Electrochemical protoplast fusion in citrus.
We report here the development of a novel protoplast fusion method for citrus somatic hybridization. This new procedure, which we have named electrochemical protoplast fusion, is based on chemically induced protoplast aggregation, using a low concentration of polyethylene glycol, and DC pulse-promoted membrane fusion. Based on the results of nucleus and mitochondria molecular analyses, we were successful in using this method to regenerate both symmetric somatic hybrids and cybrids. Various parameters, including pulse intensity, pulse length, and composition of the fusion media, were tested, and the optimum fusion condition selected consisted of two 100-micros pulses of 1,500 V cm(-1). Our conclusion is that electrochemical fusion is a reliable and reproducible method that combines the best features of both the chemical and electrical methods, thereby promoting cell division and high embryogenesis rates of the fused cells. It represents a new approach to citrus somatic hybridization. Various interesting features of this new approach are presented and discussed.


Are growth rates of Escherichia coli in batch cultures limited by respiration?
Batch cultures of Escherichia coli were grown in minimal media supplemented with various carbon sources which supported growth at specific growth rates from 0.2 to 1.3/h. The respiration rates of the cultures were measured continuously. With few exceptions, the specific rate of oxygen consumption was about 20 mmol of O2/h per g (dry weight), suggesting that the respiratory capacity was limited at this value. The adenosine triphosphate (ATP) required for the production of cell material from the different carbon sources was calculated on the basis of known ATP requirements in the biochemical pathways and routes of macromolecular synthesis. The calculated ATP requirements, together with the measured growth rates and growth yields on the different carbon sources, were used to calculate the rate of ATP synthesis by oxidative phosphorylation. This rate was closely related to the respiration rate. We suggest that aerobic growth of E. coli in batch cultures is limited by the rate of respiration and the concomitant rate of ATP generation through oxidative phosphorylation.


Fentanyl or midazolam for co-induction of anaesthesia with propofol in dogs.
Propofol may cause adverse effects (e.g. apnoea, hypotension) at induction of anaesthesia. Co-induction of anaesthesia may reduce propofol requirements. The effect of fentanyl or midazolam on propofol dose requirements and cardiorespiratory parameters was studied. Randomized, controlled, blinded clinical study. Sixty-six client owned dogs (35 male, 31 female, ASA I-II, age 6-120 months, body mass 4.7-48.0 kg) were selected. Pre-medication with acepromazine (0.025 mg kg(-1)) and morphine (0.25 mg kg(-1)) was administered by intramuscular injection. After 30 minutes group fentanyl-propofol (FP) received fentanyl (2 microg kg(-1)), group midazolam-propofol (MP) midazolam (0.2 mg kg(-1)) injected over 30 seconds via a cephalic catheter and in a third group, control-propofol (CP), the IV catheter was flushed with an equivalent volume of heparinized saline. Anaesthesia was induced 2 minutes later, with propofol (4 mg kg(-1)minute(-1)) administered to effect. After endotracheal intubation anaesthesia was maintained with a standardized anaesthetic protocol. Pulse rate, respiratory rate (RR) and mean arterial pressure (MAP) were recorded before the co-induction agent, before induction, and 0, 2 and 5 minutes after intubation. Apnoea >or=30 seconds was recorded and treated. Sedation after pre-medication, activity after the co-induction agent, quality of anaesthetic induction and endotracheal intubation were scored. Propofol dose requirement was significantly reduced in FP [2.90 mg kg(-1)(0.57)] compared to CP [3.51 mg kg(-1) (0.74)] and MP [3.58 mg kg(-1)(0.49)]. Mean pulse rate was higher in MP than in CP or FP (p = 0.003). No statistically significant difference was found between groups in mean RR, MAP or incidence of apnoea. Activity score was significantly higher (i.e. more excited) (p = 0.0001), and quality of induction score was significantly poorer (p = 0.0001) in MP compared to CP or FP. Intubation score was similar in all groups. Fentanyl decreased propofol requirement but did not significantly alter cardiovascular parameters. Midazolam did not reduce propofol requirements and caused excitement in some animals.


Oligodendrocytes in the adult rat spinal cord undergo radiation-induced apoptosis.
Mitotic-linked death is generally regarded as the mode of radiation-induced cell death, particularly in late-responding normal tissues, such as those found in the central nervous system. We have recently reported evidence for radiation-induced apoptosis in the central nervous system using the adult rat spinal cord model. Glial cells, but not neurons or vascular endothelial cells, appeared to undergo apoptosis within 24 h of irradiation. To further characterize the apoptotic process and the type of glial cells involved, a 2-cm segment of the adult rat cervical spinal cord was irradiated with single doses of 1-30 Gy and processed for detailed histological examination at 0, 4, 8, 12, 16, and 24 h after irradiation. Apoptosis was assessed using standard morphological features under the light and electron microscopes and an in situ end labeling assay. A dose response for radiation-induced apoptosis was observed over a dose range of 1-30 Gy, with the peak response at 8 h after irradiation. At 8 h after a 22-Gy irradiation, 96.1% of the apoptotic cells showed positive immunohistochemical staining with Leu-7, a specific marker for oligodendrocytes; only 4.4% of apoptotic cells were positive for Ricinus communis agglutinin-1 (a marker for microglia), and none were positive for glial fibrillary acidic protein (a marker for astrocytes). A significant decrease in the total glial cell density was observed at 24 h after irradiation with 22 (11%) or 30 Gy (14%) but not with 8 Gy. This was due primarily to a decrease in the oligodendroglial density (24%, 22 Gy, P < 0.001; 19%, 30 Gy, P = 0.001), because no decrease in the astroglial population was observed. The duration of apoptosis was estimated to be approximately 1 h. We conclude that there is a depletion of the oligodendroglial population in the adult rat spinal cord within 24 h after irradiation and that the mode of this radiation-induced cell death is apoptosis.


Changes in turnover and vacancy rates of care workers in England from 2008 to 2010: panel analysis of national workforce data.
The combination of growing demand for long-term care and higher expectations of care staff needs to be set in the context of long-standing concerns about the sustainability of recruitment and retention of front-line staff in the United Kingdom. Organisational and work environment factors are associated with vacancy levels and turnover rates. The aim of the current analysis was to investigate changes in turnover and vacancy rates over time experienced by a sample of social care employers in England. Taking a follow-up approach offers potentially more accurate estimates of changes in turnover and vacancy rates, and enables the identification of any different organisational characteristics which may be linked to reductions in these elements over time. The study constructed a panel of 2964 care providers (employers) using 18 separate data sets from the National Minimum Data Set for Social Care during 2008-2010. The findings indicate slight reductions in vacancy rates but the presence of enduring, high turnover rates among direct care workers over the study period. However, the experience of individual employers varied, with home-care providers experiencing significantly higher turnover rates than other parts of the sector. These findings raise questions around the quality and motivations of new recruits and methods of reducing specific vacancy levels. At a time of increased emphasis on care at home, it is worthwhile examining why care homes appear to have greater stability of staff and fewer vacancies than home-care agencies.


Chemical dephosphorylation for identification of multiply phosphorylated peptides and phosphorylation site determination.
We have developed a novel strategy to improve the efficiency of identification of multiply phosphorylated peptides isolated by hydroxy acid modified metal oxide chromatography (HAMMOC). This strategy consists of alkali-induced chemical dephosphorylation (beta-elimination reaction) of phosphopeptides isolated by HAMMOC prior to analysis by liquid chromatography/mass spectrometry (LC/MS). This approach identified 1.9-fold more multiply phosphorylated peptides than the conventional approach without beta-elimination from a digested mixture of three standard phosphoproteins. In addition, the accuracy of phosphorylation site determination in synthetic phosphopeptides was significantly improved. Finally, we applied this approach to a cell lysate. By combining this dephosphorylation approach with the conventional approach, we successfully identified 1649 unique phosphopeptides, including 325 multiply phosphorylated phosphopeptides, from 200 microg of cultured Arabidopsis cells. These results indicate that chemical dephosphorylation prior to LC/MS analysis increases the efficiency of identification of multiply phosphorylated peptides, as well as the accuracy of phosphorylation site determination.


Comparison of the Effects of Hysteroscopic Cold Broad Sword Play Combined with Estrogen and Progestin Sequential Therapy and Drospirenone and Ethinylestradiol Tablets in Patients with Severe Intrauterine Adhesion.
To compare the effects of hysteroscopic cold broad sword play combined with estrogen and progestin sequential therapy and drospirenone and ethinylestradiol tablets in patients with severe intrauterine adhesion. One hundred and eight patients with severe IUA admitted to our hospital from May 2019 to October 2021 were selected for the study. Patients were divided according to their treatment regimen into group A (n = 54) treated with hysteroscopic cold broad sword play + drospirenone and ethinylestradiol tablets and group B (n = 54) treated with hysteroscopic cold broad sword play + estrogen and progestin sequential therapy. The two groups were compared in terms of perioperative indicators, recovery of uterine cavity status, inflammatory factor (C-reactive protein (CRP), interleukin 6 (IL-6), and interleukin 8 (IL-8)] levels), World Health Organization Quality of Life Brief Scale (WHOQOL-BREF) score, and clinical outcome at 3 months postoperatively. After surgery, the duration of abdominal pain and vaginal bleeding was shorter in group A than in group B (P < 0.05). After surgery, the time of menstruation return was shorter in group A than in group B, and the menstrual flow score was higher than in group B (P < 0.05). At 3 months after the surgery, the uterine blood flow index, endometrial thickness, and uterine cavity volume were higher in group A than in group B, and the number of uterine readhesion was lower than in group B (P < 0.05). 3 months after the surgery, the CRP, IL-6, and IL-8 levels decreased in both groups and were lower in group A than in group B (P < 0.05). At 3 months after the surgery, the WHOQOL-BREF scores for each indicator were higher in both groups than before surgery and were higher in group A than in group B (P < 0.05). At 3 months after the surgery, the overall valid rate of group A was 94.44% better than that of group B at 79.63% (P < 0.05). The combination of hysteroscopic cold broad sword play with drospirenone and ethinylestradiol tablets has been shown to be more effective than combined estrogen and progestin sequential therapy in the treatment of patients with severe IUA, which significantly improves the post-operative menstrual status and uterine cavity morphology, significantly reduces the level of inflammatory factors in the patient's body, and significantly improves the quality of life, which is of value.


