require the joint presence of functional integration and functional differentiation, otherwise defined as brain complexity. Here we review a series of recent studies in which Transcranial Magnetic Stimulation combined with electroencephalography (TMS/EEG) has been employed to quantify brain complexity in wakefulness and during physiological (sleep), pharmacological (anesthesia) and pathological (brain injury) loss of consciousness. These studies invariably show that the complexity of the cortical response to TMS collapses when consciousness is lost during deep sleep, anesthesia and vegetative state following severe brain injury, while it recovers when consciousness resurges in wakefulness, during dreaming, in the minimally conscious state or locked-in syndrome. The present paper will also focus on how this approach may contribute to unveiling the pathophysiology of disorders of consciousness affecting brain-injured patients. Finally, we will underline some crucial methodological aspects concerning TMS/EEG measurements of brain complexity.


Kinesin-14 family proteins and microtubule dynamics define S. pombe mitotic and meiotic spindle assembly, and elongation.
To segregate the chromosomes faithfully during cell division, cells assemble a spindle that captures the kinetochores and pulls them towards opposite poles. Proper spindle function requires correct interplay between microtubule motors and non-motor proteins. Defects in spindle assembly or changes in spindle dynamics are associated with diseases, such as cancer or developmental disorders. Here, we compared mitotic and meiotic spindles in fission yeast. We show that, even though mitotic and meiotic spindles underwent the typical three phases of spindle elongation, they have distinct features. We found that the relative concentration of the kinesin-14 family protein Pkl1 is decreased in meiosis I compared to mitosis, while the concentration of the kinesin-5 family protein Cut7 remains constant. We identified the second kinesin-14 family protein Klp2 and microtubule dynamics as factors necessary for proper meiotic spindle assembly. This work defines the differences between mitotic and meiotic spindles in fission yeast Schizosaccharomyces pombe, and provides prospect for future comparative studies.This article has an associated First Person interview with the first author of the paper.


Effects of supercoiling on enhancer-promoter contacts.
Using Brownian dynamics simulations, we investigate here one of possible roles of supercoiling within topological domains constituting interphase chromosomes of higher eukaryotes. We analysed how supercoiling affects the interaction between enhancers and promoters that are located in the same or in neighbouring topological domains. We show here that enhancer-promoter affinity and supercoiling act synergistically in increasing the fraction of time during which enhancer and promoter stay in contact. This stabilizing effect of supercoiling only acts on enhancers and promoters located in the same topological domain. We propose that the primary role of recently observed supercoiling of topological domains in interphase chromosomes of higher eukaryotes is to assure that enhancers contact almost exclusively their cognate promoters located in the same topological domain and avoid contacts with very similar promoters but located in neighbouring topological domains.


The Protein Information Management System (PiMS): a generic tool for any structural biology research laboratory.
The techniques used in protein production and structural biology have been developing rapidly, but techniques for recording the laboratory information produced have not kept pace. One approach is the development of laboratory information-management systems (LIMS), which typically use a relational database schema to model and store results from a laboratory workflow. The underlying philosophy and implementation of the Protein Information Management System (PiMS), a LIMS development specifically targeted at the flexible and unpredictable workflows of protein-production research laboratories of all scales, is described. PiMS is a web-based Java application that uses either Postgres or Oracle as the underlying relational database-management system. PiMS is available under a free licence to all academic laboratories either for local installation or for use as a managed service.


The Influence of Emotional State on Learning From Reward and Punishment in Borderline Personality Disorder.
Despite preliminary evidence that individuals with borderline personality disorder (BPD) demonstrate deficits in learning from corrective feedback, no studies have examined the influence of emotional state on these learning deficits in BPD. This laboratory study examined the influence of negative emotions on learning among participants with BPD (n = 17), compared with clinical (past-year mood/anxiety disorder; n = 20) and healthy (n = 23) controls. Participants completed a reinforcement learning task before and after a negative emotion induction. The learning task involved presenting pairs of stimuli with probabilistic feedback in the training phase, and subsequently assessing accuracy for choosing previously rewarded stimuli or avoiding previously punished stimuli. ANOVAs and ANCOVAs revealed no significant between-group differences in overall learning accuracy. However, there was an effect of group in the ANCOVA for postemotion induction high-conflict punishment learning accuracy, with the BPD group showing greater decrements in learning accuracy than controls following the negative emotion induction.


Deep Learning Architecture to Assist With Steering a Powered Wheelchair.
This paper describes a novel Deep Learning architecture to assist with steering a powered wheelchair. A rule-based approach is utilized to train and test a Long Short Term Memory (LSTM) Neural Network. It is the first time a LSTM has been used for steering a powered wheelchair. A disabled driver uses a joystick to provide desired speed and direction, and the Neural Network provides a safe direction for the wheelchair. Results from the Neural Network are mixed with desired speed and direction to avoid obstacles. Inputs originate from a joystick and from three ultrasonic transducers attached to the chair. The resultant course is a blend of desired directions and directions that steer the chair to avoid collision. A rule-based approach is used to create a training and test set for the Neural Network system and applies deep learning to predict a safe route for a wheelchair. The user can over-ride the new system if necessary.


Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications.
Electron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane-derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids (AG) for use in both conventional EM and cryo-EM/ET applications. We examined the utility of AG for the selective capture of human immunodeficiency virus virus-like particles, influenza A, and measles virus. We applied nickel-nitrilotriacetic acid lipid layers in combination with molecular adaptors to selectively adhere the viruses to the AG surface. This further development of the AG method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analyses.


Non-Medical Strategies to Improve Pregnancy Outcomes of Women with Gestational Diabetes Mellitus: A literature review.
This review aimed to examine the literature related to non-medical strategies used to improve pregnancy outcomes of women with gestational diabetes mellitus (GDM) and to determine the risk of bias of the selected studies. Treatment for GDM is changing due to the increased prevalence of GDM-related maternal and neonatal complications. A growing body of evidence suggests that early detection, aggressive monitoring and management of GDM using non-medical strategies can greatly improve outcomes for pregnant women and their babies. PubMed® (National Library of Medicine, Bethesda, Maryland, USA), Cumulative Index to Nursing and Allied Health Literature® (EBSCO Information Services, Ipswich, Massachusetts, USA), SCOPUS® (Elsevier, Amsterdam, Netherlands) and other electronic databases were searched for relevant literature published between 2005-2015. A total of 15 studies on women with GDM that met the inclusion criteria were included in this review and assessment of risk of bias was performed for each study. The results of the studies were consistent with findings of significant improvement in maternal and neonatal outcomes when diet was combined with moderate exercise, self-monitoring of blood glucose and individualised health education. Future intervention studies in this area should be focussed on identifying and implementing factors that enhance and encourage adherence to the healthy behaviours mentioned above.


Replication of adenovirus and SV40 chromosomes in vitro.
As an approach to studying the mechanisms involved in the replication of eukaryotic chromosomes, we have developed and characterized cell-free replication systems for the animal viruses, adenovirus and SV40. In this report we summarize recent work on the proteins required for the initiation of DNA synthesis in these two systems. The adenovirus origin of DNA replication was shown to consist of three functionally distinct sequence domains. Cellular proteins that specifically recognize each of these domains were purified and characterized. Initiation of adenovirus DNA replication was reconstituted from two virus-encoded and three cell-encoded factors. The SV40 origin of replication consists of a 65 base pair DNA segment that contains a high affinity binding site for the viral initiation protein T antigen. Evidence is presented that the first step in initiation of SV40 DNA replication involves the specific binding of T antigen to the origin, followed by the local unwinding of the two strands of the template. The unwinding reaction is specific for DNA templates containing the SV40 origin and requires ATP hydrolysis. In addition to T antigen, efficient unwinding requires a cellular factor(s) that can be replaced by the single-stranded DNA binding protein of Escherichia coli. These results indicate that the recently discovered helicase activity of T antigen plays a central role in initiation of viral DNA synthesis.


Predictive genetic testing for BRCA1/2 in a UK clinical cohort: three-year follow-up.
This prospective multicentre study assesses long-term impact of genetic testing for breast/ovarian cancer predisposition in a clinical cohort. Areas evaluated include risk management, distress and insurance problems 3 years post-testing. Participants are adults unaffected with cancer from families with a known BRCA1/2 mutation. One hundred and ninety-three out of 285 (70% response) participants at nine UK clinical genetics centres completed assessments at 3 years: 80% female; 37% carriers of a BRCA1/2 mutation. In the 3 years, post-genetic testing carriers reported more risk management activities than non-carriers. Fifty-five per cent of female carriers opted for risk reducing surgery; 43% oophorectomy; and 34% mastectomy. Eighty-nine per cent had mammograms compared with 47% non-carriers. Thirty-six per cent non-carriers > or =50 years did not have a mammogram post-test. Twenty-two per cent male carriers had colorectal and 44% prostate screening compared with 5 and 19% non-carriers respectively. Seven per cent carriers and 1% non-carriers developed cancer. Distress levels did not differ in carriers and non-carriers at 3-year follow-up. Forty per cent of female carriers reported difficulties with life and/or health insurance. Given the return to pre-test levels of concern among female non-carriers at 3 years and a substantial minority not engaging in recommended screening, there appears to be a need to help some women understand the meaning of their genetic status.


Analysis of the volatile compounds in Senecio scandens Buch-Ham by gas chromatography-tandem mass spectrometry based on diversified scan technologies.
Static headspace gas chromatography-tandem mass spectrometry was used to identify volatile compounds from Senecio scandens Buch-Ham. The elemental composition of compounds was confirmed by exploiting the tandem mass spectra of isotopic peaks from the precursor ion. Some isomers were well distinguished by the diversified scan technologies of tandem mass spectrometry (MS/MS). The MS/MS included a product ion scan, a precursor ion scan and a neutral loss scan. The results showed that 46 volatile compounds were completely identified, and the great of majority compounds were α-pinene (11.93%), n-caproaldehyde (9.02%) and dehydrosabinene (6.22%). This qualitative method is convenient and accurate and can be considered as a complementary identification method for the qualitative analysis of volatile compounds in complex samples.


