Clonal analysis of murine graft-vs-host disease. I. Phenotypic and functional analysis of T lymphocyte clones.
Acute and chronic graft-vs-host disease (GVHD) due to non-MHC histocompatibility differences differ histopathologically. Acute GVHD is characterized by the cytotoxic destruction of recipient tissues, whereas chronic GVHD is characterized by increased collagen deposition. In an attempt to determine if acute and chronic GVHD represent two phases of the same pathophysiologic process or two distinct processes, the T lymphocytes from the C57BL/6 (B6) recipients of LP spleen cells (non-H-2 GVHD) have been cloned and compared to clones from immune mice (LP anti-B6). Acute GVHD (G) clones were established on day 10-14 posttransplant and chronic GVHD (CG) clones on day 50 from animals with clinical chronic GVHD. Immune (I) clones were established 10 to 14 days after immunization. All I clones exhibited B6-specific blastogenesis and cytotoxicity and had a Thy-1.2+, Lyt-2.2+, L3T4- phenotype. All CG clones were noncytotoxic, had I-Ab-specific blastogenesis, and had a Thy-1.2+, Lyt-2.2-, L3T4+ phenotype. The acute GVHD (G) clones were heterogeneous. Fourteen of 23 clones exhibited B6-specific blastogenesis and had a Lyt-2.2+, L3T4- phenotype (B6-G clones). Seven of 9 B6-G clones were cytotoxic for B6 targets. Nine of 23 G clones exhibited I-Ab-specific blastogenesis, and all but one clone had a Lyt-2.2-, L3T4+ phenotype as the did CG clones. Thus, the principal clonogenic T lymphocytes from mice with acute and chronic GVHD differ in terms of 1) their antigenic specificity, 2) their cytotoxic capacity, and 3) their surface phenotype. The presence of I-Ab-specific T lymphocytes with a phenotype identical to CG clones early after transplantation suggests that the immunologic events that result in chronic GVHD begin soon after transplantation. These results indicate that acute GVHD is due primarily to recipient-specific cytotoxic donor T lymphocytes, whereas chronic GVHD is due to autoreactive helper T lymphocytes.


A comparison of least squares and conditional maximum likelihood estimators under volume endpoint censoring in tumor growth experiments.
Measurements in tumor growth experiments are stopped once the tumor volume exceeds a preset threshold: a mechanism we term volume endpoint censoring. We argue that this type of censoring is informative. Further, least squares (LS) parameter estimates are shown to suffer a bias in a general parametric model for tumor growth with an independent and identically distributed measurement error, both theoretically and in simulation experiments. In a linear growth model, the magnitude of bias in the LS growth rate estimate increases with the growth rate and the standard deviation of measurement error. We propose a conditional maximum likelihood estimation procedure, which is shown both theoretically and in simulation experiments to yield approximately unbiased parameter estimates in linear and quadratic growth models. Both LS and maximum likelihood estimators have similar variance characteristics. In simulation studies, these properties appear to extend to the case of moderately dependent measurement error. The methodology is illustrated by application to a tumor growth study for an ovarian cancer cell line.


Role models' perceptions of themselves and their influence on students' specialty choices.
To examine the perceptions of faculty role models to learn whether their perceptions of role models' behaviors are congruent with those of their students. In 1996 a survey was mailed to 210 student-identified faculty role models at the University of Washington School of Medicine and the University of North Carolina at Chapel Hill School of Medicine. The nominees were asked to rate to what extent each of 32 student-identified role model characteristics was representative of their behavior. They were then asked to rank order the characteristics they felt were most important to model for students. The role models were also asked to rate how much influence they perceived themselves to have on their students' specialty choices. A final, open-ended question inquired about the single characteristics they modeled to students that most influenced the students' specialty choices. The role models' specialties were grouped as either primary care (PC) or non-primary care (NPC). Data were analyzed with several statistical methods. Of the 210 mailed surveys, a total of 177 were returned, for a response rate of 84%. The role models perceived their behaviors much like their students did; the role models' self-ratings were generally high for all of the student-defined characteristics. Although clinical reasoning was considered the most important characteristic to model for students, the role models also believed that enthusiasm and love for their work were the characteristics that most influenced their students' specialty choices. Few differences were found between the PC and the NPC role models. The role models in this study agreed with their students about what is important to model. They did not intentionally try to recruit students to join their specialties but felt that demonstrating enthusiasm and a sincere love for what they did has a strong influence toward this end.


Genome-wide association study of corticobasal degeneration identifies risk variants shared with progressive supranuclear palsy.
Corticobasal degeneration (CBD) is a neurodegenerative disorder affecting movement and cognition, definitively diagnosed only at autopsy. Here, we conduct a genome-wide association study (GWAS) in CBD cases (n=152) and 3,311 controls, and 67 CBD cases and 439 controls in a replication stage. Associations with meta-analysis were 17q21 at MAPT (P=1.42 × 10(-12)), 8p12 at lnc-KIF13B-1, a long non-coding RNA (rs643472; P=3.41 × 10(-8)), and 2p22 at SOS1 (rs963731; P=1.76 × 10(-7)). Testing for association of CBD with top progressive supranuclear palsy (PSP) GWAS single-nucleotide polymorphisms (SNPs) identified associations at MOBP (3p22; rs1768208; P=2.07 × 10(-7)) and MAPT H1c (17q21; rs242557; P=7.91 × 10(-6)). We previously reported SNP/transcript level associations with rs8070723/MAPT, rs242557/MAPT, and rs1768208/MOBP and herein identified association with rs963731/SOS1. We identify new CBD susceptibility loci and show that CBD and PSP share a genetic risk factor other than MAPT at 3p22 MOBP (myelin-associated oligodendrocyte basic protein).


Defibrotide for the prevention of hepatic veno-occlusive disease after hematopoietic stem cell transplantation: a systematic review.
Prophylactic use of defibrotide (DF) to prevent veno-occlusive disease (VOD), a relatively common and high-risk complication of hematopoietic stem cell transplantation (HSCT), may be an encouraging modality to reduce morbidity and mortality from VOD. However, conclusions remain unclear. We carried out a systematic review to summarize the state of knowledge. One randomized controlled trial (RCT), four cohort studies and eight case series studies were found, including a total of 1230 patients. The overall mean incidence of VOD in patients using DF was 4.7% (95% CI, 3.3-6.1) which was significantly lower than the data 13.7% (95% CI, 13.3-14.1) across 135 studies using no VOD prophylaxis (p < 0.005). The meta-analysis of the incidence of VOD in controlled trials revealed a statistical reduction in VOD incidence in the DF group (RR = 0.47, 95% CI, 0.31-0.73). The overall mean incidence of severe VOD was 0.8% (95% CI, 0.2-1.4). The RR was 0.31 (95% CI, 0.09-1.06). However, the lack of RCTs and the methodological weaknesses of the studies may preclude making generalizable conclusions. Our review described that DF appears promising for VOD prevention and large RCT is needed for further confirmation.


Variable clinical presentations of babesiosis: A case series.
Human babesiosis continues to spread in multiple regions of the United States. It is transmitted by Ixodes species ticks, as are Lyme disease and anaplasmosis. Its variable clinical presentations, together with serologic detection limitations, require that a high index of clinical suspicion be present for prompt diagnosis. This article discusses case presentations showing the wide range of symptoms and presentations that are possible with babesiosis.


Should sputum isolates of Haemophilus influenzae be serotyped?
In a patient with pneumonia, sputum culture revealed ampicillin-resistant Haemophilus influenzae, type b. Although nontypable H influenzae is a normal inhabitant of the upper respiratory tract and should be considered normal flora, typable H influenzae is found in less than 5% of healthy persons. Although only 1.8% of sputum cultures reveal typable H influenzae, these strains account for 98.3% of isolates from blood cultures of patients with pneumonia due to H influenzae. Serotyping of sputum isolates is recommended in patients with pneumonia to separate pathogenic typable strains from nonpathogenic nontypable strains.


[The role of social support in the relationship between job stress and job satisfaction/organizational commitment].
The purpose of this study is to investigate the role of social support in the experience of job stress among hospital nurses. This study was carefully designed to overcome methodological shortcomings found in past research, and examined two organizational effectiveness variables(job satisfaction and organizational commitment) as outcome variables. The sample used in this study consisted of 602 nurses from 5 general hospitals. Data were collected with self-administered questionnaires and analyzed using hierarchical regression and LISREL technique. It was found that: (a) three job stress variables(workload, role conflict and conflict with other medical staff) have negative effects on job satisfaction and organizational commitment; (b) social support have positive main effects on the two output variables; (c) the negative effects of job stress variables on job satisfaction and organizational commitment are not buffered by social support, and (d) social support mediates the effects of job stress on job satisfaction and organizational commitment, and the size of the mediating effects is small. Further research needs to be done to further refine this study.


Beneficial developmental acclimation in reproductive performance under cold but not heat stress.
Thermal plasticity can help organisms coping with climate change. In this study, we analyse how laboratory populations of the ectotherm species Drosophila subobscura, originally from two distinct latitudes and evolving for several generations in a stable thermal environment (18 °C), respond plastically to new thermal challenges. We measured adult performance (fecundity traits as a fitness proxy) of the experimental populations when exposed to five thermal regimes, three with the same temperature during development and adulthood (15-15 °C, 18-18 °C, 25-25 °C), and two where flies developed at 18 °C and were exposed, during adulthood, to either 15 °C or 25 °C. Here, we test whether (1) flies undergo stress at the two more extreme temperatures; (2) development at a given temperature enhances adult performance at such temperature (i.e. acclimation), and (3) populations with different biogeographical history show plasticity differences. Our findings show (1) an optimal performance at 18 °C only if flies were subjected to the same temperature as juveniles and adults; (2) the occurrence of developmental acclimation at lower temperatures; (3) detrimental effects of higher developmental temperature on adult performance; and (4) a minor impact of historical background on thermal response. Our study indicates that thermal plasticity during development may have a limited role in helping adults cope with warmer - though not colder - temperatures, with a potential negative impact on population persistence under climate change. It also emphasizes the importance of analysing the impact of temperature on all stages of the life cycle to better characterize the thermal limits.