Characterization and forensic analysis of soil samples using laser-induced breakdown spectroscopy (LIBS).
A method for the quantitative elemental analysis of surface soil samples using laser-induced breakdown spectroscopy (LIBS) was developed and applied to the analysis of bulk soil samples for discrimination between specimens. The use of a 266 nm laser for LIBS analysis is reported for the first time in forensic soil analysis. Optimization of the LIBS method is discussed, and the results compared favorably to a laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) method previously developed. Precision for both methods was <10% for most elements. LIBS limits of detection were <33 ppm and bias <40% for most elements. In a proof of principle study, the LIBS method successfully discriminated samples from two different sites in Dade County, FL. Analysis of variance, Tukey's post hoc test and Student's t test resulted in 100% discrimination with no type I or type II errors. Principal components analysis (PCA) resulted in clear groupings of the two sites. A correct classification rate of 99.4% was obtained with linear discriminant analysis using leave-one-out validation. Similar results were obtained when the same samples were analyzed by LA-ICP-MS, showing that LIBS can provide similar information to LA-ICP-MS. In a forensic sampling/spatial heterogeneity study, the variation between sites, between sub-plots, between samples and within samples was examined on three similar Dade sites. The closer the sampling locations, the closer the grouping on a PCA plot and the higher the misclassification rate. These results underscore the importance of careful sampling for geographic site characterization.


Roll-to-Roll Production of Spider Silk Nanofiber Nonwoven Meshes Using Centrifugal Electrospinning for Filtration Applications.
Filtration systems used in technical and medical applications require components for fine particle deep filtration to be highly efficient and at the same time air permeable. In high efficiency filters, nonwoven meshes, which show increased performance based on small fiber diameters (e.g., using nanofibers), can be used as fine particle filter layers. Nanofiber nonwoven meshes made by electrospinning of spider silk proteins have been recently shown to exhibit required filter properties. Needle-based electrospinning, however, is limited regarding its productivity and scalability. Centrifugal electrospinning, in contrast, has been shown to allow manufacturing of ultrathin polymer nonwoven meshes in an efficient and scalable manner. Here, continuous roll-to-roll production of nonwoven meshes made of recombinant spider silk proteins is established using centrifugal electrospinning. The produced spider silk nanofiber meshes show high filter efficiency in the case of fine particulate matter below 2.5 µm (PM2.5) and a low pressure drop, resulting in excellent filter quality.


High levels of cerebrospinal fluid free kappa chains predict conversion to multiple sclerosis.
A clinically isolated syndrome (CIS) may be the initial presentation of multiple sclerosis (MS). However, some CIS never develop MS. The identification of patients at risk of MS conversion is crucial as early treatment may improve their outcome. Free kappa chains (FKC) are increased in cerebrospinal fluid (CSF) of MS patients. We studied the accuracy of CSF FKC level measurement, using a new nephelometric test, to predict conversion of CIS patients to MS. We calculated linearity and inter-assay variability of the FKC test for CSF values and quantified this protein in CSF from 25 patients with non-inflammatory neurological diseases (NIND) and 78 consecutive CIS patients. We assessed whether high CSF FKC levels associate with CIS conversion to clinically definite MS, defined as the onset of new relapses during follow-up. Between 0.1 and 5mg/l the FKC test showed linearity of 0.98 and inter-assay correlation coefficient of =0.99. A cut-off value of 0.53 mg/l (mean+2SD of NIND group CSF FKC values) was calculated. CIS patients with CSF FKC above this value showed earlier conversion to MS in univariate and multivariate Cox analysis (HR=6.41; 95% CI=1.88-21.78, p=0.003). High CSF FKC levels accurately predict CIS patient conversion to MS.


The impact of reproductive events on the course of bipolar disorder in women.
Little is known about the impact of female reproductive hormones on the course of bipolar disorder. This study was designed to assess the influence of reproductive events and hormonal therapies on the course of bipolar disorder in women. Fifty women with DSM-IV bipolar disorder completed a structured clinical interview to assess the impact of reproductive events on the course of their illness. The onset of bipolar disorder occurred before menarche in 32% (N = 16) of women; 18% (N = 9) experienced the onset within 1 year of menarche. Most women did not receive an accurate diagnosis of nor treatment for bipolar disorder until after they had children, and therefore the majority were not treated with mood stabilizers during or immediately after pregnancies. Of women with children, 20 (67%) of 30 experienced a postpartum mood episode. Of the women who had postpartum episodes after delivery of a first child, all had episodes after subsequent pregnancies. Having a postpartum mood episode after a first pregnancy significantly increased the risk of a postpartum episode after subsequent deliveries (p = .02). Postpartum episodes were almost exclusively depressive. Increased depressive symptoms during pregnancy were significantly associated with postpartum mood episodes (p = .01). Women who were not using hormone replacement therapy (HRT) were significantly more likely than those who were using HRT to report worsening of symptoms during perimenopause/menopause (p = .02). These data suggest that hormonal fluctuations are associated with increased risk of affective dysregulation and mood episodes in women with bipolar disorder.


Growth of immigrant children in the newcomer schools of San Francisco.
A semilongitudinal study on growth and development was initiated on immigrant and refugee school-aged children in San Francisco. Anthropometric values (height, weight, arm circumference, and triceps and subscapular skinfolds) were collected soon after their arrival in the United States and repeated at 3-month intervals for 1 year. Data were analyzed by age-gender cohorts. z Score calculations for measures of height-for-age, weight-for-age, and weight-for-height demonstrated a significant overall deficiency in height-for-age and weight-for-age at the time of the first measurement. Comparisons with a US standard indicated that most of the children were between the fifth and 25th percentiles in these measures. There were fewer children who were significantly deficient in weight-for-height. Calculations for median growth rate indicated that most cohorts exhibited a median growth velocity that was close to or exceeded the median for US white children. There was also significant improvement in weight-for-age. The results indicated that these immigrant and refugee children accelerated their growth markedly in an optimum nutritional environment and were in a period of catch-up growth.


Differences in synaptic and intrinsic properties result in topographic heterogeneity of temporal processing of neurons within the inferior colliculus.
The identification and characterization of organization principals is essential for the understanding of neural function of brain areas. The inferior colliculus (IC) represents a midbrain nexus involved in numerous aspects of auditory processing. Likewise, neurons throughout the IC are tuned to a diverse range of specific stimulus features. Yet beyond a topographic arrangement of the cochlea-inherited frequency tuning, the functional organization of the IC is not well understood. Particularly, a common principle that links the diverse tuning characteristics is unknown. Here we used in vitro patch clamp recordings combined with laser-uncaging, and in vivo single cell recordings to study the spatial and functional organization principles of the central IC. We identified a topographic bias of ascending synaptic input timing that is balanced between inhibition and excitation and co-varies with in vivo first-spike latency. This bias was paralleled post-synaptically by differences in biophysical membrane properties and firing patterns, with integrating neurons predominantly found in the dorso-medial part, and coincidence-detector neurons biased to the ventro-lateral IC. Importantly, these cellular and network features translated into distinct temporal processing capabilities irrespectively of the neurons' characteristic frequency. Our data therefore imply that heterogeneity of synaptic inputs, intrinsic properties and temporal processing are functional principles that underlie the spatial organization of the central IC.


Pretreatment nomogram for predicting the outcome of three-dimensional conformal radiotherapy in prostate cancer.
Several studies have defined risk groups for predicting the outcome after external-beam radiotherapy of localized prostate cancer. However, most models formed patient risk groups, and none of these models considers radiation dose as a predictor variable. The purpose of this study was to develop a nomogram to improve the accuracy of predicting outcome after three-dimensional conformal radiotherapy. This study was a retrospective, nonrandomized analysis of patients treated at the Memorial Sloan-Kettering Cancer Center between 1988 and 1998. Clinical parameters of the 1,042 patients included stage, biopsy Gleason score, pretreatment serum prostate-specific antigen (PSA) level, whether neoadjuvant androgen deprivation therapy was administered, and the radiation dose delivered. Biochemical (PSA) treatment failure was scored when three consecutive rises of serum PSA occurred. A nomogram, which predicts the probability of remaining free from biochemical recurrence for 5 years, was validated internally on this data set using a bootstrapping method and externally using a cohort of patients treated at the Cleveland Clinic, Cleveland, OH. When predicting outcomes for patients in the validation data set from the Cleveland Clinic, the nomogram had a Somers' D rank correlation between predicted and observed failure times of 0.52. Predictions from this nomogram were more accurate (P<.0001) than the best of seven published risk stratification systems, which achieved a Somers' D coefficient of 0.47. The development process illustrated here produced a nomogram that seems to predict more accurately than other available systems and may be useful for treatment selection by both physicians and patients.


