Mutations in the WTX-gene are found in some high-grade microsatellite instable (MSI-H) colorectal cancers.
Genetically, colorectal cancers (CRCs) can be subdivided into tumors with chromosomal instability (CIN) or microsatellite instability (MSI). In both types of CRCs genes that are involved in the degradation of beta-CATENIN are frequently mutated. Whereas in CIN CRCs APC (Adenomatous Polyposis Coli) is affected in most cases, high grade MSI (MSI-H) CRCs frequently display mutations in various genes, like the APC-, AXIN2- or CTNNBI (beta-CATENIN) gene itself. Recently in Wilms tumors, WTX (Wilms tumor gene on the X-chromosome) was discovered as another gene involved in the destruction of beta-CATENIN. As the WTX-gene harbors a short T6-microsatellite in its N-terminal coding region, we hypothesized that frameshift-mutations might occur in MSI-H CRCs in the WTX gene, thus additionally contributing to the stabilization of beta-CATENIN in human CRCs. DNA was extracted from 632 formalin-fixed, paraffin-embedded metastatic CRCs (UICCIV) and analyzed for MSI-H by investigating the stability of the highly sensitive microsatellite markers BAT25 and BAT26 applying fluorescence capillary electrophoresis (FCE). Then, in the MSI-H cases, well described mutational hot spot regions from the APC-, AXIN2- and CTNNBI genes were analyzed for genomic alterations by didesoxy-sequencing while the WTX T6-microsatellite was analyzed by fragment analysis. Additionally, the PCR products of T5-repeats were subcloned and mutations were validated using didesoxy-sequencing. Furthermore, the KRAS and the BRAF proto-oncogenes were analyzed for the most common activating mutations applying pyro-sequencing. mRNA expression of WTX from MSI-H and MSS cases and a panel of colorectal cancer cell lines was investigated using reverse transcription (RT-) PCR and FCE. In our cohort of 632 metastatic CRCs (UICCIV) we identified 41 MSI-H cases (6.5%). Two of the 41 MSI-H cases (4.8%) displayed a frameshift mutation in the T6-repeat resulting in a T5 sequence. Only one case, a male patient, expressed the mutated WTX gene while being wild type for all other investigated genes. Mutations in the WTX-gene might compromise the function of the beta-CATENIN destruction complex in only a small fraction of MSI-H CRCs thus contributing to the process of carcinogenesis.


Does patient-centered care improve provision of preventive services?
While patient-centered care (PCC) is desirable for many reasons, its relationship to treatment outcomes is controversial. We evaluated the relationship between PCC and the provision of preventive services. We obtained facility-level estimates of how well each VA hospital provided PCC from the 1999 ambulatory Veterans Satisfaction Survey. PCC delivery was measured by the average percentage of responses per facility indicating satisfactory performance from items in 8 PCC domains: access, incorporating patient preferences, patient education, emotional support, visit coordination, overall coordination of care, continuity, and courtesy. Additional predictors included patient population and facility characteristics. Our outcome was a previously validated hospital-level benchmarking score describing facility-level performance across 12 U.S. Preventive Services Task Force-recommended interventions, using the 1999 Veterans Health Survey. Facility-level delivery of preventive services ranged from an overall mean of 90% compliance for influenza vaccinations to 18% for screening for seat belt use. Mean overall PCC scores ranged from excellent (>90% for the continuity of care and courtesy of care PCC domains) to modest (<70% for patient education). Correlates of better preventive service delivery included how often patients were able to discuss their concerns with their provider, the percent of visits at which patients saw their usual provider, and the percent of patients receiving >90% of care from a VA hospital. Improved communication between patients and providers, and continuity of care are associated with increased provision of preventive services, while other aspects of PCC are not strongly related to delivery of preventive services.


Brief chemotherapy, Stanford V, and adjuvant radiotherapy for bulky or advanced-stage Hodgkin's disease: a preliminary report.
Although survival rates have improved for patients with bulky and advanced-stage Hodgkin's disease (HD), current treatments entail substantial acute morbidity and risks for late effects such as infertility, second malignancies, and cardiopulmonary toxicities. A novel, brief chemotherapy regimen (doxorubicin, vinblastine, mechlorethamine, vincristine, bleomycin, etoposide, and prednisone [Stanford V]) was designed to shorten the duration of treatment, significantly reduce cumulative doses of alkylating agents, doxorubicin, and bleomycin, and maintain dose-intensity (DI). This brief chemotherapy was combined with radiation therapy (RT) to bulky disease sites. Since May 1989, 65 previously untreated patients were treated for stage II HD with bulky mediastinal involvement (n = 21) or for stage III or IV HD (n = 44). Patients received weekly chemotherapy for 12 weeks. Consolidative RT was given to the first 25 patients to sites of initial bulky disease or radiographic abnormalities that persisted after chemotherapy; in the remaining 40 patients, RT was limited to bulky disease (adenopathy > or = 5 cm and/or macroscopic splenic nodules defined by computed tomography [CT]). With a median follow-up period of 2 years, actuarial 3-year survival rate is 96% and failure-free survival (FFS) rate is 87%. The 3-year FFS rate is 100% for stage II patients with bulky mediastinal disease and 82% for patients with stage III to IV disease. There were no treatment-related deaths. In a preliminary analysis on a subset of patients, female and male fertility appears to be preserved. These preliminary results indicate that the Stanford V chemotherapy regimen with or without RT is well-tolerated and effective therapy for bulky, limited-stage, and advanced-stage HD. Less cumulative exposure to alkylating agents, doxorubicin, and bleomycin and limited use of radiation is expected to decrease risks for second neoplasms and late cardiopulmonary toxicity. Based on these results, the Stanford V chemotherapy with or without RT regimen deserves further study in the context of a randomized clinical trial.


Low bone mineral density in COPD patients with osteoporosis is related to low daily physical activity and high COPD assessment test scores.
COPD patients have an increased prevalence of osteoporosis (OP) compared with healthy people. Physical inactivity in COPD patients is a crucial risk factor for OP; the COPD assessment test (CAT) is the newest assessment tool for the health status and daily activities of COPD patients. This study investigated the relationship among daily physical activity (DPA), CAT scores, and bone mineral density (BMD) in COPD patients with or without OP. This study included 30 participants. Ambulatory DPA was measured using actigraphy and oxygen saturation by using a pulse oximeter. BMD was measured using dual-energy X-ray absorptiometry. OP was defined as a T-score (standard deviations from a young, sex-specific reference mean BMD) less than or equal to -2.5 SD for the lumbar spine, total hip, and femoral neck. We quantified oxygen desaturation during DPA by using a desaturation index and recorded all DPA, except during sleep. COPD patients with OP had lower DPA and higher CAT scores than those of patients without OP. DPA was significantly positively correlated with (lumbar spine, total hip, and femoral neck) BMD (r=0.399, 0.602, 0.438, respectively, all P<0.05) and T-score (r=0.471, 0.531, 0.459, respectively, all P<0.05), whereas CAT scores were significantly negatively correlated with (total hip and femoral neck) BMD (r=-0.412, -0.552, respectively, P<0.05) and (lumbar spine, total hip, and femoral neck) T-score (r=-0.389, -0.429, -0.543, respectively, P<0.05). Low femoral neck BMD in COPD patients was related to high CAT scores. Our results show no significant difference in desaturation index, low SpO2, and inflammatory markers (IL-6, TNF-α, IL-8/CXCL8, CRP, and 8-isoprostane) between the two groups. Chest physicians should be aware that COPD patients with OP have low DPA and high CAT scores.


Different nitrosative-induced microtubular modifications and testosterone neuroprotective effects on high-D-glucose-exposed neuroblastoma and glioma cells.
Diabetic complications can often affect the central nervous system since the chronic exposure to hyperglycemia can result in the production of high concentration of reactive oxygen species with subsequent damage of several cell structures such as the cytoskeleton. In order to antagonize the oxidative status many substances have been tested as antioxidants. In the present work attention has been focused on the possible nitrosative effect of hyperglycemia on microtubular network of neuroblastoma and glioma mortalized cell lines, testing the possible neuroprotective effect of testosterone. Neuroblastoma (C1300) and glioma (C6) cell lines were cultured in the presence of 300 mM (C1300) or 310 mM (C6) D-glucose, with or without 50 nM testosterone. After 72 hrs, morphology, growth rate, cell viability and catalase activity were evaluated. In addition, with the aim to detect any changes in the amount of tubulin isoforms, Western blot analysis was performed. In D-glucose-exposed cells, it was found a down-regulation of tubulin isoforms and an increase in 3-nitro-L-tyrosine and subsequent modifications in cell morphology, growth rate, viability and catalase activity. All these changes were more severe in neuroblastoma than in glioma cell line. When testosterone was added to the medium, all the parameters were very similar to controls. This neuroprotective action was well-detectable in C1300 cells, whereas testosterone was not able to recover significantly in C6 cells. Our results displayed: i) a selective action of high glucose on microtubules; ii) a different sensitivity to oxidative stress in neuronal and glial cells; iii) a different neuroprotective action of testosterone on neuronal and glial cells.


Impact of traumatic dental injuries on oral health-related quality of life of preschool children: A systematic review and meta-analysis.
Observational studies have suggested that traumatic dental injuries (TDI) can lead to pain, loss of function and esthetic problems, with physical, emotional and social consequences for children and their families. However, population-based studies that investigate the impact of TDI on oral health-related quality of life (OHRQoL) among preschool children are scarce and offer conflicting results. The aim of the systematic review and meta-analysis was to evaluate the impact of TDI on OHRQoL among preschool children (PROSPERO-CRD42015032513). An electronic search of six databases was performed in PubMed (MEDLINE), ISI Web of Science, Scopus, Science Direct, EMBASE and Google Scholar, with no language or publication date restrictions. The eligibility criteria were TDI as the exposure variable, OHRQoL as the outcome and a population of children up to six years of age. RevMan software was used for data analysis. Results are expressed as odds ratios with 95% confidence intervals for the total score of the Early Childhood Oral Health Impact Scale (ECOHIS) as well as the scores of the Child Impact Section (CIS) and Family Impact Section (FIS). The random effect model was chosen and heterogeneity was evaluated using the I2 test. 2,013 articles were initially retrieved; 1,993 articles were excluded based on title and abstracts; 10 articles excluded after full-text analysis. Ten studies comprising a population of 7,461 preschool children were included in the systematic review and nine studies were included in the meta-analysis. TDI caused a negative impact on OHRQoL based on the overall ECOHIS (OR: 1.24; 95% CI: 1.08-1.43) and CIS (OR: 1.23; 95% CI: 1.07-1.41), but not the FIS (OR: 1.09; 95% CI: 0.90-1.32). TDI negatively impacted on OHRQoL of preschool children. The present findings indicate the need for TDI prevention and treatment programs in early childhood.