Improving reproducibility and external validity. The role of standardization and data reporting of laboratory rat husbandry and housing.
To identify the most relevant flaws in standardization in husbandry practices and lack of transparency to report them. This review proposes some measures in order to improve transparency, reproducibility and eventually external validity in experimental surgery experiments with rat model. We performed a search of scientific articles in PUBMED data base. The survey was conducted from august 2016 to January 2017. The keywords used were "reproducibility", "external validity", "rat model", "rat husbandry", "rat housing", and the time frame was up to January 2017. Articles discarded were the ones which the abstract or the key words did not imply that the authors would discuss any relationship of husbandry and housing with the reproducibility and transparency of reporting animal experiment. Reviews and papers that discussed specifically reproducibility and data reporting transparency were laboriously explored, including references for other articles that could fulfil the inclusion criteria. A total of 246 articles were initially found but only 44 were selected. Lack of transparency is the rule and not the exception when reporting results with rat model. This results in poor reproducibility and low external validity with the consequence of considerable loss of time and financial resources. There are still much to be done to improve compliance and adherence of researchers, editors and reviewers to adopt guidelines to mitigate some of the challenges that can impair reproducibility and external validity. Authors and reviewers should avoid pitfalls of absent, insufficient or inaccurate description of relevant information the rat model used. This information should be correctly published or reported on another source easily available for readers. Environmental conditions are well known by laboratory animal personnel and are well controlled in housing facilities, but usually neglected in experimental laboratories when the rat model is a novelty for the researcher.


Determining effective interfacial tension and predicting finger spacing for DNAPL penetration into water-saturated porous media.
The difficulty in determining the effective interfacial tension limits the prediction of the wavelength of fingering of immiscible fluids in porous media. A method to estimate the effective interfacial tension using fractal concepts was presented by Chang et al. [Water Resour. Res. 30 (1994) 125]. We modified the method in that the macroscopic interface length was used instead of the system width. Methods to determine the macroscopic and the microscopic interface length are given. Lab experiments of dense nonaqueous phase liquid (DNAPL) penetrating into water-saturated glass beads were carried out in a two-dimensional (2-D) transparent chamber. The displacement processes were recorded using a 35-mm camera or a video camera, which was directly connected to and controlled by a computer. Unlike the method of Chang et al. (1994), the modified method used here gives a constant value of the effective interfacial tension over time. The predicted wavelengths of fingering are relatively close to those observed except for the fine beads.


The retentive ability of various cementing agents for polycarbonate crowns.
The following conclusions may be made within the limitations of this investigation. - A composite resin processed directly against a roughened stainless steel dye, similar in shape to a primary anterior tooth, which has been shaped to receive a polycarbonate crown, shows the highest retentive force measured in pounds per square inch (psi). - A polmethacrylate resin, when used as a cement, also shows high values, probably due to its ability to unite chemically with polycarbonated acrylic and to its low film thickness. - Composite resins of low viscosity, low film thickness, and high compressive and tensile strengths proved also to provide good retentive values and would contribute insolubility to a greater degree than the unfilled resin. - Polycarboxylate, zinc phosphate, and reinforced zinc oxide-eugenol cements are not to be recommended as agents for cementing polycarbonate crowns. - It can be recommended on the basis of retention only, composite crowns should be fabricated directly onto the tooth preparations of carious primary anterior teeth.


Postrenal azotemia due to C. albicans pyocystitis and a Candidal plug.
There are many causes of acute renal failure, with cases often being classified as pre-renal, intrarenal or post-renal based on the etiology. A post-obstructive etiology is responsible in a minority of patients. Infectious causes of post-obstructive azotemia are even more rare, but as this case illustrates, should be considered as part of the differential. This case report describes a patient who developed acute renal failure (ARF) while hospitalized. Examination showed a candidal plug at his urethral meatus. Bladder washings removed a significant amount of purulent material. Treatment for C. albicans with fluconazole resolved his urinary retention and his bilateral hydronephrosis. His acute renal failure was due to an infectious, post-obstructive pyocystitis and formation of a candidal plug in his urethral meatus. Removal of the candidal plug and treatment of his infection with fluconazole resulted in return to baseline renal function. While not common, an infectious cause of post-renal azotemia should be considered in the appropriate clinical situation.


Measurement of upper esophageal sphincter tone and relaxation during swallowing in premature infants.
Upper esophageal sphincter (UES) motor function has not been previously evaluated in premature infants. The motor patterns associated with tonic activity and swallow-related relaxation of the UES were recorded for 1 h after completion of gavage feeding in 11 healthy preterm neonates (postmenstrual age 33-37 wk) with a micromanometric assembly, which included a sleeve sensor specifically adapted for UES recordings. A clearly defined UES high-pressure zone was observed in all premature infants studied. Resting UES pressure ranged from 2.3 to 26.2 mmHg and was higher during periods of irritability and apparent discomfort. During dry swallows, UES pressure relaxed from a resting pressure of 28.2 +/- 4.0 mmHg to a nadir of 1.1 +/- 3.3 mmHg. The mean UES relaxation interval (the time from relaxation onset to relaxation offset) was 0. 31 +/- 0.11 s. We conclude that in premature infants >/=33 wk postmenstrual age the motor mechanisms regulating UES resting pressure and the onset of UES relaxation are well developed.


Relative stability of benziporphyrin and naphthiporphyrin tautomers and the emergence of macrocyclic diatropicity.
The conformations of a series of benziporphyrins, naphthiporphyrins, oxybenziporphyrins, and related structures were minimized using DFT-B3LYP/6-311++G(d,p). The relative stabilities of the tautomers for each series were calculated using M06-2X and B3LYP-D functionals, and bond lengths were obtained. The diatropic properties of each species were assessed by calculating nucleus independent chemical shifts (NICS) and the related NICSzz values. Although benziporphyrin and naphthiporphyrin tautomers essentially exhibit no aromatic properties, mono- and diprotonated species show weakly diatropic characteristics in agreement with experimental observations. Benziphlorins, intermediates in the MacDonald "3 + 1" synthesis of benziporphyrins, were also examined and tautomers with methylene units adjacent to the benzene ring were shown to be far more stable than tautomers with a CH2 bridge between two pyrrolic units. 2-Hydroxybenziporphyrin was shown to be significantly less stable than the aromatic tautomer oxybenziporphyrin, although diprotonation leads to a species with somewhat reduced diatropicity. Related systems were also analyzed and the results demonstrate that benziporphyrins and naphthiporphyrins range from structures with no measurable macrocyclic aromaticity to strongly aromatic systems that exhibit large diamagnetic ring currents.


Severe Neonatal Presentation of Progressive Familial Intrahepatic Cholestasis Type 4 in an Omani Infant.
Progressive familial intrahepatic cholestasis type 4 (PFIC4) is a relatively newly described autosomal recessive disorder caused by biallelic mutations in the gene encoding tight junction protein 2 (TJP2) which is located in chromosome 9q21. PFIC4 is characterised by cholestasis with or without other extrahepatic manifestations. Bleeding tendency due to vitamin k deficiency is a well-known complication of cholestasis. We present a neonate who presented to the Emergency Department at a tertiary care hospital in 2021 with cholestasis and multiple intracranial bleeds. He was found to have severe coagulopathy and his genetic work up revealed a homozygous variant mutation in TJP2 gene causing PFIC4. He had persistent cholestasis that necessitated an internal biliary diversion with some clinical improvement.


Delayed replantation of avulsed mature teeth with calcium hydroxide treatment.
Three avulsed teeth that were replanted and splinted after approximately a 200-min dry extraoral period in two patients are presented. In case 1 calcium hydroxide treatment was performed 1 month after replantation, because the patient did not come for endodontic treatment on the day after replantation as requested. In case 2, calcium hydroxide treatment was initiated on the day after replantation. Calcium hydroxide treatment was used to prevent or treat inflammatory root resorption. During the follow-up periods the teeth reported in these cases have remained in a stable, functional position and did not reveal clinical ankylosis or replacement resorption.


A statistics-based pitch contour model for Mandarin speech.
A statistics-based syllable pitch contour model for Mandarin speech is proposed. This approach takes the mean and the shape of a syllable log-pitch contour as two basic modeling units and considers several affecting factors that contribute to their variations. The affecting factors include the speaker, prosodic state (which essentially represents the high-level linguistic components of F0 and will be explained more clearly in Sec. I), tone, and initial and final syllable classes. The parameters of the two modeling units were automatically estimated using the expectation-maximization (EM) algorithm. Experimental results showed that the root mean squared errors (RMSEs) obtained in the closed and open tests in the reconstructed pitch period were 0.362 and 0.373 ms, respectively. This model provides a way to separate the effects of several major factors. All of the inferred values of the affecting factors were in close agreement with our prior linguistic knowledge. It also gives a quantitative and more complete description of the coarticulation effect of neighboring tones rather than conventional qualitative descriptions of the tone sandhi rules. In addition, the model can provide useful cues to determine the prosodic phrase boundaries, including those occurring at intersyllable locations, with or without punctuation marks.


Intangible Identity Theft and Intimate Partner Violence.
While tougher domestic violence laws and protective orders are frequently credited with attenuating intimate partner violence (IPV), one unexplored explanation for this observed reduction is that intimate partner abusers are shifting their abusive behavior to intangible identity theft to thwart legal mechanisms traditionally used to deter IPV. Unlike the monetary motive associated with document identity theft, intangible identity theft is committed by someone with a preexisting grievance against the victim because the theft's primary purpose is to tarnish the victim's reputation. Results from a multilevel analysis show that a woman has a lower probability of being a victim of an intimate rather than nonintimate partner crime in cities with a higher intangible identity theft rate. Such a finding suggests that intangible identity theft may be a form of intimate partner abuse with few adverse consequences for offenders because identity thieves are rarely arrested and prosecuted. Nevertheless, the current study is only preliminary. Further research is needed before our findings and conclusions can be universally accepted.


Stakeholder perspectives from 15 countries in Africa on barriers in snakebite envenoming research and the potential role of research hubs.
Snakebite envenoming is a debilitating neglected tropical disease disproportionately affecting the rural poor in low and middle-income countries in the tropics and sub-tropics. Critical questions and gaps in public health and policy need to be addressed if major progress is to be made towards reducing the negative impact of snakebite, particularly in the World Health Organisation (WHO) Africa region. We engaged key stakeholders to identify barriers to evidence-based snakebite decision making and to explore how development of research and policy hubs could help to overcome these barriers. We conducted an electronic survey among 73 stakeholders from ministries of health, health facilities, academia and non-governmental organizations from 15 countries in the WHO Africa region. The primary barriers to snakebite research and subsequent policy translation were limited funds, lack of relevant data, and lack of interest from policy makers. Adequate funding commitment, strong political will, building expert networks and a demand for scientific evidence were all considered potential factors that could facilitate snakebite research. Participants rated availability of antivenoms, research skills training and disease surveillance as key research priorities. All participants indicated interest in the development of research and policy hubs and 78% indicated their organization would be willing to actively participate. In conclusion, our survey affirms that relevant stakeholders in the field of snakebite perceive research and policy hubs as a promising development, which could help overcome the barriers to pursuing the WHO goals and targets for reducing the burden of snakebite.