Dyslipidaemia associated with antiretroviral therapy in HIV-infected patients.
Highly active antiretroviral therapy (HAART) has had a significant impact on the natural history of human immunodeficiency virus (HIV) infection, leading to a remarkable decrease in its morbidity and mortality, but is frequently associated with clinical and metabolic complications. Fat redistribution or lipodystrophy, hypertriglyceridaemia, hypercholesterolaemia, insulin resistance and diabetes mellitus have been extensively reported in subjects treated with protease inhibitor (PI)-based antiretroviral regimens. In particular, dyslipidaemia occurs in up to 70-80% of HIV-infected individuals receiving HAART and can be associated with all the available PIs, although hypertriglyceridaemia appears to be more frequent in patients treated with ritonavir, ritonavir-saquinavir, or ritonavir-lopinavir. The potential long-term consequences of HAART-associated hyperlipidaemia are not completely understood, but an increased risk of premature coronary artery disease has been reported in young HIV-positive persons receiving PIs. Dietary changes, regular aerobic exercise and switching to a PI-sparing regimen may act favourably on dyslipidaemia. Lipid-lowering therapy is often required with statins or fibrates. The choice of hypolipidaemic drugs should take into account potential pharmacological interactions with antiretroviral agents.


Betamethasone 17-valerate in childhood asthma. A double-blind crossover trial in children not taking other corticosteroid therapy.
A double-blind crossover trial of Betamethasone 17-Valerate steroid aerosol compared with placebo was carried out on 18 children with severe intermittent or chronic asthma. Seventeen of the 18 children showed a clinically obvious improvement on the active drug. The remaining child was unable to use the inhaler properly. In 14 children the response was dramatic. Improvement was shown at home by a reduction in the number of asthmatic symptoms, improvement in twice daily peak flow rates and by a lowered intake of sympathomimetic tablets. At monthly clinic visits, improvement was shown by chest examination, weight gain and by detailed respiratory function tests. By each of these criteria for improvement the benefit of Betamethasone 17-Valerate over the placebo was highly significant. There were no side effects attributable to the therapy and no evidence, as judged by short tetracosactrin tests, of adrenal suppression resulting from the drug. Betamethasone 17-Valerate appears to be a safe and effective therapy in children with severe asthma.


Concomitant use of proton pump inhibitors and clopidogrel in patients with coronary, cerebrovascular, or peripheral artery disease in the factores de Riesgo y ENfermedad Arterial (FRENA) registry.
Among patients receiving clopidogrel for coronary artery disease, concomitant therapy with proton pump inhibitors (PPIs) has been associated with an increased risk for recurrent coronary events. Factores de Riesgo y ENfermedad Arterial (FRENA) is an ongoing, multicenter, observational registry of consecutive outpatients with coronary artery disease, cerebrovascular disease, or peripheral artery disease. We retrospectively examined the influence of concomitant use of PPIs on outcome in patients receiving clopidogrel. As of March 2009, 1222 patients were using clopidogrel: 595 had coronary artery disease, 329 cerebrovascular disease, and 298 had peripheral artery disease. Of these, 519 (42%) were concomitantly using PPIs. Over a mean follow-up of 15 months, 131 patients (11%) had 139 subsequent ischemic events: myocardial infarction 44, ischemic stroke 40, and critical limb ischemia 55. Seventeen of them (13%) died within 15 days of the subsequent event. PPI users had a higher incidence of myocardial infarction (rate ratio, 2.5; 95% confidence interval [CI], 1.3-4.8), ischemic stroke (rate ratio, 1.9; 95% CI, 1.03-3.7), and a nonsignificantly higher rate of critical limb ischemia (rate ratio, 1.6; 95% CI, 0.95-2.8) than nonusers. On multivariate analysis, concomitant use of clopidogrel and PPIs was independently associated with an increased risk for subsequent ischemic events both in the whole series of patients (hazard ratio, 1.8; 95% CI, 1.1-2.7) and in those with cerebrovascular disease or peripheral artery disease (hazard ratio, 1.5; 95% CI, 1.01-2.4). In patients with established arterial disease, concomitant use of PPIs and clopidogrel was associated with a nearly doubling of the incidence of subsequent myocardial infarction or ischemic stroke. This higher incidence persisted after multivariate adjustment.


Stable establishment of wMel Wolbachia in Aedes aegypti populations in Yogyakarta, Indonesia.
The successful establishment of the wMel strain of Wolbachia for the control of arbovirus transmission by Aedes aegypti has been proposed and is being implemented in a number of countries. Here we describe the successful establishment of the wMel strain of Wolbachia in four sites in Yogyakarta, Indonesia. We demonstrate that Wolbachia can be successfully introgressed after transient releases of wMel-infected eggs or adult mosquitoes. We demonstrate that the approach is acceptable to communities and that Wolbachia maintains itself in the mosquito population once deployed. Finally, our data show that spreading rates of Wolbachia in the Indonesian setting are slow which may reflect more limited dispersal of Aedes aegypti than seen in other sites such as Cairns, Australia.


Does postoperative morphine consumption for acute surgical pain impact oncologic outcomes after colorectal cancer resection?: A retrospective cohort study.
Whether morphine used in human cancer surgery would exert tumor-promoting effects is unclear. This study aimed to investigate the effects of morphine dose on cancer prognosis after colorectal cancer (CRC) resection.In a retrospective study, 1248 patients with stage I through IV CRC undergoing primary tumor resections and using intravenous patient-controlled analgesia for acute surgical pain at a tertiary center between October 2005 and December 2014 were evaluated through August 2016. Progression-free survival (PFS) and overall survival (OS) were analyzed using proportional hazards regression models.Multivariable analysis demonstrated no dose-dependent association between the amount of morphine dose and PFS (adjusted hazard ratio, HR = 1.31, 95% confidence interval, CI = 0.85-2.03) or OS (adjusted HR = 0.86, 95% CI = 0.47-1.55). Patients were further classified into the high-dose and low-dose groups by the median of morphine consumption (49.7 mg), and the morphine doses were mean 75.5 ± standard deviation 28.8 mg and 30.1 ± 12.4 mg in high-dose and low-dose groups, respectively. Multivariable models showed no significant difference in PFS or OS between groups, either (adjusted HR = 1.24, 95% CI = 0.97-1.58 for PFS; adjusted HR = 1.01, 95% CI = 0.71-1.43 for OS).Our results did not support a definite association between postoperative morphine consumption and cancer progression or all-cause mortality in patients following CRC resection.


Moderate influenza vaccine effectiveness in Victoria, Australia, 2011.
We used a sentinel general practitioner (GP) network to conduct surveillance for laboratory-confirmed influenza amongst patients presenting with influenza-like illness (ILI) in Victoria, Australia in 2011. The test negative variation of the case control study design was used to estimate effectiveness for seasonal trivalent influenza vaccine. Cases and controls were ILI patients that tested positive and negative for influenza, respectively. Vaccination status was recorded by GPs and vaccine effectiveness (VE) was calculated as (1-adjusted odds ratio)x100%. There were 529 patients included in the study, of which 29% were influenza positive. Twelve percent of study participants were reported as vaccinated, 6% of cases and 15% of controls. Adjusted VE against all influenza was 56%, but not statistically significant. There was generally little variation in VE estimates when stratified by virus type and subtype, which is consistent with good matches between circulating strains and the vaccine strains. The VE was higher among adults of working age than among children.


Cell protrusion and retraction driven by fluctuations in actin polymerization: A two-dimensional model.
Animal cells that spread onto a surface often rely on actin-rich lamellipodial extensions to execute protrusion. Many cell types recently adhered on a two-dimensional substrate exhibit protrusion and retraction of their lamellipodia, even though the cell is not translating. Travelling waves of protrusion have also been observed, similar to those observed in crawling cells. These regular patterns of protrusion and retraction allow quantitative analysis for comparison to mathematical models. The periodic fluctuations in leading edge position of XTC cells have been linked to excitable actin dynamics using a one-dimensional model of actin dynamics, as a function of arc-length along the cell. In this work we extend this earlier model of actin dynamics into two dimensions (along the arc-length and radial directions of the cell) and include a model membrane that protrudes and retracts in response to the changing number of free barbed ends of actin filaments near the membrane. We show that if the polymerization rate at the barbed ends changes in response to changes in their local concentration at the leading edge and/or the opposing force from the cell membrane, the model can reproduce the patterns of membrane protrusion and retraction seen in experiment. We investigate both Brownian ratchet and switch-like force-velocity relationships between the membrane load forces and actin polymerization rate. The switch-like polymerization dynamics recover the observed patterns of protrusion and retraction as well as the fluctuations in F-actin concentration profiles. The model generates predictions for the behavior of cells after local membrane tension perturbations.