Out-of-hours discharge from intensive care, in-hospital mortality and intensive care readmission rates: a systematic review protocol.
Most patients are discharged from an intensive care unit with an expectation that they will survive their hospital stay, yet these patients have high subsequent in-hospital mortality. Patients are frequently discharged from an intensive care unit to a lower level of hospital care in the evenings and at night (out-of-hours). By affecting the care that patients receive, out-of-hours discharge may alter post-intensive care in-hospital mortality rates. Two searches will be conducted-the first a general search for all factors associated with post-intensive care in-hospital mortality and a second focused specifically on out-of-hours discharges. Searches will be performed in multiple databases, including Medline, Embase, Web of Knowledge, Cumulative Index of Nursing and Allied Health Literature (CINAHL) and the Cochrane Library. OpenGrey will also be searched, to ensure any unpublished 'grey' data are accessed. Language and date restrictions will not be applied. Assessment for inclusion and data extraction will be undertaken by two independent reviewers. Methodological quality will be assessed using the ACROBAT-NRSI tool. The primary outcome measure will be post-intensive care in-hospital mortality. To provide a clearer picture of this problem, studies reporting readmission to the intensive care unit (ICU) will also be included, even in the absence of report of in-hospital mortality. The primary outcome data will be synthesised and summarised using a random-effects meta-analysis. Where possible, subgroup meta-analyses will assess associated factors such as discharge destination, palliative care discharges and severity of illness scores. To the best of our knowledge, a systematic review of the association of out-of-hours discharge with in-hospital mortality has never been undertaken. Synthesis of the available information is important because out-of-hours discharge remains common and, if associated with post-intensive care unit mortality, is highly amenable to system change. PROSPERO CRD42014010321.


Etidronate disodium in postmenopausal osteoporosis.
Calcium-balance, calcium-kinetic, and physiologic observations were made in 10 osteoporotic patients treated for 6 to 12 mo with disodium etidronate (EHDP) in oral doses of 20 mg/kg/day. Absorption of EHDP averaged 10%, and effective retained dose was approximately 1.6 mg/kg/day. Patient acceptance was excellent, and there were no detectable clinical or biochemical untoward effects of treatment. Serum phosphorus was elevated on average of 1.1 mg/100 ml, and serum calcium, 0.4 mg/100 ml. EHDP reduced bone resorption by about 50% and depressed bone mineralization by almost as much. Both the total miscible calcium pool and its component compartments were smaller on EHDP, presumably because of a combination of interference with bone mineral reactivity and the decrease in mineralization rate. Urine calcium was increased by a small, but highly significant, amount, and gastrointestinal calcium absorption was also significantly increased. Calcium balance shifted slightly but significantly in the positive direction. The magnitude of these changes correlated with the urinary level of EHDP.


Complementary costimulation of human T-cell subpopulations by cluster of differentiation 28 (CD28) and CD81.
Cluster of differentiation 81 (CD81) is a widely expressed tetraspanin molecule that physically associates with CD4 and CD8 on the surface of human T cells. Coengagement of CD81 and CD3 results in the activation and proliferation of T cells. CD81 also costimulated mouse T cells that lack CD28, suggesting either a redundant or a different mechanism of action. Here we show that CD81 and CD28 have a preference for different subsets of T cells: Primary human naïve T cells are better costimulated by CD81, whereas the memory T-cell subsets and Tregs are better costimulated by CD28. The more efficient activation of naïve T cells by CD81 was due to prolonged signal transduction compared with that by CD28. We found that IL-6 played a role in the activation of the naïve T-cell subset by CD81. Combined costimulation through both CD28 and CD81 resulted in an additive effect on T-cell activation. Thus, these two costimulatory molecules complement each other both in the strength of signal transduction and in T-cell subset inclusions. Costimulation via CD81 might be useful for expansion of T cells for adoptive immunotherapy to allow the inclusion of naïve T cells with their broad repertoire.


Cell phones: modern man's nemesis?
Over the past decade, the use of mobile phones has increased significantly. However, with every technological development comes some element of health concern, and cell phones are no exception. Recently, various studies have highlighted the negative effects of cell phone exposure on human health, and concerns about possible hazards related to cell phone exposure have been growing. This is a comprehensive, up-to-the-minute overview of the effects of cell phone exposure on human health. The types of cell phones and cell phone technologies currently used in the world are discussed in an attempt to improve the understanding of the technical aspects, including the effect of cell phone exposure on the cardiovascular system, sleep and cognitive function, as well as localized and general adverse effects, genotoxicity potential, neurohormonal secretion and tumour induction. The proposed mechanisms by which cell phones adversely affect various aspects of human health, and male fertility in particular, are explained, and the emerging molecular techniques and approaches for elucidating the effects of mobile phone radiation on cellular physiology using high-throughput screening techniques, such as metabolomics and microarrays, are discussed. A novel study is described, which is looking at changes in semen parameters, oxidative stress markers and sperm DNA damage in semen samples exposed in vitro to cell phone radiation.


Association of tooth loss and oral hygiene with risk of gastric adenocarcinoma.
Poor oral health and tooth loss have been proposed as possible risk factors for some chronic diseases, including gastric cancer. However, a small number of studies have tested these associations. We conducted a case-control study in Golestan Province, Iran, that enrolled 309 cases diagnosed with gastric adenocarcinoma (118 noncardia, 161 cardia, and 30 mixed-locations) and 613 sex, age, and neighborhood matched controls. Data on oral health were obtained through physical examination and questionnaire including tooth loss, the number of decayed, missing, and filled teeth, and frequency of tooth brushing. ORs and 95% confidence intervals (95% CI) were obtained using conditional logistic regression models adjusted for potential confounders. Standard one degree-of-freedom linear trend test and a multiple degree-of-freedom global test of the effect of adding oral hygiene variables to the model were also calculated. Our results showed apparent associations between tooth loss and decayed, missing, filled teeth (DMFT) score with risk of gastric cancer, overall and at each anatomic subsite. However, these associations were not monotonic and were strongly confounded by age. The results also showed that subjects who brushed their teeth less than daily were at significantly higher risk for gastric cardia adenocarcinoma ORs (95% CI) of 5.6 (1.6-19.3). We found evidence for an association between oral health and gastric cancer, but the nonmonotonic association, the relatively strong effect of confounder adjustment, and inconsistent results across studies must temper the strength of any conclusions.


True Interindividual Variability Exists in Postprandial Appetite Responses in Healthy Men But Is Not Moderated by the FTO Genotype.
After meal ingestion, a series of coordinated hormone responses occur concomitantly with changes in perceived appetite. It is not known whether interindividual variability in appetite exists in response to a meal. The aim of this study was to 1) assess the reproducibility of appetite responses to a meal; 2) quantify individual differences in responses; and 3) explore any moderating influence of the fat mass and obesity associated (FTO) gene. Using a replicated crossover design, 18 healthy men (mean ± SD age: 28.5 ± 9.8 y; BMI: 27.0 ± 5.0 kg/m2) recruited according to FTO genotype (9 AA, 9 TT) completed 2 identical control and 2 identical standardized meal conditions (5025 kJ) in randomized sequences. Perceived appetite and plasma acylated ghrelin, total peptide YY (PYY), insulin, and glucose concentrations were measured before and after interventions as primary outcomes. Interindividual differences were explored using Pearson's product-moment correlations between the first and second replicates of the control-adjusted meal response. Within-participant covariate-adjusted linear mixed models were used to quantify participant-by-condition and genotype-by-condition interactions. The meal suppressed acylated ghrelin and appetite perceptions [standardized effect size (ES): 0.18-4.26] and elevated total PYY, insulin, and glucose (ES: 1.96-21.60). For all variables, SD of change scores was greater in the meal than in the control conditions. Moderate-to-large positive correlations were observed between the 2 replicates of control-adjusted meal responses for all variables (r = 0.44-0.86, P ≤ 0.070). Participant-by-condition interactions were present for all variables (P ≤ 0.056). FTO genotype-by-condition interactions were nonsignificant (P ≥ 0.19) and treatment effect differences between genotype groups were small (ES ≤ 0.27) for all appetite parameters. Reproducibility of postprandial appetite responses is generally good. True interindividual variability is present beyond any random within-subject variation in healthy men but we detected no moderation by the FTO genotype. These findings highlight the importance of exploring individual differences in appetite for the prevention and treatment of obesity. This trial was registered at clinicaltrials.gov as NCT03771690.


Ciliary neurotrophic factor promotes motor reinnervation of the musculocutaneous nerve in an experimental model of end-to-side neurorrhaphy.
It is difficult to repair nerve if proximal stump is unavailable or autogenous nerve grafts are insufficient for reconstructing extensive nerve damage. Therefore, alternative methods have been developed, including lateral anastomosis based on axons' ability to send out collateral sprouts into denervated nerve. The different capacity of a sensory or motor axon to send a sprout is controversial and may be controlled by cytokines and/or neurotrophic factors like ciliary neurotrophic factor (CNTF). The aim of the present study was to quantitatively assess collateral sprouts sent out by intact motor and sensory axons in the end-to-side neurorrhaphy model following intrathecal administration of CNTF in comparison with phosphate buffered saline (vehiculum) and Cerebrolysin. The distal stump of rat transected musculocutaneous nerve (MCN) was attached in an end-to-side fashion with ulnar nerve. CNTF, Cerebrolysin and vehiculum were administered intrathecally for 2 weeks, and all animals were allowed to survive for 2 months from operation. Numbers of spinal motor and dorsal root ganglia neurons were estimated following their retrograde labeling by Fluoro-Ruby and Fluoro-Emerald applied to ulnar and musculocutaneous nerve, respectively. Reinnervation of biceps brachii muscles was assessed by electromyography, behavioral test, and diameter and myelin sheath thickness of regenerated axons. Vehiculum or Cerebrolysin administration resulted in significantly higher numbers of myelinated axons regenerated into the MCN stumps compared with CNTF treatment. By contrast, the mean diameter of the myelinated axons and their myelin sheath thickness in the cases of Cerebrolysin- or CNTF-treated animals were larger than were those for rats treated with vehiculum. CNTF treatment significantly increased the percentage of motoneurons contributing to reinnervation of the MCN stumps (to 17.1%) when compared with vehiculum or Cerebrolysin treatments (at 9.9 or 9.6%, respectively). Reduced numbers of myelinated axons and simultaneously increased numbers of motoneurons contributing to reinnervation of the MCN improved functional reinnervation of the biceps brachii muscle after CNTF treatment. The present experimental study confirms end-to-side neurorrhaphy as an alternative method for reconstructing severed peripheral nerves. CNTF promotes motor reinnervation of the MCN stump after its end-to-side neurorrhaphy with ulnar nerve and improves functional recovery of the biceps brachii muscle.