Epidemiology and aetiologies of cryptococcal meningitis in Africa, 1950-2017: protocol for a systematic review.
Cryptococcal meningitis is a neglected disease and an AIDS-defining illness, responsible for 15% of all AIDS-related deaths globally. In 2014, the estimated number of incident cryptococcal meningitis cases was 223 100, with 73% of them occurring in Africa. Currently available data on the prevalence, incidence, aetiologies and mortality of cryptococcal meningitis across Africa are sparse and of limited quality. We propose to conduct the first systematic review to summarise the epidemiological data available on cryptococcal meningitis and its aetiological causes in Africa. We will search PubMed, MEDLINE, Excerpta Medica Database, ISI Web of Science, Africa Index Medicus, Cumulative Index to Nursing and Allied Health for studies on cryptococcal meningitis published between 1st January 1950 and 31st December 2017, involving adults and/or children residing in Africa. After study selection, full text paper acquisition and data extraction, we will use validated tools and checklists to assess the quality of reporting and risk of bias for each study. Heterogeneity across studies will be assessed using the χ2 test on Cochrane's Q statistic and a random effect meta-analysis will be used to estimate the overall prevalence, incidence density and mortality of cryptococcal meningitis across studies with similar characteristics. This protocol is prepared and presented in accordance with the 2015 Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols guidelines. Reporting of the results will be compliant with the Meta-Analysis Of Observational Studies in Epidemiology (MOOSE) guidelines. There is no requirement for ethical approval since we will be using data from published studies. The final report will be published in a peer-reviewed journal and further presented at conferences. This study is expected to provide useful contextual estimates needed to inform treatment policies on the African continent and assess the impact of diagnostic and prevention strategies on the burden of cryptococcal meningitis in the post antiretroviral therapy era. CRD42017081312.


Large scale screening of commonly used Iranian traditional medicinal plants against urease activity.
H. pylori infection is an important etiologic impetus usually leading to gastric disease and urease enzyme is the most crucial role is to protect the bacteria in the acidic environment of the stomach. Then urease inhibitors would increase sensitivity of the bacteria in acidic medium. 137 Iranian traditional medicinal plants were examined against Jack bean urease activity by Berthelot reaction. Each herb was extracted using 50% aqueous methanol. The more effective extracts were further tested and their IC50 values were determined. 37 plants out of the 137 crude extracts revealed strong urease inhibitory activity (more than 70% inhibition against urease activity at 10 mg/ml concentration). Nine of the whole studied plants crude extracts were found as the most effective with IC50 values less than 500 μg/ml including; Rheum ribes, Sambucus ebulus, Pistachia lentiscus, Myrtus communis, Areca catechu, Citrus aurantifolia, Myristica fragrans, Cinnamomum zeylanicum and Nicotiana tabacum. The most potent urease inhibitory was observed for Sambucus ebulus and Rheum ribes extracts with IC50 values of 57 and 92 μg/ml, respectively.


Acute effects of brexpiprazole on serotonin, dopamine, and norepinephrine systems: an in vivo electrophysiologic characterization.
Brexpiprazole, a compound sharing structural molecular characteristics with aripiprazole, is currently under investigation for the treatment of schizophrenia and depression. Using electrophysiologic techniques, the present study assessed the in vivo action of brexpiprazole on serotonin (5-HT) receptor subtypes 5-HT1A, 5-HT1B, and 5-HT2A; dopamine (DA) D2 autoreceptors, and α1- and α2-adrenergic receptors. In addition, the effects on 5-HT1A autoreceptors in the dorsal raphe nucleus (DRN) and D2 autoreceptors in the ventral tegmental area (VTA) were compared with those of aripiprazole, an agent in wide clinical use. In the DRN, brexpiprazole completely inhibited the firing of 5-HT neurons via 5-HT1A agonism and was more potent than aripiprazole (ED50 = 230 and 700 μg/kg, respectively). In the locus coeruleus, brexpiprazole reversed the inhibitory effect of the preferential 5-HT2A receptor agonist DOI (2,5-dimethoxy-4-iodoamphetamine) on norepinephrine neuronal firing (ED50 = 110 μg/kg), demonstrating 5-HT2A antagonistic action. Brexpiprazole reversed the inhibitory effect of the DA agonist apomorphine on VTA DA neurons (ED50 = 61 μg/kg), whereas it was ineffective when administered alone, indicating partial agonistic action on D2 receptors. Compared with aripiprazole, which significantly inhibited the firing activity of VTA DA neurons, brexpiprazole displayed less efficacy at D2 receptors. In the hippocampus, brexpiprazole acted as a full agonist at 5-HT1A receptors on pyramidal neurons. Furthermore, it increased 5-HT release by terminal α2-adrenergic heteroceptor but not 5-HT1B autoreceptor antagonism. In the lateral geniculate nucleus, brexpiprazole displayed α1B-adrenoceptor antagonistic action. Taken together, these results provide insight into the in vivo action of brexpiprazole on monoamine targets relevant in the treatment of depression and schizophrenia.


Birth defects in children of men exposed in utero to diethylstilbestrol (DES).
Prenatal exposure to diethylstilbestrol (DES) is associated with adverse effects, including genital anomalies and cancers in men and women. Animal studies showed birth defects and tumors in the offspring of mice prenatally exposed to DES. In humans, birth defects, such as hypospadias were observed in children of prenatally exposed women. The aim of this research was to assess the birth defects in children of prenatally exposed men. In a retrospective study conceived by a patients' association (Réseau DES France), the reports of men prenatally exposed to DES on adverse health effects in their children were compared with those of unexposed controls and general population. An increased incidence of two genital anomalies, cryptorchidism (OR=5.72; 95% CI 1.51-21.71), and hypoplasia of the penis (OR=22.92; 95% CI 3.81-137.90), was observed in the 209 sons of prenatally exposed men compared with controls, but hypospadias incidence was not increased in comparison with either the controls or the general population. No increase of genital anomalies was observed in daughters. With caution due to the methods and to the small numbers of defects observed, this work suggests an increased incidence of two male genital tract defects in sons of men prenatally exposed to DES. This transgenerational effect, already observed in animals and in the offspring of women prenatally exposed to DES, could be the result of epigenetic changes transmitted to the subsequent generation through men.


An Alternative Strategy for Trypanosome Survival in the Mammalian Bloodstream Revealed through Genome and Transcriptome Analysis of the Ubiquitous Bovine Parasite Trypanosoma (Megatrypanum) theileri.
There are hundreds of Trypanosoma species that live in the blood and tissue spaces of their vertebrate hosts. The vast majority of these do not have the ornate system of antigenic variation that has evolved in the small number of African trypanosome species, but can still maintain long-term infections in the face of the vertebrate adaptive immune system. Trypanosoma theileri is a typical example, has a restricted host range of cattle and other Bovinae, and is only occasionally reported to cause patent disease although no systematic survey of the effect of infection on agricultural productivity has been performed. Here, a detailed genome sequence and a transcriptome analysis of gene expression in bloodstream form T. theileri have been performed. Analysis of the genome sequence and expression showed that T. theileri has a typical kinetoplastid genome structure and allowed a prediction that it is capable of meiotic exchange, gene silencing via RNA interference and, potentially, density-dependent growth control. In particular, the transcriptome analysis has allowed a comparison of two distinct trypanosome cell surfaces, T. brucei and T. theileri, that have each evolved to enable the maintenance of a long-term extracellular infection in cattle. The T. theileri cell surface can be modeled to contain a mixture of proteins encoded by four novel large and divergent gene families and by members of a major surface protease gene family. This surface composition is distinct from the uniform variant surface glycoprotein coat on African trypanosomes providing an insight into a second mechanism used by trypanosome species that proliferate in an extracellular milieu in vertebrate hosts to avoid the adaptive immune response.


Characterisation of chitosan-coated vesicles encapsulating DNA suitable for gene delivery.
Encapsulation of DNA (varying type and molecular weight) in vesicles (dehydration-rehydration vesicles; DRV) prepared from zwitterionic phospholipids using the dehydration-rehydration method was determined to be in the range of 10-90%. Encapsulation was dependent on the amount of DNA added but not its molecular weight. Zeta potential measurements of the DRV suggested that DNA was associated with the vesicles exterior, while small angle neutron scattering (SANS) studies indicated that DNA was trapped in the interlamellar water space between lipid bilayers-as evidenced by a slight decrease in bilayer thickness and an increase in d-spacing. Circular dichroism measurements determined the conformation of the DNA in the DRV was not the usual B-form but rather a structure more reminiscent of the less hydrated, C-form, supporting the suggestion that DNA was present in the interlamellar water space. DRV were successfully coated with chitosan as evidenced by SANS studies and zeta potential measurements. The amount of chitosan coating the DRV [quantified by ultra violet (UV)/Vis spectroscopy] was dependent upon the starting concentration of chitosan and independent of the presence of DNA. Chitosan-coated DRV containing DNA exhibited an improved size stability while 'empty' uncoated vesicles were poorly stable. A coating of chitosan reduced the amount of DNA leaking from the vesicles.


