Relationship Reciprocation Modulates Resource Allocation in Adolescent Social Networks: Developmental Effects.
Adolescence is characterized as a period of social reorientation toward peer relationships, entailing the emergence of sophisticated social abilities. Two studies (Study 1: N = 42, ages 13-17; Study 2: N = 81, ages 13-16) investigated age group differences in the impact of relationship reciprocation within school-based social networks on an experimental measure of cooperation behavior. Results suggest development between mid- and late adolescence in the extent to which reciprocation of social ties predicted resource allocation. With increasing age group, investment decisions increasingly reflected the degree to which peers reciprocated feelings of friendship. This result may reflect social-cognitive development, which could facilitate the ability to navigate an increasingly complex social world in adolescence and promote positive and enduring relationships into adulthood.


Education Against Tobacco (EAT): a quasi-experimental prospective evaluation of a programme for preventing smoking in secondary schools delivered by medical students: a study protocol.
A survey conducted by the German Federal Centre for Health Education in 2012 showed that 35.2% of all young adults (18-25 years) and 12.0% of all adolescents (12-17 years) in Germany are regular cigarette smokers. Most smoked their first cigarette in early adolescence. We recently reported a significantly positive short-term effect of a physician-delivered school-based smoking prevention programme on the smoking behaviour of schoolchildren in Germany. However, physician-based programmes are usually very expensive. Therefore, we will evaluate and optimise Education against Tobacco (EAT), a widespread, low-cost programme delivered by about 400 medical students from 16 universities in Germany. A prospective quasi-experimental study design with two measurements at baseline (t1) and 6 months post-intervention (t2) to investigate an intervention in 10-15-year-olds in grades 6-8 at German secondary schools. The intervention programme consists of two 60-min school-based medical-student-delivered modules with (module 1) and without the involvement of patients with tobacco-related diseases and control groups (no intervention). The study questionnaire measuring smoking status (water pipe and cigarette smoking), smoking-related cognitions, and gender, social and cultural aspects was designed and pre-tested in advance. The primary end point is the prevalence of smokers and non-smokers in the two study arms at 6 months after the intervention. The percentage of former smokers and new smokers in the two groups and the measures of smoking behaviour will be studied as secondary outcome measures. In accordance with Good Epidemiologic Practice (GEP) guidelines, the study protocol was submitted for approval by the responsible ethics committee, which decided that the study does not need ethical approval (Goethe University, Frankfurt-Main, Germany). Findings will be disseminated in peer-reviewed journals, at conferences, within our scientific advisory board and through medical students within the EAT project.


Hydroxyapatite-coated tibial implants compared with cemented tibial fixation in primary total knee arthroplasty. A randomized trial of outcomes at five years.
Although excellent long-term results have been reported with cemented tibial fixation, cementless fixation as a means to improve the longevity of total knee prostheses continues to be of interest to clinicians. The purpose of this study was to compare outcomes between cementless tibial fixation with hydroxyapatite and cemented tibial fixation in the first five years following primary total knee arthroplasty. We performed a prospective, randomized clinical trial that included eighty-one patients with noninflammatory knee arthritis who underwent primary total knee arthroplasty when they were less than seventy years of age. The subjects were randomized at the time of surgery to be treated with either cementless tibial fixation with hydroxyapatite or cemented tibial fixation. Evaluations were performed preoperatively and at six months, one year, and five years postoperatively by a physical therapist who was blinded to group allocation. Self-reported pain and function, the primary outcomes, were measured with the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the RAND 36-Item Health Survey (RAND-36). Complications and revision rates were determined through a review of hospital records and at each patient evaluation. The Knee Society radiographic score was used to evaluate plain radiographs at each assessment. Seventy subjects (86%) completed the five-year assessment. Slightly more pain was reported in the hydroxyapatite group at six months as measured with both the WOMAC and the RAND-36, a difference that disappeared by one year postoperatively. No differences were seen in function, radiographic findings, or complications. No subject required revision of the tibial prosthesis during the study. At five years postoperatively, there is no difference between cementless tibial fixation with hydroxyapatite and cemented tibial fixation in terms of self-reported pain, function, health-related quality of life, postoperative complications, or radiographic scores.


The public health roots of zoning: in search of active living's legal genealogy.
Improvements in the built environment and changes in land-use policy are promising approaches to increasing physical activity among a largely sedentary population. Opportunities for walking and cycling as part of daily life are important to increasing physical activity and improving health. Yet, local zoning codes and related land-use regulations have made it difficult to create vibrant, mixed-use neighborhoods with well-connected streets and more compact development-the infrastructure necessary to support healthier rates of walking and cycling for transportation. To better understand the dynamic nature of land-use law and policy, and how policymakers might accomplish zoning reform to encourage more physically active environments, this paper traces the public health roots of zoning through a family tree of land-use legal doctrines. Zoning and public health laws evolved from the same legal ancestors-the common law of public nuisance and the expansion of state police powers, both premised on protection of the public's health. When the U.S. Supreme Court approved zoning in the 1926 case of Ambler Realty v. Village of Euclid, it nominally recognized the health basis of zoning. But it went on to craft a new legal rationale focused more on protection of property rights and residential neighborhoods. Since Euclid, court decisions have given little consideration to the public health roots of zoning. Given an emerging body of research demonstrating the importance of walking-friendly environments and the deference shown by the courts to the passage of zoning laws, the courts are likely to support policymakers as they move to change zoning systems conceived long ago. Legal, historical, and policy rationales support the modernization of zoning and land use policies that allow sensible mixes of land uses. Mixed land uses make walking an attractive alternative to driving and support a more physically active and healthy citizenry.


Deep Characterization of the Microbiomes of Calophya spp. (Hemiptera: Calophyidae) Gall-Inducing Psyllids Reveals the Absence of Plant Pathogenic Bacteria and Three Dominant Endosymbionts.
Bacteria associated with sap-feeding insect herbivores include not only symbionts that may increase their hosts' fitness but also harmful plant pathogens. Calophya spp. gall-inducing psyllids (Hemiptera: Calophyidae) are being investigated for their potential as biological control agents of the noxious weed, Brazilian peppertree (Schinus terebinthifolia), in Florida. Although there are no examples of plant pathogen transmission by members of the family Calophyidae, several insects in the superfamily Psylloidea are known to transmit pathogenic bacteria in the genera Candidatus Liberibacter and Candidatus Phytoplasma. To determine whether Calophya spp. harbor potentially harmful plant pathogenic bacteria, we sequenced small subunit (SSU) ribosomal RNA (rRNA) gene amplicons generated from individuals from four Calophya spp. populations: All microbial SSU gene sequences fell into the bacterial domain, with 98-99% belonging to the Proteobacteria. The Calophya microbiomes contained a relatively simple community, with 49-79 operational taxonomic units (OTUs; 97%) detected, and only 5-8 OTUs with greater than 1% abundance. Candidatus Carsonella showed the highest relative abundance, with OTUs from this candidate genus representing between 51-65% of all recovered sequences. The next most abundant clade observed was an unclassified Enterobacteriacae group closely related to bacteria from the genera Buchnera and Blochmannia that ranged from 20-31% in relative abundance. Wolbachia populations were the third most abundant group and represented 7-27% of the diversity in microbial OTUs. No SSU rRNA gene sequences from putative pathogenic bacteria from the genera Ca. Liberibacter or Ca. Phytoplasma were detected in the microbiomes of the four Calophya populations. The probability that infected psyllids were present in our colonies, but were not sampled, was extremley low (1.39 x 10(-10)). As far as we are aware, our study is the first to characterize the microbiome of a candidate biological control agent, and coupled with previous work demonstrating a high degree of host specificity and absence of plant viruses, suggests that releasing Calophya spp. in United States poses minimal risk to non-target plants.


Abuse of Coricidin HBP cough & cold tablets: episodes recorded by a poison center.
Cases involving ingestion of a dextromethorphan-containing product recorded at a poison control center were studied. A retrospective review of all consultations involving the ingestion of Coricidin HBP Cough & Cold tablets recorded by the California Poison Control System was conducted for the period from January 1 to October 1, 2000. Computerized charts on the consultations were reviewed to obtain data on patient age and sex, number of tablets taken, reason for tablet ingestion, symptoms, treatment, disposition, and outcome. A total of 92 charts (for 92 patients) documenting Coricidin HBP Cough & Cold tablet ingestion were reviewed. The reason for tablet ingestion was classified as abuse in 65 patients (71%), a suicide attempt in 8 (9%), misuse in 1 (1%), malicious administration in 1 (1%), and normal use (but with an adverse drug reaction) in 1 (1%); 16 patients (17%) consumed the tablets for an unknown reason. The 92 patients comprised 42 males and 50 females. Among all patients, 78 (85%) were 13-17 years old, and among those classified as having abusive intent, 58 (89%) were in the same age range. The most commonly reported signs and symptoms associated with ingestion were tachycardia (50 patients), hypertension (29), lethargy (40), mydriasis (20), agitation (15), ataxia or dizziness (20), and vomiting (9). Sixty-one patients (66%) had some alteration in mental status. Fifty-six (61%) were treated in the emergency department; 11 (12%) were admitted. All patients recovered completely. Information on the ingestion of Coricidin HBP Cough & Cold tablets recorded at a poison control center indicated a high rate of abuse of the product among teenagers.


Rec8 phosphorylation by casein kinase 1 and Cdc7-Dbf4 kinase regulates cohesin cleavage by separase during meiosis.
During meiosis, two rounds of chromosome segregation after a single round of DNA replication produce haploid gametes from diploid precursors. At meiosis I, maternal and paternal kinetochores are pulled toward opposite poles, and chiasmata holding bivalent chromosomes together are resolved by cleavage of cohesin's alpha-kleisin subunit (Rec8) along chromosome arms. This creates dyad chromosomes containing a pair of chromatids joined solely by cohesin at centromeres that had resisted cleavage. The discovery that centromeric Rec8 is protected from separase during meiosis I by shugoshin/MEI-S332 proteins that bind PP2A phosphatase suggests that phosphorylation either of separase or cohesin may be necessary for Rec8 cleavage. We show here that multiple phosphorylation sites within Rec8 as well as two different kinases, casein kinase 1delta/epsilon (CK1delta/epsilon) and Dbf4-dependent Cdc7 kinase (DDK), are required for Rec8 cleavage and meiosis I nuclear division. Rec8 with phosphomimetic mutations is no longer protected from separase at centromeres and is cleaved even when the two kinases are inhibited. Our data suggest that PP2A protects centromeric cohesion by opposing CK1delta/epsilon- and DDK-dependent phosphorylation of Rec8.