Nutrient conversions by photosynthetic bacteria in a concentrated animal feeding operation lagoon system.
A diurnal examination was conducted to determine the effect of photosynthetic bacteria on nutrient conversions in a two-stage concentrated animal feeding operation (CAFO) lagoon system in west-central Oklahoma. Changes in nutrients, microbial populations, and physical parameters were examined at three depths (0, 1.5, and 3.0 m) every 3 h over a 36-h period. The south lagoon (SL) was anaerobic (dissolved oxygen [DO] = 0.09 +/- 0.12 mg/L) while the north lagoon (NL) was facultative (DO ranged from 4.0-0.1 mg/L over 36-h period). Negative sulfide-sulfate (-0.85) and bacteriochlorophyll a (bchl a)-sulfate (-0.83) correlations, as well as positive bchl a-sulfide (0.87) and light intensity (I)-bchl a (0.89) correlations revealed that the SL was dominated by sulfur conversions driven by the photosynthetic purple sulfur bacteria (PSB). The correlation data was supported by diurnal trends for sulfate, sulfide, and bchl a. Both nitrogen and sulfur conversions played a role in the NL; however, nitrogen conversions appeared to dominate this system because of the activity of cyanobacteria. This was shown by positive chlorophyll a (chl a)-I (0.91) and chl a-nitrate (0.98) correlations and the negative correlation between ammonium and nitrite (-0.88). Correlation data was further supported by diurnal trends observed for chl a, DO, and ammonium. For both lagoons, the dominant photosynthetic microbial species determined which nutrient conversion processes were most important.


Cleaved CD44 intracellular domain supports activation of stemness factors and promotes tumorigenesis of breast cancer.
CD44 plays a role in the progression of tumors and is expressed in cancer stem cells (CSCs). However, the mechanisms underlying the crosstalk of CD44 with stemness genes in CSC maintenance remains unclear. In this study, we demonstrated how the cleaved intracellular domain of CD44 (CD44ICD) activates stemness factors such as Nanog, Sox2 and Oct4, and contributes to the tumorigenesis of breast cancer. We have found that the overexpression of CD44ICD increased mammosphere formation in breast cancer cells. Treatment with a γ-secretase inhibitor (GSI), which blocks the cleavage of CD44ICD, interfered with mammosphere formation. Interestingly, CD44ICD decreased the expression levels and nuclear localization of stemness factors, but overexpression of CD44ICD reversed these effects. In addition, we showed that nuclear localization of CD44ICD is important for transcriptional activation of the stemness factors. Furthermore, CD44ICD-overexpressed cells exhibited strong tumorigenecity and greater metastatic potential than did the control cells or CD44-depleted cells in vivo in mice models. Taken together, it was supposed that CD44 promotes tumorigenesis through the interaction and nuclear-translocation of its intracellular domain and stemness factors. We suggest that the prevention of cleavage and nuclear-translocation of CD44ICD is a potential target in treating breast cancer.


Exploring Clinical Practice Hours in Postbaccalaureate-to-Doctor of Nursing Practice Nurse Practitioner Programs.
Advanced practice nursing education is evolving to the doctor of nursing practice (DNP) degree. The American Association of Colleges of Nursing (AACN) DNP Essentials required 1000 hours of direct patient care, whereas the Report of the National Task Force on Quality Nurse Practitioner Education (NTF) Criteria for Evaluation of Nurse Practitioner Programs required 500 direct patient care hours. Indirect hours were unclear and undefined. The AACN Essentials changed the 1000-hour requirement to 500 practice hours, and the NTF increased the direct patient care hours to 750. The study sought to describe the distribution of direct and indirect hours that the National Organization of Nurse Practitioner Faculties member schools offer in their seamless postbaccalaureate-to-DNP nurse practitioner (NP) program. A quantitative survey was distributed to schools of nursing. Eighty-six surveys represented 86 distinct schools and 112 NP programs. Supervised direct patient care hours were an average of 791.31 hours. Indirect hours averaged 170.08 hours. The DNP project hours served as an indirect hour activity. Many schools have more than 750 direct hours adhering to the 2022 NTF Standards. The indirect hour allocation varies and weighs heavily on the DNP project.


Beneficial effects of statins on perioperative cardiovascular outcome.
This review evaluates clinical and experimental articles that have recently been published on the cardioprotective effect of perioperative statin therapy. Perioperative statin therapy improves short and long-term cardiac outcome following noncardiac surgery. This cardiovascular protection has been attributed to the so-called pleiotropic effects, which have a positive effect on plaque stability. With no clinical studies reporting an increased incidence of adverse effects of perioperative statin use, it appears to be a safe and helpful therapeutic option. Perioperative statin therapy may stabilize coronary plaques due to pleiotropic effects and results in a reduction of perioperative cardiovascular complications.


Antisense repression of cytosolic phosphoglucomutase in potato (Solanum tuberosum) results in severe growth retardation, reduction in tuber number and altered carbon metabolism.
The aim of this work was to investigate the role of cytosolic phosphoglucomutase (PGM; EC 18.104.22.168) in the regulation of carbohydrate metabolism. Many in vitro studies have indicated that PGM plays a central role in carbohydrate metabolism; however, until now the importance of this enzyme in plants has not been subject to reverse-genetics investigations. With this intention we cloned the cytosolic isoform of potato PGM (StcPGM) and expressed this in the antisense orientation under the control of the CaMV 35 S promoter in potato plants. We confirmed that these plants contained reduced total PGM activity and that loss in activity was due specifically to a reduction in cytosolic PGM activity. These plants were characterised by a severe phenotype: stunted aerial growth combined with limited root growth and a reduced tuber yield. Analysis of the metabolism of these lines revealed that leaves of these plants were inhibited in sucrose synthesis whereas the tubers exhibited decreased levels of sucrose and starch as well as decreased levels of glycolytic intermediates but possessed unaltered levels of adenylates. Furthermore, a broader metabolite screen utilising GC-MS profiling revealed that these lines contained altered levels of several intermediates of the TCA cycle and of amino acids. In summary, we conclude that cytosolic PGM plays a crucial role in the sucrose synthetic pathway within the leaf and in starch accumulation within the tuber, and as such is important in the maintenance of sink-source relationships.


Consideration of the pH-dependent inhibition of dihydrofolate reductase by methotrexate.
Poisson-Boltzmann calculations were used to determine the pKa of protein functional groups in the unliganded dihydrofolate reductase enzyme, and the pKa of protein and ligand groups in methotrexate-enzyme complexes. The results reported here are in conflict with two fundamental tenets of dihydrofolate reductase inhibition by methotrexate: (1) Asp27 is not expected to be protonated near pH 6.5 in the apoenzyme as previously proposed based on fitting of empirical equations to binding data, and (2) the calculated pKa for the pteridine N1 of the inhibitor while bound to the protein is significantly lower than that estimated for this group from interpretation of NMR data (>10). In fact, the electrostatic calculations and complementary quantum chemical calculations indicate that Asp27 is likely protonated when methotrexate is bound, resulting in a neutral dipole-dipole interaction rather than a salt-bridge between the enzyme and the inhibitor. Reasons for this discrepancy with the experimental data are discussed. Furthermore, His45 and Glu17 in the Escherichia coli enzyme are proposed to be in part responsible for the pH dependence of the conformational degeneracy in the inhibitor-enzyme complex.


[Violence against children: revealing the characteristics of emergency treatment, Brazil, 2006 and 2007].
The aim of this article was to describe the profile of emergency care for injuries resulting from violence against children (< 10 years of age) treated Surveillance System for Violence and Accidents (VIVA) of the Ministry of Health, for 30 consecutive days in 2006 and 2007 in selected cities of Brazil. The 518 children in the sample were predominantly male (60.6%), aged 5-9 years (52.1%), and black (71.2%). The majority (55%) of the cases occurred in the home, 34.2% involved sharp objects, and 68.7% evolved to discharge. The most frequent form of violence was physical aggression (67.4%), involving beating, sharp objects, and firearms. The other types of violence included neglect (32%), psychological abuse (9.5%), and sexual assault (3.5%). The aggressors were largely male (48.1%) and family members of the victim (36.3%). The study shows children's vulnerability to situations of violence. It is necessary to develop specific strategies for inter-sector care and social mobilization to intervene in this problem.


Protease-activated receptor-2 augments experimental crescentic glomerulonephritis.
Protease-activated receptor-2 (PAR-2) is a cellular receptor expressed prominently on epithelial, mesangial, and endothelial cells in the kidney and on macrophages. PAR-2 is activated by serine proteases such as trypsin, tryptase, and coagulation factors VIIa and Xa. It induces pleiotropic effects including vasodilatation, increasing plasminogen activator inhibitor (PAI-1) expression, mesangial cell proliferation, and cytokine production by macrophages. The role of PAR-2 in renal inflammation was studied in antiglomerular basement membrane antibody-induced crescentic glomerulonephritis (CGN) using PAR-2-deficient (PAR-2(-/-)) mice and wild-type littermate controls. PAR-2(-/-) mice had reduced crescent formation, proteinuria, and serum creatinine compared with wild-type mice 21 days after initiation of CGN. Glomerular accumulation of CD4(+) T cells and macrophages and the number of proliferating cells in glomeruli were similar in both groups. Glomerular fibrin deposition was significantly reduced in PAR-2(-/-) mice, and this was associated with reduced renal plasminogen activator inhibitor expression and increased renal matrix-metalloprotinase-9 activity. These results demonstrate a proinflammatory role for PAR-2 in CGN that is independent of effects on glomerular leukocyte recruitment and mesangial cell proliferation. PAR-2-mediated augmentation of renal plasminogen activator inhibitor expression and inhibition of matrix-metalloprotinase-9 activity may contribute to increased glomerular fibrin accumulation and glomerular injury in CGN.