Modeling and Validation of Vertical Direction Force Estimation with a Three-Dimensional Force Measurement Instrument Based on a Zero-Compliance Mechanism.
The principle of a zero-compliance mechanism was used to develop a three-dimensional force measurement instrument. In each axis, the point of force is suspended by a zero-compliance mechanism. A vertical axis force estimation operation imitates the structure of a double series magnetic suspension system. An electromagnet directly controls the movement of the first suspended object (floator), which is denoted as a detection point, and indirectly controls the motion of the second floator, which is denoted as a point of force. Indirect control of the point of force is executed by the attractive force of a permanent magnet that is fixed to the bottom part of the detection point. To achieve zero-compliance, a Proportional-Integral-Derivative (PID) control is applied to the point of force, and to make the system stable, a Proportional-Derivative (PD) control is also applied to the detection point. In such suspension conditions, when force is exerted on the point of force, the displacement of the second floator is regulated to maintain its primary position while the detection point displaces in proportion to the applied force. Thus, a zero-compliance condition is maintained at the point of force, and the external force is measured from the linear displacement of the detection point. To restrict the motions of the detection point and the point of force in translation only, they are supported with leaf springs. This paper presents the modeling of the vertical direction force measurement operation of the developed three-axis force estimation instrument, and the theoretical analyses were validated by experiments of force measurement in both the millinewton and micronewton ranges.


Surface morphologies of sputter-deposited aluminum films studied using a high-resolution phase-measuring laser interferometric microscope.
Sputter-deposited aluminum (Al) film surface morphologies were studied with a new nondestructive method that incorporates a high-resolution phase-measuring laser interferometric microscope. Good correlation is obtained between rms roughness and reflectivity for various conditions of temperature and argon gas pressure. It should be noted that the rms roughness is much more sensitive than reflectivity when reflectivity exceeds 90%. A drastic change is observed in the temperature dependence of the rms roughness and the skewness at 200 °C. As a result there are changes in Al grain sizes and surface morphologies based on concomitant scanning electron microscope observations. We found that the rms roughness value depends on the resolution of the objective especially when the Al grain sizes are comparable to the resolution.


Production of superoxide from Photosystem II in a rice (Oryza sativa L.) mutant lacking PsbS.
PsbS is a 22-kDa Photosystem (PS) II protein involved in non-photochemical quenching (NPQ) of chlorophyll fluorescence. Rice (Oryza sativa L.) has two PsbS genes, PsbS1 and PsbS2. However, only inactivation of PsbS1, through a knockout (PsbS1-KO) or in RNAi transgenic plants, results in plants deficient in qE, the energy-dependent component of NPQ. In studies presented here, under fluctuating high light, growth of young seedlings lacking PsbS is retarded, and PSII in detached leaves of the mutants is more sensitive to photoinhibitory illumination compared with the wild type. Using both histochemical and fluorescent probes, we determined the levels of reactive oxygen species, including singlet oxygen, superoxide, and hydrogen peroxide, in leaves and thylakoids. The PsbS-deficient plants generated more superoxide and hydrogen peroxide in their chloroplasts. PSII complexes isolated from them produced more superoxide compared with the wild type, and PSII-driven superoxide production was higher in the mutants. However, we could not observe such differences either in isolated PSI complexes or through PSI-driven electron transport. Time-course experiments using isolated thylakoids showed that superoxide production was the initial event, and that production of hydrogen peroxide proceeded from that. These results indicate that at least some of the photoprotection provided by PsbS and qE is mediated by preventing production of superoxide released from PSII under conditions of excess excitation energy.


Incentives for community treatment. Mental illness management services.
Serving people with mental and other chronic illnesses in community settings may improve compliance and satisfaction with treatment, but existing payment mechanisms often favor office-based treatment. This study examines the effect of a change in Medicaid payment on the location and amount of service provided by case managers. Amounts of service given by treatment providers to 185 of their clients in community settings and in mental health centers were compared before and after reimbursement changed from an all-inclusive prospective rate to a mixed prospective/retrospective payment. Clients were enrolled in two different treatment programs: continuous treatment teams with extensive training in in vivo treatment, and a case management program that emphasized office-based treatment. In-community service increased, and the amount of office-based treatment decreased. Continuous treatment teams increased in-community services more than case managers did; case managers decreased office-based treatment more. There was no change in total amount of services provided. It was concluded that mixed prospective and retrospective reimbursement can remove financial barriers to in-community treatment, but it works best in combination with a training program. Additional research is needed to determine the precise financial impact of such changes.


Acquired neuropathologic lesions associated with the hypoplastic left heart syndrome.
This study details the type, frequency, clinical presentation, and etiologic associations of acquired brain lesions in 40 infants with the hypoplastic left heart syndrome encountered during a 52-month interval. Detailed postmortem neuropathologic examinations showed that 55% of the infants were free of acquired brain lesions. However, the other 45% had combinations of hypoxic-ischemic lesions and intracranial hemorrhage. Central nervous system perfusion and glucose-oxygen delivery appeared to be important factors in the occurrence of hypoxic-ischemic lesions or intracranial hemorrhage, whereas acidosis and hypercarbia were not. Cerebral necrosis may be a predisposing factor for a major intracranial hemorrhage. A duration of cardiopulmonary bypass with hypothermic total circulatory arrest longer than 40 minutes was associated with a higher incidence of acquired neuropathology. These results indicate that the majority of infants with hypoplastic left heart syndrome are free of acquired neuropathology and suggest practical ways to reduce the risks in the others.


P2X7 receptor-dependent intestinal afferent hypersensitivity in a mouse model of postinfectious irritable bowel syndrome.
The ATP-gated P2X(7) receptor (P2X(7)R) was shown to be an important mediator of inflammation and inflammatory pain through its regulation of IL-1β processing and release. Trichinella spiralis-infected mice develop a postinflammatory visceral hypersensitivity that is reminiscent of the clinical features associated with postinfectious irritable bowel syndrome. In this study, we used P2X(7)R knockout mice (P2X(7)R(-/-)) to investigate the role of P2X(7)R activation in the in vivo production of IL-1β and the development of postinflammatory visceral hypersensitivity in the T. spiralis-infected mouse. During acute nematode infection, IL-1β-containing cells and P2X(7)R expression were increased in the jejunum of wild-type (WT) mice. Peritoneal and serum IL-1β levels were also increased, which was indicative of elevated IL-1β release. However, in the P2X(7)R(-/-) animals, we found that infection had no effect upon intracellular, plasma, or peritoneal IL-1β levels. Conversely, infection augmented peritoneal TNF-α levels in both WT and P2X(7)R(-/-) animals. Infection was also associated with a P2X(7)R-dependent increase in extracellular peritoneal lactate dehydrogenase, and it triggered immunological changes in both strains. Jejunal afferent fiber mechanosensitivity was assessed in uninfected and postinfected WT and P2X(7)R(-/-) animals. Postinfected WT animals developed an augmented afferent fiber response to mechanical stimuli; however, this did not develop in postinfected P2X(7)R(-/-) animals. Therefore, our results demonstrated that P2X(7)Rs play a pivotal role in intestinal inflammation and are a trigger for the development of visceral hypersensitivity.


Real-world evaluation of perampanel effectiveness in Japanese adolescents with epilepsy
Real-world data from adolescents treated with perampanel in a routine clinical setting are lacking in Japan. We evaluated the safety and efficacy of perampanel for adolescent patients (aged 12-17 years) with drug-resistant, refractory epilepsy in real-world settings. This was a large-scale, prospective, observational post-marketing study, with a 104-week observation period. Safety was assessed by monitoring adverse effects (adverse drug reactions). For efficacy assessments, seizure frequency was compared between the four weeks immediately prior to the last observation and the four weeks before the commencement of perampanel. In total, 519 patients were enrolled; 505 and 484 patients were included in the safety and efficacy analysis sets, respectively. The mean age was 14.4 years. The mean daily dose of perampanel was 4.4 mg/day. The main reasons for discontinuation at 104 weeks were adverse events (48.4%) and inadequate efficacy (46.8%). The retention rate at 104 weeks was 50.5%. Adverse effect and severe adverse effect incidences were 42.2% and 1.8%, respectively. The most common adverse effects were somnolence (13.5%), irritability (8.5%), dizziness (5.1%), and agitation (4.8%). There were significant differences in the occurrence of adverse effects between the initial titration interval of <2 weeks and 2-4 weeks (odds ratio=0.441, p=0.029) and 4-8 weeks (odds ratio=0.462, p=0.027). The median percent change in seizure frequency at the last observation carried forward was −50.0 for focal aware seizures with motor signs, −73.3 for focal aware seizures without motor signs, −28.6 for focal impaired awareness seizures, −62.6 for focal to bilateral tonic-clonic seizures, and −20.0 for generalized tonic-clonic seizures. In adolescent patients, perampanel was well tolerated and efficacious in reducing seizure frequency. No unexpected safety issues were observed, and slow titration may reduce the incidence of adverse effects.


Precision medicine in alcoholic and nonalcoholic fatty liver disease via modulating the gut microbiota.
Alcoholic liver disease (ALD) and nonalcoholic fatty liver disease (NAFLD) represent a major health burden in industrialized countries. Although alcohol abuse and nutrition play a central role in disease pathogenesis, preclinical models support a contribution of the gut microbiota to ALD and NAFLD. This review describes changes in the intestinal microbiota compositions related to ALD and NAFLD. Findings from in vitro, animal, and human studies are used to explain how intestinal pathology contributes to disease progression. This review summarizes the effects of untargeted microbiome modifications using antibiotics and probiotics on liver disease in animals and humans. While both affect humoral inflammation, regression of advanced liver disease or mortality has not been demonstrated. This review further describes products secreted by Lactobacillus- and microbiota-derived metabolites, such as fatty acids and antioxidants, that could be used for precision medicine in the treatment of liver disease. A better understanding of host-microbial interactions is allowing discovery of novel therapeutic targets in the gut microbiota, enabling new treatment options that restore the intestinal ecosystem precisely and influence liver disease. The modulation options of the gut microbiota and precision medicine employing the gut microbiota presented in this review have excellent prospects to improve treatment of liver disease.