Prices for Tobacco and Nontobacco Products in Pharmacies Versus Other Stores: Results From Retail Marketing Surveillance in California and in the United States.
To examine disparities in the price of tobacco and nontobacco products in pharmacies compared with other types of stores. We recorded the prices of Marlboro, Newport, the cheapest cigarettes, and bottled water in a random sample of licensed tobacco retailers (n = 579) in California in 2014. We collected comparable data from retailers (n = 2603) in school enrollment zones for representative samples of US 8th, 10th, and 12th graders in 2012. Ordinary least squares regressions modeled pretax prices as a function of store type and neighborhood demographics. In both studies, the cheapest cigarettes cost significantly less in pharmacies than other stores; the average estimated difference was $0.47 to $1.19 less in California. We observed similar patterns for premium-brand cigarettes. Conversely, bottled water cost significantly more in pharmacies than elsewhere. Newport cost less in areas with higher proportions of African Americans; other cigarette prices were related to neighborhood income and age. Neighborhood demographics were not related to water prices. Compared with other stores, pharmacies charged customers less for cigarettes and more for bottled water. State and local policies to promote tobacco-free pharmacies would eliminate an important source of discounted cigarettes.


Seventy-five years is an appropriate upper age limit for population-based mammography screening.
In this study, we assessed the results of the Dutch breast cancer screening programme for women aged 70-75, and discussed the current upper age limit of the women invited. We compared the main outcome parameters of the screening programme 1998-2000 for women aged 70-75, with those in women aged 50-69. Breast cancer detection rates were also compared with prediction from 2 variants of a simulation model of breast cancer screening, assuming the mean sojourn time, i.e., the duration of the preclinical detectable phase, to increase or not with age above the age of 65. The underlying idea is that an increase of the sojourn time with age will lead to a less favourable balance between screening benefits and harms from a certain age on. Of the 315,103 women (aged 70-75) invited, 65.6% participated. The attendance increased from 1998 to 2000. For women aged 74 and 75 years, this increase was almost 10%. As a result of the 187,207 screening examinations performed within this age group, 18.3 per 1,000 women were referred and 10.3 per 1,000 breast cancer were diagnosed. Detection rates in both initial and subsequent screens increased steadily with age and got close to those model-simulated rates, which assume a continuously increasing sojourn time with age. A major finding of this study is that the screening participation among elderly women is high. The outcomes of our study suggest a steadily increasing sojourn time of breast tumours beyond the age of 69, leading to a strong increase in detection of cancers, and therefore, disfavouring the balance with the benefits of screening. At present, 75 years of age can be regarded as an appropriate upper age limit for the Dutch programme.


Epigallocatechin gallate inhibits cell growth and regulates miRNA expression in cervical carcinoma cell lines infected with different high-risk human papillomavirus subtypes.
The aim of the present study was to investigate the inhibitory effects of the polyphenol epigallocatechin-3-gallate (EGCG) on the growth of cervical carcinoma cell lines infected with different high-risk human papillomavirus (HPV) subtypes, as well as the associated regulation of microRNA (miR) expression. Cell proliferation was measured using an MTT assay. The effects of 7 different concentrations of EGCG (100, 80, 60, 40, 20, 10 and 0 µg/ml) on HeLa cell proliferation were assessed. HeLa cell growth was significantly inhibited by EGCG in a dose- and time-dependent manner (P<0.05), and the IC50 was 90.74 and 72.74 µg/ml at 24 and 48 h, respectively. The expression of miR-210, miR-29a, miR-203 and miR-125b in HeLa (HPV16/18+), SiHa (HPV16+), CaSki (HPV16+) and C33A (HPV-) cell lines was measured using quantitative polymerase chain reaction analysis. In CA33 cells, miR-203 (all P<0.001) and miR-125b (P<0.01 and <0.0001) were significantly downregulated by EGCG, and miR-210 was significantly upregulated with 40 and 60 µg/ml EGCG (P<0.0001). miR-125b was significantly downregulated (P<0.001 and <0.0001), and miR-210 and miR-29 were significantly upregulated by ≤80 µg/ml EGCG in HeLa cells (all P<0.0001). In CaSki cells, miR-210, miR-29a (all P<0.001) and miR-125b (P<0.01-0.0001) were significantly upregulated by EGCG. In SiHa cells, miR-125b (both P<0.001) and miR-203 (P<0.01 and <0.0001) were significantly upregulated by EGCG. In conclusion, the results of the present study suggest that EGCG suppresses cervical carcinoma cell growth, possibly via regulating the expression of miRs, suggesting their potential as therapeutic targets for the control and prevention of cervical cancer. Additionally, EGCG may be considered a novel anti-cervical cancer drug in the future.


Methane and CO2 emissions from China's hydroelectric reservoirs: a new quantitative synthesis.
Controversy surrounds the green credentials of hydroelectricity because of the potentially large emission of greenhouse gases (GHG) from associated reservoirs. However, limited and patchy data particularly for China is constraining the current global assessment of GHG releases from hydroelectric reservoirs. This study provides the first evaluation of the CO2 and CH4 emissions from China's hydroelectric reservoirs by considering the reservoir water surface and drawdown areas, and downstream sources (including spillways and turbines, as well as river downstream). The total emission of 29.6 Tg CO2/year and 0.47 Tg CH4/year from hydroelectric reservoirs in China, expressed as CO2 equivalents (eq), corresponds to 45.6 Tg CO2eq/year, which is 2-fold higher than the current GHG emission (ca. 23 Tg CO2eq/year) from global temperate hydropower reservoirs. China's average emission of 70 g CO2eq/kWh from hydropower amounts to 7% of the emissions from coal-fired plant alternatives. China's hydroelectric reservoirs thus currently mitigate GHG emission when compared to the main alternative source of electricity with potentially far great reductions in GHG emissions and benefits possible through relatively minor changes to reservoir management and design. On average, the sum of drawdown and downstream emission including river reaches below dams and turbines, which is overlooked by most studies, represents the equivalent of 42% of the CO2 and 92% of CH4 that emit from hydroelectric reservoirs in China. Main drivers on GHG emission rates are summarized and highlight that water depth and stratification control CH4 flux, and CO2 flux shows significant negative relationships with pH, DO, and Chl-a. Based on our finding, a substantial revision of the global carbon emissions from hydroelectric reservoirs is warranted.


Nuclear dispositions of subtelomeric and pericentromeric chromosomal domains during meiosis in asynaptic mutants of rye (Secale cereale L.).
The nuclear dispositions of subtelomeric and pericentromeric domains in pollen mother cells (PMCs) were tracked during meiosis in wildtype and two asynaptic mutants of rye (Secale cereale L.) by means of fluorescence in situ hybridization (FISH). Homozygotes for sy1 and sy9 non-allelic mutations form axial elements during leptotene of male meiosis, but fail to form synaptonemal complexes. Consequently, recombination is severely impaired, and high univalency is observed at metaphase I. Simultaneous FISH with pSc200 subtelomeric tandem repeat and CCS1 centromeric sequence revealed that at pre-meiotic interphase the two domains are in a bipolar Rabl orientation in both the PMCs and tapetal cells. At the onset of meiotic prophase, the subtelomeric regions in PMCs of wildtype and sy9 cluster into a typical bouquet conformation. The timing of this event in rye is comparable with that in wheat, and is earlier than that observed in other organisms, such as maize, yeast and mammals. This arrangement is retained until later in leptotene and zygotene when the pericentromeric domains disperse and the subtelomeric clusters fragment. The mutant phenotype of sy9 manifests itself during leptotene to zygotene, when the pericentromeric regions become distinctly more distended than in wildtype, and largely fail to pair during zygotene. This indicates that difference in the nature or timing of chromosome condensation in this region is the cause or consequence of asynapsis. By contrast, sy1 fails to form comparable aggregates of subtelomeric regions at leptotene in only half of the nuclei studied. Instead, two to five aggregates are formed that fail to disperse at later stages of meiotic prophase. In addition, the pericentromeric regions disperse prematurely at leptotene and do not associate in pairs at any subsequent stage. It is supposed that the sy1 mutation could disrupt the nuclear disposition of centromeres and telomeres at the end of pre-meiotic interphase, which could cause, or contribute to, its asynaptic phenotype.


Fertility and developmental toxicity studies of diethylene glycol monobutyl ether (DGBE) in rats.
The solvent, dimethylene glycol monobutyl ether (DGBE), is a component of latex paints, inks; it is used as a degreasing agent, industrial detergent. The aim of the study was evaluating the effects of DGBE administered by gavage on the estrous cycle and given with drinking water on fertility in rats and early development of their progeny. Female rats were exposed to DGBE by gavage during 8 weeks at 250, 500 or 1000 mg/kg/day. Vaginal smears were collected during the exposure and 4 weeks after its cessation. Fertility studies were performed in male and female animals exposed to in drinking water. Males were exposed for 10 weeks and then mated with females exposed before mating, during pregnancy and lactation. Young animals were observed during 3 weeks after birth. DGBE does not cause disturbances of the menstrual cycle in females. Parameters used to assess the general toxicity indicate that males receiving DGBE in drinking water are more sensitive to this compound than females: significantly greater, dose-dependent relative spleen weight, significant decrease in hematological parameters from 8% to 15% depending on the dose, were observed. Clinical chemistry parameters (HDL-cholesterol, BUN) and some markers of oxidative stress differ between the exposed groups and the control one, but without adverse health effect. The microscopic examination of internal organs did not reveal morphological changes in male and female rats. The results of our study on the impact of exposure to DGBE on fertility in rats indicate that the substance administered for 9-10 weeks to females and males at a limit dose of 1000 mg/kg did not impair fertility or viability of their offspring during the first three weeks of life.


