Tomato seeds as a novel by-product feed for lactating dairy cows.
Whole tomato seeds, a novel by-product feedstuff, were fed to lactating Holstein cows to determine the nutritive value of whole tomato seeds by replacing whole cottonseed in the total mixed ration. Four primiparous and 4 multiparous Holstein cows were used in a 4×4 Latin square design and fed 1 of 4 total mixed rations. Whole tomato seeds replaced whole cottonseed on a weight-to-weight basis for lipid. The proportion of whole tomato seeds to whole cottonseed in the diets were 100:0, 50:50, 25:75, and 0:100 on a lipid basis. Thus, tomato seeds were 4.0, 2.4, 1.1, and 0% of the ration dry matter, respectively. Milk yield and the concentrations and yields of protein, lactose, and solids-not-fat did not differ for the effect of diet. However, milk fat concentration decreased and milk fat yield tended to decrease as whole tomato seeds replaced whole cottonseed. Intakes of dry matter, lipid, and crude protein did not differ. Whole-tract apparent digestibility of dry matter and ash-free neutral detergent fiber did not differ, but digestibility of total fatty acids and crude protein decreased with increasing proportion of whole tomato seeds. Urea concentration in milk and plasma both decreased with increasing whole tomato seeds. Fecal concentration of linoleic and α-linolenic acids increased with increasing whole tomato seeds, suggesting that seeds were passing out of the digestive tract undigested. The concentrations of C18:2n-6 and C18:3n-3 in milk fat had small increases, but their yields were not different, suggesting that only a small amount of whole-tomato-seed lipid might have been digested postruminally. Amounts of trans C18:1 fatty acids in milk fat were higher with increasing whole cottonseed, which might suggest a shift in rumen biohydrogenation pathways. At the level of feeding used in the current study, whole tomato seeds replaced whole cottonseed in the diet of lactating dairy cows without a change in production.


Sensitive detection of clonal immunoglobulin rearrangements in frozen and paraffin embedded tissues by polymerase chain reaction heteroduplex analysis.
Molecular detection of a clonal population of B or T cells through analysis of rearranged antigen receptor genes is an essential adjunct to the morphologic, flow cytometric, and immunohistochemical evaluation of tissue specimens for the presence of leukemia or lymphoma. Combining polymerase chain reaction (PCR) with heteroduplex annealing and polyacrylamide gel electrophoresis (PAGE) has been used to detect clonal T-cell receptor rearrangements, particularly in skin biopsy specimens. The authors have developed a similar PCR heteroduplex assay for detection of clonal VDJ immunoglobulin gene rearrangements using two sets of primers based on relatively conserved consensus regions in the J(H) and framework I and 2 regions of the immunoglobulin heavy chain V region gene. This method is able to detect a clonal rearrangement when the clone comprises as little as 1% of the population in a polyclonal B-cell background. It may be used on fresh, frozen, or paraffin-embedded tissue and detects a clonal population in a majority of lymphoma subtypes. Compared with conventional PCR analysis, this method requires only a short additional cycle of denaturation and slow renaturation before PAGE. Interpretation is simplified as the clonal PCR product migrates away from the polyclonal background products.


Self-reported drinking-game participation of incoming college students.
Drinking games are associated with excess alcohol use and alcohol-related problems, yet it is unclear whether they are unique to the college environment or whether students come to college familiar with such games. The authors queried 1,252 students attending voluntary summer orientation programs about their experiences with drinking games. A majority (63%) indicated they had played drinking games and viewed them as a means to get drunk quickly and to socialize, control others, or get someone else drunk. Logistic regression analyses revealed that familiarity with drinking situations was associated with a greater likelihood of playing drinking games. Students who reported drinking more frequently and consuming greater quantities of alcohol than others, having lifetime marijuana use, and initiating alcohol consumption between the ages of 14 and 16 years were significantly more likely to have participated in drinking games. These findings demonstrate the importance of considering students' participation in drinking games when campus officials address alcohol use.


Validation of the Dutch version of the transplant effects questionnaire in liver transplant recipients.
Little is known about the extent to which transplant recipients face emotional problems with the receipt of a transplanted organ. The Transplant Effects Questionnaire (TxEQ) enables the quantification of these problems. This study evaluates the psychometric properties of the Dutch translation of the TxEQ (TxEQ-NL) in a group of liver transplant recipients. Confirmatory factor analyses of the TxEQ-NL revealed an adequate fit with the original version. However, four items showed factor loadings <.40. Internal consistency was acceptable (.66-.79). The small correlations between the TxEQ-NL and generic measures of psychological functioning indicated that the constructs measured are related but distinguishable. Therefore, the TxEQ-NL adds a new dimension to the measurement of psychological functioning of transplant recipients.


Investigating hepatitis B immunity in patients presenting to a paediatric haematology and oncology unit in South Africa.
Hepatitis B is an important public health concern in South Africa (SA). The hepatitis B virus (HBV) vaccine was introduced into the South African Expanded Programme on Immunisation (EPI-SA) in 1995. There is no 'catch-up' programme in place. The duration of protection after hepatitis B vaccination in the SA population is unknown. Waning of vaccine-induced immunity leaves people at risk of acquiring hepatitis B infection in settings where the prevalence of infection is high and horizontal transmission is likely. To assess immunity to HBV in patients at presentation to a paediatric haematology and oncology unit. An audit of hepatitis profiles was done of all new patients seen in the unit from January 2012 to December 2013. Patients were classified as immune (antibody levels to hepatitis B surface antigen (anti-HBs) >100 mIU/ml), low immune (anti-HBs 10 - 100 mIU/ml) and not immune (anti-HBs <10 mIU/ml). Of the 210 patients included (median age 6.5 years), 84 (40.0%) had no immunity to hepatitis B despite presumed vaccination as part of the EPI schedule. Six patients tested positive for hepatitis B core antibody (anti-HBc), consistent with previous infection. No patients had active hepatitis B infection (hepatitis B surface antigen-positive). Most human immunodeficiency virus (HIV)-infected patients were not immune to HBV (80.0%). A significant number of children in SA are not immune to hepatitis B despite vaccination being part of the EPI-SA. Combined passive-active immunisation should be considered for all oncology patients in settings where exposure to HBV is possible. Consideration should also be given to offering booster vaccination to the population as a whole.


Separation of transition metals on a poly-iminodiacetic acid grafted polymeric resin column with post-column reaction detection utilising a paired emitter-detector diode system.
The selectivity, retention and separation of transition metals on a short (2 mm x 50 mm) column packed with a poly-iminodiacetic acid functionalised polymer 10 microm resin (Dionex ProPac IMAC-10) are presented. This stationary phase, typically used for the separation of proteins, is composed of long chain poly-iminodiacetic acid groups grafted to a hydrophilic layer surrounding a 10 microm polymeric bead. Through the use of a combination of a multi-step pH and picolinic acid gradient, the separation of magnesium, iron, cobalt, cadmium, zinc, lead and copper was possible, followed by post-column reaction with 4-(2-pyridylazo) resorcinol (PAR) and absorbance detection at 510 nm using a novel and inexpensive optical detector, comprised of two light emitting diodes with one acting as a light source and the other as a detector. Column efficiency for selective transition metals was in excess of N=10,000, with the baseline separation of seven metal cations in <3 min possible under optimised conditions. Detection limits of between 5 and 81 microg/L were possible based upon a 50 microL injection volume.


Characterization of human adipose tissue-resident hematopoietic cell populations reveals a novel macrophage subpopulation with CD34 expression and mesenchymal multipotency.
Adipose tissue (AT) is composed of mature adipocytes and stromal vascular fraction (SVF) cells, including adipose stem/stromal cells (ASCs). We characterized hematopoietic cells residing in human nonobese AT by analyzing the SVF isolated from human lipoaspirates and peripheral blood (PB). Flow cytometry revealed that AT-resident hematopoietic cells consisted of AT-resident macrophages (ATMs) or lymphocytes with a negligible number of granulocytes. AT-resident lymphocytes were composed of helper T cells and natural killer cells. Almost no B cells and few cytotoxic T cells were observed in nonobese AT. More than 90% of ATMs were M2 state CD206(+) macrophages (CD45(+)/CD14(+)) that were located in the periendothelium or interstitial spaces between adipocytes. We also discovered a novel subpopulation of CD34(+)/CD206(+) ATMs (11.1% of CD206(+)ATMs) that localized in the perivascular region. Microarray of noncultured CD34(+)/CD206(+) ATMs, CD34(-)/CD206(+) ATMs, CD45(-)/CD31(-)/CD34(+) ASCs, and PB-derived circulating monocytes revealed that CD34(+)/CD206(+) ATMs shared characteristics with ASCs and circulating monocytes. Unlike CD34(-)/CD206(+) ATMs, CD34(+)/CD206(+) ATMs could grow in adherent culture and were capable of differentiating into multiple mesenchymal (adipogenic, osteogenic, and chondrogenic) lineages, similar to ASCs. CD34(+)/CD206(+) ATMs grew rapidly and lost expression of CD45, CD14, and CD206 by passage 3, which resulted in a similar expression profile to ASCs. Thus, this novel ATM subpopulation (CD45(+)/CD14(+)/CD34(+)/CD206(+)) showed distinct biological properties from other ATMs and circulating monocytes/macrophages. The CD34(+)/CD206(+) ATMs possessed characteristics similar to ASCs, including adherence, localization, morphology, and mesenchymal multipotency. This AT-resident subpopulation may have migrated from the bone marrow and may be important to tissue maintenance and remolding.