Small heat shock protein activity is regulated by variable oligomeric substructure.
The alpha-crystallins are members of the small heat shock protein family of molecular chaperones that have evolved to minimize intracellular protein aggregation; however, they are also implicated in a number of protein deposition diseases. In this study, we employed novel mass spectrometry techniques to investigate the changes in quaternary structure associated with this switch from chaperone to adjuvant of aggregation. We replicated the oligomeric rearrangements observed for post-translationally modified alpha-crystallins, without altering the protein sequence, by refolding the alpha-crystallins in vitro. This refolding resulted in a loss of dimeric substructure concomitant with an augmentation of substrate affinity. We show that packaging of small heat shock proteins into dimeric units is used to control the level of chaperone function by regulating the exposure of hydrophobic surfaces. We propose that a bias toward monomeric substructure is responsible for the aberrant chaperone behavior associated with the alpha-crystallins in protein deposition diseases.


Colorectal cancer in women: an equal opportunity disease.
Colorectal cancer is the second leading cause of cancer-related deaths in developed countries. For women, it is the third leading cause of cancer-related deaths behind lung and breast cancers. Women have the same risk as men, and the lifetime risk of the development of colorectal cancer is 6%. One in 17 woman will have colorectal cancer diagnosed. There are risk factors unique to women, including gynecologic cancers, and treatment of gynecologic cancers, as well as delayed diagnosis in pregnancy. Fortunately, colorectal cancer is a preventable disease, as almost all colorectal cancers arise from premalignant polyps. Colorectal cancer screening is recommended in asymptomatic women aged 50 years and older who are at average risk. Screening and surveillance for colorectal cancer in women are important to improve the morbidity and mortality rates of this preventable disease.


Reactive polycyclic aromatic hydrocarbon dimerization drives soot nucleation.
Nucleation is an important, yet poorly understood step in soot formation. Here, the importance of reactive PAH dimerization in reducing soot nucleation reversibility is investigated by simulating soot formation in a so-called "nucleation" flame (P. Desgroux et al., Combust. Flame, 2017, 184, 153-166). There, inception of soot particles is prolonged at minimal subsequent growth. With only reversible PAH dimerization, the simulated soot concentration is negligible. Accounting however for PAH chemical bond formation after physical dimerization, stabilizes dimers by covalent bonds and increases the soot concentration by four orders of magnitude, in good agreement with Laser Induced Incandescence measurements. In particular, dimers of benzene with benzene, phenylacetylene, naphthalene, toluene, acenaphthylene and cyclopentapyrene make significant contributions to the total soot concentration. The abundance of dimers with small PAHs highlights the dominant role of PAH concentration over their size and dispersion forces on dimer formation. Higher collision factors are used for irreversible dimerization models using larger PAHs because of their lower concentrations and not their larger dispersion forces leading to reduced reversibility and more stable dimers. The qualitative trend of main peaks agrees well with stochastic simulations and aerosol mass spectra measured in the above "nucleation" as well as premixed flames highlighting the abundance of PAHs with five-membered rings and substituted aliphatic chains in incipient soot. The predicted number of trimers is very low, i.e. less than 3% of the total soot nuclei formed, indicating that covalently bonded PAH dimers can be the main contributors to soot nucleation.


Laboratory studies of transovarial transmission of La Crosse and other arboviruses by Aedes albopictus and Culex fatigans.
Transovarial transmission of La Crosse virus by experimentally infected Aedes albopictus females to 2.7% of their F1 generation offspring was demonstrated. Progeny of both sexes were infected. Mean virus titers in parent mosquitoes and infected F1 generation adults were 10(4.6) and 10(3.4) plaque forming units/insect, respectively. The La Crosse-infected offspring were randomly distributed among the female parents. After two serial passages in A. albopictus, a marked change occurred in the plaque morphology of the virus but this had no apparent effect on the subsequent vertical transmission rate. In contrast, transovarial transmission did not occur in La Crosse-infected Culex fatigans or in A. albopictus and C. fatigans infected with vesicular stomatitis-Indiana, Cache Valley, Batai, Arumowot, and Itaporanga viruses. Results of this experiment suggest that the La Crosse model might be useful in studying the mechanism of transovarial transmission in additional mosquito species.


Cervical degenerative disease at flexion-extension MR imaging: prediction criteria.
To determine if there are any neutral-position imaging criteria that can help predict functional cord impingement at flexion-extension cervical magnetic resonance (MR) imaging. Sixty-two patients with cervical degenerative disease were evaluated with regard to the dynamic changes of canal stenosis at flexion-extension MR imaging. Functional cord impingement was considered if the cord was impinged or more impinged after neck flexion or extension. Selection criteria for neutral-position MR imaging, such as cervical curvature, canal space, degenerative stage, intramedullary high signal intensity on T2-weighted images, and resting instability, were evaluated for their ability to predict functional cord impingement at flexion-extension MR imaging (Fisher exact test, logistic regression analysis). MR images in 19 (31%) of 62 patients showed functional cord impingement at extension MR imaging compared with images in two (3%) patients at flexion MR imaging. Statistically significant differences were found for the criteria cervical degeneration stage (P <.001) and spinal canal space (P =.037) for predicting functional cord impingement at extension MR imaging. In contrast, no significant differences were found among selection criteria for flexion MR imaging. Probabilities of functional cord impingement at extension MR imaging were calculated with different combinations of degenerative stages and canal spaces. Probability could increase to 79% if the patient had both stabilization degeneration (disk protrusion or osteophytic formation with hypertrophy of the ligamentum flavum) and C7 canal space of 10 mm or less. None of the selection criteria evaluated in this study has the ability to predict functional cord impingement at flexion MR imaging; however, prediction of impingement at extension MR imaging can increase from 31% to 79% if proper criteria are selected.


Distinct BK polyomavirus non-coding control region (NCCR) variants in oral fluids of HIV- associated Salivary Gland Disease patients.
HIV-associated Salivary Gland Disease (HIVSGD) is among the most common salivary gland-associated complications in HIV positive individuals and was associated with the small DNA tumorvirus BK polyomavirus (BKPyV). The BKPyV non-coding control region (NCCR) is the main determinant of viral replication and rearranges readily. This study analyzed the BKPyV NCCR architecture and viral loads of 35 immunosuppressed individuals. Throatwash samples from subjects diagnosed with HIVSGD and urine samples from transplant patients were BKPyV positive and yielded BKPyV NCCR sequences. 94.7% of the BKPyV HIVSGD NCCRs carried a rearranged OPQPQQS block arrangement, suggesting a distinct architecture among this sample set. BKPyV from HIV positive individuals without HIVSGD harbored NCCR block sequences that were distinct from OPQPQQS. Cloned HIVSGD BKPyV isolates displayed active promoters and efficient replication capability in human salivary gland cells. The unique HIVSGD NCCR architecture may represent a potentially significant oral-tropic BKPyV substrain.


Reticulocyte and haemoglobin profiles in elite triathletes over four consecutive seasons.
The World Anti-Doping Agency has implemented the Blood Passport in attempt to detect blood doping in athletes. The Blood Passport looks for uncommon changes overtime in reticulocytes percentage (Ret %), as a variable of the OFF-hr score, and haemoglobin concentration ([Hb]) reflecting potential doping violations. Few studies, however, have actually investigated the concurrent stability of Ret % and [Hb] in athletes over extended periods of time, none of which were measured in athletes who undergo strenuous and prolonged physical exercise. Measurements of Ret % and [Hb] were assessed over the course of four competitive seasons in elite triathletes (10 males and seven female). Blood was obtained at the start of the season, precompetitive period, competitive period and at the end of the competitive period. Differences (P<0.001) were observed in both [Hb] and Ret % between genders and there was a high variability between subjects. Neither males nor females exhibited differences in [Hb] across all periods within one season. Within gender, analysis revealed that Ret % varied significantly (P=0.0018) between periods only in female athletes. We conclude that Ret % and [Hb] remain stable over four consecutive seasons in elite triathletes, confirming that both parameters are valid for antidoping purposes based on the Blood Passport. In addition, Ret % fluctuations within one season require further investigation in females.


Comparison of the pharmacological characteristics of 2-[125I]iodomelatonin binding sites in the lung, spleen, brain and kidney of chicken.
We have compared the pharmacological characteristics of 2-[125I]iodomelatonin binding to crude membrane preparations of the lung, spleen, brain and kidney of chicken. Saturation studies indicated significant differences (p < 0.05) in the equilibrium dissociation constant (Kd) and maximum number of binding site (Bmax) values among the four tissues studied. The descending order of affinities was lung = spleen > brain > kidney. Competition curves of 2-[125I]iodomelatonin binding to crude membrane preparations of all four chicken tissues by melatonin were studied simultaneously to reduce individual, physiological, age and interassay variations. Similar competition experiments were also performed on 2-phenylmelatonin, 2-iodomelatonin, 6-chloromelatonin, 6-hydroxymelatonin and N-acetylserotonin (NAS). Concentrations of indoles which inhibited 50% of specific 2-[125I]iodomelatonin binding (IC50) were calculated. The IC50 of 2-[125I]iodomelatonin inhibition curves by the indole compounds in different tissues showed the following descending orders of affinity: (1) melatonin: lung = spleen > brain > kidney, (2) 2-phenylmelatonin: lung = spleen = brain = kidney, (3) 2-iodomelatonin: lung = spleen = kidney > brain, (4) 6-chloromelatonin: lung = spleen = kidney > brain, (5) 6-hydroxymelatonin: kidney > lung = spleen = brain, and (6) NAS: kidney > lung = spleen > brain. The non-hydrolizable GTP analog, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), exhibited differential effects on the saturable binding of the four tissues. GTP gamma S increased the Kd of 2-[125I]iodomelatonin binding by 2- to 3-fold in the lung and spleen, 0.5-fold in the brain and 1-fold in the kidney. Based on our findings, we would like to suggest that the 2-[125I]iodomelatonin binding sites in these four tissues may belong to three different high affinity (picomolar) subtypes of melatonin receptor. We name them cML1A represented by the lung and spleen, cML1B by the brain and cML1C by the kidney.