Approval Process: An Overview of Biosimilars in the Oncology Setting.
Following approval of the Biologics Price Competition and Innovation Act of 2009, biosimilars are gradually entering the market in the United States. With the introduction of more biosimilars into the marketplace, all healthcare providers should be familiar with the approval and evaluation process, the naming convention applied to these agents, and the importance of accurate pharmacovigilance. This article aims to describe the approval process of biosimilars, including extrapolation, and to help healthcare providers understand when a biosimilar may be interchanged for the reference biologic and the naming convention used for biosimilars. In addition, this article explores how these topics affect confidence in dispensing and pharmacovigilance. A literature review was conducted, and search terms and variation included biosimilar agents AND FDA approval, legislation, interchangeability, naming conventions, confidence in dispensing, and pharmacovigilance. Healthcare providers involved in the dispensing and administration of biosimilar and interchangeable biologics need to be continually educated to ensure confidence, familiarity, and accuracy with the processes surrounding biosimilars.


Severity of meningococcal disease associated with genomic bacterial load.
Diagnostic polymerase chain reaction (PCR) detection of Neisseria meningitidis has enabled accurate quantification of the bacterial load in patients with meningococcal disease. Quantification of the N. meningitidis DNA level by real time-PCR was conducted on whole-blood samples obtained from patients presenting with meningococcal disease to hospitals throughout England and Wales over a 3-year period. Levels were correlated with clinical outcome, infecting serogroup, and host factors including, interleukin-1 genotype (IL-1). Bacterial loads were available for 1045 patients and were not associated with the age of the patient, delay in sample submission, or administration of antibiotics prior to admission. The median log bacterial load was higher in 95 patients who died (5.29 log(10)copies/mL; interquartile range, 4.41-6.30 log(10)copies/mL) than in 950 patients who survived (3.79 log(10)copies/mL; interquartile range, 2.87-4.71 log(10)copies/mL). Logistic regression revealed that age (odds ratio, 1.04 per 1-year increase in age) and bacterial load (odds ratio, 2.04 per log(10)-copies/mL increase) had a statistically significant effect on the risk of death. Infection with N. meningitidis serogroup C was associated with increased risk of death and an increased bacterial load. Also associated with a higher bacterial load were prolonged hospitalization (duration, >10 days); digit, limb, or soft-tissue loss; and requirement of hemodialysis. Carriage of IL-1RN(+2018) was associated with increased mortality (odds ratio, 2.14; P=.07) but not with a higher bacterial load. In meningococcal disease, bacterial load is associated with likelihood of death, development of permanent disease sequelae, and prolonged hospitalization. The bacterial load was relatively higher in patients infected with N. meningitidis serogroup C than in those infected with other serogroups. The effects of age and IL-1 genotype on mortality are independent of a high genomic bacterial load.


Importin α7 is essential for zygotic genome activation and early mouse development.
Importin α is involved in the nuclear import of proteins. It also contributes to spindle assembly and nuclear membrane formation, however, the underlying mechanisms are poorly understood. Here, we studied the function of importin α7 by gene targeting in mice and show that it is essential for early embryonic development. Embryos lacking importin α7 display a reduced ability for the first cleavage and arrest completely at the two-cell stage. We show that the zygotic genome activation is severely disturbed in these embryos. Our findings indicate that importin α7 is a new member of the small group of maternal effect genes.


Absence of Azole or Echinocandin Resistance in Candida glabrata Isolates in India despite Background Prevalence of Strains with Defects in the DNA Mismatch Repair Pathway.
Candida glabrata infections are increasing worldwide and exhibit greater rates of antifungal resistance than those with other species. DNA mismatch repair (MMR) gene deletions, such as msh2Δ, in C. glabrata resulting in a mutator phenotype have recently been reported to facilitate rapid acquisition of antifungal resistance. This study determined the antifungal susceptibility profiles of 210 C. glabrata isolates in 10 hospitals in India and investigated the impact of novel MSH2 polymorphisms on mutation potential. No echinocandin- or azole-resistant strains and no mutations in FKS hot spot regions were detected among the C. glabrata isolates, supporting our in vitro susceptibility testing results. CLSI antifungal susceptibility data showed that the MICs of anidulafungin (geometric mean [GM], 0.12 μg/ml) and micafungin (GM, 0.01 μg/ml) were lower and below the susceptibility breakpoint compared to that of caspofungin (CAS) (GM, 1.31 μg/ml). Interestingly, 69% of the C. glabrata strains sequenced contained six nonsynonymous mutations in MSH2, i.e., V239L and the novel mutations E459K, R847C, Q386K, T772S, and V239/D946E. Functional analysis of MSH2 mutations revealed that 49% of the tested strains (40/81) contained a partial loss-of-function MSH2 mutation. The novel MSH2 substitution Q386K produced higher frequencies of CAS-resistant colonies upon expression in the msh2Δ mutant. However, expression of two other novel MSH2 alleles, i.e., E459K or R847C, did not confer selection of resistant colonies, confirming that not all mutations in the MSH2 MMR pathway affect its function or generate a phenotype of resistance to antifungal drugs. The lack of drug resistance prevented any correlations from being drawn with respect to MSH2 genotype.


Molecular weight of dissolved organic matter-napropamide complex transported through soil columns.
Soil-derived dissolved organic matter (DOM) has been shown to form stable complexes with the herbicide napropamide [2-(alpha-naphthoxy-N,N-diethylpropionamide] capable of enhancing the transport of napropamide through soil columns. Two soils, one containing sewage sludge-derived organic matter (SS) and the other having only natural organic matter (NoSS) were treated with napropamide and allowed to dry to promote complex formation. Soil columns were prepared by packing a 10-cm layer of untreated, dry, sieved soil followed by an overlying 5-cm layer of napropamide-treated soil. Columns were irrigated and the effluent collected and placed in dialysis chambers. After equilibration napropamide concentrations were determined on both sides of the membrane and complex and quantified based on the amount of napropamide unable to cross the membrane. it was found that for the SS soil 7% and for the NoSS 2.4% of the applied napropamide underwent facilitated transport. In addition, most of the complex transported through the columns had a molecular weight between 500 and 1000 Daltons (Da). The solutions from the SS soil were also found to have formed at least two distinct complexes that were resolved after passing through the untreated soil layer. The results obtained were in agreement with other published results and the techniques used offer a way to separate and concentrate DOM complexes from column effluents for further characterization.


Deduction of intracellular sub-systems from a topological description of the network.
Non-linear behaviour of biochemical networks, such as intracellular gene, protein or metabolic networks, is commonly represented using graphs of the underlying topology. Nodes represent abundance of molecules and edges interactions between pairs of molecules. These graphs are linear and thus based on an implicit linearization of the kinetic reactions in one or several dynamic modes of the total system. It is common to use data from different sources -- experiments conducted under different conditions or even on different species -- meaning that the graph will be a superposition of linearizations made in many different modes. The mixing of different modes makes it hard to identify functional modules, that is sub-systems that carry out a specific biological function, since the graph will contain many interactions that do not naturally occur at the same time. The ability to establish a boundary between the sub-system and its environment is critical in the definition of a module, contrary to a motif in which only internal interactions count. Identification of functional modules should therefore be done on graphs depicting the mode in which their function is carried out, i.e. graphs that only contain edges representing interactions active in the specific mode. In general, when an interaction between two molecules is established, one should always state the mode of the system in which it is active.


Human strongyloidiasis: complexities and pathways forward.
SUMMARYStrongyloidiasis is a World Health Organization neglected tropical disease usually caused by Strongyloides stercoralis, a parasitic worm with a complex life cycle. Globally, 300-600 million people are infected through contact with fecally contaminated soil. An autoinfective component of the life cycle can lead to chronic infection that may be asymptomatic or cause long-term symptoms, including malnourishment in children. Low larval output can limit the sensitivity of detection in stool, with serology being effective but less sensitive in immunocompromise. Host immunosuppression can trigger catastrophic, fatal hyperinfection/dissemination, where large numbers of larvae pierce the bowel wall and disseminate throughout the organs. Stable disease is effectively treated by single-dose ivermectin, with disease in immunocompromised patients treated with multiple doses. Strategies for management include raising awareness, clarifying zoonotic potential, the development and use of effective diagnostic tests for epidemiological studies and individual diagnosis, and the implementation of treatment programs with research into therapeutic alternatives and medication safety.


A non-redundant function of cyclin E1 in hematopoietic stem cells.
A precise balance between quiescence and proliferation is crucial for the lifelong function of hematopoietic stem cells (HSCs). Cyclins E1 and E2 regulate exit from quiescence in fibroblasts, but their role in HSCs remains unknown. Here, we report a non-redundant role for cyclin E1 in mouse HSCs. A long-term culture-initiating cell (LTC-IC) assay indicated that the loss of cyclin E1, but not E2, compromised the colony-forming activity of primitive hematopoietic progenitors. Ccne1(-/-) mice showed normal hematopoiesis in vivo under homeostatic conditions but a severe impairment following myeloablative stress induced by 5-fluorouracil (5-FU). Under these conditions, Ccne1(-/-) HSCs were less efficient in entering the cell cycle, resulting in decreased hematopoiesis and reduced survival of mutant mice upon weekly 5-FU treatment. The role of cyclin E1 in homeostatic conditions became apparent in aged mice, where HSC quiescence was increased in Ccne1(-/-) animals. On the other hand, loss of cyclin E1 provided HSCs with a competitive advantage in bone marrow serial transplantation assays, suggesting that a partial impairment of cell cycle entry may exert a protective role by preventing premature depletion of the HSC compartment. Our data support a role for cyclin E1 in controlling the exit from quiescence in HSCs. This activity, depending on the physiological context, can either jeopardize or protect the maintenance of hematopoiesis.