In-vitro development of a temporal abutment screw to protect osseointegration in immediate loaded implants.
In this study, a temporal abutment fixation screw, designed to fracture in a controlled way upon application of an occlusal force sufficient to produce critical micromotion was developed. The purpose of the screw was to protect the osseointegration of immediate loaded single implants. Seven different screw prototypes were examined by fixing titanium abutments to 112 Mozo-Grau external hexagon implants (MG Osseous®; Mozo-Grau, S.A., Valladolid, Spain). Fracture strength was tested at 30° in two subgroups per screw: one under dynamic loading and the other without prior dynamic loading. Dynamic loading was performed in a single-axis chewing simulator using 150,000 load cycles at 50 N. After normal distribution of obtained data was verified by Kolmogorov-Smirnov test, fracture resistance between samples submitted and not submitted to dynamic loading was compared by the use of Student's t-test. Comparison of fracture resistance among different screw designs was performed by the use of one-way analysis of variance. Confidence interval was set at 95%. Fractures occurred in all screws, allowing easy retrieval. Screw Prototypes 2, 5 and 6 failed during dynamic loading and exhibited statistically significant differences from the other prototypes. Prototypes 2, 5 and 6 may offer a useful protective mechanism during occlusal overload in immediate loaded implants.


Dynamics of color contrast adaptation.
Many forms of color adaptation have been found to reflect both short- and long-term adjustments. We explored the buildup and decay of adaptation to chromatic contrast (temporal modulations of color) for which the dynamics are unknown. A matching task was used to track the perceived contrast of chromatic pulses of varying physical contrast during and after adapting for 1 h to a high contrast modulation repeated over five successive days. The adaptation was characterized by rapid response changes that remained stable in both time course and form across sessions. There was no consistent evidence for long-term plasticity over the time scales we tested.


Loss of fenamate-activated K+ current from epithelial cells during corneal wound healing.
The corneal epithelium provides a barrier between the external environment and the cornea. It also serves as an ion transporting epithelium. Because of its proximity with the external environment, the corneal epithelium is frequently injured through physical or chemical insult. The purpose of this study was to determine whether corneal epithelial cell whole-cell currents change during corneal wound healing as the author of the present study has previously reported for corneal keratocytes and endothelial cells. Rabbit corneal epithelial cells were injured by scraping, heptanol exposure, or freezing. The epithelium was allowed to heal for 12 to 74 hours. Cells were dissociated from corneas, and whole-cell currents were examined using the amphotericin-perforated-patch technique. Cells from the wounded corneal groups had significantly increased capacitance values, indicating increased surface area compared with that of control cells. As previously reported, the primary control whole-cell current was a fenamate-activated K+ current. An inwardly rectifying K+ current and a Cl- current were also observed. In epithelial cells from heptanol-wounded corneas, these conductances were generally unchanged. In cells from scrape- and freeze-wounded corneas, however, the fenamate-activated current was absent or significantly attenuated. As they do in corneal keratocytes and endothelial cells, K+ channels disappear during some models of corneal epithelial wound healing. In addition, cell capacitance, a measurement of cell surface area, increases. These results suggest that substantial K+ channel activity is not required for in vivo epithelial cell proliferation during corneal wound healing.


The early achievement of measurable residual disease negativity in the treatment of adults with Philadelphia-negative B-cell acute lymphoblastic leukemia is a strong predictor for survival.
The minimal or measurable residual disease (MRD) status following induction/consolidation chemotherapy is an important prognostic endpoint in adult patients with newly diagnosed acute lymphoblastic leukemia (ALL). However, the optimal time-point (TP) of MRD assessment and its impact on outcome remains unclear. We analyzed 215 patients with newly diagnosed Philadelphia negative B-cell ALL who received intensive chemotherapy, and had available MRD assessment by multicolor flow cytometry at two separate TPs. The median time to first TP (1TP) and second TP (2TP) were 24 and 110 days, respectively. At 1TP, 148 patients (68%) were MRD negative and 67 (32%) were positive. Of the 148 patients with negative MRD at 1TP, 147 (99%) maintained it through 2TP. Patients who were MRD negative at both TPs, early MRD responders, had the 3-year event-free survival (EFS), and overall survival (OS) rates of 65% and 76%, respectively. Patients with improved MRD status from positive to negative, late MRD responders, had lower 3-year EFS and OS rates, 42% and 58%, respectively (P = .001). Multivariate analysis showed that KMT2A (MLL) rearrangement and MRD positivity at 1TP were the only factors correlated with worse OS. In conclusion, the earlier achievement of MRD negative remission is a stronger prognostic factor for survival.


Non-rheumatic atrial fibrillation as a risk factor for stroke.
The association between non-rheumatic atrial fibrillation (AF) and stroke has been studied in 402 patients consecutively admitted to a stroke unit. Brain infarction patients with sinus rhythm (n = 196) and non-rheumatic AF (n = 92) were further compared. Some findings supported an embolic origin of the stroke: half of the deceased AF patients (n = 24) at autopsy either had left atrial thrombosis or arterial embolism compared to none of the ten with sinus rhythm. Patients with AF also had a higher mortality and more severe brain lesions, findings compatible with a sudden occlusion of blood flow. However, these differences might also be explained by an atherothrombotic occlusion with impaired autoregulation in the ischaemic region in conjunction with heart failure, which was more common in the AF patients. Other findings supporting an atherothrombotic mechanism were: the prevalence of AF was higher (19-29%) in all kinds of stroke, including haemorrhage, than in age-matched controls (3-9%). Also patients with previous AF and no present embolic source resembled the whole AF group and differed from patients with sinus rhythm. Thus embolism is a plausible cause of stroke in many AF patients, whereas an atherothrombotic origin is more likely in others. Characteristics identifying the mechanism in an individual case were not found.


Liver in obesity.
We report on clinical, nutritional, and hepatic histological findings in 50 non-selected obese subjects (mean overweight +74%; range +21-138%). The pathogenesis of the liver damage was assessed with the help of multidimensional analysis of a number of clinical variables. According to the severity of the hepatic lesions, the patients have been ranged in five groups: O (normal liver) 10%; I (fatty liver) 48%; II (fatty hepatitis) 26%; III (fatty fibrosis) 8%; IV (fatty cirrhosis) 8%. The more severe changes (groups III and IV) were constantly associated with excessive alcohol intake. The multidimensional analysis was unable to find a relationship between obesity and the development of fibrosis and cirrhosis whereas it showed that: (a) there was a highly significant correlation between the daily ethanol intake and the degree of overweight, (b) severe fatty metamorphosis was significantly associated with the degree of overweight, the existence of diabetes mellitus, and the amount of alcohol and fat intake, (c) nutritional factors, in particular deficient protein intake, have only an accessory effect in the development of mild inflammation and fibrosis, (d) the consumption of potentially hepatotoxic drugs, very high in the obese (about five drugs per day) could have a role in the development of cirrhosis. In conclusion in our study, there was no evidence that obesity per se could result in severe liver damage.


Rasch analysis of the postconcussive symptom questionnaire: measuring the core construct of brain injury symptomatology.
The Postconcussive Symptom Questionnaire (PCSQ; Lees-Haley, 1992 ) is purported to measure four constructs. These include psychological, cognitive, somatic, and infrequency (i.e., items intended to reflect negative impression management) symptoms. The utility and validity of Postconcussive Syndrome (PCS) as a diagnostic condition continues to be debated. To this end, examining the instruments used to measure postconcussive symptoms can increase our understanding with respect to this issue. The aim of this study was to derive a revised PCSQ to target the core construct of subjective symptoms reported by persons with traumatic brain injury (TBI). A total of 133 people with mild to severe TBI completed the 45-item PCSQ. Items were scored dichotomously, as symptom present or absent. Rasch analysis, based on the mathematical model formulated by Rasch ( 1960 ), was used to derive the revised PCSQ. Misfitting and redundant items were removed and a second model containing 19 items was fitted. The revised PCSQ-19 had superior psychometric qualities; reliability was 0.81. The PCSQ-19 provides a more targeted, unidimensional assessment of subjective symptoms following brain injury. The findings also revealed information related to symptom hierarchy which can further our understanding of PCS.


Genome-wide survey and expression analysis of the plant-specific NAC transcription factor family in soybean during development and dehydration stress.
Plant-specific NAC transcription factors (TFs) play important roles in regulating diverse biological processes, including development, senescence, growth, cell division and responses to environmental stress stimuli. Within the soybean genome, we identified 152 full-length GmNAC TFs, including 11 membrane-bound members. In silico analysis of the GmNACs, together with their Arabidopsis and rice counterparts, revealed similar NAC architecture. Next, we explored the soybean Affymetrix array and Illumina transcriptome sequence data to analyse tissue-specific expression profiles of GmNAC genes. Phylogenetic analysis using stress-related NAC TFs from Arabidopsis and rice as seeding sequences identified 58 of the 152 GmNACs as putative stress-responsive genes, including eight previously reported dehydration-responsive GmNACs. We could design gene-specific primers for quantitative real-time PCR verification of 38 out of 50 newly predicted stress-related genes. Twenty-five and six GmNACs were found to be induced and repressed 2-fold or more, respectively, in soybean roots and/or shoots in response to dehydration. GmNAC085, whose amino acid sequence was 39%; identical to that of well-known SNAC1/ONAC2, was the most induced gene upon dehydration, showing 390-fold and 20-fold induction in shoots and roots, respectively. Our systematic analysis has identified excellent tissue-specific and/or dehydration-responsive candidate GmNAC genes for in-depth characterization and future development of improved drought-tolerant transgenic soybeans.


Pathogenesis-related protein 10 isolated from hot pepper functions as a ribonuclease in an antiviral pathway.
A hot pepper (Capsicum annuum) cDNA clone encoding pathogenesis-related protein 10 (CaPR-10) was isolated by differential screening of a cDNA library prepared from pepper leaves inoculated with tobacco mosaic virus pathotype (TMV-P0). CaPR-10 transcripts were induced in the incompatible interaction with TMV-P0 or Xanthomonas campestris pv. vesicatoria (Xcv) but not induced in the compatible interaction. Characterization of enzymatic properties of CaPR-10 indicated that the recombinant protein exhibits a ribonucleolytic activity against TMV RNA, as well as against pepper total RNA, and shows its putative antiviral activity in several conditions. The CaPR-10 protein existed at very low level in leaf tissue but was dramatically induced as soon as plants were inoculated with TMV-P0, and this was correlated with the increase of its ribonucleolytic activity. Immunoblot analysis and pull-down assays using proteins extracted from pepper leaves showed that TMV-P0 inoculation led to the phosphorylation of CaPR-10, a modification that should affect its capacity for RNase function. We present data that the induction and subsequent phosphorylation of CaPR-10 increased its ribonucleolytic activity to cleave invading viral RNAs, and this activity should be important to its antiviral pathway during viral attack in vivo.