Reduced gamma-aminobutyric acid is associated with emotional and behavioral problems in Prader-Willi syndrome.
Prader-Willi syndrome (PWS) is characterized by infantile hypotonia, hypogonadism, small hands and feet, distinct facial features and usually intellectual impairment. The disorder is associated with severe behavioral disturbances which include hyperphagia leading to morbid obesity, temper outbursts, skin-picking, and compulsive behaviors. While the brain mechanisms that underpin these disturbances are unknown these behaviors suggest a lack of inhibition and thus gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter may be implicated. In the present study, we investigated in vivo brain GABA and its relationship with emotion and behavior in individuals with PWS. Single voxel proton magnetic resonance spectroscopy (1H-MRS) was performed on 15 individuals with PWS and 15 age- and gender-matched typically developing controls. GABA levels were measured in the parieto-occipital lobe. All other metabolite levels (N-acetyl aspartate, myo-Inositol, glutathione, glutamate, and glutamine + glutamate) were measured in the anterior cingulate cortex (ACC). GABA levels were significantly lower in the participants with PWS who had clinically significant emotional and behavioral problems relative to typically developing control participants and participants with PWS who did not have emotional and behavioral problems within the clinically significant range. GABA levels were negatively correlated with total behavioral problem scores as well as temper outbursts, skin-picking, depression, social relating difficulties, and a tendency to be self-absorbed. Our data suggests that alterations of the GABAergic system may play an important role in aspects of the pathophysiology of PWS. Pathological mechanism found in PWS may be relevant to understanding the control of similar behaviors in the general population. © 2016 Wiley Periodicals, Inc.


The role of spin states in the catalytic mechanism of the intra- and extradiol cleavage of catechols by O2.
Iron-dependent enzymes and biomimetic iron complexes can catalyze the ring cleavage of very inert, aromatic compounds. The mechanisms of these transformations and the factors that lead either to extradiol cleavage or intradiol cleavage have not been fully understood. By using density functional theory we have elucidated the mechanism of the catalytic cycle for two biomimetic complexes, and explained the difference in the experimentally obtained products.


Human subthalamic nucleus in movement error detection and its evaluation during visuomotor adaptation.
Monitoring and evaluating movement errors to guide subsequent movements is a critical feature of normal motor control. Previously, we showed that the postmovement increase in electroencephalographic (EEG) beta power over the sensorimotor cortex reflects neural processes that evaluate motor errors consistent with Bayesian inference (Tan et al., 2014). Whether such neural processes are limited to this cortical region or involve the basal ganglia is unclear. Here, we recorded EEG over the cortex and local field potential (LFP) activity in the subthalamic nucleus (STN) from electrodes implanted in patients with Parkinson's disease, while they moved a joystick-controlled cursor to visual targets displayed on a computer screen. After movement offsets, we found increased beta activity in both local STN LFP and sensorimotor cortical EEG and in the coupling between the two, which was affected by both error magnitude and its contextual saliency. The postmovement increase in the coupling between STN and cortex was dominated by information flow from sensorimotor cortex to STN. However, an information drive appeared from STN to sensorimotor cortex in the first phase of the adaptation, when a constant rotation was applied between joystick inputs and cursor outputs. The strength of the STN to cortex drive correlated with the degree of adaption achieved across subjects. These results suggest that oscillatory activity in the beta band may dynamically couple the sensorimotor cortex and basal ganglia after movements. In particular, beta activity driven from the STN to cortex indicates task-relevant movement errors, information that may be important in modifying subsequent motor responses.


Treatment of infant leukemias: challenge and promise.
Leukemia in infants is rare but generates tremendous interest due to its aggressive clinical presentation in a uniquely vulnerable host, its poor response to current therapies, and its unique biology that is increasingly pointing the way toward novel therapeutic approaches. This review highlights the key clinical, pathologic, and epidemiologic features of infant leukemia, including the high frequency of mixed lineage leukemia (MLL) gene rearrangements. The state of the art with regard to current approaches to risk stratified treatment of infant leukemia in the major international cooperative groups is discussed. Finally, exciting recent discoveries elucidating the molecular biology of infant leukemia are reviewed and novel targeted therapeutic strategies, including FLT3 inhibition and modulation of aberrant epigenetic programs, are suggested.


Exposure to nitrofurantoin during the first trimester of pregnancy and the risk for major malformations.
Antibacterial drugs are among the most common medications used by pregnant women. While medical literature generally defines nitrofurantoin as an antibiotic that is safe for use during the first trimester of pregnancy, new concerns about a possible association between congenital malformations following exposure to nitrofurantoin during the first trimester of pregnancy have recently surfaced. To address these concerns, we conducted a large population-based retrospective cohort study to assess this possible association (including cases of medical terminations of pregnancy or stillbirth) and congenital malformations. A computerized database for medications dispensed to pregnant women in southern Israel was linked with records from the district hospital. Associations between exposure to nitrofurantoin during the first trimester and major malformations were assessed. Our research included a total of 105,492 pregnancies, 1,112 of which involved pregnancy terminations for medical reasons. A total of 1,329 infants and abortuses had been exposed to nitrofurantoin during the first trimester of pregnancy. Exposure to nitrofurantoin was not associated with increased risk of major malformations in general (adjusted OR = 0.85, 95% CI 0.67-1.08) or with specific malformations. First trimester exposure to nitrofurantoin was not associated with increased risk for total major congenital malformations or with specific malformations.


Impact of Endobronchial Valve Treatment on Lung Function Decline.
Endobronchial valve (EBV) treatment has been shown to be beneficial for patients with severe emphysema. The forced expiratory volume in 1 s (FEV1) was found to be significantly higher compared to baseline for up to 3 years after treatment although the magnitude of improvement gradually decreases over time. So far, it has not been investigated whether this treatment decelerates the decline in lung function. Therefore, our aim was to investigate the lung function decline before and after EBV treatment. We included patients who were treated with EBVs in our hospital, of whom pre-treatment spirometry results were available (at least 4 measurements within at least 2 years before treatment) and who had an annual FEV1 measurement up to 3 years after treatment. In total, 45 patients were included (73% female, FEV1: 28 ± 7% of predicted, residual volume: 232 ± 32% of predicted) who had a mean pre-treatment FEV1 decline of -64 mL/year. Mean FEV1 "decline" after treatment was +13 mL/year, since FEV1 was still above the baseline level at 3-year follow-up. However, the FEV1 decline between 1 and 3 years of follow-up was not significantly different compared to the pre-treatment decline (-73 mL/year, p = 0.179). Our results show that the EBV treatment does not influence the progression of disease in terms of lung function decline. However, the treatment does improve the FEV1 up to a level that is still comparable 3 years after treatment with the baseline level.


[Prevalence of malnutrition to income in a basic general hospital].
The main objective has been to evaluate and quantify the prevalence of malnutrition at admission, the degree of severity and the correlation with the nutritional parameters in a basic general hospital of the first level. Observational, transverse, and randomized study of 244 patients (59.8% male and 40.2% female) performed during the first 24 hours of hospital admission and with a nutritional assessment including nutritional screening (CONUT®), anthropometric parameters, other analytical parameters and a quantification of hospital stay. In this study, 60.7% of the sample has at least one criterion of malnutrition (anthropometric and/or analytical). When considering at least two altered nutritional parameters (one of them analytical type), the prevalence was 29.1%. Prealbumin and transferrin have been shown to be particularly sensitive to moderate to severe and mild malnutrition, respectively. CONUT® has detected nutritional risk of a mild nature in 39.8%. Malnourished patients have a longer stay, and are statistically significant in oncological versusnon-oncological patients. Anthropometric parameters, on their own, can generate some degree of controversy over the prevalence of malnutrition, so it is considered as appropriate to use two parameters, at least one of an analytical type. Prealbumin is the most sensitive and specific indicator, and CONUT® is an easy, quick and economical tool for nutritional screening. In order to greatly alleviate malnutrition at hospital admission, the objective of the study, it would be wise to strengthen its screening and treatment from Primary Health Care.


EFFICACY OF WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS) TREATMENT FOR CATTLE FEVER TICKS IN SOUTHERN TEXAS, USA.
White-tailed deer (Odocoileus virginianus) serve as a host for cattle fever ticks (Rhipicephalus [Boophilus] microplus and Rhipicephalus [Boophilus] annulatus; CFTs); therefore, deer are a concern for CFT control programs in southern Texas, US. Systemic (oral delivery of ivermectin) and topical (permethrin on pelage) treatment devices have been developed for white-tailed deer; however, the efficacy of these treatment options has not been determined for CFTs in southern Texas. Our objectives were to evaluate the effectiveness of CFT treatment strategies by 1) measuring exposure rates of deer to the acaricides permethrin and ivermectin, 2) determining the relationship between CFTs on deer and exposure to the acaricides, and 3) determining if photos from remote cameras at medicated bait sites can be used as a measure of acaricide treatment. We captured 327 deer at four sites in southern Texas. Deer visitation to medicated bait sites was monitored using remote cameras from March 2010 to February 2012. There was no relationship between the presence of permethrin and the probability of being infested with CFTs (P≥0.336). The probability of infestation with CFTs decreased as serum ivermectin levels increased for male (n=18, P=0.098) and female (n=33, P<0.001) deer. Our results indicate ivermectin may be more effective in treating CFTs than permethrin; thus it would be worthwhile to develop topical acaricides other than permethrin for treating white-tailed deer in southern Texas.