Psychometric performance of the Chichewa versions of the EQ-5D-Y-3L and EQ-5D-Y-5L among healthy and sick children and adolescents in Malawi.
The EuroQol Group has developed an extended version of the EQ-5D-Y-3L with five response levels for each of its five dimensions (EQ-5D-Y-5L). The psychometric performance has been reported in several studies for the EQ-5D-Y-3L but not for the EQ-5D-Y-5L. This study aimed to psychometrically evaluate the EQ-5D-Y-3L and EQ-5D-Y-5L Chichewa (Malawi) versions. The EQ-5D-Y-3L, EQ-5D-Y-5L and PedsQL™ 4.0 Chichewa versions were administered to children and adolescents aged 8-17 years in Blantyre, Malawi. Both of the EQ-5D-Y versions were evaluated for missing data, floor/ceiling effects, and validity (convergent, discriminant, known-group and empirical). A total of 289 participants (95 healthy, and 194 chronic and acute) self-completed the questionnaires. There was little problem with missing data (< 5%) except in children aged 8-12 years particularly for the EQ-5D-Y-5L. Ceiling effects was generally reduced in moving from the EQ-5D-Y-3L to the EQ-5D-Y-5L. For both EQ-5D-Y-3L and EQ-5D-Y-5L, convergent validity tested with PedsQL™ 4.0 was found to be satisfactory (correlation ≥ 0.4) at scale level but mixed at dimension /sub-scale level. There was evidence of discriminant validity (p > 0.05) with respect to gender and age, but not for school grade (p < 0.05). For empirical validity, the EQ-5D-Y-5L was 31-91% less efficient than the EQ-5D-Y-3L at detecting differences in health status using external measures. Both versions of the EQ-5D-Y-3L and EQ-5D-Y-5L had issues with missing data in younger children. Convergent validity, discriminant validity with respect to gender and age, and known-group validity of either measures were also met for use among children and adolescents in this population, although with some limitations (discriminant validity by grade and empirical validity). The EQ-5D-Y-3L seems particularly suited for use in younger children (8-12 years) and the EQ-5D-Y-5L in adolescents (13-17 years). However, further psychometric testing is required for test re-test reliability and responsiveness that could not be carried out in this study due to COVID-19 restrictions.


Heterogeneity of accessory cells interacting with T-helper clones.
We have investigated the comparative role of dendritic cells and macrophages as inducers of proliferative responses in antigen-specific T-helper lines and clones grown in serum-free medium. Dendritic cells are the most potent accessory cells for both soluble (ovalbumin-specific) and particulate (horse red blood cell-specific) lines. However, at the clonal level there is heterogeneity. Individual clones will respond to antigen presented on either dendritic cells or macrophages, respectively. Synergy between the two accessory cell types was not demonstrable. Depletion of cells that express Class II antigens from the macrophage population abrogated the proliferative response. These findings confirm heterogeneity of the accessory cell pool, and support the suggestion that this accessory cell heterogeneity may be one of the selective mechanisms that determine T-cell responses at the clonal level.


Simple behavioural interventions for nocturnal enuresis in children.
Nocturnal enuresis (bedwetting) is a socially disruptive and stressful condition which affects around 15% to 20% of five year olds and up to 2% of adults. Although there is a high rate of spontaneous remission, the social, emotional and psychological costs can be great. Behavioural interventions for treating bedwetting are defined as interventions that require a behaviour or action by the child which promotes night dryness and includes strategies which reward that behaviour. Behavioural interventions are further divided into:(a) simple behavioural interventions - behaviours or actions that can be achieved by the child without great effort; and(b) complex behavioural interventions - multiple behavioural interventions which require greater effort by the child and parents to achieve, including enuresis alarm therapy.This review focuses on simple behavioural interventions.Simple behavioural interventions are often used as a first attempt to improve nocturnal enuresis and include reward systems such as star charts given for dry nights, lifting or waking the children at night to urinate, retention control training to enlarge bladder capacity (bladder training) and fluid restriction. Other treatments such as medications, complementary and miscellaneous interventions such as acupuncture, complex behavioural interventions and enuresis alarm therapy are considered elsewhere. To determine the effects of simple behavioural interventions in children with nocturnal enuresis.The following comparisons were made:1. simple behavioural interventions versus no active treatment;2. any single type of simple behavioural intervention versus another behavioural method (another simple behavioural intervention, enuresis alarm therapy or complex behavioural interventions);3. simple behavioural interventions versus drug treatment alone (including placebo drugs) or drug treatment in combination with other interventions. We searched the Cochrane Incontinence Group Specialised Trials Register, which contains trials identified from the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, MEDLINE in process, and handsearching of journals and conference proceedings (searched 15 December 2011). The reference lists of relevant articles were also searched. All randomised or quasi-randomised trials of simple behavioural interventions for treating nocturnal enuresis in children up to the age of 16. Studies which included children with daytime urinary incontinence or children with organic conditions were also included in this review if the focus of the study was on nocturnal enuresis. Trials focused solely on daytime wetting and trials of adults with nocturnal enuresis were excluded. Two reviewers independently assessed the quality of the eligible trials and extracted data. Differences between reviewers were settled by discussion with a third reviewer. Sixteen trials met the inclusion criteria, involving 1643 children of whom 865 received a simple behavioural intervention. Within each comparison, outcomes were mostly addressed by single trials, precluding meta-analysis. The only exception was bladder training versus enuresis alarm therapy which included two studies and demonstrated that alarm therapy was superior to bladder training.In single small trials, rewards, lifting and waking and bladder training were each associated with significantly fewer wet nights, higher full response rates and lower relapse rates compared to controls. Simple behavioural interventions appeared to be less effective when compared with other known effective interventions (such as enuresis alarm therapy and drug therapies with imipramine and amitriptyline). However, the effect was not sustained at follow-up after completion of treatment for the drug therapies. Based on one small trial, cognitive therapy also appeared to be more effective than rewards. When one simple behavioural therapy was compared with another, there did not appear to be one therapy that was more effective than another. Simple behavioural methods may be superior to no active treatment but appear to be inferior to enuresis alarm therapy and some drug therapy (such as imipramine and amitriptyline). Simple behavioural therapies could be tried as first line treatment before considering enuresis alarm therapy or drug therapy, which may be more demanding and have adverse effects, although evidence supporting their efficacy is lacking.


Daily Protein and Energy Intake Are Not Associated with Muscle Mass and Physical Function in Healthy Older Individuals-A Cross-Sectional Study.
Dietary protein has a pivotal role in muscle mass maintenance with advancing age. However, an optimal dose and distribution of protein intake across the day as well as the interaction with energy intake for the maintenance of muscle mass and physical function in healthy older adults remain to be fully elucidated. The purpose of this study was to examine the association between muscle mass, strength, and physical function, and the total amount and distribution of protein and energy intake across the day in healthy older individuals. The research question was addressed in a cross-sectional study including 184 Danish men and woman (age: 70.2 ± 3.9 years, body mass: 74.9 ± 12.1 kg, Body Mass Index (BMI): 25.4 ± 3.7 kg/m2) where a 3-day dietary registration, muscle mass, strength, and functional measurements were collected. We found that neither daily total protein intake nor distribution throughout the day were associated with muscle mass, strength, or physical function. Consequently, we do not provide an incentive for healthy older Danish individuals who already adhere to the current internationally accepted recommended dietary protein intake (0.83 g/kg/day) to change dietary protein intake or its distribution pattern throughout the day.


The use of MolecuLight i:X device in acute hand trauma.
Early diagnosis of wound infections are crucial as they have been shown to increase patient morbidity and mortality. We evaluated the use of Moleculight i:X to identify infections in acute open wounds in hand trauma. Data were collected from patients who attended the hand trauma unit over a 4 week period prior to having surgery. Wounds were inspected for clinical signs of infection and autofluorescence images were taken using the Moleculight i:X device. Wound swabs were taken and results interpreted according to report by microbiologist. Autofluorescence images were interpreted by a clinician blinded to the microbiology results. 31 patients were included and data collected from 35 wounds. 3 wounds (8.6%) showed positive clinical signs of infection, 3 (8.6%) were positive on autofluorescence imaging and 2 (5.7%) of wound swab samples were positive for significant infection. Autofluorescence imaging correlated with clinical signs and wound swab results for 34 wounds (97.1%). In one case, the clinical assessment and autofluorescence imaging showed positive signs of infection but the wound swabs were negative. Autofluorescence imaging in acute open wounds may be useful to provide real-time confirmation of bacterial infection and therefore guide management.


Multiple immune abnormalities in tumor necrosis factor and lymphotoxin-alpha double-deficient mice.
To investigate the roles of tumor necrosis factor (TNF) and lymphotoxin (LT)-alpha in the development and function of the immune system, the Tnf and Ltalpha genes were simultaneously inactivated in mice by homologous recombination. These mutant mice are highly susceptible to Listeria monocytogenes infection and resistant to endotoxic shock induced by the combined administration of D-galactosamine (D-GaIN) and lipopolysaccharide (LPS). Their splenic microarchitecture is disorganized, characterized by the loss of the clearly defined marginal zone, ill defined T and B cell areas, and absence of MAdCAM-1 and reduced ICAM-1, VCAM-1 and Mac-1 expression. They are devoid of peripheral lymph nodes and Peyer's patches, and show a strong reduction of IgA+ plasma cells in the intestinal lamina propria. The alymphoplasia is accompanied by a marked B lymphocytosis and reduced basal lg levels. Ig depositions in the renal glomerulus and a strong up-regulation of MHC class I antigen expression on endothelial cells of different tissues are observed. The primary humoral immune response towards sheep red blood cells reveals a defective IgG isotype switch, while that against vesicular stomatitis virus is normal. The cytotoxic T cell responses are attenuated, although still effective, against vaccinia, lymphocytic choriomeningitis virus (LCMV-ARM) and LCMV-WE. In conclusion, the combined inactivation of Tnf and Ltalpha confirms their essential role in the normal development and function of the immune system.