Genomic analysis of Sparus aurata reveals the evolutionary dynamics of sex-biased genes in a sequential hermaphrodite fish.
Sexual dimorphism is a fascinating subject in evolutionary biology and mostly results from sex-biased expression of genes, which have been shown to evolve faster in gonochoristic species. We report here genome and sex-specific transcriptome sequencing of Sparus aurata, a sequential hermaphrodite fish. Evolutionary comparative analysis reveals that sex-biased genes in S. aurata are similar in number and function, but evolved following strikingly divergent patterns compared with gonochoristic species, showing overall slower rates because of stronger functional constraints. Fast evolution is observed only for highly ovary-biased genes due to female-specific patterns of selection that are related to the peculiar reproduction mode of S. aurata, first maturing as male, then as female. To our knowledge, these findings represent the first genome-wide analysis on sex-biased loci in a hermaphrodite vertebrate species, demonstrating how having two sexes in the same individual profoundly affects the fate of a large set of evolutionarily relevant genes.


Safety, pharmacokinetics, and pharmacodynamics of avoralstat, an oral plasma kallikrein inhibitor: phase 1 study.
Avoralstat is a potent small-molecule oral plasma kallikrein inhibitor under development for treatment of hereditary angioedema (HAE). This first-in-human study evaluated the safety, tolerability, pharmacokinetics, and pharmacodynamics of avoralstat. This double-blind, placebo-controlled, ascending-dose cohort trial evaluated avoralstat single doses of 50, 125, 250, 500, and 1000 mg and multiple doses up to 2400 mg daily (100, 200, 400, and 800 mg every 8 h [q8 h] up to 7 days). Avoralstat (n = 71) was generally well tolerated with no signals for a safety concern; there were no serious adverse events (AEs) or discontinuations due to AEs, and compared to placebo (n = 18), no notable difference in AEs. Four moderate severity AEs were reported in two subjects; syncope after a single 250 mg dose (one subject) and abdominal pain, back pain, and eczema after multiple doses of 800 mg avoralstat (one subject). For multiple-dose cohorts, the incidence of gastrointestinal AEs was highest at the 2400 mg/day dose. Elimination of avoralstat was bi-exponential with a terminal half-life of 12-31 h. Inhibition of plasma kallikrein was observed at all doses, and the degree of inhibition was highly correlated with avoralstat concentrations (R = 0.93). Mean avoralstat concentrations at doses ≥400 mg q8 h met or exceeded plasma kallikrein EC50 values throughout the dosing interval. Avoralstat was well tolerated, and drug exposure was sufficient to meet target levels for inhibition of plasma kallikrein. Based on these results, the 400 mg q8 h dose was selected for further evaluation in patients with HAE.


Tumor suppressor protein kinase Chk2 is a mediator of anoikis of intestinal epithelial cells.
Resistance of carcinoma cells to anoikis, apoptosis that is normally induced by detachment of nonmalignant epithelial cells from the extracellular matrix, is thought to be critical for carcinoma progression. Molecular mechanisms that control anoikis of nonmalignant and cancer cells are understood poorly. In an effort to understand them we found that detachment of nonmalignant intestinal epithelial cells triggers upregulation of Chk2, a pro-apoptotic protein kinase that has never been implicated in anoikis and has been thought to kill cells mainly under the conditions compromising genome integrity. We found that enforced downregulation of Chk2 protects intestinal epithelial cells from anoikis. Chk2 can kill cells by stabilizing p53 tumor suppressor protein or via p53-independent mechanisms, and we established that Chk2-mediated anoikis of intestinal epithelial cells is p53-independent. We further found that, unlike nonmalignant intestinal epithelial cells whose anoikis is triggered by detachment-induced Chk2 upregulation, intestinal epithelial cells carrying oncogenic ras, a known inhibitor of anoikis, remain anoikis-resistant in response to enforced Chk2 upregulation. By contrast, drugs, such as topoisomerase I inhibitors, that can kill cells via Chk2-indpendent mechanisms, efficiently triggered anoikis of ras-transformed cells. Thus, oncogenic ras can prevent Chk2 from triggering anoikis even when levels of this protein kinase are elevated in cancer cells, and the use of therapeutic agents that kill cells in a Chk-2-independent, rather than Chk-2-dependent, manner could represent an efficient strategy for overcoming ras-induced anoikis resistance of these cells. We conclude that Chk-2 is an important novel component of anoikis-promoting machinery of intestinal epithelial cells.


Increasing precipitation event size increases aboveground net primary productivity in a semi-arid grassland.
Water availability is the primary constraint to aboveground net primary productivity (ANPP) in many terrestrial biomes, and it is an ecosystem driver that will be strongly altered by future climate change. Global circulation models predict a shift in precipitation patterns to growing season rainfall events that are larger in size but fewer in number. This "repackaging" of rainfall into large events with long intervening dry intervals could be particularly important in semi-arid grasslands because it is in marked contrast to the frequent but small events that have historically defined this ecosystem. We investigated the effect of more extreme rainfall patterns on ANPP via the use of rainout shelters and paired this experimental manipulation with an investigation of long-term data for ANPP and precipitation. Experimental plots (n = 15) received the long-term (30-year) mean growing season precipitation quantity; however, this amount was distributed as 12, six, or four events applied manually according to seasonal patterns for May-September. The long-term mean (1940-2005) number of rain events in this shortgrass steppe was 14 events, with a minimum of nine events in years of average precipitation. Thus, our experimental treatments pushed this system beyond its recent historical range of variability. Plots receiving fewer, but larger rain events had the highest rates of ANPP (184 +/- 38 g m(-2)), compared to plots receiving more frequent rainfall (105 +/- 24 g m(-2)). ANPP in all experimental plots was greater than long-term mean ANPP for this system (97 g m(-2)), which may be explained in part by the more even distribution of applied rain events. Soil moisture data indicated that larger events led to greater soil water content and likely permitted moisture penetration to deeper in the soil profile. These results indicate that semi-arid grasslands are capable of responding immediately and substantially to forecast shifts to more extreme precipitation patterns.


Burnout among osteopathic otolaryngology residents: identification during formative training years.
Studies of burnout among allopathic physicians have shown many deleterious effects for both physicians and patients. To our knowledge, no studies have quantified burnout among osteopathic physicians. To determine the prevalence of burnout, mentoring, and resident training satisfaction among US osteopathic otolaryngology residents compared with previously published data for allopathic otolaryngology residents. A cross-sectional, questionnaire-based, electronic survey of US osteopathic otolaryngology residents was conducted. Residents were surveyed about demographic information, personal and professional life satisfaction, professional stressors, burnout (assessed with the Maslach Burnout Inventory-Human Services Survey), and mentor-resident interactions. Burnout was measured based on levels of emotional exhaustion (EE), depersonalization (DP), and low personal accomplishment (PA). Results were compared with previously published data for allopathic otolaryngology residents. Of the 102 osteopathic residents contacted, 48 (47%) responded and 47 provided complete responses in some categories. Burnout rates were low in 11 respondents (23%), moderate in 31 (66%), and high in 5 (11%), compared with published rates of 14%, 76%, and 10%, respectively, for allopathic residents (P=.18). The rates of EE and DP did not differ significantly from published data, but levels of PA were higher in osteopathic residents (P=.03). Sleep hours per night were significantly higher in osteopathic than in allopathic residents (mean, 6.6 vs 6.2; P=.04), and work hours per week were significantly lower (mean, 62 vs 71; P<.001). Increased EE scores were negatively associated with hours of sleep (ρ=-0.42, P=.003). Increased influence from a mentor was associated with decreased levels of burnout for all 3 components (EE: ρ=-0.54, P=.002; DP: ρ=-0.59, P<.001; PA: ρ=0.44, P=.02). To our knowledge, the current study is the first to quantify burnout among osteopathic residents, and our findings suggest that osteopathic residents have lower rates of burnout than allopathic residents. Osteopathic residents reported lower rates of low PA, increased hours of sleep, and decreased overall work hours. Further study of the relationship between mentoring and decreased burnout is needed.


Legacies of La Niña: North American monsoon can rescue trees from winter drought.
While we often assume tree growth-climate relationships are time-invariant, impacts of climate phenomena such as the El Niño Southern Oscillation (ENSO) and the North American Monsoon (NAM) may challenge this assumption. To test this assumption, we grouped ring widths (1900-present) in three southwestern US conifers into La Niña periods (LNP) and other years (OY). The 4 years following each La Niña year are included in LNP, and despite 1-2 year growth declines, compensatory adjustments in tree growth responses result in essentially equal mean growth in LNP and OY, as average growth exceeds OY means 2-4 years after La Niña events. We found this arises because growth responses in the two periods are not interchangeable: Due to differences in growth-climate sensitivities and climatic memory, parameters representing LNP growth fail to predict OY growth and vice versa (decreases in R2 up to 0.63; lowest R2 = 0.06). Temporal relationships between growth and antecedent climate (memory) show warmer springs and longer growing seasons negatively impact growth following dry La Niña winters, but that NAM moisture can rescue trees after these events. Increased importance of monsoonal precipitation during LNP is key, as the largest La Niña-related precipitation deficits and monsoonal precipitation contributions both occur in the southern part of the region. Decreases in first order autocorrelation during LNP were largest in the heart of the monsoon region, reflecting both the greatest initial growth declines and the largest recovery. Understanding the unique climatic controls on growth in Southwest conifers requires consideration of both the influences and interactions of drought, ENSO, and NAM, each of which is likely to change with continued warming. While plasticity of growth sensitivity and memory has allowed relatively quick recovery in the tree-ring record, recent widespread mortality events suggest conditions may soon exceed the capacity for adjustment in current populations.