Loss of SMARCB1/INI1 expression in poorly differentiated chordomas.
Chordomas are malignant neoplasms that typically arise in the axial spine and primarily affect adults. When chordomas arise in pediatric patients they are more likely to display unusual histological features and aggressive behavior. We noted the absence of SMARCB1/INI1 expression by immunohistochemistry in an index case of poorly differentiated chordoma of the sacrum, leading us to further examine SMARCB1/INI1 expression as well as that of brachyury, a highly specific marker of notochordal differentiation, in 3 additional poorly differentiated chordomas of the clivus, 10 typical chordomas, and 8 atypical teratoid/rhabdoid tumors (AT/RTs). All 4 poorly differentiated chordomas and all AT/RTs lacked nuclear expression of SMARCB1/INI1, while the 10 typical chordomas maintained strong nuclear SMARCB1/INI1 immunoreactivity. All 10 typical and 4 poorly differentiated chordomas expressed brachyury; all 8 AT/RTs were brachyury immunonegative. Cytogenetic evaluation utilizing FISH probes near the SMARCB1/INI1 locus on chromosome 22q was also performed in all of the poorly differentiated chordomas in this series. Three of the four poorly differentiated chordomas had evidence for deletion of this region by FISH. Analysis of the SMARCB1/INI1 gene sequence was performed using formalin-fixed paraffin-embedded tissue in all cases and no point mutations were observed. In summary, all poorly differentiated chordomas in this series showed the absence of SMARCB1/INI1 expression, and were reliably distinguished from AT/RTs, clinically by their characteristic primary sites of origin and pathologically by strong nuclear brachyury expression. Our findings reveal a likely role for SMARCB1/INI1 in a subset of chordomas with aggressive features.


The effects of topical Arnica on performance, pain and muscle damage after intense eccentric exercise.
The aim of the study was to determine if topical Arnica is effective in reducing pain, indicators of inflammation and muscle damage, and in turn improve performance in well-trained males experiencing delayed onset muscle soreness (DOMS). Twenty well-trained males matched by maximal oxygen uptake (V̇O2 Max) completed a double-blind, randomised placebo-controlled trial. Topical Arnica was applied to the skin superficial to the quadriceps and gastrocnemius muscles immediately after a downhill running protocol designed to induce DOMS. Topical Arnica was reapplied every 4 waking hours for the duration of the study. Performance measures (peak torque, countermovement and squat jump), pain assessments (visual analogue scale (VAS) and muscle tenderness) and blood analysis (interleukin-1 beta, interleukin-6, tumour necrosis factor-alpha, C-reactive protein, myoglobin and creatine kinase) were assessed at seven time points over five days (pre-, post-, 4, 24, 48, 72 and 96 hours after the downhill run). Participants in the topical Arnica group reported less pain as assessed through muscle tenderness and VAS 72 hours post-exercise. The application of topical Arnica did not affect any performance assessments or markers of muscle damage or inflammation. Topical Arnica used immediately after intense eccentric exercise and for the following 96 hours did not have an effect on performance or blood markers. It did however demonstrate the possibility of providing pain relief three days post-eccentric exercise.


Genetic risk factors in lupus nephritis and IgA nephropathy--no support of an overlap.
IgA nephropathy (IgAN) and nephritis in Systemic Lupus Erythematosus (SLE) are two common forms of glomerulonephritis in which genetic findings are of importance for disease development. We have recently reported an association of IgAN with variants of TGFB1. In several autoimmune diseases, particularly in SLE, IRF5, STAT4 genes and TRAF1-C5 locus have been shown to be important candidate genes. The aim of this study was to compare genetic variants from the TGFB1, IRF5, STAT4 genes and TRAF1-C5 locus with susceptibility to IgAN and lupus nephritis in two Swedish cohorts. We genotyped 13 single nucleotide polymorphisms (SNPs) in four genetic loci in 1252 DNA samples from patients with biopsy proven IgAN or with SLE (with and without nephritis) and healthy age- and sex-matched controls from the same population in Sweden. Genotype and allelic frequencies for SNPs from selected genes did not differ significantly between lupus nephritis patients and SLE patients without nephritis. In addition, haplotype analysis for seven selected SNPs did not reveal a difference for the SLE patient groups with and without nephritis. Moreover, none of these SPNs showed a significant difference between IgAN patients and healthy controls. IRF5 and STAT4 variants remained significantly different between SLE cases and healthy controls. In addition, the data did not show an association of TRAF1-C5 polymorphism with susceptibility to SLE in this Swedish population. Our data do not support an overlap in genetic susceptibility between patients with IgAN or SLE and reveal no specific importance of SLE associated SNPs for the presence of lupus nephritis.


A multi-dimensional search for new heavy resonances decaying to boosted W W , W Z , or Z Z boson pairs in the dijet final state at 13 Te .
A search in an all-jet final state for new massive resonances decaying to W W , W Z , or Z Z boson pairs using a novel analysis method is presented. The analysis is performed on data corresponding to an integrated luminosity of 77.3 fb - 1 recorded with the CMS experiment at the LHC at a centre-of-mass energy of 13 Te . The search is focussed on potential narrow-width resonances with masses above 1.2 Te , where the decay products of each W or Z boson are expected to be collimated into a single, large-radius jet. The signal is extracted using a three-dimensional maximum likelihood fit of the two jet masses and the dijet invariant mass, yielding an improvement in sensitivity of up to 30% relative to previous search methods. No excess is observed above the estimated standard model background. In a heavy vector triplet model, spin-1 Z ' and W ' resonances with masses below 3.5 and 3.8 Te , respectively, are excluded at 95% confidence level. In a bulk graviton model, upper limits on cross sections are set between 27 and 0.2 fb for resonance masses between 1.2 and 5.2 Te , respectively. The limits presented in this paper are the best to date in the dijet final state.


Ultrasonographic cervical length and risk of hemorrhage in pregnancies with placenta previa.
To estimate the relationship between cervical length and hemorrhage leading to preterm delivery in women with placenta previa. Between October 2007 and May 2009, transvaginal cervical-length measurements were obtained in all singleton pregnancies with placenta previa identified at or beyond 24 weeks of gestation. Only women who delivered liveborn or stillborn neonates at our hospital and had placenta previa confirmed at delivery were included. Cervical length of 30 mm or less was considered short. Clinicians were blinded to cervical-length measurements. Chi-square and logistic regression were used for analysis. Of 89 identified women with placenta previa at initial ultrasonography, 68 had placenta previa at delivery, and 29 (43%) of these had a short cervix. Gestational age at cervical-length measurement was 32+/-4 weeks in women with a short cervix and 33+/-2 weeks in those with a longer cervix (P=.4). Women with previa and a short cervix were more likely to require delivery for hemorrhage, 79% compared with 28%, and to deliver preterm, 69% compared with 21% (both P<.001). Tocodynamometer evidence of regular uterine contractions was more common with a short cervix than with a longer cervix, 69% compared with 21% (P<.001). Conversely, 64% with a cervical length greater than 30 mm had no bleeding episodes and progressed to term. In pregnancies with placenta previa, a third-trimester cervical length of 30 mm or less is associated with increased risk for hemorrhage, uterine activity, and preterm birth. II.


Current management of esophageal variceal bleeding.
Recent trends in the clinical management of bleeding esophageal varices include a shift away from endoscopic variceal sclerotherapy toward endoscopic variceal ligation. The excellent efficacy of the latter and its lower complication rate favor its increased use. Similarly, the minimally invasive nature of the TIPS procedure and its successful implementation by invasive radiologists have further reduced the need for surgical shunts in general, as well as when endoscopic intervention has failed. A multidisciplinary approach toward evaluation and treatment of variceal hemorrhage can be expected to improve the outcomes of these patients. More selective application of each of the therapies, either alone or in combination with other modalities, leads to improved results, as recently demonstrated by a series comparing outcomes of surgical shunts when used alone or as a preface to liver transplantation.


Case Report: Salmonella Enterica Serovar Paratyphi B Infection in a Febrile Ill Child during Enhanced Passive Surveillance in an Urban Slum in Mirpur, Dhaka.
Paratyphoid fever is one of the major causes of morbidity of febrile illnesses in endemic regions. We report a case of high-grade fever in an infant who was positive for Salmonella enterica serovar Paratyphi B (S. Paratyphi B) both in blood and stool cultures. The baby was enrolled in the passive surveillance of multicenter, multicomponent epidemiological study of enteric fever (Strategic Typhoid alliance across Africa and Asia; STRATAA) conducted in a population of 110,000 residents over 2 years in an urban slum, Dhaka, Bangladesh. This is the only patient who was positive for S. Paratyphi B in blood and stool among more than 6,000 febrile ill patients enrolled in the passive surveillance. The report shows the significance of surveillance to identify changes in the epidemiology of enteric fever.