Arabinoxylan rice bran (MGN-3) enhances the effects of interventional therapies for the treatment of hepatocellular carcinoma: a three-year randomized clinical trial.
This study examined the efficacy of arabinoxylan rice bran (MGN-3) in conjunction with an interventional therapy (IT) for the treatment of hepatocellular carcinoma patients. A total of sixty-eight patients with hepatocellular carcinoma (stages I and II) participated in the study. Patients were randomized to receive IT (30 patients, control group) or IT+MGN-3 (38 patients), and randomly divided into two groups using a computer-generated randomization list. Patients and investigators were blinded. IT included transarterial oily chemoembolization (TOCE) or a combination of TOCE and percutaneous ethanol injection treatment (PEIT). Patients in the IT+MGN-3 group showed: (i) lower recurrence of the disease, 31.6% (12/38), as compared to 46.7% (14/30) for the control; (ii) higher survival after the second year, 35%, as compared to 6.7% for the control; (iii) significantly lower alpha-fetoprotein level, a 38% decrease (p = 0.0001), as compared to baseline value, while the control showed no significant change; and (iv) a significant decrease in tumor volume, in contrast to the control, which showed no significant change. When the results were analyzed according to each IT modality, MGN-3+IT sub-groups displayed a greater response to treatment, in every aspect examined, than the IT sub-groups alone. However, the patients in the MGN-3+TOCE+PEIT sub-group demonstrated greater reduction in AFP levels and longer survival time than the MGN-3+TOCE sub-group. MGN-3 in conjunction with IT may be useful for the treatment of hepatocellular carcinoma and warrants further investigation in multiple clinical trials.


Plant species richness and functional composition drive overyielding in a six-year grassland experiment.
Plant diversity has been shown to increase community biomass in experimental communities, but the mechanisms resulting in such positive biodiversity effects have remained largely unknown. We used a large-scale six-year biodiversity experiment near Jena, Germany, to examine how aboveground community biomass in grasslands is affected by different components of plant diversity and thereby infer the mechanisms that may underlie positive biodiversity effects. As components of diversity we defined the number of species (1-16), number of functional groups (1-4), presence of functional groups (legumes, tall herbs, small herbs, and grasses) and proportional abundance of functional groups. Using linear models, replacement series on the level of functional groups, and additive partitioning on the level of species, we explored whether the observed biodiversity effects originated from disproportionate effects of single functional groups or species or from positive interactions between them. Aboveground community biomass was positively related to the number of species measured across functional groups as well as to the number of functional groups measured across different levels of species richness. Furthermore, increasing the number of species within functional groups increased aboveground community biomass, indicating that species within functional groups were not redundant with respect to biomass production. A positive relationship between the number of functional groups and aboveground community biomass within a particular level of species richness suggested that complementarity was larger between species belonging to different rather than to the same functional groups. The presence of legumes or tall herbs had a strong positive impact on aboveground community biomass whereas the presence of small herbs or grasses had on average no significant effect. Two- and three-way interactions between functional group presences were weak, suggesting that their main effects were largely additive. Replacement series analyses on the level of functional groups revealed strong transgressive overyielding and relative yields >1, indicating facilitation. On the species level, we found strong complementarity effects that increased over time while selection effects due to disproportionate contributions of particular species decreased over time. We conclude that transgressive overyielding between functional groups and species richness effects within functional groups caused the positive biodiversity effects on aboveground community biomass in our experiment.


Towards global consensus on outcome measures for atopic eczema research: results of the HOME II meeting.
The use of nonstandardized and inadequately validated outcome measures in atopic eczema trials is a major obstacle to practising evidence-based dermatology. The Harmonising Outcome Measures for Eczema (HOME) initiative is an international multiprofessional group dedicated to atopic eczema outcomes research. In June 2011, the HOME initiative conducted a consensus study involving 43 individuals from 10 countries, representing different stakeholders (patients, clinicians, methodologists, pharmaceutical industry) to determine core outcome domains for atopic eczema trials, to define quality criteria for atopic eczema outcome measures and to prioritize topics for atopic eczema outcomes research. Delegates were given evidence-based information, followed by structured group discussion and anonymous consensus voting. Consensus was achieved to include clinical signs, symptoms, long-term control of flares and quality of life into the core set of outcome domains for atopic eczema trials. The HOME initiative strongly recommends including and reporting these core outcome domains as primary or secondary endpoints in all future atopic eczema trials. Measures of these core outcome domains need to be valid, sensitive to change and feasible. Prioritized topics of the HOME initiative are the identification/development of the most appropriate instruments for the four core outcome domains. HOME is open to anyone with an interest in atopic eczema outcomes research.


Congenital adrenal hyperplasia and Leydig cell tumor of testis. Case report and review of literature.
Adrenogenital Syndrome, more properly defined as Congenital Adrenal Hyperplasia (CAH), is related to the enzyme 21-beta-hydroxylase deficiency, with impaired glucocorticoids and aldosterone syntheses and increased ACTH synthesis. This report describes a case of a monorchid patient suffering from Adrenogenital Syndrome and Leydig cell tumor of his testis. A right orchidectomy with implantation of testis prosthesis was performed, after informing the patient on the consequences of his castration and obtaining his consent. Histology showed a testis measuring 4 x 3 x 2.5 cm with a 6 cm long spermatic cord; there was a yellowish, well-defined nodule measuring 3.5 x 1.5 cm, surrounded by normal parenchyma. This nodule had morphologic and immunohistochemical characteristics of a Leydig cell tumor, even found in the spermatic cord; those cells showed positivity to inibine, MART-1 and vimentine.


Abnormal uterine bleeding.
Concerns about abnormal menstrual bleeding are a common reason for women to consult a primary care physician. The first step in the evaluation is to determine the patient's ovulatory status. Women with heavy bleeding but normal ovulatory cycles should be evaluated for coagulopathies, structural lesions, and hypothyroidism. In the absence of a systemic or structural cause, menorrhagia can be treated with OCPs or NSAIDs. Intermenstrual bleeding in OCP users may be due to noncompliance or the use of low-dose pills. Encouraging patient compliance and adjustment of the estrogen dose can often solve the problem. If the patient is not on OCPs, intermenstrual bleeding is usually due to a structural or inflammatory lesion. The differential diagnosis for anovulatory bleeding is extensive. Pregnancy, systemic illnesses, and structural lesions should be ruled out by history, physical examination, and laboratory evaluation. Endometrial biopsy is indicated in patients over age 35 and younger patients with risk factors for endometrial cancer, such as chronic anovulation and obesity. Dysfunctional uterine bleeding is a nonspecific term for abnormal uterine bleeding in the absence of systemic or structural disease. It is usually associated with anovulation. Adolescents frequently have dysfunctional uterine bleeding owing to immaturity of the hypothalamic-pituitary-ovarian axis. Perimenopausal women have an increased incidence of irregular bleeding secondary to decreased estrogen production by the ovary. Obesity, polycystic ovary syndrome, stress, crash diets, and vigorous exercise can all disrupt normal ovulatory function. Treatment options for dysfunctional uterine bleeding include oral contraceptives, cyclic progesterone, or hormone replacement with estrogen and progesterone. Patients with structural lesions or those who do not resume normal withdrawal bleeding patterns on hormone therapy should be referred to a gynecologist for further evaluation and treatment.


Effect of Chicory-derived Inulin on Abdominal Sensations and Bowel Motor Function.
To determine the effect of a prebiotic chicory-derived inulin-type fructan on the tolerance of intestinal gas. Subjects with gas-related complaints exhibit impaired handling of intestinal gas loads and we hypothesized that inulin would have a beneficial effect. Placebo-controlled, parallel, randomized, double-blind trial. Subjects with abdominal symptoms and reduced tolerance of intestinal gas (selected by a pretest) received either inulin (8 g/d, n=18) or maltodextrin as a placebo (8 g/d, n=18) for 4 weeks. A gas challenge test (4 h jejunal gas infusion at 12 mL/min while measuring abdominal symptoms and gas retention for 3 h) was performed before and at the end of the intervention phase. Gastrointestinal symptoms and bowel habits (using daily questionnaires for 1 wk) and fecal bifidobacteria counts were measured before and at the end of the intervention. Inulin decreased gas retention during the gas challenge test (by 22%; P=0.035 vs. baseline), while the placebo did not, but the intergroup difference was not statistically significant (P=0.343). Inulin and placebo reduced the perception of abdominal sensations in the gas challenge test to a similar extent (by 52% and 43%, respectively). Participants reported moderate gastrointestinal symptoms and normal bowel habits during baseline examination, and these findings remained unchanged in both groups during the intervention. Inulin led to a higher relative abundance of bifidobacteria counts (P=0.01 vs. placebo). A daily dose of inulin that promotes bifidobacteria growth and may improve gut function, is well tolerated by subjects with gastrointestinal complaints.


Endoskeletal structure in Cheirolepis (Osteichthyes, Actinopterygii), An early ray-finned fish.
As the sister lineage of all other actinopterygians, the Middle to Late Devonian (Eifelian-Frasnian) Cheirolepis occupies a pivotal position in vertebrate phylogeny. Although the dermal skeleton of this taxon has been exhaustively described, very little of its endoskeleton is known, leaving questions of neurocranial and fin evolution in early ray-finned fishes unresolved. The model for early actinopterygian anatomy has instead been based largely on the Late Devonian (Frasnian) Mimipiscis, preserved in stunning detail from the Gogo Formation of Australia. Here, we present re-examinations of existing museum specimens through the use of high-resolution laboratory- and synchrotron-based computed tomography scanning, revealing new details of the neuro-cranium, hyomandibula and pectoral fin endoskeleton for the Eifelian Cheirolepis trailli. These new data highlight traits considered uncharacteristic of early actinopterygians, including an uninvested dorsal aorta and imperforate propterygium, and corroborate the early divergence of Cheirolepis within actinopterygian phylogeny. These traits represent conspicuous differences between the endoskeletal structure of Cheirolepis and Mimipiscis. Additionally, we describe new aspects of the parasphenoid, vomer and scales, most notably that the scales display peg-and-socket articulation and a distinct neck. Collectively, these new data help clarify primitive conditions within ray-finned fishes, which in turn have important implications for understanding features likely present in the last common ancestor of living osteichthyans.


The future of cell therapies in the treatment of Parkinson's disease.
Parkinson's disease (PD) is a common neurological disorder of the brain which has as a part of its core pathology the progressive degeneration of the dopaminergic nigrostriatal pathway. Therefore, cell therapies that aim to restore this degenerated dopaminergic network represent a promising strategy in helping to cure PD. In this review, the authors start by discussing the progress of research on the use of fetal ventral mesencephalic (VM) tissue in transplantation therapies in PD, both from the clinical and experimental perspectives. Then the issues pertinent to its adoption in the clinic are discussed, including the ethical and practical problems with its use, the varied composition of VM tissue that is implanted with the graft and how this may account for some of the problems seen in the clinical trials using this tissue, especially graft-induced dyskinesia. Finally other promising sources of tissue for PD cell therapy are described, including mesenchymal and embryonic stem cells, before concluding on what is the best approach to the cellular repair of the parkinsonian brain.