Dendritic arborization and spine dynamics are abnormal in the mouse model of MECP2 duplication syndrome.
MECP2 duplication syndrome is a childhood neurological disorder characterized by intellectual disability, autism, motor abnormalities, and epilepsy. The disorder is caused by duplications spanning the gene encoding methyl-CpG-binding protein-2 (MeCP2), a protein involved in the modulation of chromatin and gene expression. MeCP2 is thought to play a role in maintaining the structural integrity of neuronal circuits. Loss of MeCP2 function causes Rett syndrome and results in abnormal dendritic spine morphology and decreased pyramidal dendritic arbor complexity and spine density. The consequences of MeCP2 overexpression on dendritic pathophysiology remain unclear. We used in vivo two-photon microscopy to characterize layer 5 pyramidal neuron spine turnover and dendritic arborization as a function of age in transgenic mice expressing the human MECP2 gene at twice the normal levels of MeCP2 (Tg1; Collins et al., 2004). We found that spine density in terminal dendritic branches is initially higher in young Tg1 mice but falls below control levels after postnatal week 12, approximately correlating with the onset of behavioral symptoms. Spontaneous spine turnover rates remain high in older Tg1 animals compared with controls, reflecting the persistence of an immature state. Both spine gain and loss rates are higher, with a net bias in favor of spine elimination. Apical dendritic arbors in both simple- and complex-tufted layer 5 Tg1 pyramidal neurons have more branches of higher order, indicating that MeCP2 overexpression induces dendritic overgrowth. P70S6K was hyperphosphorylated in Tg1 somatosensory cortex, suggesting that elevated mTOR signaling may underlie the observed increase in spine turnover and dendritic growth.


Health reform: a community experience using design research as a guide.
Meaningful health reform in the United States must improve the health of the population while lowering costs. In an effort to provide a framework for doing so, the Institute of Health Care Improvement created the triple aim, which encompasses the goals of (1) improving individual health and experience with the health care system, (2) improving population health, and (3) decreasing the rate of per capita health care costs. Current reform efforts have focused on the development of Patient-Centered Medical Homes (an innovative team-based model of care that facilitates a partnership between the patient's personal physician coordinating care throughout a patient's lifetime to maximize health outcomes), but these relatively narrow efforts are focused on office practice and payment methods and are not generally oriented toward community needs. We sought to apply design research in assessing a community opportunity to apply the triple aim as a strategy to transform health care delivery. Mixed methodology provides greater insight into the unexpressed health needs of individuals and into the creation of delivery systems more likely to achieve the triple aim. In a small, midwestern town, a mixed methods approach was used to assess community health needs to facilitate design and implementation of care delivery systems. The research findings suggest that health system design concepts should focus on the creation of health, not health care; foster simplicity; create nurturing relationships; eliminate user fear; and contain costs. These observations can be helpful to health care professionals who are developing new methods of care delivery and policymakers and payers contemplating new payment systems to achieve the goals of the triple aim.


Does distrust in providers affect health-care utilization in China?
How trust affects health-care utilization is not well-understood, especially in low- and middle-income countries. This article focuses on China, a middle-income country where low trust in health-care settings has become a prominent issue, but actual levels of distrust and their implications for utilization are unknown. We conducted a nationally representative survey of the Chinese population (November 2012 to January 2013), which resulted in a sample of 3680 adult men and women. Respondents rated their trust in different types of health-care providers. Using multivariate logistic and negative binomial regression models, we estimated the association between distrust in clinics and respondents' hospital visits in the last year; whether they had sought hospital treatment first for two common symptoms (headache, cold) in the last 2 months; and whether they said they would go first to a hospital if they had a minor or major illness. We analysed these associations before and after adjusting for performance evaluations of clinics and hospitals, controlling for sex, age, education, income, insurance status, household registration and self-assessed health. We found that distrust in hospitals is low, but distrust in clinics is high and strongly associated with increased hospital utilization, especially for minor symptoms and illnesses. Further research is needed to understand the reasons for distrust in clinics because its effects are not fully accounted for by poor evaluations of their competence.


Mutational analysis of a nucleosidase involved in quorum-sensing autoinducer-2 biosynthesis.
5'-Methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTAN) is important in a number of cellular functions such as polyamine biosynthesis, methionine salvaging, biological methylation, and quorum sensing. The nucleosidase is found in many microbes but not in mammalian systems, thus making MTAN a broad-spectrum antimicrobial drug target. Substrate binding and catalytic residues were identified from the crystal structure of MTAN complexed with 5'-methylthiotubercidin [Lee, J. E., Cornell, K. A., Riscoe, M. K. and Howell, P. L. (2003) J. Biol. Chem. 278 (10) 8761-8770]. The roles of active site residues Met9, Glu12, Ile50, Ser76, Val102, Phe105, Tyr107, Phe151, Met173, Glu174, Arg193, Ser196, Asp197, and Phe207 have been investigated by site-directed mutagenesis and steady-state kinetics. Mutagenesis of residues Glu12, Glu174, and Asp197 completely abolished activity. The location of Asp197 and Glu12 in the active site is consistent with their having a direct role in enzyme catalysis. Glu174 is suggested to be involved in catalysis by stabilizing the transition state positive charge at the O3', C2', and C3' atoms and by polarizing the 3'-hydroxyl to aid in the flow of electrons to the electron withdrawing purine base. This represents the first indication of the importance of the 3'-hydroxyl in the stabilization of the transition state. Furthermore, mutation of Arg193 to alanine shows that the nucleophilic water is able to direct its attack without assistance from the enzyme. This mutagenesis study has allowed a reevaluation of the catalytic mechanism.


A survey of well conserved families of C2H2 zinc-finger genes in Daphnia.
A recent comparative genomic analysis tentatively identified roughly 40 orthologous groups of C2H2 Zinc-finger proteins that are well conserved in "bilaterians" (i.e. worms, flies, and humans). Here we extend that analysis to include a second arthropod genome from the crustacean, Daphnia pulex. Most of the 40 orthologous groups of C2H2 zinc-finger proteins are represented by just one or two proteins within each of the previously surveyed species. Likewise, Daphnia were found to possess a similar number of orthologs for all of these small orthology groups. In contrast, the number of Sp/KLF homologs tends to be greater and to vary between species. Like the corresponding mammalian Sp/KLF proteins, most of the Drosophila and Daphnia homologs can be placed into one of three sub-groups: Class I-III. Daphnia were found to have three Class I proteins that roughly correspond to their Drosophila counterparts, dSP1, btd, CG5669, and three Class II proteins that roughly correspond to Luna, CG12029, CG9895. However, Daphnia have four additional KLF-Class II proteins that are most similar to the vertebrate KLF1/2/4 proteins, a subset not found in Drosophila. Two of these four proteins are encoded by genes linked in tandem. Daphnia also have three KLF-Class III members, one more than Drosophila. One of these is a likely Bteb2 homolog, while the other two correspond to Cabot and KLF13, a vertebrate homolog of Cabot. Consistent with their likely roles as fundamental determinants of bilaterian form and function, most of the 40 groups of C2H2 zinc-finger proteins are conserved in kind and number in Daphnia. However, the KLF family includes several additional genes that are most similar to genes present in vertebrates but missing in Drosophila.


[Healthcare, European Stability Mechanism and public funding following the Covid-19 pandemic].
The aim of this editorial is to illustrate the new public funding framework of the Italian National Health System following the Covid-19 pandemic. The document reviews the measures put in place by the Italian Government and European Institutions such as the European Commission (EC), the European Central Bank (ECB) and the European Stability Mechanism (ESM) to deal with this health crisis and subsequent severe economic recession, with particular reference to sources and uses of resources. The use of new budgetary financial spaces in deficit entails greater attention to the assessment of interventions and makes it necessary to keep expenditure under strict control. At the same time, the remodeling of expenditure within its aggregates, public investment in innovation, and the removal of administrative obstacles can strengthen the capacity of the healthcare system to meet the extraordinary needs deriving from the spread of Covid-19 and its resilience to future health shocks.


Functional defects in NOD2 signaling in experimental and human Crohn disease.
Increasing evidence suggests that a deficit in innate immunity may play a causative role in the pathogenesis of inflammatory bowel disease. The most compelling support for this hypothesis comes from the genetic association of Crohn disease (CD) with carriage of polymorphisms within the NOD2 gene, which represent the most frequent genetic defect in CD. Our findings suggest that SAMP1/YitFc mice, which develop CD-like ileitis in the absence of NOD2 genetic mutations, fail to respond to MDP administration by displaying decreased innate cytokine production and impaired bacterial clearance before the onset of disease. This provides evidence that dysregulated NOD2 signaling, genetic or functional in nature, predisposes to chronic intestinal inflammation, and supports a new paradigm that CD may occur from a deficit in innate immunity as opposed to an overly aggressive immune response. This new paradigm could lead to potential development of new preventative or therapeutic modalities for patients with CD.


Beyond the Hox: how widespread is homeobox gene clustering?
The arrangement of Hox genes into physical clusters is fundamental to the patterning of animal body plans, through the phenomenon of colinearity. Other homeobox genes are often described as dispersed, implying they are not arranged into clusters. Contrary to this view, however, two clusters of non-Hox homeobox genes have been reported: the amphioxus ParaHox gene cluster and the Drosophila 93D/E cluster (referred to here as the NKL cluster). Here I examine the antiquity of these gene clusters, their conservation and their pattern of evolution in vertebrate genomes. I argue that the ParaHox gene cluster arose early in animal evolution, and duplicated in vertebrates to give the four clusters in human and mouse genomes. The NKL cluster is also ancient, and also duplicated to yield four descendent clusters in mammalian genomes. The NKL and Hox gene clusters were originally chromosomal neighbours, within an ancient and extensive array of at least 30 related homeobox genes. There is no necessary relationship between clustering and colinearity, although it is argued that the ParaHox gene cluster does show modified spatial colinearity. A novel hypothesis for the evolution of ParaHox gene expression in deuterostomes is presented.


Multivariate linkage analysis of specific language impairment (SLI).
Specific language impairment (SLI) is defined as an inability to develop appropriate language skills without explanatory medical conditions, low intelligence or lack of opportunity. Previously, a genome scan of 98 families affected by SLI was completed by the SLI Consortium, resulting in the identification of two quantitative trait loci (QTL) on chromosomes 16q (SLI1) and 19q (SLI2). This was followed by a replication of both regions in an additional 86 families. Both these studies applied linkage methods to one phenotypic trait at a time. However, investigations have suggested that simultaneous analysis of several traits may offer more power. The current study therefore applied a multivariate variance-components approach to the SLI Consortium dataset using additional phenotypic data. A multivariate genome scan was completed and supported the importance of the SLI1 and SLI2 loci, whilst highlighting a possible novel QTL on chromosome 10. Further investigation implied that the effect of SLI1 on non-word repetition was equally as strong on reading and spelling phenotypes. In contrast, SLI2 appeared to have influences on a selection of expressive and receptive language phenotypes in addition to non-word repetition, but did not show linkage to literacy phenotypes.


Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogenesis and tumor growth.
Recent studies of tissue culture cells have defined a widespread system of oxygen-regulated gene expression based on the activation of a heterodimeric transcription factor termed hypoxia-inducible factor-1 (HIF-1). To determine whether the HIF-1 transcriptional response is activated within solid tumors and to define the consequences, we have studied tumor xenografts of a set of hepatoma (Hepa-1) cells that are wild type (wt), deficient (c4), and revertant (Rc4) for an obligatory component of the HIF-1 heterodimer, HIF-1beta. Because HIF-1beta is also essential for the xenobiotic response (in which it is termed the aryl hydrocarbon receptor nuclear translocator), we also studied c31 cells, which have a different defect in the xenobiotic response and form the HIF-1 complex normally. Two genes that show different degrees of HIF-1-dependent hypoxia-inducible expression in cell culture were selected for analysis-the glucose transporter, GLUT3, and vascular endothelial growth factor (VEGF). In situ hybridization showed intense focal induction of gene expression in tumors derived from wt, Rc4, and c31 cells, which was reduced (VEGF) or not seen (GLUT3) in those derived from c4 cells. In association with these changes, tumors of c4 cells had reduced vascularity and grew more slowly. These findings show that HIF-1 activation occurs in hypoxic regions of tumors and demonstrate a major influence on gene expression, tumor angiogenesis, and growth.


The tragic 1824 journey of the Hawaiian king and queen to London: history of measles in Hawaii.
The susceptibility of isolated island-based populations to acute infections like measles is well documented, most clearly in Fiji and the Faröe Islands. We review the remarkably tragic 1824 journey of King Kamehameha II and Queen Kamamalu of Hawaii to London and the later enormous impact of measles on Hawaii on first arrival in 1848. The young royalty came to seek an audience with King George IV to negotiate an alliance with England. Virtually the entire royal party developed measles within weeks of arrival, 7 to 10 days after visiting the Royal Military Asylum housing hundreds of soldiers' children. Within the month the king (27) and queen (22) succumbed to measles complications. Their bodies were transported to Hawaii by Right Honorable Lord Byron (Captain George Anson, the poet's cousin). Before 1848 measles was unknown in Hawaii. Several epidemics struck Hawaii in late 1848, beginning with measles and pertussis, then diarrhea and influenza. Measles arrived at this time from California, spreading from Hilo, Hawaii, through all the islands; 10% to 33% of the population died. Subsequent measles epidemics occurred in 1861, 1889 to 1890, 1898, and 1936 to 1937, the latter with 205 deaths. The imported epidemics of infections including measles diminished Hawaii's population from approximately 300,000 at Captain Cook's arrival in 1778 to 135,000 in 1820 and 53,900 in 1876. The measles deaths of the king and queen in London in 1824, likely acquired visiting a large children's home, was a harbinger of the devastating impact of measles upon Hawaiians 24 years later with its first arrival to the Sandwich (Hawaiian) Islands.


Mechanism of inhibition of microsomal mixed-function oxidation by the gut-contents inhibitor of the southern armyworm (Prodenia eridania).
A potent inhibitor of microsomal mixed-function oxidation reactions in insects had previously been isolated and partially purified from the gut contents of Prodenia eridania and shown to be associated with proteinase activity. Incubation of rat liver microsomal fraction with low concentrations of this inhibitor led to solubilization of NADPH-cytochrome c reductase, which was paralleled by the inactivation of reduction of cytochrome P-450 by NADPH and by the inhibition of NADPH-linked benzo[3,4]pyrene hydroxylation and aminopyrine demethylation. There was little or no effect on cytochromes b(5) and P-450, nor was the capacity of the latter catalyst to combine with exogenous substrates decreased. Contrary to the findings with NADPH, preincubation of microsomal fraction with the inhibitor did not cause a significant decrease in the rate of cytochrome P-450 reduction by NADH, supporting the assumption that different catalysts are involved in the electron transfer from NADH and NADPH to cytochrome P-450. The findings indicate the importance of taking the possible presence of endogenous inhibitors into consideration when evaluating low or absent mixed-function oxidation activities found in insect systems in vitro.


T cell lines mediating experimental autoimmune uveoretinitis (EAU) in the rat.
Long-term S-antigen (S-Ag)-specific T lymphocyte lines were derived from the lymph nodes of immunized Lewis rats that had been pretreated with low-dose cyclophosphamide. The protocol consisted of functional selection by alternating cycles of stimulation with S-Ag presented on syngeneic accessory cells and proliferation in IL 2-containing spleen-conditioned medium, coupled with early phenotypic selection for cells bearing the helper/inducer membrane marker (W3/25), by panning on antibody-coated plastic dishes. This protocol consistently resulted in the rapid generation of in vivo functional cell lines capable of mediating experimental autoimmune uveoretinitis (EAU) when transferred into naive rats at 5 to 10 X 10(6) cells/rat systemically or 1 to 2 X 10(6) cells/rat intravitreally. The disease appeared within 6 to 8 days, usually with minimal anterior chamber involvement, and was often unilateral. Pathologic changes resembled those seen in EAU induced by active immunization. The disease could be transferred without concomitant formation of serum antibodies. The uveitogenic line cells were negative for Ia antigen and positive for the W3/25 membrane marker, which appeared to be stable in long-term culture. They proliferated vigorously in vitro and produced IL 2 in response to S-Ag or Con A, but not to unrelated antigens. The establishment of uveitogenic T helper lymphocyte lines will permit the analysis of the cellular mechanisms involved in EAU in a more defined system than has been available.


Feeling Valued and Adding Value: A Participatory Action Research Project on Co-creating Practices of Social Inclusion in Kindergartens and Communities.
Background: Contemporary public health problems connect to the social determinants of health, with a growing recognition of social inclusion as imperative to sustainable development. In this quest for social inclusion, early childhood and families are of particular interest. Although co-creation is suggested as viable path to support well-being, less is known how social inclusion might be co-created in practice. The aim of this study was to explore how Participatory Action Research (PAR) can be a tool for transformative practices in a local community, pointing to kindergartens as meeting places for recognizing social inclusion as a common value in early childhood. Methods: A qualitative PAR study was embedded in a Norwegian municipality as an integrated part of their local public health work. The study involved a wide range of participants and stakeholders in three kindergartens and the wider community. Together, we explored potentials for co-creating social inclusion to achieve well-being through cycles of transformative actions and reflections. Reflexive thematic analysis was applied to generate patterns and themes in the data. Results: The participants formulated and took on ownership to an inclusive agenda through the PAR-process. Acts of inclusion was framed by an intersection between political aims of achieving health and well-being for all and public value co-creation unfolding at the level of the place, in the context of the Norwegian welfare regime. To feel valued and adding value was seen as important aspects for social inclusion. Four themes were generated from analysis; (1) Co-creating a shared vision of inclusive communities, (2) Becoming aware and empowered through caring, sharing and collaboration, (3) Places and spaces of inclusiveness in kindergartens and beyond, and (4) Valuing and practicing inclusion, and signs of transformative change. Conclusions: Through the PAR process, parents, kindergartens employees, community members and policy makers appear to have opened a creative toolbox for inclusive and transformational change through formulating and co-creating inclusion and well-being as public values. The results suggest that local actors might support adaptive social systems to taking on relational responsibility for inclusive processes and outcomes in the pursuit of well-being for all.


Genetic parameters of linear conformation type traits and their relationship with milk yield throughout lactation in mixed-breed dairy goats.
Conformation traits are of interest to many dairy goat breeders not only as descriptive traits in their own right, but also because of their influence on production, longevity, and profitability. If these traits are to be considered for inclusion in future dairy goat breeding programs, relationships between them and production traits such as milk yield must be considered. With the increased use of regression models to estimate genetic parameters, an opportunity now exists to investigate correlations between conformation traits and milk yield throughout lactation in more detail. The aims of this study were therefore to (1) estimate genetic parameters for conformation traits in a population of crossbred dairy goats, (2) estimate correlations between all conformation traits, and (3) assess the relationship between conformation traits and milk yield throughout lactation. No information on milk composition was available. Data were collected from goats based on 2 commercial goat farms during August and September in 2013 and 2014. Ten conformation traits, relating to udder, teat, leg, and feet characteristics, were scored on a linear scale (1-9). The overall data set comprised data available for 4,229 goats, all in their first lactation. The population of goats used in the study was created using random crossings between 3 breeds: British Alpine, Saanen, and Toggenburg. In each generation, the best performing animals were selected for breeding, leading to the formation of a synthetic breed. The pedigree file used in the analyses contained sire and dam information for a total of 30,139 individuals. The models fitted relevant fixed and random effects. Heritability estimates for the conformation traits were low to moderate, ranging from 0.02 to 0.38. A range of positive and negative phenotypic and genetic correlations between the traits were observed, with the highest correlations found between udder depth and udder attachment (0.78), teat angle and teat placement (0.70), and back legs and back feet (0.64). The genetic correlations estimated between conformation traits and milk yield across the first lactation demonstrated changes during this period. The majority of correlations estimated between milk yield and the udder and teat traits were negative. Therefore, future breeding programs would benefit from including these traits to ensure that selection for increased productivity is not accompanied by any unwanted change in functional fitness.


Learning Efficient Sparse and Low Rank Models.
Parsimony, including sparsity and low rank, has been shown to successfully model data in numerous machine learning and signal processing tasks. Traditionally, such modeling approaches rely on an iterative algorithm that minimizes an objective function with parsimony-promoting terms. The inherently sequential structure and data-dependent complexity and latency of iterative optimization constitute a major limitation in many applications requiring real-time performance or involving large-scale data. Another limitation encountered by these modeling techniques is the difficulty of their inclusion in discriminative learning scenarios. In this work, we propose to move the emphasis from the model to the pursuit algorithm, and develop a process-centric view of parsimonious modeling, in which a learned deterministic fixed-complexity pursuit process is used in lieu of iterative optimization. We show a principled way to construct learnable pursuit process architectures for structured sparse and robust low rank models, derived from the iteration of proximal descent algorithms. These architectures learn to approximate the exact parsimonious representation at a fraction of the complexity of the standard optimization methods. We also show that appropriate training regimes allow to naturally extend parsimonious models to discriminative settings. State-of-the-art results are demonstrated on several challenging problems in image and audio processing with several orders of magnitude speed-up compared to the exact optimization algorithms.