A comparison of neuronal population dynamics measured with calcium imaging and electrophysiology.
Calcium imaging with fluorescent protein sensors is widely used to record activity in neuronal populations. The transform between neural activity and calcium-related fluorescence involves nonlinearities and low-pass filtering, but the effects of the transformation on analyses of neural populations are not well understood. We compared neuronal spikes and fluorescence in matched neural populations in behaving mice. We report multiple discrepancies between analyses performed on the two types of data, including changes in single-neuron selectivity and population decoding. These were only partially resolved by spike inference algorithms applied to fluorescence. To model the relation between spiking and fluorescence we simultaneously recorded spikes and fluorescence from individual neurons. Using these recordings we developed a model transforming spike trains to synthetic-imaging data. The model recapitulated the differences in analyses. Our analysis highlights challenges in relating electrophysiology and imaging data, and suggests forward modeling as an effective way to understand differences between these data.


Racial/ethnic differences in serum sex steroid hormone concentrations in US adolescent males.
Contrary to the hypothesis that the racial/ethnic disparity in prostate cancer has a hormonal basis, we did not observe a difference in serum testosterone concentration between non-Hispanic black and white men in the Third National Health and Nutrition Examination Survey (NHANES III), although non-Hispanic black men had a higher estradiol level. Unexpectedly, Mexican-American men had the highest testosterone level. Next, we evaluated whether the same patterns are observed during adolescence, the time of prostate maturation. We measured serum testosterone, estradiol, and sex hormone-binding globulin (SHBG) by immunoassay in 134 males aged 12-19 in NHANES III. Mean concentrations were compared by race/ethnicity adjusting for age, Tanner stage, percent body fat, waist, physical activity, tobacco smoke, and the other hormones. After multivariable adjustment, in the 12-15-year-old males, testosterone concentration was lower in non-Hispanic blacks than whites (p = 0.043), SHBG concentration did not significantly differ between the two groups. Mexican-Americans had the highest testosterone (versus non-Hispanic black: p = 0.002) and lowest SHBG (versus non-Hispanic white: p = 0.010; versus non-Hispanic black: p = 0.047) concentrations. Estradiol concentration was lower in non-Hispanic blacks (p = 0.11) and Mexican-Americans (p = 0.033) compared with non-Hispanic whites. After multivariable adjustment, in the 16-19-year-old males, testosterone, estradiol, and SHBG concentrations did not differ between non-Hispanic blacks and whites. Mexican-Americans had the highest testosterone concentration (versus non-Hispanic white: p = 0.08), but did not differ from the other groups on estradiol and SHBG concentrations. In both age groups, these patterns were generally present, but less pronounced after adjusting for age and Tanner stage only. In adolescent males, non-Hispanic blacks did not have a higher testosterone concentration than non-Hispanic whites, and Mexican-Americans had the highest testosterone concentration, patterns similar to adult males.


Effect of vision, proprioception, and the position of the vestibular organ on postural sway.
When measured together, it seems that vision and proprioception as well as position of the vestibular organ affect postural sway, vision the most. Mediolateral (ML) sway does not seem to be influenced by the position of the vestibular organ. To investigate how postural sway was affected by provocation of vision, by the position of the vestibular organ, and by provocation of proprioception, when measured together. Postural sway was measured by using a force plate. Tests were performed with eyes open and eyes closed, with head in neutral position and rotated to the right and to the left and with head maximally extended, both standing on firm surface and on foam. Measures of ML speed (mm/s), anteriorposterior (AP) speed (mm/s), and sway area (SA) (mm(2)/s) were analyzed using a multilevel approach. The multilevel analysis revealed how postural sway was significantly affected by closed eyes and standing on foam, and by the position of the vestibular organ. Closed eyes and standing on foam both significantly prolonged the dependent measurement, irrespective of whether it was ML, AP or SA. However, only AP and SA were significantly affected by vestibular position, i.e. maximal head movement to the right and extension of the head.


Character trees from transcriptome data: Origin and individuation of morphological characters and the so-called "species signal".
We elaborate a framework for investigating the evolutionary history of morphological characters. We argue that morphological character trees generated by phylogenetic analysis of transcriptomes provide a useful tool for identifying causal gene expression differences underlying the development and evolution of morphological characters. They also enable rigorous testing of different models of morphological character evolution and origination, including the hypothesis that characters originate via divergence of repeated ancestral characters. Finally, morphological character trees provide evidence that character transcriptomes undergo concerted evolution. We argue that concerted evolution of transcriptomes can explain the so-called "species signal" found in several recent comparative transcriptome studies. The species signal is the phenomenon that transcriptomes cluster by species rather than character type, even though the characters are older than the respective species. We suggest the species signal is a natural consequence of concerted gene expression evolution resulting from mutations that alter gene regulatory network interactions shared by the characters under comparison. Thus, character trees generated from transcriptomes allow us to investigate the variational independence, or individuation, of morphological characters at the level of genetic programs.


[Seroprevalence of Leptospira infection in habitants of peripheral neighborhoods in Cali, Colombia].
Leptospirosis is a zoonotic disease with varying modes of transmission and clinical expression that affects humans inhabiting urban and rural areas worldwide. In Colombia, little is known regarding leptospirosis in urban settings. To establish the seroprevalence of leptospiral infection in humans, to assess the serological reactivity and to identify factors associated with seropositivity. We conducted an infection prevalence study in 259 inhabitants of peripheral neighborhoods in the city of Cali, Colombia, stratified by sex and age. Sociodemographic information, time residing in the same neighborhood, occupation, exposure to potentially infectious sources like water or animals, and serum used in a Microagglutination test against 19 Leptospira serovars were obtained. The population was composed mainly of students (37.1%), housewives (32%) and workers in non-formal activities (24.6%). The prevalence of anti-Leptospira antibodies was 23.3% (C195% 18.3-28.3), with high frequency in people older than 57 years of age and significantly higher in men than women (p=0.045). There was an association between the presence of antibodies and contact with animals (p=0.038). Reactivity was observed against 16 of the 19 serovars evaluated, but the antibody titers were low. The high infection seroprevalence found in this study suggests frequent contact with Leptospira in putatively high risk neighborhoods of Cali. Contact with animals was the most significant factor associated with the presence of anti-Leptospira antibodies.


Theory and Algorithms for Shapelet-Based Multiple-Instance Learning.
We propose a new formulation of multiple-instance learning (MIL), in which a unit of data consists of a set of instances called a bag. The goal is to find a good classifier of bags based on the similarity with a "shapelet" (or pattern), where the similarity of a bag with a shapelet is the maximum similarity of instances in the bag. In previous work, some of the training instances have been chosen as shapelets with no theoretical justification. In our formulation, we use all possible, and thus infinitely many, shapelets, resulting in a richer class of classifiers. We show that the formulation is tractable, that is, it can be reduced through linear programming boosting (LPBoost) to difference of convex (DC) programs of finite (actually polynomial) size. Our theoretical result also gives justification to the heuristics of some previous work. The time complexity of the proposed algorithm highly depends on the size of the set of all instances in the training sample. To apply to the data containing a large number of instances, we also propose a heuristic option of the algorithm without the loss of the theoretical guarantee. Our empirical study demonstrates that our algorithm uniformly works for shapelet learning tasks on time-series classification and various MIL tasks with comparable accuracy to the existing methods. Moreover, we show that the proposed heuristics allow us to achieve the result in reasonable computational time.


Effect of Lipopolysaccharide-Induced Apical Periodontitis in Diabetes Mellitus Rats on Periapical Inflammation.
To evaluate periapical inflammation through immunohistochemical analysis of interleukin 6 (IL-6) and tumor necrosis factor α (TNF-a) expression resulting from lipopolysaccharide (LPS)-induced apical periodontitis in diabetes mellitus rats, observed at 14, 28, and 42 days. Diabetes model on rats was induced by streptozotocin (STZ). Fifteen rats were injected with low-dose STZ for 5 days and waited for 5 days until the blood glucose level was stable and measured above 300 mg/dL confirmed by a digital glucometer. LPS was used to induce apical periodontitis. After performing access cavity, pulpal and root canal extirpation was done on the right mandibular first molar's root canal space of rats, under anesthesia. LPS of 1 mg/mL dose was induced in the pulpal and root canal space. Apical periodontitis was expected 14 days afterward and then, the rats were randomly allocated to three groups. The first group was terminated 14 days after induction and used as control. The second group was observed 28 days after induction, and the third group was observed 42 days after induction. IL-6 and TNF-a expression was analyzed by immunohistochemistry on macrophages in the periapical area. Data were analyzed using one-way ANOVA and continued with the post hoc Tukey HSD test. Significance was considered if p < 0.05. LPS induced apical periodontitis in diabetes mellitus rats at control (14 days), 28 and 42 days observation showed a significant increase in the expression of IL-6 and TNF-a. There were significant differences between the control and observed groups (p < 0.05). The expression of IL-6 in the apical area was not significant at 14 and 28 days (p > 0.05) but increased significantly at 42 days (p < 0.05). The expression of TNF-a in the apical area was significantly increased after 14 days (p < 0.05) and remained stable at 28 and 42 days (p > 0.05). The periapical inflammation of LPS-induced apical periodontitis in diabetes mellitus rats increased macrophages' expression of IL-6 at 42 days and TNF-a at 28 days.


DNA origami nanopores: developments, challenges and perspectives.
DNA nanotechnology has enabled the construction of DNA origami nanopores; synthetic nanopores that present improved capabilities for the area of single molecule detection. Their extraordinary versatility makes them a new and powerful tool in nanobiotechnology for a wide range of important applications beyond molecular sensing. In this review, we briefly present the recent developments in this emerging field of research. We discuss the current challenges and possible solutions that would enhance the sensing capabilities of DNA origami nanopores. Finally, we anticipate novel avenues for future research and highlight a range of exciting ideas and applications that could be explored in the near future.