Psychopathology and endorphins.
Suggestive evidence exists linking endorphins to the schizophrenic syndrome; narcotic antagonists appear to slightly attenuate some symptoms, attentional performance is improved and CSF opiate-binding substances are reported to be elevated in a sub-group of patients. Many fewer affectively ill patients have been studied and little evidence has accumulated suggesting a relationship between symptoms of affective illness and endorphins although CSF endorphins appear elevated in some manic-depressive patients and "pain patients" with depression have higher CSF endorphins than pain patients without depression. Catatonic symptoms as well as other psychomotor functions remain promising areas for study. Opioid effects on manic symptoms have been reported by only a few research groups and would benefit from study with longer-acting antagonists administered daily.


Scanning tunneling microscopy reveals single-molecule insights into the self-assembly of amyloid fibrils.
Many severe diseases are associated with amyloid fibril deposits in the body caused by protein misfolding. Structural information on amyloid fibrils is accumulating rapidly, but little is known about the assembly of peptides into fibrils at the level of individual molecules. Here we investigate self-assembly of the fibril-forming tetrapeptides KFFE and KVVE on a gold surface under ultraclean vacuum conditions using scanning tunneling microscopy. Combined with restrained molecular dynamics modeling, we identify peptide arrangements with interesting similarities to fibril structures. By resolving individual peptide residues and revealing conformational heterogeneities and dynamics, we demonstrate how conformational correlations may be involved in cooperative fibril growth. Most interestingly, intermolecular interactions prevail over intramolecular interactions, and assembly of the phenyl-rich KFFE peptide appears not to be dominated by π-π interactions. This study offers interesting perspectives for obtaining fundamental single-molecule insights into fibril formation using a surface science approach to study idealized model systems.


Joint associations of depression, genetic susceptibility and the area of residence for coronary heart disease incidence.
Depression is a risk factor for coronary heart disease (CHD), but less is known whether genetic susceptibility to CHD or regional-level social indicators modify this association. Risk factors of CHD including a Polygenic Risk Score (PRS) were measured for 19 999 individuals residing in Finland in 1997, 2002, 2007 and 2012 (response rates 60%-75%). During the register-based follow-up until 2015, there were 1381 fatal and non-fatal incident CHD events. Unemployment rate, degree of urbanisation and crime rate of the municipality of residence were used as regional level social indicators. HRs were calculated using register-based antidepressant purchases as a non-reversible time-dependent covariate. Those having depression and in the highest quartile of PRS had somewhat higher CHD risk than predicted only by the main effects of depression and PRS (HR for interaction 1.53, 95% CI 0.95 to 2.45). Depression was moderately associated with CHD in high crime (HR 1.51, 95% CI 1.20 to 1.90) and weakly in low crime regions (HR 1.07, 95% CI 0.86 to 1.33; p value of interaction=0.087). Otherwise, we did not found evidence for interactions. Those having both depression and high genetic susceptibility need a special attention in healthcare for CHD.


The trabecular meshwork in acute and chronic angle closure glaucoma.
To determine the effect of acute and chronic primary angle closure glaucoma (PACG) on the trabecular meshwork. Trabecular specimens of 16 consecutive patients with primary angle closure glaucoma (PACG)--6 acute PACG eyes, and 10 chronic PACG eyes without an acute attack--were studied by light and electron microscopy. Acute PACG: The trabecular meshwork revealed a generalised oedema and an accumulation of pigment in the widened trabecular spaces and Schlemm's canal. Attenuated trabecular endothelial cells appeared to be devoid of subcellular components. Chronic PACG: In chronic PACG eyes the trabecular architecture had lost its regular arrangement, with fewer and narrower trabecular spaces and fusion of the trabecular beams in areas. There were numerous electron-dense bodies in the trabecular tissues, both within the trabecular beams and in the extracellular spaces, which had a banded fibrillar structure. An overall loss of endothelial cells was noted; the remaining cells were crowded together and were polymorphic. Melanin pigment was present both within the stroma and in the endothelial cells. Pigment accumulation in the trabecular spaces and within the cells and a noninflammatory degeneration appeared to be the primary changes in the trabecular meshwork after acute angle closure glaucoma. In chronic PACG eyes, there was evidence of loss of endothelial cells and reactive repair processes. These changes were present in areas away from visible peripheral anterior synechiae. A gonioscopic evaluation of the extent of peripheral anterior synechiae alone may not reflect the extent of trabecular meshwork damage in acute and chronic PACG. Patients experiencing an acute attack of PACG require a long-term follow up, because the intraocular pressure (IOP) may rise later, due to ongoing changes compromising the outflow facility, or due to the effects of aging in the trabecular meshwork.


Evidence for paternal age-related alterations in meiotic chromosome dynamics in the mouse.
Increasing age in a woman is a well-documented risk factor for meiotic errors, but the effect of paternal age is less clear. Although it is generally agreed that spermatogenesis declines with age, the mechanisms that account for this remain unclear. Because meiosis involves a complex and tightly regulated series of processes that include DNA replication, DNA repair, and cell cycle regulation, we postulated that the effects of age might be evident as an increase in the frequency of meiotic errors. Accordingly, we analyzed spermatogenesis in male mice of different ages, examining meiotic chromosome dynamics in spermatocytes at prophase, at metaphase I, and at metaphase II. Our analyses demonstrate that recombination levels are reduced in the first wave of spermatogenesis in juvenile mice but increase in older males. We also observed age-dependent increases in XY chromosome pairing failure at pachytene and in the frequency of prematurely separated autosomal homologs at metaphase I. However, we found no evidence of an age-related increase in aneuploidy at metaphase II, indicating that cells harboring meiotic errors are eliminated by cycle checkpoint mechanisms, regardless of paternal age. Taken together, our data suggest that advancing paternal age affects pairing, synapsis, and recombination between homologous chromosomes--and likely results in reduced sperm counts due to germ cell loss--but is not an important contributor to aneuploidy.


A bispecific immunotoxin (DTAT13) targeting human IL-13 receptor (IL-13R) and urokinase-type plasminogen activator receptor (uPAR) in a mouse xenograft model.
A bispecific immunotoxin (IT) called DTAT13 was synthesized in order to target simultaneously the urokinase-type plasminogen activator receptor (uPAR)-expressing tumor neovasculature and IL-13 receptor expressing glioblastoma cells with the goal of intratumoral administration for brain tumors. The recombinant hybrid was created using the non-internalizing N-terminal fragment (ATF) of uPA and the IL-13 molecule for binding plus the catalytic and translocation portion of diphtheria toxin (DT) for killing. The 71 kDa protein was highly selective for human glioblastoma in vitro showing no loss on binding compared with DTAT and DTIL13 controls. In vivo, DTAT13 caused the regression of small tumors when administered at 10 micro g/day given on a five-dose schedule every other day. DTAT13 was able to target both overexpressed uPAR and the vasculature, as demonstrated by its ability to kill HUVEC cells. Also, mortality studies indicated that DTAT13 was less toxic than DTAT or DTIL13. These findings indicate that bispecific IT may allow treatment of a broader subset of antigenically diverse patients while simultaneously reducing the exposure to toxin required than if two separate agents were employed.


Neuropeptide Y1 receptor antagonist but not neuropeptide Y itself increased bone mineral density when locally injected with hyaluronic acid in male Wistar rats
The nervous system controls bone mass via both the central (CNS) and the peripheral (PNS) nervous systems. Intriguingly, neuropeptide Y (NPY) signaling occurs in both. Less is known on how the PNS stimulated NPY signaling controls bone metabolism. The objective of this study was to evaluate whether NPY or NPY1 receptor antagonist changes local bone mineral density (BMD) when injected into a Wistar rat tibia. Tibial intramedullary area of 24 wild type male Wistar rats (average weight = 350 ± 50 g, average age = 4 ± 0.5 months) were injected with NPY (1 × 10-5 M and 1 × 10-6 M) and NPY1 receptor antagonist (1 × 10-4 M) dissolved in hyaluronic acid (HA) separately. Tibiae were collected after one and two weeks. BMD was measured with dual-energy X-ray absorptiometry (DXA) and micro quantitative computer tomography (QCT). Histological changes were analyzed with light microscopy, Goldner's Masson trichrome (MT), and hematoxylin-eosin staining. According to DXA, the mean BMD of NPY dose 1 (1 × 10-5 M) was significantly lower than that of the control (HA applied) group and not significantly but still lower than that of the NPY dose 2 and NPY1 antagonist applied groups. QCT results indicated the same pattern statistically insignificantly in the trabecular area but not in the cortex of the bones. Histologically, only NPY1 antagonist applied tibiae revealed young spongiosis bone trabeculae formed in the borderline of the cortical bones. HA was remarkably biocompatible and late degrading in the tissues. Local administration of NPY and NPY1 antagonists may hold regulating potential of BMD and bone formation. NPY1 antagonist caused new bone formation in trabecular bone when applied locally. NPY dissolved in HA however can be used to suppress bone formation.