Dual respiratory virus infections.
We retrospectively reviewed eight prospective epidemiological studies conducted between 1991 and 1995 for dual respiratory virus infection (DRVI) to determine the frequency, associated comorbid conditions, clinical presentations, and morbidity related to DRVI among immunocompetent persons. Two viruses were identified as the cause of 67 (5.0%) of 1,341 acute respiratory virus infections. DRVI was detected in patients from < 1 year to 79 years of age, in both sexes, and in many races. Forty-two percent of patients with DRVI were < or = 4 years old. Fifty-eight percent of patients with DRVI had underlying chronic lung disease. DRVI was associated with upper respiratory tract illness; lower respiratory tract illness, including pneumonia; systemic influenza-like illnesses; and exacerbations of asthma or chronic obstructive pulmonary disease. All of the common acute respiratory viruses were identified; picornaviruses and influenzavirus A were the most common. The rate of DRVI (11.6%) was highest in the epidemiological studies in which cell culture, serology, and polymerase chain reaction were used together. Patients with DRVI were hospitalized significantly more often than those with respiratory infection due to a single virus (46.3% vs. 21.7%; P < .01). The percentage of DRVIs increased proportionally with the number of diagnostic methods used.


Validation of the Malignant Wound Assessment Tool - Research (MWAT-R) using cognitive interviewing.
Malignant wounds as a result of cancer are under-recognized for the physical and emotional distress they cause patients and their families. Unfortunately, there is a lack of valid and reliable screening and assessment tools to aid in the management of malignant wounds. This study aims to validate a patient-reported outcome measurement tool, Malignant Wound Assessment Tool - Research (MWAT-R). Eight patients were recruited and interviewed using the cognitive interviewing methodology to validate this tool. Patients' understanding and overall impression of the MWAT-R were explored. Our findings showed that the wording and response options posed challenges for patients in completing the tool. Overall, participants felt that questions captured the key issues related to dealing with a malignant wound and accounted for the patients' perspective. Establishing the content and face validity of the MWAT-R from the patients' perspectives using cognitive interviews has provided further evidence to the validity of this tool.


Quality of life assessment in patients with cystic fibrosis by means of the Cystic Fibrosis Questionnaire.
To assess the quality of life (QoL) of patients with cystic fibrosis (CF) followed at a university referral center for CF. A cross-sectional study involving application of the Cystic Fibrosis Questionnaire (CFQ) and Shwachman score in CF patients between April of 2008 and June of 2009. The sample consisted of 75 patients. The mean age was 12.5 ± 5.1 years (range, 6.1-26.4 years). The patients were divided into three groups by age in years: group I (< 12), II (12-14), and III (> 14). The highest and lowest CFQ scores were for the nutrition domain in group III (89.3 ± 16.2) and the social domain in group II (59.5 ± 22.3), respectively. Groups I and III differed significantly regarding the treatment domain (p = 0.001). Regarding Shwachman scores, there were significant differences between patients scoring < 70 and those scoring > 70 in the social (group I; p = 0.045), respiratory (group II; p = 0.053), and digestive (p = 0.042) domains. In group III, severity did not correlate with QoL. In groups I and II, patients with an FEV1 < 80% of predicted did not differ from other patients for any CFQ domain. However, in group III, values for the following domains were significantly lower in patients with an FEV1 < 80%: physical (p = 0.012); body image (p = 0.031); respiratory (p = 0.023), emotional (p = 0.041); and social role (p = 0.024). It is important to assess QoL in CF patients, because it can improve treatment compliance.


Cosavirus, Salivirus and Bufavirus in Diarrheal Tunisian Infants.
Three newly discovered viruses have been recently described in diarrheal patients: Cosavirus (CosV) and Salivirus (SalV), two picornaviruses, and Bufavirus (BuV), a parvovirus. The detection rate and the role of these viruses remain to be established in acute gastroenteritis (AGE) in diarrheal Tunisian infants. From October 2010 through March 2012, stool samples were collected from 203 children <5 years-old suffering from AGE and attending the Children's Hospital in Monastir, Tunisia. All samples were screened for CosV, SalV and BuV as well as for norovirus (NoV) and group A rotavirus (RVA) by molecular biology. Positive samples for the three screened viruses were also tested for astrovirus, sapovirus, adenovirus, and Aichi virus, then genotyped when technically feasible. During the study period, 11 (5.4%) samples were positive for one of the three investigated viruses: 2 (1.0%) CosV-A10, 7 (3.5%) SalV-A1 and 2 (1.0%) BuV-1, whereas 71 (35.0%) children were infected with NoV and 50 (24.6%) with RVA. No mixed infections involving the three viruses were found, but multiple infections with up to 4 classic enteric viruses were found in all cases. Although these viruses are suspected to be responsible for AGE in children, our data showed that this association was uncertain since all infected children also presented infections with several enteric viruses, suggesting here potential water-borne transmission. Therefore, further studies with large cohorts of healthy and diarrheal children will be needed to evaluate their clinical role in AGE.


Glycomyces fuscus sp. nov. and Glycomyces albus sp. nov., actinomycetes isolated from a hypersaline habitat.
Two actinomycete strains, designated TRM 49117(T) and TRM 49136(T), were isolated from a hypersaline habitat in Xinjiang Province, north-west China and were characterized taxonomically by using a polyphasic study. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain TRM 49117(T) had 93.93% similarity with the type strain Glycomyces halotolerans TRM 40137(T) (GenBank accession no. HQ651156) and TRM 49136(T) had 94.32% similarity with G. halotolerans TRM 40137(T). The 16S rRNA gene sequence similarity between the two new isolates was 93%. The isolates contained meso-diaminopimelic acid as the diagnostic diamino acid and anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0 as major cellular fatty acids. The predominant menaquinones of the isolates were MK-9(H4) and MK-9(H6). The whole-cell sugar patterns of these strains contained xylose and ribose, and strain TRM 49136(T) also contained arabinose. The polar lipid pattern of strain TRM 49117(T) comprised phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol mannosides, phosphatidylcholine, phosphatidylinositol and three additional unknown phospholipids. The polar lipid pattern of strain TRM 49136(T) comprised phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, glycolipids and two phosphoglycolipids of unknown composition. Genotypic and phenotypic data confirmed that strains TRM 49117(T) and TRM 49136(T) represent two novel species, clearly different from related species of the genus Glycomyces, for which the names Glycomyces fuscus sp. nov. (type strain TRM 49117(T) = CCTCC AA 2013003(T) = NRRL B-59998(T) = KACC 17682(T)) and Glycomyces albus sp. nov. (type strain TRM 49136(T) = CCTCC AA 2013004(T) = NRRL B-24927(T) = KACC 17681(T)) are proposed.


Trends of Status of Hypertension in Southern China, 2012-2019.
Little is known about the recent trends of hypertension in southern China. The aim of the study was to investigate the trends of hypertension in Guangdong Province between 2012 and 2019. We conducted two cross-sectional surveys in 2012 and 2019 in southern China, which included 10,970 and 27,483 participants, respectively, aged 35 to 75 years old using a method of stratified, multistage, and cluster sampling. Hypertension was defined as a mean systolic/diastolic blood pressure (SBP/DBP) ≥140/90mmHg, or a self-reported condition, or any pharmacological treatment in the last 2 weeks. In addition, according to the 2017 ACC/AHA guideline for high blood pressure, we estimated the prevalence and control rate of hypertension. According to the 2010 Chinese guideline, the age-standardized rate of hypertension prevalence was 34.7% in 2012 and 36.9% in 2019 with a slight increase, while the prevalence of prehypertension was stable (14.5% vs 14.3%). Over the period of our study, 45.6% and 60.7% of hypertensive patients knew their diagnosis in 2012 and 2019, and 40.8% and 51.5% were using antihypertensive medications, respectively. The control rates increased from 15.1% to 23.6%. Hypertension prevalence was 64.5% in 2012 and 63.2% in 2019, and the control rate increased from 3.0% to 4.8% during the study period under the 2017 ACC/AHA guideline. Although the past 7 years have seen some progress in hypertension management, the rates of hypertension awareness, treatment, and control in southern Chinese remained regrettably low, and the prevalence rate was still high.


COMP-Angiopoietin-1 accelerates muscle regeneration through N-cadherin activation.
Angiopoietin-1 modulates vascular stability via Tie2 on endothelial cells. In our previous study, we also showed it acts as an inhibitor of cardiomyocyte death. However, it remains poorly understood how Ang1 regulates myogenesis during muscle regeneration. Here we found that COMP-Ang1 (cAng1) enhances muscle regeneration through N-cadherin activation. Muscle fiber regeneration after limb muscle damage by ischemic injury was enhanced with cAng1 treatment. Mechanistically cAng1 directly bound to N-cadherin on the myoblast surface in a Ca2+ dependent manner. The interaction enhanced N-cadherin activation via N-cadherin/p120-catenin complex formation, which in turn activated p38MAPK (but not AKT or ERK) and myogenin expression (but not myoD) as well as increasing myogenin+ cells in/ex vivo. After transplantation of GFP-expressing myoblasts (GFP-MB), we showed an increased generation of GFP+ myotubes with adenovirus cAng1 (Adv-cAng1) injection. Adv-cAng1, however, could not stimulate myotube formation in N-cadherin-depleted GFP-MB. Taken together, this study uncovers the mechanism of how cAng1 promotes myoblast differentiation and muscle regeneration through the N-cadherin/p120-catenin/p38MAPK/myogenin axis.


Cutaneous responses to tests with influenza vaccines.
Responses to the intracutaneous infection of influenza vaccines were examined at 24 and 48 hours. Whole virus and subunit virus vaccines of comparable potency induced local reactions of comparable magnitude. Cutaneous reactions to the influenza vaccines were present in almost all adults but absent in young children. Reactions were greatly reduced by ingested prednisone. Responses to a subunit commercial trivalent influenza vaccine were compared with responses to tuberculin in tuberculin-sensitive subjects. In each subject the tuberculin reaction had greater duration than the influenza reaction. In separate tuberculin-sensitive individuals the flu vaccine and tuberculin reactions were biopsied at 24 hours. There was mild to moderate cellularity in the responses to both reactants. The cellular infiltration was mainly mononuclear cells except in one case in whom both agents elicited an infiltrate in which neutrophils and eosinophils predominated. The microscopic findings in reactions to tuberculin and influenza vaccine were comparable. Immunofluorescence studies revealed the deposition of complement (C3) in most reactions and of immunoglobulin (particularly IgM) in some reactions of both types. The findings support the view that local reactions to influenza vaccines are acquired and probably immunologic; the difference in duration from tuberculin reactions points to a different pathogenesis which is not yet entirely characterized.