Early strains of multidrug-resistant Salmonella enterica serovar Kentucky sequence type 198 from Southeast Asia harbor Salmonella genomic island 1-J variants with a novel insertion sequence.
Salmonella genomic island 1 (SGI1) is a 43-kb integrative mobilizable element that harbors a great diversity of multidrug resistance gene clusters described in numerous Salmonella enterica serovars and also in Proteus mirabilis. The majority of SGI1 variants contain an In104-derivative complex class 1 integron inserted between resolvase gene res and open reading frame (ORF) S044 in SGI1. Recently, the international spread of ciprofloxacin-resistant S. enterica serovar Kentucky sequence type 198 (ST198) containing SGI1-K variants has been reported. A retrospective study was undertaken to characterize ST198 S. Kentucky strains isolated before the spread of the epidemic ST198-SGI1-K population in Africa and the Middle East. Here, we characterized 12 ST198 S. Kentucky strains isolated between 1969 and 1999, mainly from humans returning from Southeast Asia (n = 10 strains) or Israel (n = 1 strain) or from meat in Egypt (n = 1 strain). All these ST198 S. Kentucky strains did not belong to the XbaI pulsotype X1 associated with the African epidemic clone but to pulsotype X2. SGI1-J subgroup variants containing different complex integrons with a partial transposition module and inserted within ORF S023 of SGI1 were detected in six strains. The SGI1-J4 variant containing a partially deleted class 1 integron and thus showing a narrow resistance phenotype to sulfonamides was identified in two epidemiologically unrelated strains from Indonesia. The four remaining strains harbored a novel SGI1-J variant, named SGI1-J6, which contained aadA2, floR2, tetR(G)-tetA(G), and sul1 resistance genes within its complex integron. Moreover, in all these S. Kentucky isolates, a novel insertion sequence related to the IS630 family and named ISSen5 was found inserted upstream of the SGI1 complex integron in ORF S023. Thus, two subpopulations of S. Kentucky ST198 independently and exclusively acquired the SGI1 during the 1980s and 1990s. Unlike the ST198-X1 African epidemic subpopulation, the ST198-X2 subpopulation mainly from Asia harbors variants of the SGI1-J subgroup that are encountered mainly in the Far East, as previously described for S. enterica serovars Emek and Virchow.


Immunoregulation in normal pregnancy and pre-eclampsia: an overview.
Pre-eclampsia is a major disorder of human pregnancy, which may have an immunological basis. It is a disease of two stages. The first stage concerns the relative failure of early trophoblast invasion and remodelling of the spiral arteries, leading to a poor blood supply to the placenta, exposing it to oxidative stress. The inadequate trophoblast invasion may result from decreased expression of human leukocyte antigen-G (HLA-G) leading to an abnormal interaction with decidual natural killer (NK) cells, which are believed to play a major role in these processes through the production of immunoregulatory cytokines and angiogenic factors. Recent evidence suggests that the interaction between trophoblast human leukocyte antigen-C (HLA-C) molecules and decidual NK cell receptors may be the point at which the apparent partner specificity of the disease originates. The second stage is the maternal syndrome, which is characterized by a generalized systemic inflammatory response involving both leukocytes and endothelium. The inflammatory stimulus is believed to come from the placenta. In pre-eclampsia, placental oxidative stress may lead to increased shedding of apoptotic and/or necrotic syncytiotrophoblast debris into the maternal circulation. There is evidence that such trophoblast debris interacts with maternal leukocytes and endothelial cells to stimulate the release of proinflammatory cytokines, which could then trigger the maternal disease.


Gender Difference in Internet Use and Internet Problems among Quebec High School Students.
There are presently no data available concerning Internet addiction (IA) problems among adolescents in Canada and the province of Quebec. The goal of this study is thus to document and compare the influence of gender on Internet use and addiction. The study data were collected from a larger research project on gambling among adolescents. Activities conducted online (applications used and time spent) as well as answers to the Internet Addiction Test (IAT) were collected from 3938 adolescents from grades 9 to 11. The two most often employed cut-off points for the IAT in the literature were documented: (40-69 and 70+) and (50+). Boys spent significantly more time on the Internet than did girls. A greater proportion of the girls made intense use of social networks, whereas a greater proportion of the boys made intense use of massively multiplayer online role-playing games, online games, and adult sites. The proportion of adolescents with a potential IA problem varied according to the cut-off employed. When the cut-off was set at 70+, 1.3% of the adolescents were considered to have an IA, while 41.7% were seen to be at risk. At a 50+ cut-off, 18% of the adolescents were considered to have a problem. There was no significant difference between the genders concerning the proportion of adolescents considered to be at risk or presenting IA problems. Finally, analysis of the percentile ranks would seem to show that a cut-off of 50+ better describes the category of young people at risk. The results of this study make it possible to document Internet use and IA in a large number of Quebec adolescents.


Low-density lipoprotein, collagen, and thrombin models reveal that Rosemarinus officinalis L. exhibits potent antiglycative effects.
Using the low-density lipoprotein (LDL), collagen, and thrombin models, we report here that the rosemary extracts (REs), either the aqueous (REw) or the acetonic (REA), all possessed many antiglycation-related features, and the effective concentrations required were as follows: 0.1 mg/mL for suppressing the relative electrophoretic mobility, 1.3 microg/mL for anticonjugated diene induction, 0.5 mg/mL for inhibition of thiobarbituric acid reactive substances production, 0.1 mg/mL for AGEs (advanced glycation end products) formation, 0.1 mg/mL to block glucose incorporation, and 0.05 mg/mL as an effective anti-antithrombin III. Using high-performance liquid chromatography/mass spectrometry, we identified five major constituents among eight major peaks, including rosmarinic acid, carnosol, 12-methoxycarnosic acid, carnosic acid, and methyl carnosate. In the LDL model, REA was proven to be more efficient than REw; yet, the reverse is true for the collagen and the thrombin III models, the reason of which was ascribed to the higher lipid-soluble antioxidant content (such as rosmarinic acid, carnosol, carnosic acid, 12-methoxycarnosic acid and methyl carnosate) in REA than in REw and the different surface lipid characteristics between LDL and collagen; although to act as anti-AGEs, both extracts were comparable. To assist the evidence, a larger 2,2-diphenyl-1-picrylhydrazyl radical scavenging capability with less total polyphenolic content was found in REA. We conclude that rosemary is an excellent multifunctional therapeutic herb; by looking at its potential potent antiglycative bioactivity, it may become a good adjuvant medicine for the prevention and treatment of diabetic, cardiovascular, and other neurodegenerative diseases.


Psychological therapies (remotely delivered) for the management of chronic and recurrent pain in children and adolescents.
Chronic pain is common during childhood and adolescence and is associated with negative outcomes such as increased severity of pain, reduced function (e.g. missing school), and low mood (e.g. high levels of depression and anxiety). Psychological therapies, traditionally delivered face-to-face with a therapist, are efficacious at reducing pain intensity and disability. However, new and innovative technology is being used to deliver these psychological therapies remotely, meaning barriers to access to treatment such as distance and cost can be removed or reduced. Therapies delivered with technological devices, such as the Internet, computer-based programmes, smartphone applications, or via the telephone, can be used to deliver treatment to children and adolescents with chronic pain. To determine the efficacy of psychological therapies delivered remotely compared to waiting-list, treatment-as-usual, or active control treatments, for the management of chronic pain in children and adolescents. We searched four databases (CENTRAL, MEDLINE, EMBASE, and PsycINFO) from inception to June 2014 for randomised controlled trials of remotely delivered psychological interventions for children and adolescents (0 to 18 years of age) with chronic pain. We searched for chronic pain conditions including, but not exclusive to, headache, recurrent abdominal pain, musculoskeletal pain, and neuropathic pain. We also searched online trial registries for potential trials. A citation and reference search for all included studies was conducted. All included studies were randomised controlled trials that investigated the efficacy of a psychological therapy delivered remotely via the Internet, smartphone device, computer-based programme, audiotapes, or over the phone in comparison to an active, treatment-as-usual, or waiting-list control. We considered blended treatments, which used a combination of technology and face-to-face interaction. We excluded interventions solely delivered face-to-face between therapist and patient from this review. Children and adolescents (0 to 18 years of age) with a primary chronic pain condition were the target of the interventions. Each comparator arm, at each extraction point had to include 10 or more participants. For the analyses, we combined all psychological therapies. We split pain conditions into headache and mixed (non-headache) pain and analysed them separately. Pain, disability, depression, anxiety, and adverse events were extracted as primary outcomes. We also extracted satisfaction with treatment as a secondary outcome. We considered outcomes at two time points: first immediately following the end of treatment (known as 'post-treatment'), and second, any follow-up time point post-treatment between 3 and 12 months (known as 'follow-up'). We assessed all included studies for risk of bias. Eight studies (N = 371) that delivered treatment remotely were identified from our search; five studies investigated children with headache conditions, one study was with children with juvenile idiopathic arthritis, and two studies included mixed samples of children with headache and mixed (i.e. recurrent abdominal pain, musculoskeletal pain) chronic pain conditions. The average age of children receiving treatment was 12.57 years.For headache pain conditions, we found one beneficial effect of remotely delivered psychological therapy. Headache severity was reduced post-treatment (risk ratio (RR) = 2.65, 95% confidence interval (CI) 1.56 to 4.50, z = 3.62,p < 0.01, number needed to treat to benefit (NNTB) = 2.88). For mixed pain conditions, we found only one beneficial effect: psychological therapies reduced pain intensity post-treatment (standardised mean difference (SMD) = -0.61, 95% CI -0.96 to -0.25, z = 3.38, p < 0.01). No effects were found for reducing pain at follow-up in either analysis. For headache and mixed conditions, there were no beneficial effects of psychological therapies delivered remotely for disability post-treatment and a lack of data at follow-up meant no analyses could be run. Only one analysis could be conducted for depression outcomes. We found no beneficial effect of psychological therapies in reducing depression post-treatment for headache conditions. Only one study presented data in children with mixed pain conditions for depressive outcomes and no data were available for either condition at follow-up. Only one study presented anxiety data post-treatment and no studies reported follow-up data, therefore no analyses could be run. Further, there were no data available for adverse events, meaning that we are unsure whether psychological therapies are harmful to children who receive them. Satisfaction with treatment is described qualitatively.'Risk of bias' assessments were low or unclear. We judged selection, detection, and reporting biases to be mostly low risk for included studies. However, judgements made on performance and attrition biases were mostly unclear. Psychological therapies delivered remotely, primarily via the Internet, confer benefit in reducing the intensity or severity of pain after treatment across conditions. There is considerable uncertainty around these estimates of effect and only eight studies with 371 children contribute to the conclusions. Future studies are likely to change the conclusions reported here. All included trials used either behavioural or cognitive behavioural therapies for children with chronic pain, therefore we cannot generalise our findings to other therapies. However, satisfaction with these treatments was generally positive. Larger trials are needed to increase our confidence in all conclusions regarding the efficacy of remotely delivered psychological therapies. Implications for practice and research are discussed.