Immediate histologic correlation in patients with different HPV genotypes and ages: a single center analysis in China.
Human papillomavirus (HPV) has been confirmed as a major causative factor for malignant transformation of cervical epithelial cells and for the development of cervical intraepithelial neoplasia (CIN) and invasive cervical cancer. We carried out this study to investigate the association of different HPV genotypes and ages with immediate histological cervical lesions in opportunistic screening patients in a single center. A total of 1,661 samples with biopsy-confirmed histologic findings were collected from the gynecological clinic of our hospital between October 2017 and May 2020 for analysis. The distribution of single-type HPV genotypes in CIN of different severities and the age-dependent prevalence for single-type HPV infection were analyzed. In both CIN2 and CIN3 group, HPV16, 58, 52, 33 and 31/18 were detected as top 5 high-risk human papillomavirus (hrHPV) types, which accounts for 89.25% and 88.54% of single HPV infection incidence respectively. Besides, not a single case of HPV45 was found in CIN2 and CIN3. HPV16 was the dominant genotype in both CIN2 and CIN3, accounted for 46.24% and 55.21%, respectively. The prevalence of HPV16 was the most frequent in all the age groups, except ≥ 65 years group in CIN3, and almost one in three HPV16-positive patients were diagnosed with high grade CIN. The peak of the incidence of CIN3 was observed at 25 ~ 34 years (33.68%), followed by 35 ~ 44 years (31.58%). High grade CIN peak at 25 ~ 44 years, women of this age are recommended for normative screening if conditions permit. HPV16-positive patients should be given high priority in opportunistic screening, while the single-center data suggesting a low risk of CIN2/3 in HPV45-positive patients. For women ≥ 65 years old, patients infected with other HPV types should be also taken seriously. In general, HPV16, 58, 52, 33, 31 and 18 were the most common genotypes in CIN2/3, and a vaccine including these predominant genotypes might be of great significance for cervical cancer prevention in China.


Insights into the cellular responses to hypoxia in filamentous fungi.
Most eukaryotes require molecular oxygen for growth. In general, oxygen is the terminal electron acceptor of the respiratory chain and represents an important substrate for the biosynthesis of cellular compounds. However, in their natural environment, such as soil, and also during the infection, filamentous fungi are confronted with low levels of atmospheric oxygen. Transcriptome and proteome studies on the hypoxic response of filamentous fungi revealed significant alteration of the gene expression and protein synthesis upon hypoxia. These analyses discovered not only common but also species-specific responses to hypoxia with regard to NAD(+) regeneration systems and other metabolic pathways. A surprising outcome was that the induction of oxidative and nitrosative stress defenses during oxygen limitation represents a general trait of adaptation to hypoxia in many fungi. The interplay of these different stress responses is poorly understood, but recent studies have shown that adaptation to hypoxia contributes to virulence of pathogenic fungi. In this review, results on metabolic changes of filamentous fungi during adaptation to hypoxia are summarized and discussed.


Alternative primer sets for PCR detection of genotypes involved in bacterial aerobic BTEX degradation: distribution of the genes in BTEX degrading isolates and in subsurface soils of a BTEX contaminated industrial site.
Eight new primer sets were designed for PCR detection of (i) mono-oxygenase and dioxygenase gene sequences involved in initial attack of bacterial aerobic BTEX degradation and of (ii) catechol 2,3-dioxygenase gene sequences responsible for meta-cleavage of the aromatic ring. The new primer sets allowed detection of the corresponding genotypes in soil with a detection limit of 10(3)-10(4) or 10(5)-10(6) gene copies g(-1) soil, assuming one copy of the gene per cell. The primer sets were used in PCR to assess the distribution of the catabolic genes in BTEX degrading bacterial strains and DNA extracts isolated from soils sampled from different locations and depths (vadose, capillary fringe and saturated zone) within a BTEX contaminated site. In both soil DNA and the isolates, tmoA-, xylM- and xylE1-like genes were the most frequently recovered BTEX catabolic genes. xylM and xylE1 were only recovered from material from the contaminated samples while tmoA was detected in material from both the contaminated and non-contaminated samples. The isolates, mainly obtained from the contaminated locations, belonged to the Actinobacteria or Proteobacteria (mainly Pseudomonas). The ability to degrade benzene was the most common BTEX degradation phenotype among them and its distribution was largely congruent with the distribution of the tmoA-like genotype. The presence of tmoA and xylM genes in phylogenetically distant strains indicated the occurrence of horizontal transfer of BTEX catabolic genes in the aquifer. Overall, these results show spatial variation in the composition of the BTEX degradation genes and hence in the type of BTEX degradation activity and pathway, at the examined site. They indicate that bacteria carrying specific pathways and primarily carrying tmoA/xylM/xylE1 genotypes, are being selected upon BTEX contamination.


A Biomimetic Multiparametric Assay to Characterise Anti-Amyloid Drugs.
Alzheimer's disease (AD) is the most widespread form of senile dementia worldwide and represents a leading socioeconomic problem in healthcare. Although it is widely debated, the aggregation of the amyloid β peptide (Aβ) is linked to the onset and progression of this neurodegenerative disease. Molecules capable of interfering with specific steps in the fibrillation process remain of pharmacological interest. To identify such compounds, we have set up a small molecule screening process combining multiple experimental methods (UV and florescence spectrometry, ITC, and ATR-FTIR) to identify and characterise potential modulators of Aβ1-42 fibrillation through the description of the biochemical interactions (molecule-membrane Aβ peptide). Three known modulators, namely bexarotene, Chicago sky blue and indomethacin, have been evaluated through this process, and their modulation mechanism in the presence of a biomembrane has been described. Such a well-adapted physico-chemical approach to drug discovery proves to be an undeniable asset for the rapid characterisation of compounds of therapeutic interest for Alzheimer's disease. This strategy could be adapted and transposed to search for modulators of other amyloids such as tau protein.


MR angiography compared to conventional selective angiography in acute stroke.
Accuracy of intracranial magnetic resonance angiography (MRA) and reliability of interpretation are not well established compared to conventional selective catheter angiography. The purpose of this study was to determine the accuracy of MRA in evaluation of intracranial vessels in acute stroke and transient ischemic attack (TIA) patients Twenty-nine patients (seven females, 22 males; median age 53) with acute stroke or TIA were enrolled into the study. All patients underwent both MRA using a 3 T clinical magnet and conventional angiography within 48 hours. Median time between MRA and angiography was 263 minutes. Conventional angiography preceded MRA in 15 cases. Fourteen patients received thrombolysis during MRA or angiography. National Institutes of Health Stroke Scale scores were obtained prior to the MR exam. One neuroradiologist rated all conventional angiograms, which were used as gold standard. Five observers, blinded to conventional angiography results and all clinical information except symptom side, rated the MR angiograms. Kappa statistics were used to assess reliability; contingency tables were used to assess accuracy of non-enhanced and enhanced MRA. Two hundred and fifty two intracranial vessels were assessed. Agreement between raters was good for both non-enhanced (kappa = 0.50) and gadolinium-enhanced (kappa = 0.46) images. There were a total of 26 vessels occluded by DSA. Overall, the non-enhanced MRA showed sensitivity of 84.2% (95% CI 60.4-96.6) and specificity of 84.6% (95% CI 78.6-89.4). The enhanced MRA showed sensitivity of 69.2 (95% CI 38.6-90.9) and specificity of 73.6 (95% CI 65.5-80.7). Magnetic resonance angiography is a good non-invasive screening tool for assessing intracranial vessel status in acute ischemic stroke. Angiography remains the gold standard for definitive assessment of the intracranial circulation.


MADS8 is indispensable for female reproductive development at high ambient temperatures in cereal crops.
Temperature is a major factor that regulates plant growth and phenotypic diversity. To ensure reproductive success at a range of temperatures, plants must maintain developmental stability of their sexual organs when exposed to temperature fluctuations. However, the mechanisms integrating plant floral organ development and temperature responses are largely unknown. Here, we generated barley and rice loss-of-function mutants in the SEPALLATA-like MADS-box gene MADS8. The mutants in both species form multiple carpels that lack ovules at high ambient temperatures. Tissue-specific markers revealed that HvMADS8 is required to maintain floral meristem determinacy and ovule initiation at high temperatures, and transcriptome analyses confirmed that temperature-dependent differentially expressed genes in Hvmads8 mutants predominantly associate with floral organ and meristem regulation. HvMADS8 temperature-responsive activity relies on increased binding to promoters of downstream targets, as revealed by a cleavage under targets and tagmentation (CUT&Tag) analysis. We also demonstrate that HvMADS8 directly binds to two orthologs of D-class floral homeotic genes to activate their expression. Overall, our findings revealed a new, conserved role for MADS8 in maintaining pistil number and ovule initiation in cereal crops, extending the known function of plant MADS-box proteins in floral organ regulation.