Synthesis of arylpyrazole linked benzimidazole conjugates as potential microtubule disruptors.
In an attempt to develop potent and selective anticancer agents, a series of twenty arylpyrazole linked benzimidazole conjugates (10a-t) were designed and synthesized as microtubule destabilizing agents. The joining of arylpyrazole to the benzimidazole moiety resulted in a four ring (A, B, C and D) molecular scaffold that comprises of polar heterocyclic rings in the middle associated with rotatable single bonds and substituted aryl rings placed in the opposite directions. These conjugates were evaluated for their ability to inhibit the growth of sixty cancer cell line panel of the NCI. Among these some conjugates like 10a, 10b, 10d, 10e, 10p and 10r exhibited significant growth inhibitory activity against most of the cell lines ranging from 0.3 to 13μM. Interestingly, the conjugate 10b with methoxy group on D-ring expressed appreciable cytotoxic potential. A549 cells treated with some of the potent conjugates like 10a, 10b and 10d arrested cells at G2/M phase apart from activating cyclin-B1 protein levels and disrupting microtubule network. Moreover, these conjugates effectively inhibited tubulin polymerization with IC50 values of 1.3-3.8μM. Whereas, the caspase assay revealed that they activate the casepase-3 leading to apoptosis. Particularly 10b having methoxy substituent induced activity almost 3 folds higher than CA-4. Furthermore, a competitive colchicine binding assay and molecular modeling analysis suggests that these conjugates bind to the tubulin successfully at the colchicine binding site. These investigations reveal that such conjugates having pyrazole and benzimidazole moieties have the potential in the development of newer chemotherapeutic agents.


Therapeutic monitoring and variability of atazanavir in HIV-infected patients, with and without HCV coinfection, receiving boosted or unboosted regimens.
Adequate plasma trough concentrations (Ctrough) of protease inhibitors are required to maintain antiviral activity throughout the dosing interval. Therapeutic drug monitoring is used in clinical practice to optimize dosage and avoid toxic or subtherapeutic drug exposure. The pharmacokinetic variability of Atazanavir (ATV) can be relatively large, as a result of several factors. One of the affecting factors may be hepatic impairment due to hepatitis C virus (HCV) coinfection. We collected trough plasma samples from human immunodeficiency virus (HIV)-1-infected outpatients, with and without HCV coinfection and/or cirrhosis, receiving stable highly active antiretroviral therapy containing ATV. In the total population, we mainly compared the 2 regimens: 300ATV + 100RTV OD [ritonavir (RTV), once daily (OD)] versus 400ATV OD. We used a threshold value of 0.15 μg/mL, based on the proposed therapeutic range (0.15-0.85 μg/mL). Plasma concentrations of ATV were determined by a validated assay using high-performance liquid chromatography with ultraviolet detection. A total of 214 HIV-infected outpatients were included. For each regimen, we compared 3 groups of subjects: HIV+/HCV-, HIV+/HCV+, and HIV+/HCV+ with cirrhosis. In the whole study population, we observed a large variability and found suboptimal Ctrough levels (<0.15 μg/mL) in 23 subjects (2 belonging to the 300/100 OD group and 21 to the 400 OD group). For the standard dosage regimen of 300ATV + 100RTV OD, we did not find a statistical difference between HIV-infected patients without HCV coinfection versus HIV-infected patients with HCV coinfection: median 0.85 (interquartile range 0.53-1.34) and 0.95 (0.70-1.36) μg/mL, respectively. In HIV+/HCV+-infected patients with cirrhosis, we found a median Ctrough of 0.70 (0.43-1.0) μg/mL, with no statistical difference when compared with HIV+/HCV- infected patients. For the 400ATV OD (n=90) dosage regimen, the total median ATV Ctrough was 0.40 (0.23-1.0) μg/mL. In this group, we found a statistically significant difference between HIV+/HCV- and HIV+/HCV+-infected patients: median Ctrough was 0.23 (0.11-0.42) and 0.52 (0.20-1.0) μg/mL, respectively. In HIV+/HCV+ subjects with cirrhosis, the Ctrough median value was 0.42 (0.13-0.75) μg/mL, and there was a significant difference when compared with HIV patients without coinfection. Therapeutic drug monitoring of ATV in patients receiving unboosted regimen may be useful to identify those HIV-infected subjects, with or without HCV coinfection, who may benefit from adding low RTV doses, or the subset of patients in whom removal of RTV could be attempted without the risk of suboptimal plasma ATV exposure.


Noise in acute patient care areas.
This study was designed to describe the level of sound in acute patient care areas. A convenience sample of 25 subjects from four intensive care and two general care units within three hospitals in a large metropolitan area was studied. Continuous decibel levels [dB(A)] and equivalent continuous sound pressure levels [LEQ] were measured for 24 continuous hours. Two 3-hour observations were made to assess the sources of sound, and patients were interviewed to determine their perceptions of sound effects. Continuous high noise levels were found in the larger hospital's open heart recovery room and intensive care units [48.5-68.5 dB(A), 15-minute LEQ], lower levels in the smaller hospital's intensive care units [32.5-57 dB(A)] and varying levels in the general ward areas [34.25-62.5 dB(A)]. Decibel levels generated from equipment reached 90 dB(A) in some instances. Patients' perceptions ranged from content to highly perturbed. Although some sources of noise were not immediately changeable, other were adaptable, preventable, or reducible.


A rapid SPE-based analytical method for UPLC/MS/MS determination of aminoglycoside antibiotic residues in bovine milk, muscle, and kidney.
An SPE-based cleanup protocol was developed for ultra-performance LC (UPLC)/MS/MS determination of residues of the common aminoglycoside antibiotics streptomycin, dihydrostreptomycin, neomycin, and gentamicin in bovine milk, kidney, and muscle. Recoveries for all compounds except neomycin ranged from 80 to 104% for all matrixes studied; recoveries for neomycin ranged from 71 to 84%. Intraday and interday precision data were under 15% for all sample matrixes. Compared with other recently reported cleanup methods, less sample is required, the use of potentially dangerous reagents is minimized, and fewer manipulations are required by the analyst. A high throughput 96-well plate format was used for SPE cleanup and UPLC/MS analysis.


Inequalities in health: evidence from Denmark of the interaction of circumstances and health-related behaviour.
It is well known that the experience of poor health depends on adverse social and material circumstances and on unhealthy behaviour. In 1990 a British study asserted that privileged people gain more health benefit from a healthy lifestyle than do deprived people. In the present study this assertion was taken as a hypothesis, assuming statistical interaction between circumstances and health behaviour. The combined effect of these variables was studied in Denmark. Data was obtained from a 1987 national Health and Morbidity Survey; variables were selected to correspond closely with those in the earlier British study. The analysis included a multivariate analysis of variance. The results show that health-related behaviour has a positive effect on health both for those who are deprived and those who are not deprived, which is at variance with the findings of the British study. It confirms earlier analyses of Danish and Dutch data.


Freezing African elephant semen as a new population management tool.
The captive elephant population is not self-sustaining and with a limited number of breeding bulls, its genetic diversity is in decline. One way to overcome this is to import young and healthy animals from the wild. We introduce here a more sustainable alternative method - importation of semen from wild bulls without removing them from their natural habitat. Due to the logistics involved, the only practical option would be to transport cryopreserved sperm. Despite some early reports on African elephant semen cryopreservation, the utility of this new population management tool has not been evaluated. Semen was collected by electroejaculation from 14 wild African savanna elephant (Loxodonta africana) bulls and cryopreserved using the directional freezing technique. Sperm treatments evaluated included the need for centrifugation, the use of hen or quail yolk, the concentration of glycerol (3%, 5% or 7%) in the extender, and maintenance of motility over time after thawing. Our results suggest that dilution in an extender containing hen yolk and 7% glycerol after centrifugation best preserved post-thaw sperm motility when compared to all other treatments (P≤0.012 for all). Using this approach we were able to achieve after thawing (mean ± SD) 54.6±3.9% motility, 85.3±2.4% acrosome integrity, and 86.8±4.6% normal morphology with no decrease in motility over 1 h incubation at 37°C. Sperm cryopreserved during this study has already lead to a pregnancy of a captive female elephant following artificial insemination. With working techniques for artificial insemination and sperm cryopreservation of both African and Asian elephants in hand, population managers can now enrich captive or isolated wild elephant populations without removing valuable individuals from their natural habitat.


EBprot: Statistical analysis of labeling-based quantitative proteomics data.
Labeling-based proteomics is a powerful method for detection of differentially expressed proteins (DEPs). The current data analysis platform typically relies on protein-level ratios, which is obtained by summarizing peptide-level ratios for each protein. In shotgun proteomics, however, some proteins are quantified with more peptides than others, and this reproducibility information is not incorporated into the differential expression (DE) analysis. Here, we propose a novel probabilistic framework EBprot that directly models the peptide-protein hierarchy and rewards the proteins with reproducible evidence of DE over multiple peptides. To evaluate its performance with known DE states, we conducted a simulation study to show that the peptide-level analysis of EBprot provides better receiver-operating characteristic and more accurate estimation of the false discovery rates than the methods based on protein-level ratios. We also demonstrate superior classification performance of peptide-level EBprot analysis in a spike-in dataset. To illustrate the wide applicability of EBprot in different experimental designs, we applied EBprot to a dataset for lung cancer subtype analysis with biological replicates and another dataset for time course phosphoproteome analysis of EGF-stimulated HeLa cells with multiplexed labeling. Through these examples, we show that the peptide-level analysis of EBprot is a robust alternative to the existing statistical methods for the DE analysis of labeling-based quantitative datasets. The software suite is freely available on the Sourceforge website http://ebprot.sourceforge.net/. All MS data have been deposited in the ProteomeXchange with identifier PXD001426 (http://proteomecentral.proteomexchange.org/dataset/PXD001426/).


Amygdala integrates emotional expression and gaze direction in response to dynamic facial expressions.
Neuroimaging studies suggest that the amygdala integrates emotional expression and gaze direction, but the findings are inconsistent. We hypothesized that the dynamic facial expressions, which are more salient stimuli than static facial expressions are, would reveal the integration of emotional expression and gaze direction in amygdala activity. To test this hypothesis, we presented dynamic and static facial expressions of anger and happiness looking toward and away from the subject, and we visualized brain activity during these presentations using functional magnetic resonance imaging (fMRI). After the image acquisition phase of the study, the subject's experienced emotion in response to each stimulus was investigated. The left amygdala showed an interaction between presentation condition and gaze direction, indicating higher activity in response to dynamic, but not static, expressions looking toward the subjects than to expressions looking away from them. This was true for both angry and happy expressions. This pattern corresponded with the intensity of emotional arousal. These results suggest that the amygdala integrates emotional expression and gaze direction in dynamic facial expressions and modulates emotional arousal.