Elucidating the structural basis of diphenyl ether derivatives as highly potent enoyl-ACP reductase inhibitors through molecular dynamics simulations and 3D-QSAR study.
Diphenyl ether derivatives are good candidates for anti-tuberculosis agents that display a promising potency for inhibition of InhA, an essential enoyl-acyl carrier protein (ACP) reductase involved in fatty acid biosynthesis pathways in Mycobacterium tuberculosis. In this work, key structural features for the inhibition were identified by 3D-QSAR CoMSIA models, constructed based on available experimental binding properties of diphenyl ether inhibitors, and a set of four representative compounds was subjected to MD simulations of inhibitor-InhA complexes for the calculation of binding free energies. The results show that bulky groups are required for the R1 substituent on the phenyl A ring of the inhibitors to favor a hydrophobic pocket formed by residues Phe149, Met155, Pro156, Ala157, Tyr158, Pro193, Met199, Val203, Leu207, Ile215, and Leu218. Small substituents with a hydrophilic property are required at the R3 and R4 positions of the inhibitor phenyl B rings to form hydrogen bonds with the backbones of Gly96 and Met98, respectively. For the R2 substituent, small substituents with simultaneous hydrophilic or hydrophobic properties are required to favor the interaction with the pyrophosphate moiety of NAD(+) and the methyl side chain of Ala198, respectively. The reported data provide structural guidance for the design of new and potent diphenyl ether-based inhibitors with high inhibitory activities against M. tuberculosis InhA.


Leptin receptor of Zucker fatty rat performs reduced signal transduction.
Zucker fatty (fa/fa) rats exhibit overt obesity, hypercholesterolemia, hyperlipidemia, and hyperglycemia as recessive traits. The fa mutation has been determined to be a missense mutation in the extracellular domain of the leptin receptor. We report herein the construction of CHO cells that stably express the fa-type leptin receptor and the characterization of this receptor using mRNA expression levels of the immediate early genes, c-fos, c-jun, and jun-B, which are induced by leptin as a criterion of signal transduction. The fa-type receptor not only exhibits a slightly reduced leptin-binding affinity, but also performs reduced signal transduction.


Transgenic mice expressing mutant forms VCP/p97 recapitulate the full spectrum of IBMPFD including degeneration in muscle, brain and bone.
Inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia (IBMPFD) is a dominantly inherited degenerative disorder caused by mutations in the valosin-containing protein (VCP) gene. VCP (p97 in mouse, TER94 in Drosophila melanogaster and CDC48 in Saccharomyces cerevisiae) is a highly conserved AAA(+)-ATPase that regulates a wide array of cellular processes. The mechanism of IBMPFD pathogenesis is unknown. Towards elucidating the pathogenic mechanism we have developed and characterized transgenic mice with ubiquitous expression of wild-type and disease-causing versions of human VCP/p97. Here, we report that mice expressing VCP/p97 harboring the mutations R155H or A232E develop pathology that is limited to muscle, brain and bone, recapitulating the spectrum of disease in humans with IBMPFD. The mice exhibit progressive muscle weakness and pathological examination of muscle shows classic characteristics of inclusion body myopathy including rimmed vacuoles and TDP-43 pathology. The mice exhibit abnormalities in behavioral testing and pathological examination of the brain shows widespread TDP-43 pathology. Furthermore, radiological examination of the skeleton reveals that mutant mice develop severe osteopenia accompanied by focal lytic and sclerotic lesions in vertebrae and femur. In vitro studies indicate that mutant VCP causes inappropriate activation of the NF-kappaB signaling cascade, which could contribute to the mechanism of pathogenesis in multiple tissues including muscle, bone and brain.


The presenting features of brain tumours: a review of 200 cases.
To determine the presenting features of brain tumours in children. Retrospective case note review. Paediatric and neurosurgical services at the Wessex Neurology Centre and Southampton General Hospital, UK. 200 patients presenting with a CNS tumour between 1988 and 2001. The commonest first presenting symptoms were headache (41%), vomiting (12%), unsteadiness (11%), visual difficulties (10%), educational or behavioural problems (10%), and seizures (9%). The commonest symptoms occurring at any time were headache (56%), vomiting (51%), educational or behavioural problems (44%), unsteadiness (40%), and visual difficulties (38%). Neurological signs were present at diagnosis in 88%: 38% had papilloedema, 49% cranial nerve abnormalities, 48% cerebellar signs, 27% long tract signs, 11% somatosensory abnormalities, and 12% a reduced level of consciousness. The median symptom interval was 2.5 months (range 1 day to 120 months). A short symptom interval was significantly associated with high grade tumours and patient age of 3 years or younger. The well known predominance of headache in children with CNS tumours is confirmed. Visual, behavioural, and educational symptoms were also prominent. With the exception of seizures, every initial symptom was accompanied by other symptoms or signs by the time of diagnosis. Questions about visual symptoms and educational or behavioural difficulties, as well as the more widely recognised symptoms of raised intracranial pressure and motor dysfunction, are important in the diagnosis of brain tumours, as are vision assessment and the appropriate plotting of growth and head size.


Capsular polysaccharide of Cryptococcus neoformans induces proinflammatory cytokine release by human neutrophils.
Human polymorphonuclear leukocytes from normal subjects produced proinflammatory cytokines in response to stimulation with Cryptococcus neoformans yeast cells. The cytokines released after stimulation of neutrophils included interleukin-1beta (IL-1beta), IL-6, IL-8 and tumor necrosis factor alpha. The magnitude of the cytokine response was related to the yeast capsule size. Cells of a large-capsule isolate stimulated release of greater amounts of cytokine than did a thinly encapsulated isolate, which, in turn, stimulated release of greater amounts of cytokine than an acapsular isolate. Cytokine release was also stimulated by supernatant fluids from cryptococcal cells that were preincubated with 10% human serum, suggesting the generation of a soluble mediator. The major capsular polysaccharide, glucuronoxylomannan, stimulated release of tumor necrosis factor alpha, IL-1beta, and IL-8 in a dose-dependent fashion. These results differ from previous studies of cytokine secretion by human monocytes in several important respects, including the importance of encapsulation in stimulation of cytokine secretion and the ability of purified glucuronoxylomannan to induce cytokine secretion.


Sodium/myo-inositol cotransporter 1 and myo-inositol are essential for osteogenesis and bone formation.
myo-Inositol (MI) plays an essential role in several important processes of cell physiology, is involved in the neural system, and provides an effective treatment for some psychiatric disorders. Its role in osteogenesis and bone formation nonetheless is unclear. Sodium/MI cotransporter 1 (SMIT1, the major cotransporter of MI) knockout (SMIT1(-/-)) mice with markedly reduced tissue MI levels were used to characterize the essential roles of MI and SMIT1 in osteogenesis. SMIT1(-/-) embryos had a dramatic delay in prenatal mineralization and died soon after birth owing to respiratory failure, but this could be rescued by maternal MI supplementation. The rescued SMIT1(-/-) mice had shorter limbs, decreased bone density, and abnormal bone architecture in adulthood. Deletion of SMIT1 resulted in retarded postnatal osteoblastic differentiation and bone formation in vivo and in vitro. Continuous MI supplementation partially restored the abnormal bone phenotypes in adult SMIT1(-/-) mice and strengthened bone structure in SMIT1(+/+) mice. Although MI content was much lower in SMIT1(-/-) mesenchymal cells (MSCs), the I(1,4,5)P(3) signaling pathway was excluded as the means by which SMIT1 and MI affected osteogenesis. PCR expression array revealed Fgf4, leptin, Sele, Selp, and Nos2 as novel target genes of SMIT1 and MI. SMIT1 was constitutively expressed in multipotential C3H10T1/2 and preosteoblastic MC3T3-E1 cells and could be upregulated during bone morphogenetic protein 2 (BMP-2)-induced osteogenesis. Collectively, this study demonstrated that deficiency in SMIT1 and MI has a detrimental impact on prenatal skeletal development and postnatal bone remodeling and confirmed their essential roles in osteogenesis, bone formation, and bone mineral density (BMD) determination.


Experimental evidence for the role of domain swapping in the evolution of the histone fold.
The histone fold forms the fundamental endoskeleton of the protein core of the nucleosome and is also found in several transcription factors. We have investigated the evolutionary origins of this ubiquitous protein motif, which is found soluble exclusively as an antiparallel (handshake motif) dimer. We introduced a three amino acid insertion into the middle of a homodimeric archaeal histone fold motif. The engineered molecule was found to be a soluble and stable monomer with properties consistent with a four-helix-bundle protein. The experimental evidence presented here support the hypothesis that the handshake association motif characteristic of present-day histone dimers is the evolutionary product of domain swapping between two four-helix bundle domains, each of which derived from the tandem duplication of a primitive helix-strand-helix unit.


Von Hippel-Lindau disease maps to the region of chromosome 3 associated with renal cell carcinoma.
Von Hippel-Lindau disease (VHL) is an autosomal dominant disorder with inherited susceptibility to various forms of cancer, including hemangioblastomas of the central nervous system, phaeochromocytomas, pancreatic malignancies, and renal cell carcinomas. Renal cell carcinomas constitute a particularly frequent cause of death in this disorder, occurring as bilateral and multifocal tumours, and presenting at an earlier age than in sporadic, non-familial cases of this tumour type. We report here that the VHL gene is linked to the locus encoding the human homologoue of the RAF1 oncogene, which maps to chromosome 3p25 (ref. 4). Crossovers with the VHL locus suggest that the defect responsible for the VHL phenotype is not a mutation in the RAF1 gene itself. An alternative or prior event to oncogene activation in tumour formation may be the inactivation of a putative 'tumour suppressor' which can be associated with both the inherited and sporadic forms of the cancer. Sporadic renal cell carcinomas have previously been associated with the loss of regions on chromosome 3p (refs 5, 6). Consequently, sporadic and VHL-associated forms of renal cell carcinoma might both result from alterations causing loss of function of the same 'tumour suppressor' gene on this chromosome.