Brief Report: Miscarriages in Female Rheumatoid Arthritis Patients: Associations With Serologic Findings, Disease Activity, and Antirheumatic Drug Treatment.
To study the association between miscarriage in rheumatoid arthritis (RA) patients and serologic findings, disease activity, and antirheumatic drug treatment, and to study disease activity and reproductive outcomes after a miscarriage. Within a nationwide prospective cohort study (Pregnancy-Induced Amelioration of RA study), patients with RA were followed up from preconception until 6 months after delivery or miscarriage. Univariate and logistic regression analyses were performed to assess variables of interest, with covariates included in the models if the P value for association with miscarriage was <0.20 and subsequently excluded if the P value was >0.10. Among 162 pregnancies, 28 miscarriages occurred (17.3%; 95% confidence interval 12.2-24.0%). Women who miscarried were older than women with an ongoing pregnancy. Women who miscarried tended to be more often positive for anti-citrullinated protein antibodies (ACPAs), to have higher disease activity scores, and to have more often received methotrexate (MTX) therapy in the past. Logistic regression showed a tendency toward a higher likelihood of miscarriage in association with increasing age (P = 0.065) and presence of ACPAs (P = 0.092). After miscarriage, 33% of women had a flare of RA. Within 1 year, 68% of women became pregnant again, 14% stopped trying to conceive, and 11% were lost to followup. The live birth rate of the subsequent pregnancy was 90%. The miscarriage rate in the PARA cohort is comparable to that in the general population. Due to the low frequency of miscarriages in this study, the associations between miscarriage in RA and the presence of ACPAs, disease activity, and MTX use did not reach statistical significance. Within 1 year after miscarriage, the majority of patients who continued trying to conceive achieved a pregnancy resulting in a live birth.


Non-resonant wavelength modulation saturation spectroscopy in acetylene-filled hollow-core photonic bandgap fibres applied to modulation-free laser diode stabilisation.
In this paper the application of Wavelength Modulation (WM) techniques to non-resonant saturation spectroscopy in acetylene-filled Hollow-Core Photonic Bandgap Fibres (HC-PBFs) and modulation-free Laser Diode (LD) frequency stabilisation is investigated. In the first part WM techniques are applied to non-resonant pump-probe saturation of acetylene overtone rotational transitions in a HC-PBF. A high-power DFB chip-on-carrier mounted LD is used in conjunction with a tuneable External Cavity Laser (ECL) and the main saturation parameters are characterized. In the second part a novel feedback system to stabilize the DFB emission wavelength based on the WM saturation results is implemented. Modulation-free locking of the DFB laser frequency to the narrow linewidth saturation feature is achieved for both constant and variable LD temperatures.


Outcome of drug-eluting balloon angioplasty versus endarterectomy in common femoral artery occlusive disease.
Common femoral artery (CFA) occlusive disease remains a debatable site for endovascular therapy, and the outcome of drug-eluting balloon (DEB) angioplasty in treating CFA occlusive disease is largely unknown. This study compared the efficacy, safety, and short-term patency rate of DEB angioplasty and femoral endarterectomy for treatment of CFA occlusive disease. From March 2013 to June 2016, there were 100 patients with symptomatic CFA occlusive disease who were retrospectively reviewed. Forty patients were treated with DEB angioplasty and 60 were treated with femoral endarterectomy. Each patient received regular follow-up. Patency rate, ankle-brachial index, target lesion revascularization, and adverse events were assessed. Technical success was 100% in all patients. The DEB group had a lower 1-year primary patency rate (75.0% vs 96.7%; P = .003), but the secondary patency rate was similar between the two groups (97.5% vs 98.3%; P = 1.000). At 2-year follow-up, the primary patency was lower in the DEB group (57.1%) than in the endarterectomy group (94.1%; P = .001), whereas the secondary patency rate had no significant difference (90.5% vs 97.1%; P = 1.000). Both groups had significant improvement in ankle-brachial index. Freedom from target lesion revascularization was lower in the DEB group both at 1 year (75.0% vs 96.7%; P = .003) and at 2 years (57.1% vs 94.1%; P = .001). There was no significant difference in the incidence of complications and adverse events. Femoral endarterectomy has a better primary patency rate compared with DEB angioplasty in treating CFA occlusive disease without significant increase in complications. In patients not suitable for endarterectomy, DEB angioplasty provides a similar secondary patency rate and could be considered an alternative treatment.


Radiolabeling strategies and pharmacokinetic studies for metal based nanotheranostics.
Radiolabeled metal-based nanoparticles (MNPs) have drawn considerable attention in the fields of nuclear medicine and molecular imaging, drug delivery, and radiation therapy, given the fact that they can be potentially used as diagnostic imaging and/or therapeutic agents, or even as theranostic combinations. Here, we present a systematic review on recent advances in the design and synthesis of MNPs with major focuses on their radiolabeling strategies and the determinants of their in vivo pharmacokinetics, and together how their intended applications would be impacted. For clarification, we categorize all reported radiolabeling strategies for MNPs into indirect and direct approaches. While indirect labeling simply refers to the use of bifunctional chelators or prosthetic groups conjugated to MNPs for post-synthesis labeling with radionuclides, we found that many practical direct labeling methodologies have been developed to incorporate radionuclides into the MNP core without using extra reagents, including chemisorption, radiochemical doping, hadronic bombardment, encapsulation, and isotope or cation exchange. From the perspective of practical use, a few relevant examples are presented and discussed in terms of their pros and cons. We further reviewed the determinants of in vivo pharmacokinetic parameters of MNPs, including factors influencing their in vivo absorption, distribution, metabolism, and elimination, and discussed the challenges and opportunities in the development of radiolabeled MNPs for in vivo biomedical applications. Taken together, we believe the cumulative advancement summarized in this review would provide a general guidance in the field for design and synthesis of radiolabeled MNPs towards practical realization of their much desired theranostic capabilities. This article is categorized under: Nanotechnology Approaches to Biology > Nanoscale Systems in Biology Diagnostic Tools > Diagnostic Nanodevices Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease.


Comparison of in-sight and handheld navigation devices toward supporting industry 4.0 supply chains: First and last mile deliveries at the human level.
Last (and First) mile deliveries are an increasingly important and costly component of supply chains especially those that require transport within city centres. With reduction in anticipated manufacturing and delivery timescales, logistics personnel are expected to identify the correct location (accurately) and supply the goods in appropriate condition (safe delivery). Moving towards more environmentally sustainable supply chains, the last/first mile of deliveries may be completed by a cyclist courier which could result in significant reductions in congestion and emissions in cities. In addition, the last metres of an increasing number of deliveries are completed on foot i.e. as a pedestrian. Although research into new technologies to support enhanced navigation capabilities is ongoing, the focus to date has been on technical implementations with limited studies addressing how information is perceived and actioned by a human courier. In the research reported in this paper a comparison study has been conducted with 24 participants evaluating two examples of state-of-the-art navigation aids to support accurate (right time and place) and safe (right condition) navigation. Participants completed 4 navigation tasks, 2 whilst cycling and 2 whilst walking. The navigation devices under investigation were a handheld display presenting a map and instructions and an in-sight monocular display presenting text and arrow instructions. Navigation was conducted in a real-world environment in which eye movements and device interaction were recorded using Tobii-Pro 2 eye tracking glasses. The results indicate that the handheld device provided better support for accurate navigation (right time and place), with longer but less frequent gaze interactions and higher perceived usability. The in-sight display supported improved situation awareness with a greater number of hazards acknowledged. The benefits and drawbacks of each device and use of visual navigation support tools are discussed.


Self-doping processes between planes and chains in the metal-to-superconductor transition of YBa2Cu3O6.9.
The interplay between the quasi 1-dimensional CuO-chains and the 2-dimensional CuO2 planes of YBa(2)Cu(3)O(6+x) (YBCO) has been in focus for a long time. Although the CuO-chains are known to be important as charge reservoirs that enable superconductivity for a range of oxygen doping levels in YBCO, the understanding of the dynamics of its temperature-driven metal-superconductor transition (MST) remains a challenge. We present a combined study using x-ray absorption spectroscopy and resonant inelastic x-ray scattering (RIXS) revealing how a reconstruction of the apical O(4)-derived interplanar orbitals during the MST of optimally doped YBCO leads to substantial hole-transfer from the chains into the planes, i.e. self-doping. Our ionic model calculations show that localized divalent charge-transfer configurations are expected to be abundant in the chains of YBCO. While these indeed appear in the RIXS spectra from YBCO in the normal, metallic, state, they are largely suppressed in the superconducting state and, instead, signatures of Cu trivalent charge-transfer configurations in the planes become enhanced. In the quest for understanding the fundamental mechanism for high-Tc-superconductivity (HTSC) in perovskite cuprate materials, the observation of such an interplanar self-doping process in YBCO opens a unique novel channel for studying the dynamics of HTSC.


Passive seismic experiment.
Seismometer operation for 21 days at Tranquillity Base revealed, among strong signals produced by the Apollo 11 lunar module descent stage, a small proportion of probable natural seismic signals. The latter are long-duration, emergent oscillations which lack the discrete phases and coherence of earthquake signals. From similarity with the impact signal of the Apollo 12 ascent stage, they are thought to be produced by meteoroid impacts or shallow moonquakes. This signal character may imply transmission with high Q and intense wave scattering, conditions which are mutually exclusive on earth. Natural background noise is very much smaller than on earth, and lunar tectonism may be very low.