Assessment of Hydration Thermodynamics at Protein Interfaces with Grid Cell Theory.
Molecular dynamics simulations have been analyzed with the Grid Cell Theory (GCT) method to spatially resolve the binding enthalpies and entropies of water molecules at the interface of 17 structurally diverse proteins. Correlations between computed energetics and structural descriptors have been sought to facilitate the development of simple models of protein hydration. Little correlation was found between GCT-computed binding enthalpies and continuum electrostatics calculations. A simple count of contacts with functional groups in charged amino acids correlates well with enhanced water stabilization, but the stability of water near hydrophobic and polar residues depends markedly on its coordination environment. The positions of X-ray-resolved water molecules correlate with computed high-density hydration sites, but many unresolved waters are significantly stabilized at the protein surfaces. A defining characteristic of ligand-binding pockets compared to nonbinding pockets was a greater solvent-accessible volume, but average water thermodynamic properties were not distinctive from other interfacial regions. Interfacial water molecules are frequently stabilized by enthalpy and destabilized entropy with respect to bulk, but counter-examples occasionally occur. Overall detailed inspection of the local coordinating environment appears necessary to gauge the thermodynamic stability of water in protein structures.


Notch signaling is involved in human articular chondrocytes de-differentiation during osteoarthritis.
During osteoarthritis (OA), chondrocytes undergo de-differentiation, resulting in the acquisition of a fibroblast-like morphology, decreased expression of collagen type II (colII) and aggrecan, and increased expression of collagen type I (colI), metalloproteinase 13 (MMP13) and nitric oxide synthase (eNOS). Notch signaling plays a crucial role during embryogenesis. Several studies showed that Notch is expressed in adulthood. The aim of our study was to confirm the involvement of Notch signaling in human OA at in vitro and ex vivo levels. Normal human articular chondrocytes were cultured during four passages either treated or not with a Notch inhibitor: DAPT. Human OA cartilage was cultured with DAPT for five days. Chondrocytes secreted markers and some Notch pathway components were analyzed using Western blotting and qPCR. Passaging chondrocytes induced a decrease in the cartilage markers: colII and aggrecan. DAPT-treated chondrocytes and OA cartilage showed a significant increase in healthy cartilage markers. De-differentiation markers, colI, MMP13 and eNOS, were significantly reduced in DAPT-treated chondrocytes and OA cartilage. Notch1 expression was proportional to colI, MMP13 and eNOS expression and inversely proportional to colII and aggrecan expression in nontreated cultured chondrocytes. Notch ligand: Jagged1 increased in chondrocytes culture. DAPT treatment resulted in reduced Jagged1 expression. Notch target gene HES1 increased during chondrocyte culture and was reduced when treated with DAPT. Targeting Notch signaling during OA might lead to the restitution of the typical chondrocyte phenotype and even to chondrocyte redifferentiation during the pathology.


Assessing Medical Students', Residents', and the Public's Perceptions of the Uses of Personal Digital Assistants.
Although medical schools are encouraging the use of personal digital assistants (PDAs), there have been few investigations of attitudes toward their use by students or residents and only one investigation of the public's attitude toward their use by physicians. In 2006, the University of Louisville School of Medicine surveyed 121 third- and fourth-year medical students, 53 residents, and 51 members of the non-medical public about their attitudes toward PDAs. Students were using either the Palm i705 or the Dell Axim X50v; residents were using devices they selected themselves (referred to in the study generically as PDAs). Three survey instruments were designed to investigate attitudes of (a) third- and fourth-year medical students on clinical rotations, (b) Internal Medicine and Pediatrics residents, and (c) volunteer members of the public found in the waiting rooms of three university practice clinics. Both residents and medical students found their devices useful, with more residents (46.8%) than students (16.2%) (p < 0.001) rating PDAs "very useful." While students and residents generally agreed that PDAs improved the quality of their learning, residents' responses were significantly higher (p < 0.05) than students'. Residents also responded more positively than students that PDAs made them more effective as clinicians. Although members of the public were generally supportive of PDA use, they appeared to have some misconceptions about how and why physicians were using them. The next phase of research will be to refine the research questions and survey instruments in collaboration with another medical school.


Alcohol and sleep I: effects on normal sleep.
This review provides a qualitative assessment of all known scientific studies on the impact of alcohol ingestion on nocturnal sleep in healthy volunteers. At all dosages, alcohol causes a reduction in sleep onset latency, a more consolidated first half sleep and an increase in sleep disruption in the second half of sleep. The effects on rapid eye movement (REM) sleep in the first half of sleep appear to be dose related with low and moderate doses showing no clear trend on REM sleep in the first half of the night whereas at high doses, REM sleep reduction in the first part of sleep is significant. Total night REM sleep percentage is decreased in the majority of studies at moderate and high doses with no clear trend apparent at low doses. The onset of the first REM sleep period is significantly delayed at all doses and appears to be the most recognizable effect of alcohol on REM sleep followed by the reduction in total night REM sleep. The majority of studies, across dose, age and gender, confirm an increase in slow wave sleep (SWS) in the first half of the night relative to baseline values. The impact of alcohol on SWS in the first half of night appears to be more robust than the effect on REM sleep and does not appear to be an epiphenomenon REM sleep reduction. Total night SWS is increased at high alcohol doses across gender and age groups.


Copper-dependent transcriptional regulation by Candida albicans Mac1p.
We have previously shown that copper uptake and regulation in the opportunistic pathogen Candida albicans has some similarities to those in Saccharomyces cerevisiae, including the activation of the copper transporter gene CaCTR1 under low-copper conditions by the transcription factor CaMac1p. However, in this study, further analysis has shown that the actual mechanism of regulation by CaMac1p is different from that of its S. cerevisiae homologue. We demonstrate for the first time, to our knowledge, that the CaMAC1 gene is transcriptionally autoregulated in a copper-dependent manner, in contrast to ScMAC1, which is constitutively transcribed. We also demonstrate that the presence of one copper response element in the promoters of CaCTR1, CaMAC1 and the ferric/cupric reductase gene CaFRE7 is sufficient for normal levels of copper-responsive transcription. In contrast, two promoter elements are essential for normal levels of copper-dependent transcriptional activation by ScMac1p. CaMac1p is also involved in the regulation of the iron-responsive transcriptional repressor gene SFU1 and the alternative oxidase gene AOX2. This work describes a key feature of the copper uptake system in C. albicans that distinguishes it from similar processes in the model yeast S. cerevisiae. The importance of copper uptake in the environment of the human host and the implications for the disease process are discussed.


Effect of ethanol on vascular prostacyclin (prostaglandin I2) synthesis, platelet aggregation, and platelet thromboxane release.
A series of experiments with platelets from healthy volunteers showed a concentration related inhibitory effect of ethanol on platelet aggregation and release of thromboxane A2. This effect was observed at blood alcohol concentrations ranging between 66 and 132 mg/dl (14.3 and 28.6 mmol/l), which are commonly found in alcoholics. Investigations carried out by incubating ethanol with platelet rich plasma in vitro also showed an inverse linear correlation between ethanol concentration and platelet thromboxane synthesis. In contrast, the incubation of a wide range of concentrations of ethanol with human endothelial cells and rat aortic rings did not alter the ability of these systems to synthesise prostacyclin (prostaglandin I2). This finding of a selective inhibition of thromboxane A2 synthesis and platelet aggregation without an alteration of prostaglandin I2 synthesis may provide an explanation for the reported ethanol mediated protection against vascular disease. This effect of ethanol may also be relevant to the induction of acute gastrointestinal haemorrhage that occurs after bouts of excessive alcohol consumption.


Lead mine waste: hazards to livestock.
Waste from disused lead mines is used for a variety of purposes and has been implicated in outbreaks of lead poisoning in animals. Three outbreaks of lead poisoning, two in cattle and one in farm dogs, occurred in the south of Scotland. In the first outbreak, three housed suckler cows died and two were clinically affected but recovered after treatment. In the second, four heifers died and two recovered after therapy. Blood samples taken from cattle in both outbreaks contained lead in concentrations from 0 . 59 to 2 . 00 ppm (normal blood lead level 0 . 05 to 0 . 25 ppm). Post mortem examinations revealed signs of lead poisoning. Metallic particles in the rumens and abomasums of affected animals suggested mine waste as the source of the lead and subsequently it was found that the material had been used in the construction of a ramp over which silage was pushed (outbreak 1) and to fill potholes on the farm (outbreak 2). Covering the relevant areas with concrete prevented further occurrences. Farm dogs were affected over a period where mine waste was used to resurface a road. Two dogs died, six others showed signs of lead poisoning and others, with no clinical signs, had elevated blood lead levels.


A First Report on Experience in Managing Infants with Congenital Toxoplasmosis in Ethiopia: Case Reports and a Review of Evaluation and Treatment.
Background. Congenital toxoplasmosis is a major sequela of untreated primary maternal infection. With or without symptoms, untreated infections eventually lead to multiple neurologic complications. Despite the high Toxoplasma gondii seroprevalence in the Ethiopian population, there are no reports of newborns diagnosed and treated for congenital toxoplasmosis. Presentation of Cases. The clinical presentation, evaluation, and management of three infants with congenital toxoplasmosis are described. Two were symptomatic at birth. All three had confirmed diagnoses using Toxoplasma serologic tests. Two completed their treatment with one infant developing complications of strabismus and seizure disorder. Discussion and Conclusions. There is little experience in managing congenital toxoplasmosis in Ethiopia due to constraints in diagnostics and therapy. The description of this first such report underscores the need for risk assessment and evaluation during antenatal care to obtain favorable fetal outcomes.