Elevated plasma ferritin in elderly individuals with high neocortical amyloid-β load.
Ferritin, an iron storage and regulation protein, has been associated with Alzheimer's disease (AD); however, it has not been investigated in preclinical AD, detected by neocortical amyloid-β load (NAL), before cognitive impairment. Cross-sectional analyses were carried out for plasma and serum ferritin in participants in the Kerr Anglican Retirement Village Initiative in Aging Health cohort. Subjects were aged 65-90 years and were categorized into high and low NAL groups via positron emission tomography using a standard uptake value ratio cutoff=1.35. Ferritin was significantly elevated in participants with high NAL compared with those with low NAL, adjusted for covariates age, sex, apolipoprotein E ɛ4 carriage and levels of C-reactive protein (an inflammation marker). Ferritin was also observed to correlate positively with NAL. A receiver operating characteristic curve based on a logistic regression of the same covariates, the base model, distinguished high from low NAL (area under the curve (AUC)=0.766), but was outperformed when plasma ferritin was added to the base model (AUC=0.810), such that at 75% sensitivity, the specificity increased from 62 to 71% on adding ferritin to the base model, indicating that ferritin is a statistically significant additional predictor of NAL over and above the base model. However, ferritin's contribution alone is relatively minor compared with the base model. The current findings suggest that impaired iron mobilization is an early event in AD pathogenesis. Observations from the present study highlight ferritin's potential to contribute to a blood biomarker panel for preclinical AD.


Making birthing safe for Pakistan women: a cluster randomized trial.
Two out of three neonatal deaths occur in just 10 countries and Pakistan stands third among them. Maternal mortality is also high with most deaths occurring during labor, birth, and first few hours after birth. Enhanced access and utilization of skilled delivery and emergency obstetric care is the demonstrated strategy in reducing maternal and neonatal mortality. This trial aims to compare reduction in neonate mortality and utilization of available safe birthing and Emergency Obstetric and Neonatal Care services among pregnant mothers receiving 'structured birth planning', and/or 'transport facilitation' compared to routine care. A pragmatic cluster randomized trial, with qualitative and economic studies, will be conducted in Jhang, Chiniot and Khanewal districts of Punjab, Pakistan, from February 2011 to May 2013. At least 29,295 pregnancies will be registered in the three arms, seven clusters per arm; 1) structured birth planning and travel facilitation, 2) structured birth planning, and 3) control arm. Trial will be conducted through the Lady Health Worker program. Main outcomes are difference in neonatal mortality and service utilization; maternal mortality being the secondary outcome. Cluster level analysis will be done according to intention-to-treat. A nationwide network of about 100,000 lady health workers is already involved in antenatal and postnatal care of pregnant women. They also act as "gatekeepers" for the child birthing services. This gate keeping role mainly includes counseling and referral for skill birth attendance and travel arrangements for emergency obstetric care (if required). The review of current arrangements and practices show that the care delivery process needs enhancement to include adequate information provision as well as informed "decision" making and planned "action" by the pregnant women. The proposed three-year research is to develop, through national technical working group process, and then test a set of arrangements for achieving the enhanced utilization of safe birthing services. Current Controlled Trials ISRCTN86264432.


Fluid-phase pinocytosis of native low density lipoprotein promotes murine M-CSF differentiated macrophage foam cell formation.
During atherosclerosis, low-density lipoprotein (LDL)-derived cholesterol accumulates in macrophages to form foam cells. Macrophage uptake of LDL promotes foam cell formation but the mechanism mediating this process is not clear. The present study investigates the mechanism of LDL uptake for macrophage colony-stimulating factor (M-CSF)-differentiated murine bone marrow-derived macrophages. LDL receptor-null (LDLR-/-) macrophages incubated with LDL showed non-saturable accumulation of cholesterol that did not down-regulate for the 24 h examined. Incubation of LDLR-/- macrophages with increasing concentrations of (125)I-LDL showed non-saturable macrophage LDL uptake. A 20-fold excess of unlabeled LDL had no effect on (125)I-LDL uptake by wild-type macrophages and genetic deletion of the macrophage scavenger receptors CD36 and SRA did not affect (125)I-LDL uptake, showing that LDL uptake occurred by fluid-phase pinocytosis independently of receptors. Cholesterol accumulation was inhibited approximately 50% in wild-type and LDLR-/- mice treated with LY294002 or wortmannin, inhibitors of all classes of phosphoinositide 3-kinases (PI3K). Time-lapse, phase-contrast microscopy showed that macropinocytosis, an important fluid-phase uptake pathway in macrophages, was blocked almost completely by PI3K inhibition with wortmannin. Pharmacological inhibition of the class I PI3K isoforms alpha, beta, gamma or delta did not affect macrophage LDL-derived cholesterol accumulation or macropinocytosis. Furthermore, macrophages from mice expressing kinase-dead class I PI3K beta, gamma or delta isoforms showed no decrease in cholesterol accumulation or macropinocytosis when compared with wild-type macrophages. Thus, non-class I PI3K isoforms mediated macropinocytosis in these macrophages. Further characterization of the components necessary for LDL uptake, cholesterol accumulation, and macropinocytosis identified dynamin, microtubules, actin, and vacuolar type H(+)-ATPase as contributing to uptake. However, Pak1, Rac1, and Src-family kinases, which mediate fluid-phase pinocytosis in certain other cell types, were unnecessary. In conclusion, our findings provide evidence that targeting those components mediating macrophage macropinocytosis with inhibitors may be an effective strategy to limit macrophage accumulation of LDL-derived cholesterol in arteries.


Nucleotide binding switches the information flow in ras GTPases.
The Ras superfamily comprises many guanine nucleotide-binding proteins (G proteins) that are essential to intracellular signal transduction. The guanine nucleotide-dependent intrinsic flexibility patterns of five G proteins were investigated in atomic detail through Molecular Dynamics simulations of the GDP- and GTP-bound states (S(GDP) and S(GTP), respectively). For all the considered systems, the intrinsic flexibility of S(GDP) was higher than that of S(GTP), suggesting that Guanine Exchange Factor (GEF) recognition and nucleotide switch require higher amplitude motions than effector recognition or GTP hydrolysis. Functional mode, dynamic domain, and interaction energy correlation analyses highlighted significant differences in the dynamics of small G proteins and Gα proteins, especially in the inactive state. Indeed, S(GDP) of Gα(t), is characterized by a more extensive energy coupling between nucleotide binding site and distal regions involved in GEF recognition compared to small G proteins, which attenuates in the active state. Moreover, mechanically distinct domains implicated in nucleotide switch could be detected in the presence of GDP but not in the presence of GTP. Finally, in small G proteins, functional modes are more detectable in the inactive state than in the active one and involve changes in solvent exposure of two highly conserved amino acids in switches I and II involved in GEF recognition. The average solvent exposure of these amino acids correlates in turn with the rate of GDP release, suggesting for them either direct or indirect roles in the process of nucleotide switch. Collectively, nucleotide binding changes the information flow through the conserved Ras-like domain, where GDP enhances the flexibility of mechanically distinct portions involved in nucleotide switch, and favors long distance allosteric communication (in Gα proteins), compared to GTP.


Genetic variations in androgen metabolism genes and associations with prostate cancer in South African men.
Background. In South Africa white men have the highest incidence of prostate cancer (PCa), coloured (mixed ancestry) men have an intermediate incidence, and low incidences are reported for black and Asian men. It has been suggested that ethnic differences in incidence and mortality of PCa are related to genetic variations in genes that regulate androgen metabolism. We investigated the role of genetic variants in the androgen metabolism genes and the probability of developing PCa in South African coloured and white men. Methods. Genotype and allele counts and frequencies of single nucleotide polymorphisms (SNPs) in CYP3A5, CYP3A4 and CYP3A43 were assessed in coloured men (160 case individuals, 146 control individuals) and white men (121 case individuals, 141 control individuals). Results. A genetic association indicating an increased probability of developing PCa was observed with the G allele of the SNP rs2740574 in CYP3A4 in coloured men, the A allele of rs776746 (CYP3A5) and the G allele of rs2740574 (CYP3A4) in white men, and the G allele of rs2740574 and the C allele of rs501275 (CYP3A43) in the combined ethnic groups analysis. In addition, we identified allele combinations (termed haplotypes) with significantly higher frequencies in the PCa case individuals than in the control individuals. Conclusions. The findings support the role of variants in genes that regulate androgen metabolism and the probability of developing PCa. The study paves the way to identify other genetic associations in South African men, and to establish genetic profiles that could be used to determine disease progression and prognosis.


Age and sharing of needle injection equipment in a cohort of Massachusetts injection drug users: an observational study.
Hepatitis C infection (HCV) among individuals aged 15-24 years has increased in Massachusetts, likely due to injection drug use. The prevalence of injection equipment sharing (sharing) and its association with age was examined in a cohort of out-of-treatment Massachusetts substance users. This analysis included baseline data from a behavioral intervention with substance users. Younger and older (<25 versus ≥ 25 years) injection drug users were compared on demographic characteristics, substance use practices, including factors present during the most recent sharing event ("event-level factors"), and HCV testing history. Sharing was reported by 41% of the 484 individuals who reported injection drug use in the past 30 days. Prevalence of sharing varied by age (50% <25 years old versus 38% ≥ 25 years, p=0.02). In a multivariable logistic regression model younger versus older individuals had twice the odds of sharing (95% CI=1.26, 3.19). During their most recent sharing event, fewer younger individuals than older had their own drugs available (50% versus 75%, p<0.001); other injection event-level factors did not vary by age. In the presence of PTSD, history of exchanging sex for money, or not being US born, prevalence of sharing by older users was higher and was similar to that of younger users, such that there was no association between age and sharing. In this cohort of injection drug users, younger age was associated with higher prevalence of sharing, but only in the absence of certain stressors. Harm reduction efforts might benefit from intervening on mental health and other stressors in addition to substance use. Study findings suggest a particular need to address the dangers of sharing with young individuals initiating injection drug use.