Predictive Value of Arterial Stiffness and Subclinical Carotid Atherosclerosis for Cardiovascular Disease in Patients with Rheumatoid Arthritis.
We evaluated the predictive value of these vascular biomarkers for cardiovascular disease (CVD) events in patients with rheumatoid arthritis (RA): aortic pulse wave velocity (aPWV), augmentation index (AIx), carotid intima-media thickness (cIMT), and carotid plaques (CP). They are often used as risk markers for CVD. In 2007, 138 patients with RA underwent clinical examination, laboratory tests, blood pressure testing, and vascular biomarker measurements. Occurrence of CVD events was recorded in 2013. Predictive values were assessed in Kaplan-Meier plots, log-rank, and crude and adjusted Cox proportional hazard (PH) regression analyses. Baseline median age and disease duration was 59.0 years and 17.0 years, respectively, and 76.1% were women. CVD events occurred in 10 patients (7.2%) during a mean followup of 5.4 years. Compared with patients with low aPWV, AIx, cIMT, and without CP, patients with high aPWV (p < 0.001), high AIx (p = 0.04), high cIMT (p = 0.01), and CP (p < 0.005) at baseline experienced more CVD events. In crude Cox PH regression analyses, aPWV (p < 0.001), cIMT (p < 0.001), age (p = 0.01), statin (p = 0.01), and corticosteroid use (p = 0.01) were predictive of CVD events, while AIx was nonsignificant (p = 0.19). The Cox PH regression estimates for vascular biomarkers were not significantly altered when adjusting individually for demographic variables, traditional CVD risk factors, RA disease-related variables, or medication. All patients who developed CVD had CP at baseline. CP, aPWV, and cIMT were predictive of CVD events in this cohort of patients with RA. Future studies are warranted to examine the additive value of arterial stiffness and carotid atherosclerosis markers in CVD risk algorithms. Regional Ethical Committee approval numbers 2009/1582 and 2009/1583.


High light induced disassembly of photosystem II supercomplexes in Arabidopsis requires STN7-dependent phosphorylation of CP29.
Photosynthetic oxidation of water and production of oxygen by photosystem II (PSII) in thylakoid membranes of plant chloroplasts is highly affected by changes in light intensities. To minimize damage imposed by excessive sunlight and sustain the photosynthetic activity PSII, organized in supercomplexes with its light harvesting antenna, undergoes conformational changes, disassembly and repair via not clearly understood mechanisms. We characterized the phosphoproteome of the thylakoid membranes from Arabidopsis thaliana wild type, stn7, stn8 and stn7stn8 mutant plants exposed to high light. The high light treatment of the wild type and stn8 caused specific increase in phosphorylation of Lhcb4.1 and Lhcb4.2 isoforms of the PSII linker protein CP29 at five different threonine residues. Phosphorylation of CP29 at four of these residues was not found in stn7 and stn7stn8 plants lacking the STN7 protein kinase. Blue native gel electrophoresis followed by immunological and mass spectrometric analyses of the membrane protein complexes revealed that the high light treatment of the wild type caused redistribution of CP29 from PSII supercomplexes to PSII dimers and monomers. A similar high-light-induced disassembly of the PSII supercomplexes occurred in stn8, but not in stn7 and stn7stn8. Transfer of the high-light-treated wild type plants to normal light relocated CP29 back to PSII supercomplexes. We postulate that disassembly of PSII supercomplexes in plants exposed to high light involves STN7-kinase-dependent phosphorylation of the linker protein CP29. Disruption of this adaptive mechanism can explain dramatically retarded growth of the stn7 and stn7stn8 mutants under fluctuating normal/high light conditions, as previously reported.


Choosing parameters of kernel subspace LDA for recognition of face images under pose and illumination variations.
This paper addresses the problem of automatically tuning multiple kernel parameters for the kernel-based linear discriminant analysis (LDA) method. The kernel approach has been proposed to solve face recognition problems under complex distribution by mapping the input space to a high-dimensional feature space. Some recognition algorithms such as the kernel principal components analysis, kernel Fisher discriminant, generalized discriminant analysis, and kernel direct LDA have been developed in the last five years. The experimental results show that the kernel-based method is a good and feasible approach to tackle the pose and illumination variations. One of the crucial factors in the kernel approach is the selection of kernel parameters, which highly affects the generalization capability and stability of the kernel-based learning methods. In view of this, we propose an eigenvalue-stability-bounded margin maximization (ESBMM) algorithm to automatically tune the multiple parameters of the Gaussian radial basis function kernel for the kernel subspace LDA (KSLDA) method, which is developed based on our previously developed subspace LDA method. The ESBMM algorithm improves the generalization capability of the kernel-based LDA method by maximizing the margin maximization criterion while maintaining the eigenvalue stability of the kernel-based LDA method. An in-depth investigation on the generalization performance on pose and illumination dimensions is performed using the YaleB and CMU PIE databases. The FERET database is also used for benchmark evaluation. Compared with the existing PCA-based and LDA-based methods, our proposed KSLDA method, with the ESBMM kernel parameter estimation algorithm, gives superior performance.


Cyclic AMP-Induced p53 Destabilization is Independent of CREB in Pre-B Acute Lymphoblastic Leukemia Cells.
Elevated cAMP levels in B-cell precursor acute lymphoblastic leukemia (BCP-ALL) cells attenuate the doxorubicin-induced p53 accumulation and protect cells against apoptosis. cAMP responsive element binding protein (CREB) is a cAMP-stimulated transcription factor that regulates genes whose deregulated expression cooperate in oncogenesis. In the present study, we investigated the role of CREB on inhibitory effect of cAMP on apoptosis and p53 accumulation in BCP-ALL NALM-6 cells. To determine whether targeting CREB can modulate the effect of cAMP on doxorubicin-induced apoptosis, we knocked down CREB in NALM-6 cells using lentiviral CREB shRNA. Knocked down cells were treated with doxorubicin in the presence or absence of cAMP-elevating agents. p53 protein level and apoptosis were assessed by western blot analysis and flow cytometry, respectively. p53 protein expression was reduced in cells treated with combination of cAMP-elevating agents and doxorubicin in contrast to cells treated with doxorubicin alone even in CREB-knocked down cells. Apoptosis assay showed that the cAMP-elevating agents decreased doxorubicin-induced apoptosis in CREB-knocked down and control cells. Although, CREB plays a particularly important role in cAMP signaling pathway our data suggest that CREB does not mediate the inhibitory effect of cAMP on doxorubicin-induced apoptosis and p53 accumulation in BCP-ALL NALM-6 cells.


Change in settings for early-season influenza vaccination among US adults, 2012 to 2013.
Vaccination in non-medical settings is recommended as a strategy to increase access to seasonal influenza vaccine. To evaluate change in early-season influenza vaccination setting, we analyzed data from the National Internet Flu Survey. Bivariate comparison of respondent characteristics by location of vaccination was assessed using chi-square tests. Multinomial logistic regression was performed to compare the predicted probability of being vaccinated in medical, retail, and mobile settings in 2012 vs 2013. In both 2012 and 2013, vaccination in medical settings was more likely among elderly adults, those with chronic conditions, and adults with a high school education or less. Adults 18-64 without a chronic condition had a lower probability of vaccination in the medical setting, and higher probability of vaccination in a retail or mobile setting, in 2013 compared to 2012. Adults 18-64 with a chronic condition had no change in their location of flu vaccination. Elderly adults had a lower probability of vaccination in the medical setting, and higher probability of vaccination in a retail setting, in 2013 compared to 2012. Non-medical settings continue to play an increasing role in influenza vaccination of adults, particularly for adults without a chronic condition and elderly adults. Retail and mobile settings should continue to be viewed as important mechanisms to ensure broad access to influenza vaccination.


Context-dependent effects of parental effort on malaria infection in a wild bird population, and their role in reproductive trade-offs.
Although trade-offs between reproductive effort and other fitness components are frequently documented in wild populations, the underlying physiological mechanisms remain poorly understood. Parasitism has been suggested to mediate reproductive trade-offs, yet only a limited number of parasite taxa have been studied, and reproductive effort-induced changes in parasitism are rarely linked to trade-offs observed in the same population. We conducted a brood size manipulation experiment in blue tits (Cyanistes caeruleus) infected with malaria (Plasmodium) parasites, and used quantitative PCR to measure changes in parasitaemia. In one of two years investigated, parasitaemia increased as a result of brood enlargement, and was also positively associated with two other indicators of reproductive effort: clutch size and single parenthood. These associations between both experimental and naturally varying reproductive effort and parasitaemia suggest that immune control of chronic malaria infections can be compromised when parents are working hard. Brood size manipulation significantly affected the number of independent offspring produced, which was maximised when brood size was unchanged. Moreover, when parents were infected with one of two common Plasmodium species, the shape of this trade-off curve was more pronounced, suggesting that parasitic infection may exacerbate the trade-off between quantity and quality of offspring. Although the involvement of parasites in survival costs of reproduction has received much attention, these results suggest their role in other commonly documented reproductive trade-offs, such as that between number and quality of offspring, warrants further study.