Genetic diversity of Mycobacterium tuberculosis in Peru and exploration of phylogenetic associations with drug resistance.
There is limited available data on the strain diversity of M tuberculosis in Peru, though there may be interesting lessons to learn from a setting where multidrug resistant TB has emerged as a major problem despite an apparently well-functioning DOTS control programme. Spoligotyping was undertaken on 794 strains of M tuberculosis collected between 1999 and 2005 from 553 community-based patients and 241 hospital-based HIV co-infected patients with pulmonary tuberculosis in Lima, Peru. Phylogenetic and epidemiologic analyses permitted identification of clusters and exploration of spoligotype associations with drug resistance. Mean patient age was 31.9 years, 63% were male and 30.4% were known to be HIV+. Rifampicin mono-resistance, isoniazid mono-resistance and multidrug resistance (MDR) were identified in 4.7%, 8.7% and 17.3% of strains respectively. Of 794 strains from 794 patients there were 149 different spoligotypes. Of these there were 27 strains (3.4%) with novel, unique orphan spoligotypes. 498 strains (62.7%) were clustered in the nine most common spoligotypes: 16.4% SIT 50 (clade H3), 12.3% SIT 53 (clade T1), 8.3% SIT 33 (LAM3), 7.4% SIT 42 (LAM9), 5.5% SIT 1 (Beijing), 3.9% SIT 47 (H1), 3.0% SIT 222 (clade unknown), 3.0% SIT1355 (LAM), and 2.8% SIT 92 (X3). Amongst HIV-negative community-based TB patients no associations were seen between drug resistance and specific spoligotypes; in contrast HIV-associated MDRTB, but not isoniazid or rifampicin mono-resistance, was associated with SIT42 and SIT53 strains. Two spoligotypes were associated with MDR particularly amongst patients with HIV. The MDR-HIV association was significantly reduced after controlling for SIT42 and SIT53 status; residual confounding may explain the remaining apparent association. These data are suggestive of a prolonged, clonal, hospital-based outbreak of MDR disease amongst HIV patients but do not support a hypothesis of strain-specific propensity for the acquisition of resistance-conferring mutations.


Methods to assess breathlessness in healthy subjects: a critical evaluation and application to analyse the acute effects of diazepam and promethazine on breathlessness induced by exercise or by exposure to raised levels of carbon dioxide.
1. Methods were devised and evaluated for inducing breathlessness by submaximal graded exercise in healthy subjects while objective measurements of cardiorespiratory function were made. Breathlessness was assessed with serial visual analogue scales (VAS), but with various measures to enhance repeatability. 2. A high level of reproducibility was obtained in spite of the subjective nature of the assessment. Individual responses were described by the relationship between breathlessness and ventilation. The sensitivity of the method was demonstrated by the use of inspiratory resistances which disturbed this relationship and caused greater breathlessness for a given level of ventilation. 3. These methods were applied to six healthy subjects to analyse the effects of acute doses of diazepam and promethazine on breathlessness induced by graded exercise or by rebreathing carbon dioxide in a double-blind study. 4. During exercise, diazepam and promethazine did not reduce breathlessness, although there was a minor trend with promethazine. During exposure to elevated levels of carbon dioxide, diazepam and promethazine had no effect on breathlessness. Diazepam and promethazine produced similar levels of sedation, but neither drug had significant effects on the ventilatory response to carbon dioxide. These preliminary findings contrast with those reported for chronic diazepam in 'pink puffers'. 5. Raised levels of carbon dioxide caused greater breathlessness in relation to ventilation than did exercise.


Inhibition of calcium uptake via the sarco/endoplasmic reticulum Ca2+-ATPase in a mouse model of Sandhoff disease and prevention by treatment with N-butyldeoxynojirimycin.
Gangliosides are found at high levels in neuronal tissues where they play a variety of important functions. In the gangliosidoses, gangliosides accumulate because of defective activity of the lysosomal proteins responsible for their degradation, usually resulting in a rapidly progressive neurodegenerative disease. However, the molecular mechanism(s) leading from ganglioside accumulation to neurodegeneration is not known. We now examine the effect of ganglioside GM2 accumulation in a mouse model of Sandhoff disease (one of the GM2 gangliosidoses), the Hexb-/- mouse. Microsomes from Hexb-/- mouse brain showed a significant reduction in the rate of Ca2+-uptake via the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA), which was prevented by feeding Hexb-/- mice with N-butyldeoxynojirimycin (NB-DNJ), an inhibitor of glycolipid synthesis that reduces GM2 storage. Changes in SERCA activity were not due to transcriptional regulation but rather because of a decrease in Vmax. Moreover, exogenously added GM2 had a similar effect on SERCA activity. The functional significance of these findings was established by the enhanced sensitivity of neurons cultured from embryonic Hexb-/- mice to cell death induced by thapsigargin, a specific SERCA inhibitor, and by the enhanced sensitivity of Hexb-/- microsomes to calcium-induced calcium release. This study suggests a mechanistic link among GM2 accumulation, reduced SERCA activity, and neuronal cell death, which may be of significance for delineating the neuropathophysiology of Sandhoff disease.


Analysis and neuronal modeling of the nonlinear characteristics of a local cardiac reflex in the rat.
Previous experimental results have suggested the existence of a local cardiac reflex in the rat. In this study, the putative role of such a local reflex in cardiovascular regulation is quantitatively analyzed. A model for the local reflex is developed from anatomical experimental results and physiological data in the literature. Using this model, a systems-level analysis is conducted. Simulation results indicate that the neuromodulatory mechanism of the local reflex attenuates the nonlinearity of the relationship between cardiac vagal drive and arterial pressure. This behavior is characterized through coherence analysis. Furthermore, the modulation of phase-related characteristics of the cardiovascular system is suggested as a plausible mechanism for the nonlinear attenuation. Based on these results, it is plausible that the functional role of the local reflex is highly robust nonlinear compensation at the heart, which results in less complex dynamics in other parts of the reflex.


Whatever does not kill us: cumulative lifetime adversity, vulnerability, and resilience.
Exposure to adverse life events typically predicts subsequent negative effects on mental health and well-being, such that more adversity predicts worse outcomes. However, adverse experiences may also foster subsequent resilience, with resulting advantages for mental health and well-being. In a multiyear longitudinal study of a national sample, people with a history of some lifetime adversity reported better mental health and well-being outcomes than not only people with a high history of adversity but also than people with no history of adversity. Specifically, U-shaped quadratic relationships indicated that a history of some but nonzero lifetime adversity predicted relatively lower global distress, lower self-rated functional impairment, fewer posttraumatic stress symptoms, and higher life satisfaction over time. Furthermore, people with some prior lifetime adversity were the least affected by recent adverse events. These results suggest that, in moderation, whatever does not kill us may indeed make us stronger.


Renal type II Na/Pi-cotransporter is strongly impaired whereas the Na/sulphate-cotransporter and aquaporin 1 are unchanged in cadmium-treated rats.
The cellular mechanisms of cadmium (Cd) nephrotoxicity are poorly understood. In this study we investigated the cellular causes of the Cd-induced phosphaturia in the rat. Compared to controls, Cd-treated rats (2 mg Cd/kg body weight, s.c. for 14 days) showed a marked polyuria, proteinuria and phosphaturia. As studied by the rapid filtration technique in isolated cortical brush-border membrane vesicles (BBMV), Na+-gradient-driven uptake of phosphate ([32Pi]) and of [3H] glucose were markedly decreased in Cd-treated rats, whereas uptake of sulphate ([35S]) remained unchanged. By Western blotting of BBMV proteins and by indirect immunocytochemistry in 4-micron thick frozen fixed kidney sections, using an antibody against the type II Na/Pi-cotransporter (NaPi-2), we found a diminished expression of this protein in the brush-border membrane from Cd-treated rats. How ever, the expression of the water channel aquaporin 1, estimated from the specific antibody staining in brush-border membranes, remained unchanged by Cd. Northern blot analysis showed a strong reduction of 2.7 kb NaPi-2-related mRNA in Cd-affected kidneys. Our data indicate that: (1) Cd may reduce reabsorption of Pi in proximal tubules by affecting the expression of the functional Na/Pi-cotransporters in the luminal membrane, and (2) Cd effects on brush-border transporters are selective.


Canonical Wnt signaling promotes early hematopoietic progenitor formation and erythroid specification during embryonic stem cell differentiation.
The generation of hematopoietic stem cells (HSCs) during development is a complex process linked to morphogenic signals. Understanding this process is important for regenerative medicine applications that require in vitro production of HSC. In this study we investigated the effects of canonical Wnt/β-catenin signaling during early embryonic differentiation and hematopoietic specification using an embryonic stem cell system. Our data clearly demonstrates that following early differentiation induction, canonical Wnt signaling induces a strong mesodermal program whilst maintaining a degree of stemness potential. This involved a complex interplay between β-catenin/TCF/LEF/Brachyury/Nanog. β-catenin mediated up-regulation of TCF/LEF resulted in enhanced brachyury levels, which in-turn lead to Nanog up-regulation. During differentiation, active canonical Wnt signaling also up-regulated key transcription factors and cell specific markers essential for hematopoietic specification, in particular genes involved in establishing primitive erythropoiesis. This led to a significant increase in primitive erythroid colony formation. β-catenin signaling also augmented early hematopoietic and multipotent progenitor (MPP) formation. Following culture in a MPP specific cytokine cocktail, activation of β-catenin suppressed differentiation of the early hematopoietic progenitor population, with cells displaying a higher replating capacity and a propensity to form megakaryocytic erythroid progenitors. This bias towards erythroid lineage commitment was also observed when hematopoietic progenitors were directed to undergo myeloid colony formation. Overall this study underscores the importance of canonical Wnt/β-catenin signaling in mesodermal specification, primitive erythropoiesis and early hematopietic progenitor formation during hematopoietic induction.