Epithelial NAIPs protect against colonic tumorigenesis.
NLR family apoptosis inhibitory proteins (NAIPs) belong to both the Nod-like receptor (NLR) and the inhibitor of apoptosis (IAP) families. NAIPs are known to form an inflammasome with NLRC4, but other in vivo functions remain unexplored. Using mice deficient for all NAIP paralogs (Naip1-6(Δ/Δ)), we show that NAIPs are key regulators of colorectal tumorigenesis. Naip1-6(Δ/Δ) mice developed increased colorectal tumors, in an epithelial-intrinsic manner, in a model of colitis-associated cancer. Increased tumorigenesis, however, was not driven by an exacerbated inflammatory response. Instead, Naip1-6(Δ/Δ) mice were protected from severe colitis and displayed increased antiapoptotic and proliferation-related gene expression. Naip1-6(Δ/Δ) mice also displayed increased tumorigenesis in an inflammation-independent model of colorectal cancer. Moreover, Naip1-6(Δ/Δ) mice, but not Nlrc4-null mice, displayed hyper-activation of STAT3 and failed to activate p53 18 h after carcinogen exposure. This suggests that NAIPs protect against tumor initiation in the colon by promoting the removal of carcinogen-elicited epithelium, likely in a NLRC4 inflammasome-independent manner. Collectively, we demonstrate a novel epithelial-intrinsic function of NAIPs in protecting the colonic epithelium against tumorigenesis.


Induction without methanol: novel regulated promoters enable high-level expression in Pichia pastoris.
Inducible high-level expression is favoured for recombinant protein production in Pichia pastoris. Therefore, novel regulated promoters are desired, ideally repressing heterologous gene expression during initial growth and enabling it in the production phase. In a typical large scale fed-batch culture repression is desired during the batch phase where cells grow on a surplus of e.g. glycerol, while heterologous gene expression should be active in the feed phase under carbon (e.g. glucose) limitation. DNA microarray analysis of P. pastoris wild type cells growing in glycerol-based batch and glucose-based fed batch was used for the identification of genes with both, strong repression on glycerol and high-level expression in the feed phase. Six novel glucose-limit inducible promoters were successfully applied to express the intracellular reporter eGFP. The highest expression levels together with strong repression in pre-culture were achieved with the novel promoters P(G1) and P(G6). Human serum albumin (HSA) was used to characterize the promoters with an industrially relevant secreted protein. A P(G1) clone with two gene copies reached about 230% of the biomass specific HSA titer in glucose-based fed batch fermentation compared to a P(GAP) clone with identical gene copy number, while P(G6) only achieved 39%. Two clones each carrying eleven gene copies, expressing HSA under control of P(G1) and P(G6) respectively were generated by post-transformational vector amplification. They produced about 1.0 and 0.7 g L(-1) HSA respectively in equal fed batch processes. The suitability in production processes was also verified with HyHEL antibody Fab fragment for P(G1) and with porcine carboxypeptidase B for P(G6). Moreover, the molecular function of the gene under the control of P(G1) was determined to encode a high-affinity glucose transporter and named GTH1. A set of novel regulated promoters, enabling induction without methanol, was successfully identified by using DNA microarrays and shown to be suitable for high level expression of recombinant proteins in glucose-based protein production processes.


Glucose-regulated protein 78 mediates the anticancer efficacy of shikonin in hormone-refractory prostate cancer cells.
Glucose-regulated protein 78 (GRP78) is a key modulator of prostate cancer progression and therapeutic resistance. Prostate cancer is a worldwide health problem, and therapeutic resistance is a critical obstacle for the treatment of hormone-refractory prostate cancer patients. Shikonin inhibits prostate cancer proliferation and metastasis. However, the role of GRP78 in the cytotoxic effect of shikonin in prostate cancer cells remains unclear. GRP78 expression was abolished using small interfering RNA (siRNA), and the anticancer effects of shikonin were assessed using MTT assays, the XCELLigence biosensor, flow cytometric cell cycle analysis, and Annexin V-PI apoptotic assays. PC-3 cells expressed more GRP78 than DU-145 cells, and the MTT assays revealed that DU-145 cells were more sensitive to shikonin than PC-3 cells. GRP78 knockdown (GRP78KD) PC-3 cells were more sensitive to shikonin treatment than scrambled siRNA control cells. Based on cell cycle analysis and AnnexinV-PI apoptotic assays, apoptosis dramatically increased in GRP78KD cells compared with the control PC-3 in response to shikonin. Finally, in response to shikonin treatment, Mcl-1 and Bcl-2 levels increased in the scrambled control cells treated with shikonin, whereas Bcl-2 decreased and Mcl-1 slightly increased in the GRP78KD PC-3 cells. The levels of Bax and Bad did not change in the scrambled control or GRP78KD cells after shikonin treatment. These results are consistent with the increased sensitivity to shikonin after knockdown of GRP78. GRP78 expression may determine the therapeutic efficacy of shikonin against prostate cancer cells.


Unexpected high pressure effects on the structural properties of condensed whey protein systems.
We show that application of high hydrostatic pressure (600 MPa for 15 min) on condensed whey protein (WP) systems (e.g., 80% w/w solids content) results in unexpected structure-function behavior when compared with conventional thermal treatment. Unraveling the relaxation properties in first-order thermodynamic transitions, the manifestation of glass transition phenomena and the preservation of native conformation in condensed preparations were recorded using small-deformation dynamic oscillation in shear, modulated differential scanning calorimetry, and infrared spectroscopy. Informed temperature application results in the formation of continuous networks at the denaturation temperature, which undergo vitrification at subzero temperatures. In contrast, high-pressure-treated WPs resist physicochemical denaturation, hence preserving the native conformation of secondary and tertiary structures. This was rationalized on the basis of a critical concentration threshold where transfer of water molecules to nonpolar residues in the protein interior is minimized because of low moisture content and restricted molecular mobility. The physical state and morphology of these high-solid preparations were further examined by the combined framework of reduced variables and Williams, Landel, and Ferry equation/free volume theory. Theoretical treatment of experimental observations unveils the dynamic range of the mechanical manifestation of the glass transition region in samples subjected to heat or pressure. In addition to preserving native conformation, WPs subjected to high pressure form glassy systems at parity with the structural functionality of the thermally treated counterparts.


The effects of age and dietary restriction without nutritional supplementation on whole bone structural properties in C57BL/6J mice.
While caloric restriction is a proven means to extend longevity, its effects on bone are not well understood. This study examined the effects of dietary restriction without vitamin or mineral supplementation on bone in female 60- and 120-day-old C57BL/6J mice. Baseline controls were sacrificed at 60 or 120 days, while diet-restricted animals ate approximately 72.9-78.6% of the ad libitum fed animals for thirty days. 60-day-old ad libitum animals experienced normal growth with average increases of 6.4% in bone length, 23.5% in bone mass, 9.4% in %mineralization, 36.4% in maximum strength, 59.2% in stiffness, 22.3% in cortical thickness, 12.9% in %cortical area, and 11.3% in microhardness. Growth in 120-day-old ad libitum animals followed a trend but with more modest increases. Diet-restricted mice matured very little from baseline levels in 60-day-old animals. There were no significant changes from baseline levels in the parameters indicated above, except for a 8.3% decrease in %cortical area attributable to increased resorption. 120-day-old diet-restricted animals also evidenced little deviation from baseline levels except for significant decreases in %mineralization (2.1%) and %cortical area (6.7%). The effects of diet restriction on bone properties decreased with age. Bone from 60-day-old diet-restricted mice showed diminished mechanical and compositional properties, resulting from little growth and excess resorption. Bone from 120-day-old diet-restricted mice showed little growth and some resorption. Increased resorption, localized on the endosteal surfaces, likely minimized the negative impact of structural degradation of the long bones. Resorption may have also provided minerals to compensate for nutritional deficiencies.


Profibrotic potential of prominin-1+ epithelial progenitor cells in pulmonary fibrosis.
In idiopathic pulmonary fibrosis loss of alveolar epithelium induces inflammation of the pulmonary tissue followed by accumulation of pathogenic myofibroblasts leading eventually to respiratory failures. In animal models inflammatory and resident cells have been demonstrated to contribute to pulmonary fibrosis. Regenerative potential of pulmonary and extra-pulmonary stem and progenitor cells raised the hope for successful treatment option against pulmonary fibrosis. Herein, we addressed the contribution of lung microenvironment and prominin-1(+) bone marrow-derived epithelial progenitor cells in the mouse model of bleomycin-induced experimental pulmonary fibrosis. Prominin-1(+) bone marrow-derived epithelial progenitors were expanded from adult mouse lungs and differentiated in vitro by cytokines and growth factors. Pulmonary fibrosis was induced in C57Bl/6 mice by intratracheal instillation of bleomycin. Prominin-1(+) progenitors were administered intratracheally at different time points after bleomycin challenge. Green fluorescence protein-expressing cells were used for cell tracking. Cell phenotypes were characterized by immunohistochemistry, flow cytometry and quantitative reverse transcription-polymerase chain reaction. Prominin-1(+) cells expanded from healthy lung represent common progenitors of alveolar type II epithelial cells, myofibroblasts, and macrophages. Administration of prominin-1(+) cells 2 hours after bleomycin instillation protects from pulmonary fibrosis, and some of progenitors differentiate into alveolar type II epithelial cells. In contrast, prominin-1(+) cells administered at day 7 or 14 lose their protective effects and differentiate into myofibroblasts and macrophages. Bleomycin challenge enhances accumulation of bone marrow-derived prominin-1(+) cells within inflamed lung. In contrast to prominin-1(+) cells from healthy lung, prominin-1(+) precursors isolated from inflamed organ lack regenerative properties but acquire myofibroblast and macrophage phenotypes. The microenvironment of inflamed lung impairs the regenerative capacity of bone marrow-derived prominin-1(+) progenitors and promotes their differentiation into pathogenic phenotypes.