Optimal robust non-unique probe selection using Integer Linear Programming.
Besides their prevalent use for analyzing gene expression, microarrays are an efficient tool for biological, medical and industrial applications due to their ability to assess the presence or absence of biological agents, the targets, in a sample. Given a collection of genetic sequences of targets one faces the challenge of finding short oligonucleotides, the probes, which allow detection of targets in a sample. Each hybridization experiment determines whether the probe binds to its corresponding sequence in the target. Depending on the problem, the experiments are conducted using either unique or non-unique probes and usually assume that only one target is present in the sample. The problem at hand is to compute a design, i.e. a minimal set of probes that allows to infer the targets in the sample from the result of the hybridization experiment. If we allow to test for more than one target in the sample, the design of the probe set becomes difficult in the case of non-unique probes. Building upon previous work on group testing for microarrays, we describe the first approach to select a minimal probe set for the case of non-unique probes in the presence of a small number of multiple targets in the sample. The approach is based on an ILP formulation and a branch-and-cut algorithm. Our preliminary implementation greatly reduces the number of probes needed while preserving the decoding capabilities. http://www.inf.fu-berlin.de/inst/ag-bio


Characterization of a KCNQ1/KVLQT1 polymorphism in Asian families with LQT2: implications for genetic testing.
Congenital long QT syndrome (LQTS) is a genetic disease that predisposes affected individuals to arrhythmias, syncope, and sudden death. Mutations in several ion channel genes have been discovered in different families with LQTS: KCNQ1 (KVLQT1, LQT1), KCNH2 (HERG, LQT2), SCN5A (LQT3), KCNE1 (minK, LQT5), and KCNE2 (MiRP1, LQT6). Previously, the P448R-KVLQT1 missense mutation has been reported as an LQT1-causing mutation. In this report, we demonstrate the presence of the P448R polymorphism in two, unrelated Chinese LQTS families. Although absent from 500 reference alleles derived from 150 white and 100 African-American subjects, P448R was present in 14% of healthy Chinese volunteers. Given the inconsistencies between the genotype (LQT1) and clinical phenotype (LQT2) in our two LQTS families, together with the finding that the P448R appears to be a common, ethnic-specific polymorphism, mutational analysis was extended to the other LQTS-causing genes resulting in the identification of distinct HERG missense mutations in each of these two families. Heterologous expression of P448R-KVLQT1 yielded normal, wild-type (WT) currents. In contrast, the two unique HERG mutations resulted in dominant-negative suppression of the WT HERG channel. Our study has profound implications for those engaged in genetic research. Importantly, one child of the original proband was initially diagnosed with LQT1 based upon the presence of P448R-KVLQT1 and was treated with beta-blockers. However, he did not possess the subsequently determined LQT2-causing mutation. On the other hand, his untreated P448R-negative brother harbored the true, disease-causing HERG mutation. These findings underscore the importance of distinguishing channel polymorphisms from mutations pathogenic for LQTS and emphasize the importance of using appropriate ethnically matched controls in the genotypic analysis of LQTS.


From radiotherapy to Targeted therapy: 20 years in the management of non-small-cell lung cancer.
Non-small-cell lung cancer (NSCLC) is the leading cause of cancer death worldwide. Before 1980, radiotherapy was considered the only real recourse in advanced disease. In 1995, a landmark meta-analysis of trials conducted in the 1980s and early 1990s demonstrated a survival benefit with platinum-based chemotherapy. Newer chemotherapy agents and improved supportive care measures have allowed more patients to benefit from chemotherapy with reduced toxicity. Concurrent platinum-based chemotherapy and radiotherapy has improved the survival in stage III disease, and recently chemotherapy has also demonstrated improved survival in resected early-stage disease. The majority of patients still present with advanced unresectable disease for whom the prognosis remains poor, but for key subpopulations the outlook has improved markedly since the emergence of targeted therapies directed against the epidermal growth factor receptor and vascular endothelial growth factor receptor pathways. Patient selection and the incorporation of targeted therapies with cytotoxic chemotherapy are the focus of many ongoing studies, and there is an abundance of new agents undergoing clinical trials. Together, these developments have moved us away from the nihilism of 20 years ago into an era of unprecedented optimism in taking on the many remaining challenges of managing NSCLC in the 21st century.


Endovascular treatment of ruptured posterior circulation aneurysms using electrolytically detachable coils.
The authors report their experience using electrolytically detachable coils for the treatment of ruptured posterior circulation aneurysms. Twenty-six patients with 28 posterior circulation aneurysms were treated. All patients were referred for endovascular treatment by experienced vascular neurosurgeons. Patients underwent follow-up angiography immediately after treatment, 1 to 6 weeks posttreatment, and 6 months posttreatment. Six-month follow-up angiograms obtained in 19 patients with 20 aneurysms demonstrated that 18 (90%) of the 20 aneurysms were 99 to 100% occluded, one aneurysm (5%) was approximately 90% occluded, and one aneurysm (5%) was approximately 75% occluded. The patient with the aneurysm that was approximately 75% occluded needed additional treatment, consisting of parent artery balloon occlusion, and was considered a treatment failure (3.8% of patients). There was one treatment-associated mortality (3.8%) but no treatment-associated serious neurological or nonneurological morbidity in the patient group. There was no recurrent aneurysm rupture during treatment or during the mean 27-month follow-up period. Endovascular treatment of ruptured posterior circulation aneurysms with electrolytically detachable coils can be accomplished with low morbidity and mortality rates. The primary goal of treatment--preventing recurrent aneurysm--can be achieved over the short term. Endovascular coil occlusion will play an important role in the treatment of ruptured posterior circulation aneurysms, particularly if long-term efficacy in preventing recurrent aneurysm hemorrhage can be documented.


Food prices and marketing margins during the COVID-19 pandemic: Evidence from vegetable value chains in Ethiopia.
It is widely feared that the shock of the COVID-19 pandemic will lead to a significant worsening of the food security situation in low and middle-income countries. One reason for this is the disruption of food marketing systems and subsequent changes in farm and consumer prices. Based on primary data in Ethiopia collected just before the start and a few months into the pandemic, we assess changes in farm and consumer prices of four major vegetables and the contribution of different segments of the rural-urban value chain in urban retail price formation. We find large, but heterogeneous, price changes for different vegetables with relatively larger changes seen at the farm level, compared to the consumer level, leading to winners and losers among local vegetable farmers due to pandemic-related trade disruptions. We further note that despite substantial hurdles in domestic trade reported by most value chain agents, increases in marketing-and especially transportation-costs have not been the major contributor to overall changes in retail prices. Marketing margins even declined for half of the vegetables studied. The relatively small changes in marketing margins overall indicate the resilience of these domestic value chains during the pandemic in Ethiopia.


First reports evaluating the effectiveness of strategies for preventing violence: firearms laws. Findings from the Task Force on Community Preventive Services.
During 2000-2002, the Task Force on Community Preventive Services (the Task Force), an independent nonfederal task force, conducted a systematic review of scientific evidence regarding the effectiveness of firearms laws in preventing violence, including violent crimes, suicide, and unintentional injury. The following laws were evaluated: bans on specified firearms or ammunition, restrictions on firearm acquisition, waiting periods for firearm acquisition, firearm registration and licensing of firearm owners, "shall issue" concealed weapon carry laws, child access prevention laws, zero tolerance laws for firearms in schools, and combinations of firearms laws. The Task Force found insufficient evidence to determine the effectiveness of any of the firearms laws or combinations of laws reviewed on violent outcomes. (Note that insufficient evidence to determine effectiveness should not be interpreted as evidence of ineffectiveness.) This report briefly describes how the reviews were conducted, summarizes the Task Force findings, and provides information regarding needs for future research.


Life Expectancy after Inpatient Rehabilitation for Traumatic Brain Injury in the United States.
This study characterized life expectancy after traumatic brain injury (TBI). The TBI Model Systems (TBIMS) National Database (NDB) was weighted to represent those ≥16 years of age completing inpatient rehabilitation for TBI in the United States (US) between 2001 and 2010. Analyses included Standardized Mortality Ratios (SMRs), Cox regression, and life expectancy. The US mortality rates by age, sex, race, and cause of death for 2005 and 2010 were used for comparison purposes. Results indicated that a total of 1325 deaths occurred in the weighted cohort of 6913 individuals. Individuals with TBI were 2.23 times more likely to die than individuals of comparable age, sex, and race in the general population, with a reduced average life expectancy of 9 years. Independent risk factors for death were: older age, male gender, less-than-high school education, previously married at injury, not employed at injury, more recent year of injury, fall-related TBI, not discharged home after rehabilitation, less functional independence, and greater disability. Individuals with TBI were at greatest risk of death from seizures; accidental poisonings; sepsis; aspiration pneumonia; respiratory, mental/behavioral, or nervous system conditions; and other external causes of injury and poisoning, compared with individuals in the general population of similar age, gender, and race. This study confirms prior life expectancy study findings, and provides evidence that the TBIMS NDB is representative of the larger population of adults receiving inpatient rehabilitation for TBI in the US. There is an increased risk of death for individuals with TBI requiring inpatient rehabilitation.


Strength and anaerobic responses of elite young female sprint and distance runners.
The purpose of this study was to assess the strength and anaerobic responses of elite young female runners, in order to determine the relationship of these characteristics to age and high proficiency in sprint events. Thirty-one national junior-level sprint and middle distance runners were evaluated for isokinetic peak torque for leg extension at velocities ranging for 30 degrees to 300 degrees X s-1, anaerobic power and capacity based on the Wingate test, and body composition based on underwater weighing. Age-related and event-related differences were noted for both the peak torque and anaerobic response measures. Peak torques were greater for the older (greater than 11.67 yr) runners and greater for the sprinters when measured at the higher testing velocities. Anaerobic power and capacity were also greater for the older runners, but event-related differences only appeared when anaerobic power was adjusted for fat-free weight (adjusted anaerobic power was greater in the older sprinters). These results indicate that both age and fat-free weight influence strength and anaerobic responses in young female runners and that exceptional levels of such characteristics only appear for sprinters within the upper age groups of youth competition.


Development and validation of the short-form Adolescent Health Promotion Scale.
Health-promoting lifestyle choices of adolescents are closely related to current and subsequent health status. However, parsimonious yet reliable and valid screening tools are scarce. The original 40-item adolescent health promotion (AHP) scale was developed by our research team and has been applied to measure adolescent health-promoting behaviors worldwide. The aim of our study was to examine the psychometric properties of a newly developed short-form version of the AHP (AHP-SF) including tests of its reliability and validity. The study was conducted in nine middle and high schools in southern Taiwan. Participants were 814 adolescents randomly divided into two subgroups with equal size and homogeneity of baseline characteristics. The first subsample (calibration sample) was used to modify and shorten the factorial model while the second subsample (validation sample) was utilized to validate the result obtained from the first one. The psychometric testing of the AHP-SF included internal reliability of McDonald's omega and Cronbach's alpha, convergent validity, discriminant validity, and construct validity with confirmatory factor analysis (CFA). The results of the CFA supported a six-factor model and 21 items were retained in the AHP-SF with acceptable model fit. For the discriminant validity test, results indicated that adolescents with lower AHP-SF scores were more likely to be overweight or obese, skip breakfast, and spend more time watching TV and playing computer games. The AHP-SF also showed excellent internal consistency with a McDonald's omega of 0.904 (Cronbach's alpha 0.905) in the calibration group. The current findings suggest that the AHP-SF is a valid and reliable instrument for the evaluation of adolescent health-promoting behaviors. Primary health care providers and clinicians can use the AHP-SF to assess these behaviors and evaluate the outcome of health promotion programs in the adolescent population.