Quality of life: the continent ileostomy.
Ten patients are presented in whom a continent ileostomy (internal ileal reservoir or Kock Pouch) was constructed. The series is unique in that it includes the first such ileostomy successfully constructed in this country. The other 9 patients, for a variety of reasons, sought conversion from a standard ileostomy with an external appliance, to a continent ileostomy. Eight of 9 were successfully converted. Subsequent loss of continence has occurred in 2 of the patients due to reduction or intususception of the nipple valve. In one of the two, a satisfactory level of continence appears related to adequate pouch size. The second patient is seriously inconvenienced by a varying level of continence in part related to slow development of pouch size. Seven of the 10 attest to a dramatic improvement in life style, and illustrate the high level of patient satisfaction with a continent ileostomy.


Current treatment practice and outcomes. Report of the hyponatremia registry.
Current management practices for hyponatremia (HN) are incompletely understood. The HN Registry has recorded diagnostic measures, utilization, efficacy, and outcomes of therapy for eu- or hypervolemic HN. To better understand current practices, we analyzed data from 3087 adjudicated adult patients in the registry with serum sodium concentration of 130 mEq/l or less from 225 sites in the United States and European Union. Common initial monotherapy treatments were fluid restriction (35%), administration of isotonic (15%) or hypertonic saline (2%), and tolvaptan (5%); 17% received no active agent. Median (interquartile range) mEq/l serum sodium increases during the first day were as follows: no treatment, 1.0 (0.0-4.0); fluid restriction, 2.0 (0.0-4.0); isotonic saline, 3.0 (0.0-5.0); hypertonic saline, 5.0 (1.0-9.0); and tolvaptan, 4.0 (2.0-9.0). Adjusting for initial serum sodium concentration with logistic regression, the relative likelihoods for correction by 5 mEq/l or more (referent, fluid restriction) were 1.60 for hypertonic saline and 2.55 for tolvaptan. At discharge, serum sodium concentration was under 135 mEq/l in 78% of patients and 130 mEq/l or less in 49%. Overly rapid correction occurred in 7.9%. Thus, initial HN treatment often uses maneuvers of limited efficacy. Despite an association with poor outcomes and availability of effective therapy, most patients with HN are discharged from hospital still hyponatremic. Studies to assess short- and long-term benefits of correction of HN with effective therapies are needed.


Investigating the Clinical Effect of Kinesio Tape on Muscle Performance in Healthy Young Soccer Players - A Prospective Cohort Study.
Kinesio tape (KT) is a visible adhesive restorative tape that has typically been utilized for injury prevention, recovery, and even performance improvement, but limited studies have assessed the effect of KT on muscle performance. The purpose of this study was to investigate the clinical impact of KT on muscle performance in healthy young soccer players. Between 25 March and 21 April 2017, sixteen healthy soccer players with a mean age of 20±2.17 were enrolled in this prospective cohort study. All participants were selected from the college football team of Prince Sattam Bin Abdulaziz University. The muscle performance of the players was evaluated with an isokinetic dynamometer for the following three conditions: without tape, immediately after applying KT, and 8 hrs post-KT application while the tape remained on the same site. The differences in peak torque and total work among the three conditions were nonsignificant (p>0.05). Additionally, applying KT to the thigh muscles did not decrease or increase the performance of non injured healthy soccer players (p>0.05). KT does not lead to beneficial outcomes of muscle performance in healthy young soccer players.


Diffusion tensor brain imaging findings at term-equivalent age may predict neurologic abnormalities in low birth weight preterm infants.
Low birth weight preterm infants are at high risk of brain injury, particularly injury to the white matter. Diffusion tensor imaging is thought to be more sensitive than conventional MR imaging for detecting subtle white matter abnormalities. The objective of this study was to examine whether diffusion tensor imaging could detect abnormalities that may be associated with later neurologic abnormalities in infants with otherwise normal or minimally abnormal conventional MR imaging findings. We prospectively studied 137 low birth weight (<1800 g) preterm infants. Neonatal conventional MR imaging and diffusion tensor imaging were performed near term-equivalent age before discharge, and neurologic development of the infants was later followed up at 18 to 24 months of age. Among the preterm infants who were fully studied, 63 underwent normal conventional MR imaging. Three of these infants developed cerebral palsy, and 10 others showed abnormal neurologic outcome. Diffusion tensor imaging results for these infants showed a significant reduction of fractional anisotropy in the posterior limb of the internal capsule in neurologically abnormal infants (including those with cerebral palsy) compared with control preterm infants with normal neurologic outcomes. These results suggest that neonatal diffusion tensor imaging may allow earlier detection of specific anatomic findings of microstructural abnormalities in infants at risk for neurologic abnormalities and disability. The combination of conventional MR imaging and diffusion tensor imaging may increase the predictive value of neonatal MR imaging for later neurologic outcome abnormalities and may become the basis for future interventional clinical studies to improve outcomes.


Identification of testis-specific (Limk2t) and brain-specific (Limk2c) isoforms of mouse LIM-kinase 2 gene transcripts.
LIM-kinase 1 (LIMK1) and LIM-kinase 2 (LIMK2) are members of a novel serine/threonine kinase subfamily with structural features composed of N-terminal two LIM domains, an internal PDZ-like domain, and a C-terminal protein kinase domain. We recently identified and characterized the mouse Limk2 gene and two Limk2 transcripts (Limk2a and Limk2b) coding for proteins with distinct N-terminal LIM structures. Here we describe two additional transcripts of the mouse Limk2 gene. One is a 1.7-kb transcript, termed Limk2t, which is specifically expressed in the testis and codes for an N-terminally truncated form of LIMK2 consisting of only a part of a PDZ-like domain and a protein kinase domain. The other is a transcript, termed Limk2c, which is specifically expressed in the brain and codes for a protein with a 6-amino-acid insert within the protein kinase domain. Exons specific to the 5'-terminal extra sequence of Limk2t and the insert sequence of Limk2c locate between exons 5-6 and exons 8-9 in the mouse Limk2 gene, respectively. Testis- and brain-specific expression of Limk2t and Limk2c suggests specific roles in these tissues.


Assessment of dermal exposure and skin condition of workers exposed to nickel at a South African base metal refinery.
The objectives of this study were to assess dermal exposure of cell workers to nickel at a South African base metal refinery and to characterize their skin condition by measuring the skin hydration and trans epidermal water loss (TEWL) indices. The skin hydration index of the index finger, palm, neck, and forehead was measured before, during and at the end of the shift. The TEWL index was measured before and at the end of the shift. Dermal exposure samples were collected with Ghostwipes from the index finger and palm of the dominant hand, before, during, and at the end of the shift. Neck and forehead samples were collected before and at the end of the shift. Wipe samples of various surfaces in the workplace were also collected. Wipes were analyzed for nickel according to NIOSH method 9102, using inductively coupled plasma-atomic emission spectrometry. Hydration indices measured on the hands decreased significantly during the shift, but recovered to normal levels by the end of the shift. TEWL indices for the index finger and palm of the hands are indicative of a low barrier function even before commencement of the shift, which further deteriorated significantly during the shift. During the shift, substantial nickel skin loading occurred on the index finger and palm of the hand. Levels on the neck and forehead were much lower. Various workplace surfaces, which workers come into contact with, were also contaminated with nickel. The skin condition and high levels of nickel on the skin were most probably caused by inadequate chemical protection provided by protective gloves. Although, the permeability of nickel through intact skin is considered to be low, a decreased barrier function of dehydrated or slightly damaged skin will increase its permeability for nickel. The ethnicity of these exposed workers may contribute significantly toward the low incidence of allergic contact dermatitis observed. Several measures to lower dermal exposure to nickel are also recommended.


Supplementation of complex milk lipid concentrate (CMLc) improved the memory of aged rats.
The socio-economic impact from age-related mental decline is escalating. Supplementation of functional foods for sustaining mental health is desirable. We examined the effect of long-term supplementation of complex milk lipid concentrate (CMLc), mixed dairy phospholipids, on memory and associated vascular and neuronal changes in aged rats. Fisher/Norway Brown rats were used. Two groups of aged rats (24 months) were fed with either gelatin-formulated CMLc or blank gelatin as the control, for 4 months. To determine age-related changes, a young group (5 months) was also fed with blank gelatin. Morris water maze tests were carried out after the supplementation and brain tissues were collected for biological analysis. The aged control rats learnt to locate the platform slower than the young control rats during acquisition trials (*P < 0.05), and made fewer entries to and more initial heading errors from the platform zone during testing trials (*P < 0.05). The CMLc supplementation improved memory by showing the reduced initial heading errors in a delayed probe trial ((#)P < 0.05). We also found that the aged rats with CMLc supplementation improved vascular density, dopamine output, and neuroplasticity ((#)P < 0.05) in the brain regions involved in memory compared with that of the aged control rats. The data suggested that the supplementation of CMLc during the early stage of brain aging may prevent memory decline possibly through improving vascular and neuronal function.


In vivo and ex vivo MR imaging of slowly cycling melanoma cells.
Slowly cycling cells are believed to play a critical role in tumor progression and metastatic dissemination. The goal of this study was to develop a method for in vivo detection of slowly cycling cells. To distinguish these cells from more rapidly proliferating cells that constitute the vast majority of cells in tumors, we used the well-known effect of label dilution due to division of cells with normal cycle and retention of contrast agent in slowly dividing cells. To detect slowly cycling cells, melanoma cells were labeled with iron oxide particles. After labeling, we observed dilution of contrast agent in parallel with cell proliferation in the vast majority of normally cycling cells. A small and distinct subpopulation of iron-retaining cells was detected by flow cytometry after 20 days of in vitro proliferation. These iron-retaining cells exhibited high expression of a biological marker of slowly cycling cells, JARID1B. After implantation of labeled cells as xenografts into immunocompromised mice, iron-retaining cells were detected in vivo and ex vivo by magnetic resonance imaging that was confirmed by Prussian Blue staining. Magnetic resonance imaging detects not only iron retaining melanoma cells but also iron positive macrophages. Proposed method opens up opportunities to image subpopulation of melanoma cells, which is critical for continuous tumor growth.