An Outbreak of Bacterial Speck Caused by Pseudomonas syringae pv. tomato on Tomato Transplants Grown in Commercial Seedling Companies Located in the Western Mediterranean Region of Turkey.
A serious outbreak of a leaf spot disease was observed on tomato transplants grown in commercial seedling companies in southwestern Turkey (Antalya) during the springs of 2002 and 2003. Disease incidence was more severe in the western Mediterranean Region of Turkey. Occurrence of the outbreak resulted in approximately 20 and 25% seedling losses in the springs of 2002 and 2003, respectively. The initial symptoms consisted of pronounced water-soaked, dark brown-to-black spots on young expanding leaves that were 1 to 2 mm in diameter. Later, a number of leaf spots on older leaves enlarged and coalesced, causing leaf desiccation and finally, seedling death. In addition, in 2003 the disease incidence was approximately 5% in 142 commercial greenhouses. Tomato production was unaffected since significant outbreaks did not occur on greenhouse plants. No fruit symptoms were observed. Twenty-six strains were isolated from diseased tomato seedlings and plants from different greenhouses located in different places and all were gram negative and fluorescent on King's B medium. All strains were levan and gelatin liquefaction positive and oxidase and arginine dihydrolase negative. None of the 26 strains utilized erythritol and l-lactate as the sole carbon source (1,2). Fatty acid analysis identified the strains as Pseudomonas syringae pv. tomato with similarity indices ranging from 0.876 to 0.932%. Pathogenicity of the isolates was confirmed on 4-week-old tomato seedlings (cv. Biotek Selin) sprayed with the bacterial suspensions containing 108 CFU/ml of sterile water. Later, a number of leaf spots on the leaves enlarged and coalesced, causing leaf desiccation. Inoculated and control tomato seedlings were covered with polyethylene bags and placed in a growth chamber at 25°C for 48 h and then the bags were removed. Small (1 to 2 mm), water-soaked, dark brown-to-black spots similar to those observed in the greenhouses of commercial seedling companies and commercial greenhouses that produce tomato developed on the young expanding leaves of inoculated plants within 7 to 10 days. No symptoms developed on control plants. The bacterium was reisolated from inoculated plants and identified as a strain of Pseudomonas syringae pv. tomato. Koch's postulates were fulfilled. To our knowledge, this is the first report for the occurrence and outbreak of the bacterial speck disease on tomato transplants in greenhouses of commercial tomato seedling production companies in Turkey. References: (1) D. C. Hildebrand et al. Pages 60-80 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. N. D. Schaad, ed. The American Phytopathological Society, 1988. (2) J. B. Jones et al. Plant Dis. 70:151, 1986.


The efficacy of trichlorphon and naphthalophos against multiple anthelmintic-resistant nematodes of naturally infected sheep in Argentina.
An anthelmintic efficacy trial was conducted in sheep harbouring anthelmintic-resistant worms in Argentina. Seventy lambs were selected from a flock that had been grazed on pastures infected with trichostrongyles previously shown to be resistant to the main anthelmintic groups. Lambs were allocated to comparable groups of ten animals each and treated with trichlorphon (50 mg/kg body weight (b.w.) orally); naphthalophos (50 mg/kg b.w. orally); ivermectin (0.2 mg/kg b.w. subcutaneously); fenbendazole (5 mg/kg b.w. orally); levamisole (8 mg/kg b.w. subcutaneously) and closantel (10 mg/kg b.w. orally). There was also an untreated group. The dose selection was based on manufacturer's recommendations.Faecal samples were collected 0 and 10 days post treatment to estimate efficacy (faecal egg count reduction). Six animals from each group were necropsied at day 10 for enumeration/identification of worms from the abomasum, small and large intestines to determine the absolute efficacy of each agent (controlled efficacy test). Trichlorphon and naphthalophos were effective (> 99 %) against Haemonchus contortus (p < 0.05).Naphthalophos also showed efficacy against Trichostrongylus axei (99.3 %), Teladorsagia circumcincta (97.8 %), Trichostrongylus colubriformis (99.2 %), Cooperia punctata/curticei/pectinata (90.4 %), Nematodirus spathiger (89.2 %) and Oesophagostomum venulosum/columbianum (93.7 %). Fenbendazole and levamisole showed efficacy (> 95 %) against all nematodes except T. colubriformis. The efficacy of ivermectin was low against H. contortus (23 %) and Cooperia spp. (46.3 %). Closantel showed low efficacy against T. axei (64.4 %), H. contortus (80.6 %) and T. colubriformis (59.5 %).When anthelmintic resistance is widespread, trichlorphon treatment is appropriate if H. contortus is present; however, naphthalophos represents an effective therapeutic alternative for incorporation into worm control programmes.


A framework for allocating conservation resources among multiple threats and actions.
Land managers decide how to allocate resources among multiple threats that can be addressed through multiple possible actions. Additionally, these actions vary in feasibility, effectiveness, and cost. We sought to provide a way to optimize resource allocation to address multiple threats when multiple management options are available, including mutually exclusive options. Formulating the decision as a combinatorial optimization problem, our framework takes as inputs the expected impact and cost of each threat for each action (including do nothing) and for each overall budget identifies the optimal action to take for each threat. We compared the optimal solution to an easy to calculate greedy algorithm approximation and a variety of plausible ranking schemes. We applied the framework to management of multiple introduced plant species in Australian alpine areas. We developed a model of invasion to predict the expected impact in 50 years for each species-action combination that accounted for each species' current invasion state (absent, localized, widespread); arrival probability; spread rate; impact, if present, of each species; and management effectiveness of each species-action combination. We found that the recommended action for a threat changed with budget; there was no single optimal management action for each species; and considering more than one candidate action can substantially increase the management plan's overall efficiency. The approximate solution (solution ranked by marginal cost-effectiveness) performed well when the budget matched the cost of the prioritized actions, indicating that this approach would be effective if the budget was set as part of the prioritization process. The ranking schemes varied in performance, and achieving a close to optimal solution was not guaranteed. Global sensitivity analysis revealed a threat's expected impact and, to a lesser extent, management effectiveness were the most influential parameters, emphasizing the need to focus research and monitoring efforts on their quantification.


Comparison of Tumor Enucleation and Standard Partial Nephrectomy According to Trifecta Outcomes: A Multicenter Study by the Turkish Academy of Urology, Uro-Oncology Working Group.
We aimed to evaluate the impact of the resection technique (tumor enucleation (TE) or standard partial nephrectomy (SPN)) on trifecta outcomes in patients having undergone partial nephrectomy (PN). We retrospectively analyzed the clinical and pathologic parameters in patients with localized renal cell carcinoma (pT1-2N0M0) who had undergone PN between January 2001-December 2018 at one of 15 different tertiary referral centers. Multivariable logistic regression analysis was applied to investigate independent predictors of trifecta failure, decreased postoperative renal functions (decreased estimated glomerular filtration rate (eGFR) > 10%), perioperative complications (Clavien-Dindo > 1), and positive surgical margins. A total of 1070 patients with a mean age 56.11 ± 11.88 years were included in our study. PN was performed with TE in 848 (79.25%) and SPN in 222 (20.75%) patients. Trifecta failure rate was 56.2% for TE and 64.4% for SPN (p = 0.028). On multivariable analysis, TE was associated with less trifecta failure (p = 0.025) and eGFR decrease >10% rates (p = 0.024). On the other hand, there was no statistically significant difference between TE and SPN according to positive surgical margins (p = 0.450) and complication > Clavien-Dindo grade 1 (p = 0.888) rates. The only independent predictive factor for complications > Clavien-Dindo 1 was the Charlson comorbidity index (CCI) (p = 0.001). TE is associated with less trifecta failure than SPN. This result is mainly due to better preservation of renal function with TE.


Effect of auricular pellet acupressure on antioxidative systems in high-risk diabetes mellitus.
Free radicals and lipid peroxides, both of which are easily formed in the diabetic state, play an important role in the development of diabetic complications. Antioxidative therapy may help prevent diabetic complications caused by lipoperoxidation and free-radical formation in diabetes mellitus (DM). A number of findings suggest that oxidative stress exists in persons with high-risk DM. Auricular pellet acupressure has reportedly been an effective treatment method for a variety of medical conditions, including anxiety, juvenile myopia, essential hypertension, and senile vascular dementia. However, its effects on antioxidative enzymes have not been elucidated. We therefore evaluated the impact of auricular pellet acupressure on antioxidative status in persons with high-risk DM. Our study involved 69 persons with high-risk DM, who were allocated either to undergo acupressure as active treatment for the experimental group or to a control group. The experimental group in the study received auricular pellet acupressure three times daily for 5 consecutive days. After a 2-day rest period, the procedure was performed on the contralateral ear. Acupressure was performed twice on each ear, with each application followed by its application to the contralateral ear, over a total treatment period of 20 days. The control groups did not undergo auricular pellet acupressure. At the end of the 20-day period of treatment of the experimental group, blood was collected from all of the study participants for assay of serum superoxide dismutase (SOD) and catalase concentrations, as was also done for the control group. Serum concentrations of SOD (p < 0.05) and catalase (p < 0.0001) were significantly higher in the experimental group than in the control group. Our findings suggest that auricular pellet acupressure can increase the concentration of antioxidative enzymes in persons with high-risk DM.


L1 (LINE-1) retrotransposon evolution and amplification in recent human history.
L1 (LINE-1) elements constitute a large family of mammalian retrotransposons that have been replicating and evolving in mammals for more than 100 Myr and now compose 20% or more of the DNA of some mammals. Here, we investigated the evolutionary dynamics of the active human Ta L1 family and found that it arose approximately 4 MYA and subsequently differentiated into two major subfamilies, Ta-0 and Ta-1, each of which contain additional subsets. Ta-1, which has not heretofore been described, is younger than Ta-0 and now accounts for at least 50% of the Ta family. Although Ta-0 contains some active elements, the Ta-1 subfamily has replaced it as the replicatively dominant subfamily in humans; 69% of the loci that contain Ta-1 inserts are polymorphic for the presence or absence of the insert in human populations, as compared with 29% of the loci that contain Ta-0 inserts. This value is 90% for loci that contain Ta-1d inserts, which are the youngest subset of Ta-1 and now account for about two thirds of the Ta-1 subfamily. The successive emergence and amplification of distinct Ta L1 subfamilies shows that L1 evolution has been as active in recent human history as it has been found to be for rodent L1 families. In addition, Ta-1 elements have been accumulating in humans at about the same rate per generation as recently evolved active rodent L1 subfamilies.


Pediatric Aggressive Mature B-Cell Lymphomas, Version 3.2022, NCCN Clinical Practice Guidelines in Oncology.
NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines) for Pediatric Aggressive Mature B-Cell Lymphomas include recommendations for the diagnosis and management of pediatric patients with primary mediastinal large B-cell lymphoma (PMBL) and sporadic variants of Burkitt lymphoma and diffuse large B-cell lymphoma. PMBL is now considered as a distinct entity arising from mature thymic B-cells accounting for 2% of mature B-cell lymphomas in children and adolescents. This discussion section includes the recommendations outlined in the NCCN Guidelines for the diagnosis and management of pediatric patients with PMBL.