Clinical Reasoning Tasks and Resident Physicians: What Do They Reason About?
A framework of clinical reasoning tasks thought to occur in a clinical encounter was recently developed. It proposes that diagnostic and therapeutic reasoning comprise 24 tasks. The authors of this current study used this framework to investigate what internal medicine residents reason about when they approach straightforward clinical cases. Participants viewed three video-recorded clinical encounters portraying common diagnoses. After each video, participants completed a post encounter form and think-aloud protocol. Two authors analyzed transcripts from the think-aloud protocols using a constant comparative approach. They conducted iterative coding of the utterances, classifying each according to the framework of clinical reasoning tasks. They evaluated the type, number, and sequence of tasks the residents used. Ten residents participated in the study in 2013-2014. Across all three cases, the residents employed 14 clinical reasoning tasks. Nearly all coded tasks were associated with framing the encounter or diagnosis. The order in which residents used specific tasks varied. The average number of tasks used per case was as follows: Case 1, 4.4 (range 1-10); Case 2, 4.6 (range 1-6); and Case 3, 4.7 (range 1-7). The residents used some tasks repeatedly; the average number of task utterances was 11.6, 13.2, and 14.7 for, respectively, Case 1, 2, and 3. Results suggest that the use of clinical reasoning tasks occurs in a varied, not sequential, process. The authors provide suggestions for strengthening the framework to more fully encompass the spectrum of reasoning tasks that occur in residents' clinical encounters.


Protective and ameliorative effect of sea buckthorn leaf extract supplementation on lead induced hemato-biochemical alterations in Wistar rats.
To evaluate the protective and ameliorative effect of aqueous sea buckthorn leaf extract (SLE) on hemato-biochemical profile in lead intoxicated Wistar rats. An experiment was conducted for 60 days. 36 adult male Wistar rats with a mean body weight of 177.8±12.6 g were divided into five groups and were subjected to various daily oral treatment regimens. Group I served as a negative control receiving only feed and water, Group II (positive control for lead) received lead acetate at 250 ppm in drinking water, and Group III (positive control for SLE) received SLE at 100 mg/kg b.wt. Animals in Group IV received a combination of lead acetate at 250 ppm in drinking water for the first 45 days and SLE at 100 mg/kg b.wt. throughout the experimental period of 60-day, and in Group V for the last 15 days of the trial after the administration of lead acetate until the first 45 days of the trial to study the protective and ameliorating effects of SLE, respectively. Blood samples were collected from retro-orbital fossa of each rat on 0th, 45th, and 60th day of the experiment for hemato-biochemical analysis including hemoglobin (Hb), packed cell volume (PCV), serum total protein, albumin, globulin, albumin:globulin ratio, cholesterol, urea, and creatinine. Significantly (p<0.01) lower levels of serum total proteins and albumin, and a significantly (p<0.01) higher serum cholesterol, urea and creatinine levels were observed in Group II (lead intoxicated group) in comparison to Group I (negative control). Administration of SLE at 100 mg/kg body wt. to lead intoxicated Wistar rats resulted in normalization of almost all the biochemical parameters studied in both the treatment Groups, i.e., IV and V (protective and ameliorative). However, the effects were more pronounced in the protective group. No effects of SLE supplementation were observed on Hb levels. PCV levels improved in protective groups, but no effect was observed in ameliorative group in comparison to lead intoxicated groups. SLE administration at 100 mg/kg b.wt. to lead intoxicated Wistar rats may be used to protect/ameliorate lead induced biochemical alterations in Wistar rats.


Bat-cave catchment areas: using stable isotopes (deltaD) to determine the probable origins of hibernating bats.
The application of stable hydrogen isotope (deltaD) techniques has swiftly advanced our understanding of animal movements, but this progression is dominated by studies of birds and relatively long-distance, north-south migrants. This dominance reflects the challenge of incorporating multiple sources of error into geographic assignments and the nature of spatially explicit deltaD models, which possess greater latitudinal than longitudinal resolution. However, recent progress in likelihood-based assignments that incorporate multiple sources of isotopic error and Bayesian approaches that include additional sources of information may advance finer-scale understanding of animal movements. We develop a stable-isotope method for determining probable origins of bats within hibernacula and show that this method produces spatially explicit, continuous assignments with regional resolution. We outline how these assignments can be used to infer hibernacula connectivity, an application that could inform spatial modeling of white-nose syndrome. Additionally, estimates of seasonal and annual flight distances for many cave-dwelling bat species can be derived from this approach. We also discuss how this application can be used in general to provide insights into variable migratory and foraging strategies within bat populations.


Dietary influences on tissue concentrations of phytanic acid and AMACR expression in the benign human prostate.
Alpha-methylacyl-CoA racemase (AMACR) is an enzyme involved in fatty acid metabolism that is markedly over-expressed in virtually all prostate cancers (PCa), relative to benign tissue. One of AMACR's primary substrates, phytanic acid, is derived predominately from red meat and dairy product consumption. Epidemiological evidence suggests links between dairy/red meat intake, as well as phytanic acid levels, and elevated PCa risk. This study investigates the relationships among dietary intake, serum and tissue concentrations of phytanic acid, and AMACR expression (mRNA and protein) in the histologically benign human prostate. Men undergoing radical prostatectomy for the treatment of localized disease provided a food frequency questionnaire (n = 68), fasting blood (n = 35), benign fresh frozen prostate tissue (n = 26), and formalin-fixed paraffin-embedded (FFPE) sections (n = 67). Serum and tissue phytanic acid concentrations were obtained by gas chromatography-mass spectrometry. We extracted RNA from epithelial cells using laser capture microdissection and quantified mRNA expression of AMACR and other genes involved in the peroxisomal phytanic acid metabolism pathway via qRT-PCR. Immunohistochemistry for AMACR was performed on FFPE sections and subsequently quantified via digital image analysis. Associations between diet, serum, and tissue phytanic acid levels, as well as AMACR and other gene expression levels were assessed by partial Spearman correlation coefficients. High-fat dairy intake was the strongest predictor of circulating phytanic acid concentrations (r = 0.35, P = 0.04). Tissue phytanic acid concentrations were not associated with any dietary sources and were only weakly correlated with serum levels (r = 0.29, P = 0.15). AMACR gene expression was not associated with serum phytanic acid (r = 0.13, P = 0.47), prostatic phytanic acid concentrations (r = 0.03, P = 0.88), or AMACR protein expression (r = -0.16, P = 0.20). Our data underscore the complexity of the relationship between AMACR and its substrates and do not support the unifying hypothesis that excess levels of dietary phytanic acid are responsible for both the overexpression of AMACR in prostate cancer and the potential association between PCa risk and intake of dairy foods and red meat.


A clear and present danger: tick-borne diseases in Europe.
Ticks can transmit a variety of viruses, bacteria or parasites that can cause serious infections or conditions in humans and animals. While tick-borne diseases are becoming an increasing and serious problem in Europe, tick-borne diseases are also responsible for major depressions in livestock production and mortality in sub-Saharan Africa, Latin America and Asia. This review will focus on the most important circulating tick-transmitted pathogens in Europe (Borrelia spp., Anaplasma phagocytophilum, Babesia spp., tick-borne encephalitis virus, Rickettsia spp. and Crimean-Congo hemorrhagic fever virus).


How the global layout of the mask influences masking strength.
In visual backward masking, the perception of a target is influenced by a trailing mask. Masking is usually explained by local interactions between the target and the mask representations. However, recently it has been shown that the global spatial layout of the mask rather than its local structure determines masking strength (Hermens & Herzog, 2007). Here, we varied the mask layout by spatial, luminance, and temporal cues. We presented a vernier target followed by a mask with 25 elements. Performance deteriorated when the length of the two mask elements neighboring the target vernier was doubled. However, when the length of every second mask element was doubled, performance improved. When the luminance of the neighboring elements was doubled, performance also deteriorated but no improvement in performance was observed when every second element had a double luminance. For temporal manipulations, a complex nonmonotonic masking function was observed. Hence, changes in the mask layout by spatial, luminance, and temporal cues lead to highly different results.


Failure of digital echocardiography to accurately diagnose intracardiac shunts.
Intravenous agitated saline injection during transthoracic echocardiography assists in the detection of right to left intracardiac and intrapulmonary shunts. Whether digital echocardiography offers comparable sensitivity and specificity to analog tape recording to assess shunts is unknown. Technical differences between methods could lead to significant differences in shunt detection. Agitated saline was injected intravenously at rest and with Valsalva in 189 consecutive patient studies (406 injections). Echocardiographers assessed presence and degree of left ventricle contrast on simultaneously recorded analog tape and digital echocardiography images in blinded fashion. Digital echocardiography had low overall sensitivity (rest 0.50, valsalva 0.63, late 0.39) compared to analog tape. Longer clip lengths improved sensitivity for detection of late contrast passage (rest 0.50, valsalva 0.67, late 0.46). Digital echocardiography saline contrast studies have poor sensitivity for assessment of intracardiac shunts versus analog tape, and increasing clip length only modestly increases sensitivity. Joint Photographic Experts Group digital compression losses may be an important cause of failure to detect intracardiac shunts, including patent foramen ovale.