Mifepristone and misoprostol compared with misoprostol alone for second-trimester abortion: a randomized controlled trial.
To estimate the clinical benefit of pretreatment with mifepristone followed by misoprostol compared with misoprostol alone for second-trimester abortion. Two hundred sixty women with live fetuses of gestational ages 14-21 weeks were enrolled in a randomized, placebo-controlled, double-blind trial in Vietnam. Eligible consenting women received either mifepristone or placebo to take on their own at home and were scheduled to return to the hospital the next day. At the hospital, women were given 400 micrograms buccal misoprostol every 3 hours, up to five doses, until both the fetus and placenta were expelled. Efficacy was evaluated 15 hours after misoprostol dosing began and the procedure was considered complete if uterine evacuation was achieved without recourse to surgical evacuation. Pretreatment with mifepristone resulted in more than twice the chance of complete uterine evacuation in 15 hours (79.8% compared with 36.9%, relative risk 2.16, 95% confidence interval 1.70-2.75). The mean induction-to-abortion interval for complete uterine evacuations was statistically significantly shorter among participants pretreated with mifepristone compared with those given misoprostol alone (8.1, standard deviation 2.8, range 2.5-14.8; and 10.6, standard deviation 2.5, range 6.5-15.5 hours, respectively; P<.001). The side-effect profiles for the two regimens did not differ significantly and acceptability of the treatments was high. Mifepristone-misoprostol is more efficacious and faster than misoprostol alone. Services offering home administration of mifepristone as pretreatment could optimize efficacy and acceptability of medical abortions for women with gestations 14-21 weeks since the last menstrual period. ClinicalTrials.gov, www.clinicaltrials.gov, NCT00784186. I.


Super-selective transarterial chemoembolization of hepatocellular carcinoma with doxorubicin-eluting beads sized 40-75 microns: assessment of efficacy and safety.
We aimed to evaluate the effectiveness and safety of super-selective transarterial chemoembolization (TACE) with doxorubicin-loaded drug-eluting beads (DEB) sized 40-75 μm for hepatocellular carcinoma (HCC) in early and intermediate stages according to Barcelona Clinic Liver Cancer (BCLC) staging system. This single-center retrospective study was conducted with 45 consecutive HCC patients treated by 72 sessions of DEB-TACE during the 2012-2017 period. Thirty-seven patients (82.2%) had single tumor staged BCLC A and B (53.3% and 46.7%, respectively). All procedures were performed by super-selective approach using 1.7-2.0 F microcatheters. Cone beam CT was performed to detect all tumor-feeding arteries and assess the treatment results immediately. Dynamic MRI and laboratory tests were obtained at 1-month follow-up and every 3 months thereafter. Response to treatment according to modified Response Evaluation Criteria in Solid Tumors, demographic and clinical status, laboratory tests, time-to-event durations and rates, complications according to the National Cancer Institute Criteria for Adverse Events were evaluated. A total of 45 patients (median age, 65.6 years; range, 35-88 years; 33 men, 73.4%) were included. Eight patients (17.7%) underwent liver transplantation after DEB-TACE, and 20 (44.4%) died during the follow-up period. Median follow-up was 22 months (range, 13-31), and 42 (93.3%) patients were followed up for more than 1 year. Overall complete response, partial response, and progressive disease rates were 53.3%, 33.3%, and 13.4% at one year and 22.2%, 26.7%, and 13.3% at three years, respectively. For target lesions, these rates were 60.0%, 26.7%, and 13.3% at one year and 28.9%, 6.7%, and 4.4% at three years, respectively. Median overall survival (OS) duration was 24 months (95% CI, 20.9-31.9 months). At one year and three years, OS rates were 71.0% and 44.4%, respectively. The only statistically significant relationship with OS was presence of chronic liver disease, which worsened the OS rate (P = 0.031). Time-to-progression (TTP) was 23 months (95% CI, 15.1-40.0), and progression-free survival (PFS) was 28 months (95% CI, 6.2-39.8). Post-embolization syndrome occurred in 10 patients (22.2%). Transient grade I/II bilirubin and aminotransferase elevation was observed in 26 (57.7%) and 18 (40%) patients, respectively. Super-selective DEB-TACE with doxorubicin-loaded beads sized 40-75 μm is an effective and safe treatment method with prolonged TTP and PFS in early and intermediate stages of HCC. Presence of chronic liver disease is the only significant factor that worsened OS ratios after DEB-TACE.


Perioperative cost-finding analysis of the routine use of intraoperative forced-air warming during general anesthesia.
Despite the well-documented ability of forced-air warming (FAW) to maintain normothermia, it is unclear whether this technique results in a net increase or decrease in costs. The authors did a prospective cost-finding study comparing FAW with routine thermal care in patients at low risk for perioperative complications who were undergoing general anesthesia. After institutional review board approval was received, 100 patients were studied who were having elective surgery scheduled for more than 2 h during general endotracheal anesthesia. Patients were randomly assigned to one of two groups: FAW or routine thermal care. All patients received a standardized anesthetic. Anesthesia providers were blinded to core temperatures and the use of FAW. Primary outcomes were those associated with perioperative costs. The time from completion of surgical dressing until tracheal extubation was significantly reduced in the FAW group (10 +/- 1 min compared with 14 +/- 1 min; mean +/- SEM; P < 0.01). There was no demonstrable difference in attainment of postanesthesia care unit discharge criteria between the two groups, although the FAW group used one less cotton blanket there. The net savings related to the use of the FAW depends on the percentage of the intraoperative costs that are fixed rather than variable ($15 additional for FAW if all costs are fixed compared with $29 savings if all costs were variable). Routine intraoperative FAW significantly reduced time until extubation and use of cotton blankets in the postanesthesia care unit. These results suggest that the influence of FAW on net total perioperative costs depends on patient and surgical characteristics and institutional factors related to cost accounting.


Reduced astrocyte density underlying brain volume reduction in activity-based anorexia rats
Severe grey and white matter volume reductions were found in patients with anorexia nervosa (AN) that were linked to neuropsychological deficits while their underlying pathophysiology remains unclear. For the first time, we analysed the cellular basis of brain volume changes in an animal model (activity-based anorexia, ABA). Female rats had 24 h/day running wheel access and received reduced food intake until a 25% weight reduction was reached and maintained for 2 weeks. In ABA rats, the volumes of the cerebral cortex and corpus callosum were significantly reduced compared to controls by 6% and 9%, respectively. The number of GFAP-positive astrocytes in these regions decreased by 39% and 23%, total astrocyte-covered area by 83% and 63%. In neurons no changes were observed. The findings were complemented by a 60% and 49% reduction in astrocyte (GFAP) mRNA expression. Volumetric brain changes in ABA animals mirror those in human AN patients. These alterations are associated with a reduction of GFAP-positive astrocytes as well as GFAP expression. Reduced astrocyte functioning could help explain neuronal dysfunctions leading to symptoms of rigidity and impaired learning. Astrocyte loss could constitute a new research target for understanding and treating semi-starvation and AN.


Predictors of neuropsychological recovery in treatment for anorexia nervosa.
Previous research indicates that individuals with Anorexia Nervosa (AN) often experience some degree of neuropsychological dysfunction. Although most aspects of cognition improve with treatment, factors that predict neuropsychological improvement remain elusive. The present study investigated whether cognitive reserve, the estimated level of premorbid cognitive functioning, and AN subtype predicted neuropsychological improvement during inpatient treatment for AN. Neuropsychological functioning was assessed pre- and post-hospitalization in 28 women with AN (18 with restricting type and 10 with binge-eating/purging type), and cognitive reserve was estimated at admission using a word reading test. Level of cognitive reserve and AN subtype were both significant predictors of neuropsychological improvement in this sample. Cognitive reserve was significantly associated with improvements in verbal memory, semantic fluency, basic auditory attention and visuospatial construction. Participants with AN binge-eating/purging type demonstrated significantly greater neuropsychological improvement during treatment than did participants with AN restricting type. Information about cognitive reserve and AN subtype may provide clinicians with valuable prognostic information and help guide treatment.


Prevalence of Collateral Typology in Coronary Chronic Total Occlusion and Its Impact on Percutaneous Intervention Performance.
The presence of collateral channels providing distal blood supply is a distinctive characteristic of chronic total occlusion (CTO) lesions. However, data about the distinct baseline and procedural characteristics of each collateral subset are scarce. Accordingly, we sought to explore the procedural aspects specific for each collateral typology (ipsilateral collaterals [ICs], contralateral collaterals [CCs] or mixed) in CTO-percutaneous coronary intervention (PCI). A retrospective analysis of our CTO-PCI registry was performed to investigate the prevalence, procedural characteristics, and outcomes specific for each CTO-PCI subset, defined according to the inter-arterial connection anatomy. A total of 209 cases were included. Of the included cases, 45 (22%) and 92 (44%) patients displayed solely IC or CC, respectively, whereas in 72 (34%) both IC and CC were present (mixed). The procedural success rate was high (91.1%) and comparable among the different groups, despite greater lesion complexity in the CC group. The most frequent target vessel was the left circumflex in the IC group (51% of cases) and the right coronary artery in the CC (63%) and mixed (57%) groups. Among the IC cases, 42% showed a poor collateral connection function (2% and 10% for the CC and mixed group, respectively), and 46% showed a suboptimal collateral recipient artery filling (21% and 20% for the CC and mixed group, respectively). Most of the IC cases were performed using a single access (96%). In conclusion, the success and complication rates were comparable among the collateral typology groups, irrespective of the differences in the baseline and procedural characteristics. Phenotyping CTO as hereby proposed might be helpful for targeted procedural considerations.