An integrated and comparative approach towards identification, characterization and functional annotation of candidate genes for drought tolerance in sorghum (Sorghum bicolor (L.) Moench).
Drought is the most disastrous abiotic stress that severely affects agricultural productivity worldwide. Understanding the biological basis of drought-regulated traits, requires identification and an in-depth characterization of genetic determinants using model organisms and high-throughput technologies. However, studies on drought tolerance have generally been limited to traditional candidate gene approach that targets only a single gene in a pathway that is related to a trait. In this study, we used sorghum, one of the model crops that is well adapted to arid regions, to mine genes and define determinants for drought tolerance using drought expression libraries and RNA-seq data. We provide an integrated and comparative in silico candidate gene identification, characterization and annotation approach, with an emphasis on genes playing a prominent role in conferring drought tolerance in sorghum. A total of 470 non-redundant functionally annotated drought responsive genes (DRGs) were identified using experimental data from drought responses by employing pairwise sequence similarity searches, pathway and interpro-domain analysis, expression profiling and orthology relation. Comparison of the genomic locations between these genes and sorghum quantitative trait loci (QTLs) showed that 40% of these genes were co-localized with QTLs known for drought tolerance. The genome reannotation conducted using the Program to Assemble Spliced Alignment (PASA), resulted in 9.6% of existing single gene models being updated. In addition, 210 putative novel genes were identified using AUGUSTUS and PASA based analysis on expression dataset. Among these, 50% were single exonic, 69.5% represented drought responsive and 5.7% were complete gene structure models. Analysis of biochemical metabolism revealed 14 metabolic pathways that are related to drought tolerance and also had a strong biological network, among categories of genes involved. Identification of these pathways, signifies the interplay of biochemical reactions that make up the metabolic network, constituting fundamental interface for sorghum defence mechanism against drought stress. This study suggests untapped natural variability in sorghum that could be used for developing drought tolerance. The data presented here, may be regarded as an initial reference point in functional and comparative genomics in the Gramineae family.


Constituents of cinnamon inhibit bacterial acetyl CoA carboxylase.
Cinnamon bark ( CINNAMOMUM ZEYLANICUM) is used extensively as an antimicrobial material and currently is being increasingly used in Europe by people with type II diabetes to control their glucose levels. In this paper we describe the action of cinnamon oil, its major component, TRANS-cinnamaldehyde, and an analogue, 4-hydroxy-3-methoxy- TRANS-cinnamaldehyde against bacterial acetyl-CoA carboxylase in an attempt to elucidate the mechanism of action of this well-known antimicrobial material. These natural products inhibited the carboxyltransferase component of ESCHERICHIA COLI acetyl-CoA carboxylase but had no effect on the activity of the biotin carboxylase component. The inhibition patterns indicated that these products bound to the biotin binding site of carboxyltransferase with TRANS-cinnamaldehyde having a K (i) value of 3.8 ± 0.6 mM. The inhibition of carboxyltransferase by 4-hydroxy-3-methoxy- TRANS-cinnamaldehyde was analyzed with a new assay for this enzyme based on capillary electrophoresis. These results explain, in part, the antibacterial activity of this well-known antimicrobial material.


Secondary syphilis and the nephrotic syndrome.
A case of nephrotic syndrome in a 21-yr-old black man with secondary syphilis and diabetes mellitus is described. A renal biopsy was performed, which showed membranous glomerulopathy stage I associated with mesangial hyperplasia and mesangial deposits. The clinical course and the histologic findings, compatible with syphilitic nephropathy, are offered to remind internists (nephrologists) that sexually transmitted diseases, like syphilis or hepatitis B, in addition to human immunodeficiency virus, can have important renal manifestations.


Mechanisms of action of antireflux surgery: theory and fact.
Despite the absence of definitive explanations regarding either the physiologic or surgical factors which curtail gastro-esophageal reflux, effective antireflux operations exist. This article explores the theoretical factors relevant to the surgical control of reflux. These theoretical features include: (1) the pressure, length, and location of the manometrically defined lower esophageal sphincter (LES); (2) the gastro-esophageal valve as defined by the angle of His and a mucosal flap valve; and (3) the mechanical effects of a fundoplication which serve to increase the opening pressure of the cardia and optimize the physical relationships described by the law of La Place. Finally, the relation of these theoretical factors to actual operations is discussed.


Utilization of nitrogen in cows consuming wet distillers grains with solubles in alfalfa and corn silage-based dairy rations.
The objectives of this experiment were to determine the effects of forage type on nutrient digestibility, purine derivative excretion, nitrogen utilization, and milk production in dairy cattle consuming rations containing high levels of wet distillers grains with solubles (WDGS). Primiparous (n=8) and multiparous (n=20) Holstein cows were used in a replicated 4 x 4 Latin square. Animals were fed 1 of 4 treatments during each 21-d period: 1) CONT-CS, 0% WDGS and high corn silage; 2) CONT-AS, 0% WDGS and high alfalfa silage; 3) WDGS-CS, 25% WDGS and high corn silage; and 4) WDGS-AS, 25% WDGS and high alfalfa silage (dry matter basis). Intake and milk data were collected daily and averaged for d 15 to 21 of each period. Dry matter intake was lower for CONT-CS than for CONT-AS, WDGS-CS, and WDGS-AS (22.5, 24.6, 24.6, and 24.8 kg/d, respectively). Digestibility of dry matter, organic matter, neutral detergent fiber, and N were not affected by treatment, averaging 59.6, 62.3, 40.1, and 58.6%, respectively. Excretion of urinary purine derivatives was greatest for WDGS-AS, followed by WDGS-CS, and then CONT-CS and CONT-AS. Thus, by calculation, estimated microbial protein flow was highest for WDGS-AS (2,189.9 g/d) followed by WDGS-CS (1,996.2 g/d), CONT-AS (1,640.0 g/d), and CONT-CS (1,627.0 g/d). Mass of fecal N was not different among treatments (averaging 287.1+/-14.8 g/d), but urinary and manure N were reduced for rations with WDGS compared with those not including WDGS. Observed 4% fat-corrected milk was greatest for WDGS-AS, followed by WDGS-CS, and then CONT-CS and CONT-AS (30.7, 29.7, 28.3, and 27.2 kg/d, respectively). Milk protein yield was greatest for WDGS-AS (1.00 kg/d), followed by WDGS-CS, and then CONT-AS and CONT-CS (0.96, 0.91, 0.86 kg/d, respectively). This research demonstrated that rations can be balanced for dairy cattle to include up to 25% WDGS and result in increased microbial protein synthesis, milk production, and milk protein yield.


Prominent transcription of zebrafish N-myc (nmyc1) in tectal and retinal growth zones during embryonic and early larval development.
Because of its oncogenic capacity and expression restricted to embryonic and newborn tissues, the N-myc proto-oncogene is suggested to play a key role in vertebrate organogenesis as well as in the control of cell proliferation and differentiation. To further approach the developmental function of N-myc, we cloned full-length zebrafish N-myc (nmyc1) and analyzed its expression in the embryo and early larva. nmyc1 transcription is initiated at the mid-blastula stage. At somitogenesis stages, its expression was detected in the retina, midbrain, posterior hindbrain and presumptive spinal cord. nmyc1 was also transcribed in the endoderm and its derivatives as well as in branchial arches. At later developmental stages, posterior neural expression of nmyc1 was switched off, but expression remained intense in the brain, mainly in the optic tectum, cerebellar plate and dorsal rhombomere 2. Comparison of nmyc1 transcription with proliferation zones using a M phase mitotic marker revealed that nmyc1 expression is specifically associated with mitosis in the optic tectum and the retina. This result contrasts with previous studies in other vertebrates where N-myc expression can persist in differentiating cells.


Lost to translation: How design factors of the mouse-tracking procedure impact the inference from action to cognition.
From an embodiment perspective, action and cognition influence each other constantly. This interaction has been utilized in mouse-tracking studies to infer cognitive states from movements, assuming a continuous manifestation of cognitive processing into movement. However, it is mostly unknown how this manifestation is affected by the variety of possible design choices in mouse-tracking paradigms. Here we studied how three design factors impact the manifestation of cognition into movement in a Simon task with mouse tracking. We varied the response selection (i.e., with or without clicking), the ratio between hand and mouse cursor movement, and the location of the response boxes. The results show that all design factors can blur or even prevent the manifestation of cognition into movement, as reflected by a reduction in movement consistency and action dynamics, as well as by the adoption of unsuitable movement strategies. We conclude that deliberate and careful design choices in mouse-tracking experiments are crucial to ensuring a continuous manifestation of cognition in movement. We discuss the importance of developing a standard practice in the design of mouse-tracking experiments.