Effects of acupuncture on the heart rate variability, cortisol levels and behavioural response induced by thunder sound in beagles.
Sound stimuli such as fireworks, firearms, and claps of thunder have been used as a stress reactivity model for dogs. Acupuncture has been widely used to treat and prevent physiological and behavioural disorders induced by stress. Our study aims to evaluate the effects of acupuncture on cardiac autonomic modulation (heart rate variability - HRV), behavioural (reactivity) and endocrine (cortisol levels) responses in dogs exposed to sounds of thunder. Twenty-four laboratory beagles (12 males and 12 females, 1-6years old) with no history of phobia to thunder were subjected to a sound stimulus that consisted of a standardized recording of thunder over a 150s period with a maximum intensity of 103-104dB. Before the sound, the dogs underwent a 20-minute session of needle insertion at acupuncture points Yintang, GV20, HT7, PC6 and ST36 (ACUP), in non-points (NP) or left undisturbed (CTL). Cardiac intervals were recorded using a frequency meter (RS 800cx, Polar, Kempele, Finland) to evaluate the HRV, and the data were later analysed using CardioSeries v2.4.1 software. Acupuncture (ACUP) changed the sympathovagal balance with a shift towards parasympathetic modulation, reducing the prompt sound-induced increase in LF/HF (low frequency/high frequency) ratio and in the power of the LF band of the cardiac interval spectrum, and decreased the power of the HF band of the cardiac interval spectrum (p<0.05); however there was no change in the heart rate. Acupuncture reduced the behavioural response induced by sounds of thunder (when all behavioural parameters were considered together) and the behaviours hiding, restlessness, bolting and running around (when the parameters were analysed separately (p<0.05). There were no changes in cortisol levels due to the sound stimulus or acupuncture. Our results demonstrate that a session of acupuncture prior to sound stimulus can reduce cardiac autonomic and behavioural responses, without changing cortisol levels in beagles.


Adaptive responses of the maternal hypothalamic-pituitary-adrenal axis during pregnancy and lactation.
Over the past 40 years, it has been recognised that the maternal hypothalamic-pituitary-adrenal (HPA) axis undergoes adaptations through pregnancy and lactation that might contribute to avoidance of adverse effects of stress on the mother and offspring. The extent of the global adaptations in the HPA axis has been revealed and the underlying mechanisms investigated within the last 20 years. Both basal, including the circadian rhythm, and stress-induced adrenocorticotrophic hormone and glucocorticoid secretory patterns are altered. Throughout most of pregnancy, and in lactation, these changes predominantly reflect reduced drive by the corticotropin-releasing factor (CRF) neurones in the parvocellular paraventricular nucleus (pPVN). An accompanying profound attenuation of HPA axis responses to a wide variety of psychological and physical stressors emerges after mid-pregnancy and persists until the end of lactation. Central to this suppression of stress responsiveness is reduced activation of the pPVN CRF neurones. This is consequent on the reduced effectiveness of the stimulation of brainstem afferents to these CRF neurones (for physical stressors) and of altered processing by limbic structures (for emotional stressors). The mechanism of reduced CRF neurone responses to physical stressors in pregnancy is the suppression of noradrenaline release in the PVN by an up-regulated endogenous opioid mechanism, which is induced by neuroactive steroid produced from progesterone. By contrast, in lactation suckling the young provides a neural stimulus that dampens the HPA axis circadian rhythm and reduces stress responses. Reduced noradrenergic input activity is involved in reduced stress responses in lactation, although central prolactin action also appears important. Such adaptations limit the adverse effects of excess glucocorticoid exposure on the foetus(es) and facilitate appropriate metabolic and immune responses.


The prognostic value of vitamin D level for in-hospital mortality in patients with acute pulmonary embolism.
The aim of this study was to investigate the prognostic value of the serum vitamin D (Vit-D) level on admission in patients with acute pulmonary embolism (APE) to determine in-hospital mortality. Ninety-nine patients who were diagnosed with APE between January 2015 and January 2018 and had a record of an admission serum Vit-D level were enrolled in the study. The serum Vit-D level was measured using an immune-based assay in all cases. The primary outcome of the study was in-hospital all-cause mortality. The study population was divided into 2 groups according to the median value of serum Vit-D level: Vit-D level ≤7.36 ng/mL in 49 patients and Vit-D level >7.36 ng/mL in 50 patients. The patients with a serum Vit-D level ≤7.36 ng/mL had a higher of incidence of in-hospital death compared with those whose serum Vit-D level was >7.36 ng/mL (6 [12.2%] vs. 1 [2%]; p=0.048). In Cox regression analysis, the serum Vit-D level (Hazard ratio: 0.82, 95% confidence interval: 0.68-0.98; p=0.043) was found to be independently associated with in-hospital mortality. The optimal value of serum Vit-D level for the prediction of in-hospital mortality was ≤6.47 ng/mL, with a sensitivity of 71.4% and a specificity 86.9% (area under the curve: 0.81, 95% CI: 0.72-0.88; p=0.004). The findings demonstrated that the serum Vit-D level on admission may be an independent predictor for in-hospital mortality in patients with APE.


Antigenic properties of cultured tumor cell lines derived from spleens of Friend virus-infected BALB/c and BALB/c-H-2b mice.
BALB/c-H-2b (BALB.B) mice are less susceptible to the Friend virus (FV) disease syndrome than congenic BALB/c (H-2d) mice, and spleen cells from FV-infected BALB.B mice are markedly less tumorigenic on transplantation to syngeneic hosts than those from FV-infected BALB/c mice. For these reasons we investigated the expression of FV-associated cell surface antigens on cultured, FV-trnasformed cell lines of BALB.B and BALB/c origin. Both cell lines induced transplantation immunity in syngeneic hosts toward further implantations of the same tumor, BALB.B cells being significantly more potent in this respect than BALB/c cells. BALB.B tumor cells, which produce complete, infectious FV, expressed both the cell surface antigen, FMR (corresponding to the cytotoxic antibodies in anti-FV antisera), and virus envelope antigen (VEA, corresponding to the virus-neutralizing antibodies in the anti-FV antisera). BALB/c tumor cells, on the other hand, which are FV-nonproducers, expressed no FMR antigen, but did express VEA on their surfaces for at least 100 passages in culture. These cells could induce FV-neutralizing but not cytotoxic anti-FMR antibodies when used to immunize syngeneic hosts. The absence of FMR antigen may be the basis for the reduced capacity of BALB/c tumor cells, by comparison with BALB.B tumor cells, to induce transplantation immunity. After about the 125th serial transfer in culture, BALB/c tumor cells spontaneously ceased to express VEA and simultaneously became very weak inducers of transplantation immunity in BALB/c hosts. This loss of VEA did not stem from the loss of either the spleen focus-forming virus or the helper virus genomes from these cells, since both viruses could still be recovered from the cell line.


Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield: the use of a respiratory strain as a microbial cell factory.
Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass properties do lead to increased volumetric yields without the need to resort to complex control or cultivation schemes. This is anticipated to be of particular value in the production of challenging targets such as membrane proteins.


Identification of an effective early signaling signature during neo-vasculogenesis in vivo by ex vivo proteomic profiling.
Therapeutic neo-vasculogenesis in vivo can be achieved by the co-transplantation of human endothelial colony-forming progenitor cells (ECFCs) with mesenchymal stem/progenitor cells (MSPCs). The underlying mechanism is not completely understood thus hampering the development of novel stem cell therapies. We hypothesized that proteomic profiling could be used to retrieve the in vivo signaling signature during the initial phase of human neo-vasculogenesis. ECFCs and MSPCs were therefore either transplanted alone or co-transplanted subcutaneously into immune deficient mice. Early cell signaling, occurring within the first 24 hours in vivo, was analyzed using antibody microarray proteomic profiling. Vessel formation and persistence were verified in parallel transplants for up to 24 weeks. Proteomic analysis revealed significant alteration of regulatory components including caspases, calcium/calmodulin-dependent protein kinase, DNA protein kinase, human ErbB2 receptor-tyrosine kinase as well as mitogen-activated protein kinases. Caspase-4 was selected from array results as one therapeutic candidate for targeting vascular network formation in vitro as well as modulating therapeutic vasculogenesis in vivo. As a proof-of-principle, caspase-4 and general caspase-blocking led to diminished endothelial network formation in vitro and significantly decreased vasculogenesis in vivo. Proteomic profiling ex vivo thus unraveled a signaling signature which can be used for target selection to modulate neo-vasculogenesis in vivo.


Use of tamoxifen and raloxifene for breast cancer chemoprevention in 2010.
Two selective estrogen receptor modulators, tamoxifen and raloxifene, have been shown in randomized clinical trials to reduce the risk of developing primary invasive breast cancer in high-risk women. In 1998, the U.S. Food and Drug Administration (FDA) used these studies as a basis for approving tamoxifen for primary breast chemoprevention in both premenopausal and postmenopausal women at high risk. In 2007, the FDA approved raloxifene for primary breast cancer chemoprevention for postmenopausal women. Data from the year 2010 National Health Interview Survey were analyzed to estimate the prevalence of tamoxifen and raloxifene use for chemoprevention of primary breast cancers among U.S. women. Prevalence of use of chemopreventive agents for primary tumors was 20,598 (95 % CI, 518-114,864) for U.S. women aged 35-79 for tamoxifen. Prevalence was 96,890 (95 % CI, 41,277-192,391) for U.S. women aged 50-79 for raloxifene. Use of tamoxifen and raloxifene for prevention of primary breast cancers continues to be low. In 2010, women reporting medication use for breast cancer chemoprevention were primarily using the more recently FDA approved drug raloxifene. Multiple possible explanations for the low use exist, including lack of awareness and/or concern about side effects among primary care physicians and patients.


Fungal infections in intestinal and multivisceral transplant recipients.
Patients who undergo intestinal and multivisceral transplantation are at increased risk for infectious complications. Fungal infections are major causes of morbidity and mortality in these patients. The current review highlights key diagnostic and management issues in this population. Invasive infections caused by Candida spp. remain the most common invasive fungal infections in intestinal and multivisceral transplant recipients. Aspergillus is an emerging pathogen but data are limited to case reports or case series. Other fungi including the mucorales, Cryptococcus and endemic mycoses are emerging pathogens but data regarding incidence and timing of disease in intestinal and multivisceral transplant recipients are lacking. Invasive candidiasis is the most common fungal infection in patients with intestinal and multivisceral transplants. Experience for diagnosis and management comes from case series and single centers. Diagnosis and management of infections caused by other pathogens such as Aspergillus, Cryptococcus, Mucor, and endemic mycoses is usually extrapolated from other solid organ transplant recipients.