Isolation of UV-B resistant bacteria from two high altitude Andean lakes (4,400 m) with saline and non saline conditions.
Laguna (L.) Negra and L. Verde are high altitude Andean lakes located at the 4,400 m altitude in the Andean desert (Puna) in the Argentine northwest. Both lakes are exposed to extreme weather conditions but differ in salinity contents (salinity 6.7% for L. Negra and 0.27% for L. Verde). The aim of this work was to isolate ultraviolet B fraction (UV-B) resistant bacteria under UV-stress in order to determine, a possible connection, between resistance to UV-B and tolerance to salinity. DNA damage was determined by measuring CPDs accumulation. Connection among pigmentation production and UV resistance was also studied. Water samples were exposed to artificial UV-B radiation for 24 h. Water aliquots were plated along the exposition on different media, with different salinity and carbon source content (Lake medium (LM) done with the lake water plus agar and LB). CFU were counted and DNA damage accumulation was determined. Isolated bacteria were identified by 16S rDNA sequence. Their salinity tolerance, were measured at 1, 5 and 10% NaCl and their pigment production in both media was determined. In general it was found that UV resistance and pigment production were the optimum in Lake Medium done with lake water which maintained similar salinity. The most resistant bacteria in L. Negra were different strains of Exiguobacterium sp. and, in L. Verde, Staphylococcus sp. and Stenotrophomonas maltophilia. These bacteria showed the production and increase of UV-Vis absorbing compounds under UV stress and in LM. Bacterial communities from both lakes were well adapted to high UV-B exposure under the experimental conditions, and in many cases UV-B even stimulated growth. The idea that resistance to UV-B could be related to adaptation to high salinity is still an open question that has to be answered with future experiments.


Inflammation associated ethanolamine facilitates infection by Crohn's disease-linked adherent-invasive Escherichia coli.
The predominance of specific bacteria such as adherent-invasive Escherichia coli (AIEC) within the Crohn's disease (CD) intestine remains poorly understood with little evidence uncovered to support a selective pressure underlying their presence. Intestinal ethanolamine is however readily accessible during periods of intestinal inflammation, and enables pathogens to outcompete the host microbiota under such circumstances. Quantitative RT-PCR (qRT-PCR) to determine expression of genes central to ethanolamine metabolism; transmission electron microscopy to detect presence of bacterial microcompartments (MCPs); in vitro infections of both murine and human macrophage cell lines examining intracellular replication of the AIEC-type strain LF82 and clinical E. coli isolates in the presence of ethanolamine; determination of E. coli ethanolamine utilization (eut) operon transcription in faecal samples from healthy patients, patients with active CD and the same patients in remission following treatment. Growth on the intestinal short chain fatty acid propionic acid (PA) stimulates significantly increased transcription of the eut operon (fold change relative to glucose: >16.9; p-value <.01). Additionally ethanolamine was accessible to intra-macrophage AIEC and stimulated significant increases in growth intracellularly when it was added extracellularly at concentrations comparable to those in the human intestine. Finally, qRT-PCR indicated that expression of the E. coli eut operon was increased in children with active CD compared to healthy controls (fold change increase: >4.72; P < .02). After clinical remission post-exclusive enteral nutrition treatment, the same CD patients exhibited significantly reduced eut expression (Pre vs Post fold change decrease: >15.64; P < .01). Our data indicates a role for ethanolamine metabolism in selecting for AIEC that are consistently overrepresented in the CD intestine. The increased E. coli metabolism of ethanolamine seen in the intestine during active CD, and its decrease during remission, indicates ethanolamine use may be a key factor in shaping the intestinal microbiome in CD patients, particularly during times of inflammation. FUND: This work was funded by Biotechnology and Biological Sciences Research Council (BBSRC) grants BB/K008005/1 & BB/P003281/1 to DMW; by a Tenovus Scotland grant to MJO; by Glasgow Children's Hospital Charity, Nestle Health Sciences, Engineering and Physical Sciences Research Council (EPSRC) and Catherine McEwan Foundation grants awarded to KG; and by a Natural Environment Research Council (NERC) fellowship (NE/L011956/1) to UZI. The IBD team at the Royal Hospital for Children, Glasgow are supported by the Catherine McEwan Foundation and Yorkhill IBD fund. RKR and RH are supported by NHS Research Scotland Senior fellowship awards.


Forecasting incidence of dengue in Rajasthan, using time series analyses.
To develop a prediction model for dengue fever/dengue haemorrhagic fever (DF/DHF) using time series data over the past decade in Rajasthan and to forecast monthly DF/DHF incidence for 2011. Seasonal autoregressive integrated moving average (SARIMA) model was used for statistical modeling. During January 2001 to December 2010, the reported DF/DHF cases showed a cyclical pattern with seasonal variation. SARIMA (0,0,1) (0,1,1) 12 model had the lowest normalized Bayesian information criteria (BIC) of 9.426 and mean absolute percentage error (MAPE) of 263.361 and appeared to be the best model. The proportion of variance explained by the model was 54.3%. Adequacy of the model was established through Ljung-Box test (Q statistic 4.910 and P-value 0.996), which showed no significant correlation between residuals at different lag times. The forecast for the year 2011 showed a seasonal peak in the month of October with an estimated 546 cases. Application of SARIMA model may be useful for forecast of cases and impending outbreaks of DF/DHF and other infectious diseases, which exhibit seasonal pattern.


Additively Manufactured Device for Dynamic Culture of Large Arrays of 3D Tissue Engineered Constructs.
The ability to test large arrays of cell and biomaterial combinations in 3D environments is still rather limited in the context of tissue engineering and regenerative medicine. This limitation can be generally addressed by employing highly automated and reproducible methodologies. This study reports on the development of a highly versatile and upscalable method based on additive manufacturing for the fabrication of arrays of scaffolds, which are enclosed into individualized perfusion chambers. Devices containing eight scaffolds and their corresponding bioreactor chambers are simultaneously fabricated utilizing a dual extrusion additive manufacturing system. To demonstrate the versatility of the concept, the scaffolds, while enclosed into the device, are subsequently surface-coated with a biomimetic calcium phosphate layer by perfusion with simulated body fluid solution. 96 scaffolds are simultaneously seeded and cultured with human osteoblasts under highly controlled bidirectional perfusion dynamic conditions over 4 weeks. Both coated and noncoated resulting scaffolds show homogeneous cell distribution and high cell viability throughout the 4 weeks culture period and CaP-coated scaffolds result in a significantly increased cell number. The methodology developed in this work exemplifies the applicability of additive manufacturing as a tool for further automation of studies in the field of tissue engineering and regenerative medicine.


Tolerance mechanisms in murine autoimmune diabetes induced by anti-ICAM-1/LFA-1 mAb and anti-CD8 mAb.
A short-term administration of antibodies against ICAM-1/LFA-1 or CD8 molecules during a critical period of younger age resulted in complete protection of autoimmune diabetes in NOD mice. In this study, we attempted to elucidate the tolerance mechanisms. Transfer of splenocytes from both antibody-treated NOD mice to NOD-SCID mice failed to develop diabetes. On the other hand, when splenocytes from diabetic mice were transferred to the antibody-treated mice, 40% of both mAb-treated recipients became diabetic. In vitro response of T cells from these protected mice exhibited strong proliferation against syngeneic islet cells or ConA. Furthermore, semiquantitative RT-PCR analysis of cytokines showed that T cells from anti-CD8-treated mice could express IFN-gamma, IL-4, IL-10 and TGF-beta1 in response to islet antigen. In contrast, T cells from anti-ICAM-1/LFA-1-treated mice expressed IFN-gamma, IL-10 and TGF-beta1 but not IL-4. These results suggest that tolerance mechanisms like clonal deletion, anergy, immunoregulatory T cells or Th1 to Th2/Th3 cytokine shifting are not responsible for the tolerance induction, indicating the presence of other unrevealed mechanism responsible for the loss of capability of autoreactive T cells to infiltrate and destroy the pancreatic beta-cells in vivo.


Weight control strategies of overweight adolescents who successfully lost weight.
The purpose of this pilot study was to evaluate the differences in weight control behaviors, dietary intake, and physical activity between overweight adolescents who lost weight and overweight adolescents who did not lose weight. This cross-sectional study compared 62 overweight adolescents who lost weight in the past with 68 overweight adolescents who did not lose weight. Youth responded to questions regarding weight control behaviors during the past year, physical activity, sedentary activity, and dietary intake. Results showed that adolescents who lost weight were more likely to report using healthful weight control behaviors such as drinking less soda and increasing their exercise level, self-weighing, consuming diets higher in protein, and spending less time watching television compared to overweight adolescents who did not lose weight. Unhealthful weight control behaviors and specific weight loss plans were not associated with weight loss in these teenagers. Adolescents would benefit from hearing this information to prevent the development of these behaviors. Providers should advise adolescents to engage in healthful weight control behaviors, such as increasing physical activity and decreasing the amount of time spent watching television, to assist with weight management.


A concise genetic and clinical guide to multiple endocrine neoplasias and related syndromes.
Several familial neoplastic syndromes are associated with endocrine gland oncogenesis. The main ones are: multiple endocrine neoplasia type 1 (MEN 1), which affects primarily the pituitary, pancreas, and parathyroid glands; MEN 2A and MEN 2B, which involve mainly the thyroid and parathyroid glands and the adrenal medulla; familial medullary thyroid carcinoma (FMTC), which affects only the thyroid gland; and, finally, Carney complex, which affects the adrenal cortex, pituitary, thyroid gland, and the gonads. Carney complex is also associated with pigmentation abnormalities and myxoid and other neoplasms of mesenchymal origin. Thus, this syndrome also belongs to another group of genetic disorders, those associated with pigmentation defects and multiple tumors, including tumors of the endocrine glands. Peutz-Jeghers syndrome and Cowden disease are just two of these disorders that have recently been elucidated at the molecular level. von Hippel-Lindau disease is another condition that affects the pancreas and adrenal medulla and its gene is also known. The inheritance of the MENs, Carney complex, and related syndromes is autosomal dominant. Clinical recognition of these syndromes at a young age improves clinical outcome and prognosis of the various tumors and decreases associated morbidity and mortality. This review considers a wider, more inclusive view of the MEN syndromes, summarizes their clinical features and presents the newest information on their molecular elucidation.


Odor Perception by Dogs: Evaluating Two Training Approaches for Odor Learning of Sniffer Dogs.
In this study, a standardized experimental set-up with various combinations of herbs as odor sources was designed. Two training approaches for sniffer dogs were compared; first, training with a pure reference odor, and second, training with a variety of odor mixtures with the target odor as a common denominator. The ability of the dogs to identify the target odor in a new context was tested. Six different herbs (basil, St. John's wort, dandelion, marjoram, parsley, ribwort) were chosen to produce reference materials in various mixtures with (positive) and without (negative) chamomile as the target odor source. The dogs were trained to show 1 of 2 different behaviors, 1 for the positive, and 1 for the negative sample as a yes/no task. Tests were double blind with one sample presented at a time. In both training approaches, dogs were able to detect chamomile as the target odor in any presented mixture with an average sensitivity of 72% and a specificity of 84%. Dogs trained with odor mixture containing the target odor had more correct indications in the transfer task.