Sequencing of a 'mouse azoospermia' gene panel in azoospermic men: identification of RNF212 and STAG3 mutations as novel genetic causes of meiotic arrest.
What is the diagnostic potential of next generation sequencing (NGS) based on a 'mouse azoospermia' gene panel in human non-obstructive azoospermia (NOA)? The diagnostic performance of sequencing a gene panel based on genes associated with mouse azoospermia was relatively successful in idiopathic NOA patients and allowed the discovery of two novel genes involved in NOA due to meiotic arrest. NOA is a largely heterogeneous clinical entity, which includes different histological pictures. In a large proportion of NOA, the aetiology remains unknown (idiopathic NOA) and yet, unknown genetic factors are likely to play be involved. The mouse is the most broadly used mammalian model for studying human disease because of its usefulness for genetic manipulation and its genetic and physiological similarities to man. Mouse azoospermia models are available in the Mouse Genome Informatics database (MGI: http://www.informatics.jax.org/). The first step was to design of a 'mouse azoospermia' gene panel through the consultation of MGI. The second step was NGS analysis of 175 genes in a group of highly selected NOA patients (n = 33). The third step was characterization of the discovered gene defects in human testis tissue, through meiotic studies using surplus testicular biopsy material from the carriers of the RNF212 and STAG3 pathogenic variants. The final step was RNF212 and STAG3 expression analysis in a collection of testis biopsies. From a total of 1300 infertile patients, 33 idiopathic NOA patients were analysed in this study, including 31 unrelated men and 2 brothers from a consanguineous family. The testis histology of the 31 unrelated NOA patients was as follows: 20 Sertoli cell-only syndrome (SCOS), 11 spermatogenic arrest (6 spermatogonial arrest and 5 spermatocytic arrest). The two brothers were affected by spermatocytic arrest. DNA extracted from blood was used for NGS on Illumina NextSeq500 platform. Generated sequence data was filtered for rare and potentially pathogenic variants. Functional studies in surplus testicular tissue from the carriers included the investigation of meiotic entry, XY body formation and metaphases by performing fluorescent immunohistochemical staining and immunocytochemistry. mRNA expression analysis through RT-qPCR of RNF212 and STAG3 was carried out in a collection of testis biopsies with different histology. Our approach was relatively successful, leading to the genetic diagnosis of one sporadic NOA patient and two NOA brothers. This relatively high diagnostic performance is likely to be related to the stringent patient selection criteria i.e. all known causes of azoospermia were excluded and to the relatively high number of patients with rare testis histology (spermatocytic arrest). All three mutation carriers presented meiotic arrest, leading to the genetic diagnosis of three out of seven cases with this specific testicular phenotype. For the first time, we report biallelic variants in STAG3, in one sporadic patient, and a homozygous RNF212 variant, in the two brothers, as the genetic cause of NOA. Meiotic studies allowed the detection of the functional consequences of the mutations and provided information on the role of STAG3 and RNF212 in human male meiosis. All genes, with the exception of 5 out of 175, included in the panel cause azoospermia in mice only in the homozygous or hemizygous state. Consequently, apart from the five known dominant genes, heterozygous variants (except compound heterozygosity) in the remaining genes were not taken into consideration as causes of NOA. We identified the genetic cause in approximately half of the patients with spermatocytic arrest. The low number of analysed patients can be considered as a limitation, but it is a very rare testis phenotype. Due to the low frequency of this specific phenotype among infertile men, our finding may be considered of low clinical impact. However, at an individual level, it does have relevance for prognostic purposes prior testicular sperm extraction. Our study represents an additional step towards elucidating the genetic bases of early spermatogenic failure, since we discovered two new genes involved in human male meiotic arrest. We propose the inclusion of RNF212 and STAG3 in a future male infertility diagnostic gene panel. Based on the associated testis phenotype, the identification of pathogenic mutations in these genes also confers a negative predictive value for testicular sperm retrieval. Our meiotic studies provide novel insights into the role of these proteins in human male meiosis. Mutations in STAG3 were first described as a cause of female infertility and ovarian cancer, and Rnf212 knock out in mice leads to male and female infertility. Hence, our results stimulate further research on shared genetic factors causing infertility in both sexes and indicate that genetic counselling should involve not only male but also female relatives of NOA patients. This work was funded by the Spanish Ministry of Health Instituto Carlos III-FIS (grant number: FIS/FEDER-PI14/01250; PI17/01822) awarded to CK and AR-E, and by the European Commission, Reproductive Biology Early Research Training (REPROTRAIN, EU-FP7-PEOPLE-2011-ITN289880), awarded to CK, WB, and AE-M. The authors have no conflict of interest.


African American experiences in healthcare: "I always feel like I'm getting skipped over".
Although African Americans perceive discrimination in health care settings, experience higher levels of medical mistrust compared with European Americans, and experience poorer communication with health care providers, little is known as to how these barriers to quality patient-provider relationships arise and manifest themselves. This study examined experiences of African American community members regarding these barriers and additionally explored participants' perspectives on race in the patient-provider relationship. Focus groups were conducted as part of a study exploring participants' experiences and relationships in health care settings. Sixty African American adults were recruited through community settings and activities to participate in 1 of 9 focus groups segmented by gender. Transcripts were reviewed for content related to perceived discrimination, mistrust, poor communication, and race discordance. Themes providing insight into participants' subjective experience of these potential relationship barriers were derived through qualitative coding (using NVivo 10) and iterative discussion. Perceived discrimination arose when African American patients, particularly women, felt their symptoms or problems were discredited. Medical mistrust occurred when clinicians did not convey respect to patients, leaving patients to wonder whether their clinician's treatment was discriminatory or not. Poor communication arose when clinicians did not acknowledge patients' perspectives during interactions. Patients often viewed these actions as discriminatory. African Americans experience poor communication with their health care providers, medical mistrust, and perceived discrimination when accessing health care in numerous and sometimes interrelated ways. The investigators recommend ways to reduce the experience of such barriers and to improve patient-provider relationships for African Americans in health care. (PsycINFO Database Record


A controlled-flow vacuum-free bottle system enhances preterm infants' nutritive sucking skills.
We have shown that a controlled-flow vacuum-free bottle system (CFVFB) vs. a standard bottle (SB) facilitates overall transfer and rate of milk transfer, and shortens oral feeding duration in very-low-birth-weight (VLBW) infants. We aimed to understand the basis by which this occurs. Thirty infants (19 males; 27 +/- 1 weeks gestation) were randomized to a CFVFB or SB. Outcomes monitored at 1-2 and 6-8 oral feedings/day when infants were around 34 and 36 weeks postmenstrual age, respectively, included: overall transfer (% volume taken/volume prescribed), rate of milk transfer (ml/min), sucking stage, frequency of suction (#S/s) and expression (#E/s), suction amplitude (mmHg), and sucking burst duration (s). At both periods we confirmed that infants using a CFVFB vs. SB demonstrated greater overall transfer and rate of milk transfer, along with more mature sucking stages. Suction and expression frequencies were decreased with CFVFB vs. SB at 1-2 oral feeding/day; only that of suction was reduced at 6-8 oral feedings/day. No group differences in suction amplitude and burst duration were observed. We speculate that oral feeding performance improves without significant change in sucking effort with a CFVFB vs. SB. In addition, we have shown that VLBW infants can tolerate faster milk flow than currently presumed. Finally, the use of a CFVFB may reduce energy expenditure as it enhances feeding performance without increasing sucking effort.


Estimating the cost of increasing retention in care for HIV-infected patients: results of the CDC/HRSA retention in care trial.
Retaining HIV patients in medical care promotes access to antiretroviral therapy, viral load suppression, and reduced HIV transmission to partners. We estimate the programmatic costs of a US multisite randomized controlled trial of an intervention to retain HIV patients in care. Six academically affiliated HIV clinics randomized patients to intervention (enhanced personal contact with patients across time coupled with basic HIV education) and control [standard of care (SOC)] arms. Retention in care was defined as 4-month visit constancy, that is, at least 1 primary care visit in each 4-month interval over a 12-month period. We used microcosting methods to collect unit costs and measure the quantity of resources used to implement the intervention in each clinic. All fixed and variable labor and nonlabor costs of the intervention were included. Visit constancy was achieved by 45.7% (280/613) of patients in the SOC arm and by 55.8% (343/615) of patients in the intervention arm, representing an increase of 63 patients (relative improvement 22.1%; 95% confidence interval: 9% to 36%; P < 0.01). The total annual cost of the intervention at the 6 clinics was $241,565, the average cost per patient was $393, and the estimated cost per additional patient retained in care beyond SOC was $3834. Our analyses showed that a retention in care intervention consisting of enhanced personal contact coupled with basic HIV education may be delivered at fairly low cost. These results provide useful information for guiding decisions about planning or scaling-up retention in care interventions for HIV-infected patients.


Effect of hormone replacement therapy on cardiac autonomic modulation.
To investigate the effects of hormone replacement therapy (HRT) on heart rate variability (HRV) in healthy postmenopausal women. Two groups were evaluated: group 1 (G1): 20 women not undergoing HRT (60 ± 5.89 years), group 2 (G2): 20 women undergoing HRT (59 ± 5.70 years). The HRTs involved were either conjugated equine estrogen with or without medroxyprogesterone, synthetic estrogen hormone, estradiol associated with norethisterone acetate or isoflavonoids. Electrocardiogram was recorded in the supine position for 10 min. Spectral analysis included low and high frequencies in absolute (LF and HF) and normalized units (LFnu and HFnu), which are predominantly cardiac sympathetic modulation (CSM) and cardiac vagal modulation (CVM) indicators, respectively. The LF/HF ratio was also calculated. Symbolic analysis involved the following indexes: 0V % (CSM indicator), 1V % (CSM and CVM indicators), 2LV % (predominantly CVM indicator) and 2UV % (CVM indicator). Shannon and conditional entropies were also calculated. Spectral analysis demonstrated that HRT affected HRV. LF, LFnu and LF/HF ratio were higher (showing increased CSM), while HFnu was lower (representing decreased CVM) in G2 than in G1. Correlations between complexity indices and HFnu were significant and positive only in G1. Women undergoing HRT presented higher CSM and lower CVM than those who were not. Moreover, the expected positive relationship between CVM and complexity of HRV was found only in control group, thus indicating that CVM in women under therapy drop below a minimal value necessary to the association to become apparent, suggesting an unfavorable cardiac autonomic modulation in spite of HRT.


Derepression of MicroRNA-138 Contributes to Loss of the Human Articular Chondrocyte Phenotype.
To investigate the function of microRNA-138 (miR-138) in human articular chondrocytes (HACs). The expression of miR-138 in intact cartilage and cultured chondrocytes and the effects of miR-138 overexpression on chondrocyte marker genes were investigated. Targets of miR-138 relevant to chondrocytes were identified and verified by overexpression of synthetic miRNA mimics and inhibitors, luciferase assays, chromatin immunoprecipitation, and RNA immunoprecipitation of native argonaute 2, using quantitative polymerase chain reaction, Western blotting, and luciferase assays. Expression levels of miR-138 were maintained at relatively low levels in intact human cartilage but were greatly increased upon loss of the differentiated phenotype in culture, with a concomitant decrease in the major cartilage extracellular matrix component COL2A1. We showed that miR-138 is able to repress the expression of COL2A1 by directly targeting Sp-1 and hypoxia-inducible factor 2α (HIF-2α), 2 transcription factors that are essential for COL2A1 transcription. We further demonstrated a direct association of these targets with miR-138 in the RNA-induced silencing complex and confirmed binding of Sp-1 to the COL2A1 promoter region in HACs. We propose that an evolutionary pressure helps to suppress expression levels of miR-138 in human cartilage, thus enabling expression of appropriate tissue-specific matrix genes. Inhibition of miR-138 may serve as a potential therapeutic strategy to maintain the chondrocyte phenotype or reduce the progression of dedifferentiation in cultured HACs.


[Benefit assessment of diagnostic procedures: quo vadimus?].
Not only therapeutic procedures, but also diagnostic procedures, have to demonstrate their patient-relevant benefits if they are to be reimbursed by public health insurance. Randomized trials comparing two diagnostic procedures allow us to assess these benefits directly if appropriate outcomes are used. However, owing to the widespread lack of such studies, it is now necessary to use the "linked evidence" approach as well, trying to predict the patient-relevant benefits from the results of comparative accuracy studies. Such a prediction is based on explicitly specifying our expectations with regard to the consequences of a change in diagnosis at the level of a single patient. We discuss the basic properties of these two approaches, which are relevant to the understanding of their possible role in the benefit assessment of diagnostic procedures. We try to predict the future roles of the two approaches and outline some of the issues on which a consensus is required to allow their successful use in benefit assessment. Furthermore, we indicate some of the developments related to the paradigm of individualized care that may influence the use of benefit assessments for diagnostic studies in the future.


Etanercept improves lipid profile and oxidative stress measures in patients with juvenile idiopathic arthritis.
To investigate the effect of 1-year treatment with the anti-tumor necrosis factor-α (TNF-α) drug etanercept on lipid profile and oxidative stress in children and adolescents with juvenile idiopathic arthritis (JIA). Thirty children with JIA (22 females; mean age 12.3 ± SD 5.7 yrs), all eligible for anti-TNF-α treatment, were assessed at baseline and after 6- and 12-month treatment with etanercept. Disease activity was determined using the Juvenile Arthritis Disease Activity Score (JADAS). Blood samples were drawn to measure the acute-phase reactants C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR), lipids, and the proinflammatory cytokines TNF-α, interleukin-1β (IL-1β), IL-6 and interferon-γ. To measure the oxidative stress marker 8-iso-prostaglandin F2α, 24-h urine samples were collected. Inflammatory indicators (CRP and ESR) and JADAS scores improved significantly after 1 year of etanercept treatment (all p < 0.001). Proinflammatory cytokines showed significant reduction during the study period (all p < 0.001). Similar reductions were detected in total cholesterol (p < 0.001), low-density lipoprotein cholesterol (p = 0.04), and triglycerides (p < 0.001), whereas no significant change was found in high-density lipoprotein cholesterol. No side effects were observed during the treatment period. This study shows for the first time that anti-TNF-α therapy for JIA is associated not only with a beneficial effect on clinical disease activity and inflammatory indexes, but also with improved lipid profile and oxidative stress. These findings suggest that TNF-α blockers might reduce atherosclerotic risk in children with JIA.


Blood collection from the sublingual vein in mice and hamsters: a suitable alternative to retrobulbar technique that provides large volumes and minimizes tissue damage.
Blood examination is a key element in studies of laboratory animals. In rodents, retrobulbar venous plexus puncture is a commonly used method for obtaining a blood sample. Although this technique yields large volumes of blood, the disadvantage is that it can lead to severe tissue damage. The aim of the present study was to develop the puncture of V. sublingualis as a suitable alternative technique for drawing blood in mice and other rodents. In rats, this method has been established for collecting large blood volumes. During the first part of the study, the sublingual bleeding technique was developed for use in mice and hamsters. Guinea pigs, however, do not have a sublingual vein; therefore, in this species the method is not possible. In the second part of the study, retrobulbar and sublingual methods were compared using male CD-1 mice. When compared with the retrobulbar method, sublingual venepuncture showed less tissue destruction in mice, with a decreased mean severity in the histological examination. In conclusion, sublingual venepuncture can be recommended as a suitable, alternative blood collection technique, because of the reduced risk of tissue damage in mice and hamsters.


Some implications of driver training for road accidents in Gaborone.
In many African countries, including Botswana, the driving schools and instruction given to pupils are uncontrolled. Thus, this paper presents the results of an investigation into the level of professional training offered by the driving schools in Gaborone. A total of 400 randomly selected drivers completed questionnaires on their training, accident records, and L-tests. Male trainee drivers received more instruction than females, and there was some indication that the pass rate on L-tests was lower for female drivers. Also, the pickup van forms the majority of the national vehicle population. It accounts for almost 40% of the accidents but is presently not used for training in the commercial driving schools. It is concluded that the quality of professional instruction in driving schools needs improvement so that learners can be encouraged to take training from these institutes.


Obesity and periodontitis are not associated in pregnant women.
The objective of this study was to investigate an association between obesity (exposure) and periodontitis (outcome) in pregnant women. This association was investigated and only five studies were identified as showing a positive association. However, some of these studies had limitations such as reduced sample sizes, inadequate exposure criteria and outcome measures, which question the internal validity of these investigations. A cross-sectional study was carried out with a sample of 644 pregnant women of the public health service of the municipality of Santo Antônio de Jesus, Bahia, Brazil. Data were obtained by collecting of socioeconomic-demographic information, health behavior, health conditions, and reproductive history through an interview. Obesity was evaluated using body mass index adjusted for gestational age and expected weight gain. The diagnosis of periodontitis followed two criteria: (a) Center for Disease Prevention and Control and American Academy of Periodontology (CDC/AAP); (b) Gomes-Filho et al (2018) using criterion that also evaluated bleeding upon probing; Prevalence ratios and respective 95% confidence intervals were obtained by Poisson regression analysis. In accordance with the outcome diagnostic criterion, the frequency of periodontitis was 17.24% (Gomes-Filho et al) and 66.92% (CDC/AAP). The participants were classified as low weight (19.72%), adequate weight (42.39%), overweight (24.84%), and obesity (13.04%), based on the exposure diagnostic criterion. The low weight and overweight groups were excluded from the data analysis, giving a final sample of 357 pregnant women. The association between obesity in pregnant women and periodontitis was not statistically significant, after adjusting for confounders such as age, schooling level, alcoholic beverage consumption, alimentary and nutritional orientation, urinary infection, and dental flossing. The findings showed a high frequency of periodontitis, obesity, and overweight in the studied population but no association between obesity and periodontitis in pregnant women was found.


Estimates of disease effects on soybean yields in the United States 2003 to 2005.
Research must focus on management of diseases that cause extensive losses, especially when funds for research are limited. Knowledge of the losses caused by various soybean diseases is essential when prioritizing research budgets. The objective of this project was to compile estimates of soybean yields suppressed due to diseases for each soybean-producing state in the US from 2003 to 2005. The goal was to provide this information to help funding agencies and scientists prioritize research objectives and budgets. Yield suppression due to individual diseases varied among regions in the US, and the total of soybean yields suppressed due to diseases in the US varied among years. Soybean cyst nematode suppressed US soybean yield more during 2003 to 2005 than any other disease. Phytophthora root and stem rot, sudden death syndrome, and seedling diseases ranked in the top four on the list of diseases that suppressed soybean yield during these years. This is the first report of soybean yield suppression due to Asian soybean rust in the United States.


Defined anaerobic growth medium for studying Candida albicans basic biology and resistance to eight antifungal drugs.
The polymorphic fungus Candida albicans is one of the most versatile opportunistic pathogens in humans. Many organs of the human body are potential targets for infection by this pathogen, but infection is commonly localized in the gastrointestinal tract, an environment providing anaerobic growth conditions. We describe a chemically defined anaerobic growth medium for four strains of Candida albicans (A72, SC5314, MEN, and 10261). It is a defined liquid glucose-phosphate-proline growth medium supplemented with oleic acid, nicotinic acid, and ammonium chloride. The cells did not require or respond to added ergosterol. Oleic acid and nicotinic acid are growth factors which are required only for the anaerobic growth of C. albicans. An important technical feature of this study was the use of anaerobically grown inocula to study anaerobic growth. Anaerobically, the cells grew exclusively as mycelia at 25, 30, and 37 degrees C. The doubling time at 30 degrees C was ca. 20 h. The cells did not produce farnesol and did not respond to exogenous farnesol, and they were resistant to the highest tested levels of amphotericin B and four of the azole antifungals. We suggest that the anaerobic growth of C. albicans may contribute to the trailing end point phenomenon and the resistance of C. albicans biofilms to antifungal drugs.


Inhibition of Intestinal OATP2B1 by the Calcium Receptor Antagonist Ronacaleret Results in a Significant Drug-Drug Interaction by Causing a 2-Fold Decrease in Exposure of Rosuvastatin.
Rosuvastatin is a widely prescribed antihyperlipidemic which undergoes limited metabolism, but is an in vitro substrate of multiple transporters [organic anion transporting polypeptide 1B1 (OATP1B1), OATP1B3, OATP1A2, OATP2B1, sodium-taurocholate cotransporting polypeptide, breast cancer resistance protein (BCRP), multidrug resistance protein 2 (MRP2), MRP4, organic anion transporter 3]. It is therefore frequently used as a probe substrate in clinical drug-drug interaction (DDI) studies to investigate transporter inhibition. Although each of these transporters is believed to play a role in rosuvastatin disposition, multiple pharmacogenetic studies confirm that OATP1B1 and BCRP play an important role in vivo. Ronacaleret, a drug-development candidate for treatment of osteoporosis (now terminated), was shown to inhibit OATP1B1 in vitro (IC50 = 11 µM), whereas it did not inhibit BCRP. Since a DDI risk through inhibition of OATP1B1 could not be discharged, a clinical DDI study was performed with rosuvastatin before initiation of phase II trials. Unexpectedly, coadministration with ronacaleret decreased rosuvastatin exposure by approximately 50%, whereas time of maximal plasma concentration and terminal half-life remained unchanged, suggesting decreased absorption and/or enhanced first-pass elimination of rosuvastatin. Of the potential in vivo rosuvastatin transporter pathways, two might explain the observed results: intestinal OATP2B1 and hepatic MRP4. Further investigations revealed that ronacaleret inhibited OATP2B1 (in vitro IC50 = 12 µM), indicating a DDI risk through inhibition of absorption. Ronacaleret did not inhibit MRP4, discharging the possibility of enhanced first-pass elimination of rosuvastatin (reduced basolateral secretion from hepatocytes into blood). Therefore, a likely mechanism of the observed DDI is inhibition of intestinal OATP2B1, demonstrating the in vivo importance of this transporter in rosuvastatin absorption in humans.