Patient Reported Outcomes After Radiation Therapy for Bone Metastases as a Function of Age: A Secondary Analysis of the NCIC CTG SC-Twenty-Three Randomized Trial.
To explore the age difference in response and patient-reported outcomes in patients with cancer having bone metastases undergoing palliative radiotherapy. Patients completed the European Organisation for Research and Treatment of Cancer (EORTC) Quality-of-Life (QOL) Bone Metastases module (QLQ-BM22), EORTC QOL Core-15-Palliative (QLQ-C15-PAL), and Dexamethasone Symptom Questionnaire (DSQ) before a single 8-Gy radiation treatment, on days 10 and 42 after treatment. Patient demographics, performance status, analgesic consumption, BM22, C15, and DSQ were compared with multivariant analysis between patients under 75 years and 75 years and older. Multiple linear regression models were used to assess the differences between age-groups, adjusting for baseline demographics and primary disease sites. There were 298 patients (170 male) with 209 (70%) less than 75 years of age. Most common primary cancer sites include lung, prostate, and breast. At baseline, younger patients had better performance status, consumed more analgesic, and reported worse scores in nausea, insomnia, and functional interference, while older patients more commonly had prostate cancer. There were no significant differences in the incidence of radiation-induced pain flare; response to radiation; changes from baseline for BM22, C15-PAL; and DSQ, nor overall survival at day 42 between the 2 groups. Responders to radiation in the elderly group reported better improvement in physical and emotional domains when compared with nonresponders. In patients with cancer having bone metastases undergoing palliative radiotherapy, there was no significant difference in general with age in response to radiation and patient-reported outcomes. Palliative radiotherapy should be offered to elderly patients when needed.


Rethinking Context in Continuing Professional Development: From Identifying Barriers to Understanding Social Dynamics.
For continuing professional development (CPD) to reach its potential to improve outcomes requires an understanding of the role of context and the influencing conditions that enable interventions to succeed. We argue that the heuristic use of frameworks to design and implement interventions tends to conceptualize context as defined lists of barriers, which may obscure consideration of how different contextual factors interact with and intersect with each other. We suggest a framework approach that would benefit from postmodernist theory that explores how ideologies, meanings, and social structures in health care settings shape social practices. As an illustrative example, we conducted a Foucauldian discourse analysis of diabetes care to make visible how the social, historical, and political conditions in which clinicians experience, practice, and shape possibilities for behavior change. The discursive construction of continuing education as a knowledge translation mechanism assumes and is contingent on family physicians to implement guidelines. However, they enact other discursively constituted roles that may run in opposition. This paradoxical position creates a tension that must be navigated by family physicians, who may perceive it possible to provide good care without necessarily implementing guidelines. We suggest marrying "framework" thinking with postmodernist theory that explores how ideologies, meanings, and social structures shape practice behavior change. Such a proposed reconceptualization of context in the design of continuing professional development interventions could provide a more robust and nuanced understanding of how the dynamic relationships and interactions between clinicians, patients, and their work environments shape educational effectiveness.


Historical Overview of Transitional Feeding Recommendations and Vegetable Feeding Practices for Infants and Young Children.
Although recommendations for introducing solid foods to infants and young children have changed significantly since the beginning of the 20th century, vegetable consumption recommendations have always been an important part of the child-feeding repertoire. In 1958, the first report of the American Academy of Pediatrics (AAP) Committee on Nutrition stated that developmental maturity of the gut and neuromuscular system, growth rate, and activity level were good indicators for determining when to introduce solid foods to infants than age. All 7 editions of the AAP Pediatric Nutrition Handbook use an evidence-based model for recommendations concerning the complementary feeding of infants and young children. The model includes developmental readiness principles, feeding practices, and age-appropriate nutrient requirements. Dietary patterns and nutrient consumption among infants and young children have been analyzed using data from the 2002 and 2008 Feeding Infants and Toddlers Study (FITS). The 2008 FITS also collected information concerning participation in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). Since 1972, WIC has been a cost-effective means of improving the diets and health of infants and young children from low-income families. Data from the 2008 FITS showed that many young children did not consume recommended amounts of fiber or potassium, and vegetable and fruit intakes continued to be lower than recommended. Low vegetable consumption and limited variety were also seen among WIC participants and nonparticipants aged 6 months to 4 years prior to changes in the WIC food package. Increasing children's consumption of all vegetables should continue to be a focus going forward.


Analysis of the frequencies and of the memory T cell phenotypes of human CD8+ T cells specific for influenza A viruses.
We characterized the human CD8+ T cell response against influenza A viruses by a flow cytometry-based assay. Peripheral blood mononuclear cells (PBMCs) were incubated with inactivated influenza virus preparation, for 17 h, and were stained for intracellular interferon-gamma. Major histocompatibility complex class I-restricted memory CD8+ T cells specific for influenza antigens were detected in PBMCs from all 19 adult donors, at an average frequency of 0.39%. On average, 83% of influenza virus-specific CD8+ T cells expressed the differentiation-associated marker CD27, a percentage that is significantly higher than that of CD8+ T cells specific for pp65 of human cytomegalovirus (53%). These observations indicate that class I-restricted immunity against influenza A viruses is characterized by the persistence, after clearance of infection, of circulating antigen-specific CD8+ T cells. The different patterns of CD27 expression in influenza virus- and cytomegalovirus-specific CD8+ T cells suggest that influenza virus-specific memory and effector CD8+ T cells can be differentiated by phenotypic analysis.


Storage study and quality evaluation of coconut protein powder.
Coconut skim milk and insoluble protein are 2 major byproducts in the production of virgin coconut oil. Coconut skim milk was homogenized along with insoluble protein and spray dried to obtain a value-added product, namely, coconut protein powder (CPP). This study deals with the storage study of CPP under different conditions (refrigerated [control], ambient and accelerated). CPP samples were withdrawn periodically at designated intervals of 15 d for accelerated and control, and 30 d for ambient condition. CPP stored at different conditions exhibited marginal moisture uptake (by 0.74 % w/w for control, 0.76 % w/w for ambient, and 1.26 % w/w for accelerated condition) and as a result, had very little effect on the functional properties of the powder. Withdrawn CPP was tested for sensory quality aspects and subjected to instrumental analysis as well. Withdrawn CPP was incorporated as a milk substitute in dessert (Kheer). Quantitative descriptive analysis of the powder and product (Kheer) showed no significant difference in attributes of CPP during the storage period of 2 mo. Electronic nose analysis revealed that CPP samples were not much different with respect to aroma pattern matching, respectively.


Fractures and alcohol abuse - patient opinion of alcohol intervention.
To clarify patient opinions about alcohol intervention in relation to surgery before investigating the effect in a Scandinavian multi-centre randomized trial. A qualitative study. Thirteen consecutive alcohol patients with fractures participated after informed consent. They were interviewed during their hospital stay. The number of participants was based on the criteria of data-saturation. The analysis followed the applied qualitative framework model aimed at evaluation of specific participant needs within a larger overall project. All patients regarded alcohol intervention in relation to surgery as a good idea. They did not consider quit drinking as a major problem during their hospital stay and had all remained abstinent in this period. About half of the patients were ready or partly ready to participate in an alcohol intervention. Patient opinions and their readiness to participate were expressed in four groups, which also reflected their readiness to stop drinking in the perioperative period, their general acceptance of supportive disulfiram as part of an alcohol intervention as well as their awareness of postoperative complications. This study clarified that the patients found alcohol intervention relevant in relation to surgery.


Does Orientation of the Femoral Head Affect Acetabular Development? An Experimental Study in Lamb.
Derotational osteotomy of the proximal femur has proved to be effective in the treatment of residual acetabular dysplasia. However, the reason why this osteotomy is effective remains debatable. The purpose of this study is to investigate if an alteration of femoral head orientation affects acetabular growth. A proximal femoral osteotomy was performed in 21 lambs aged 3 months: 5 varus osteotomies (110 degrees), 4 valgus osteotomies (150 degrees), and 12 derotation osteotomies. Results were compared with a control group (5 animals). Osteotomy was fixed with a screw-plate device. Version was controlled intraoperatively with K-wires. Animals were killed 3 months after surgical procedure. A morphometric study of both proximal femur and acetabulum was performed, including deepness, volume and diameters of the acetabulum, neck-shaft angle and femoral version. The average neck-shaft angle for the normal, anteversion, and retroversion groups was 129 degrees, whereas it was 110 degrees for the varus group and 149 degrees for the valgus group. The average femoral version for the normal, valgus, and varus groups was 21 degrees of anteversion, whereas it was 38 degrees of anteversion for the so-called anteversion group and 17 degrees of retroversion for the retroversion group. Nor the neck-shaft angle, nor the femoral version correlated with the acetabular anteroposterior diameter (P=0.698, 0.6, respectively), the acetabular inferosuperior diameter (P=0.083, 0.451, respectively) or the acetabular deepness (P=0.14, 0.371, respectively). The neck-shaft angle correlated significantly with acetabular volume (P=0.023), so that the lower the neck-shaft angle, the higher the acetabular volume (r=-0.453). The femoral version did not correlated with acetabular volume (P=0.381). Decreasing the neck-shaft angle provokes an increase in acetabular volume, whereas changes in femoral version do not affect the acetabular growth. Extra-articular osteotomies that alter femoral orientation affect intra-articular gross morphology. Level II-therapeutic study.


MALDI-FTICR imaging mass spectrometry of drugs and metabolites in tissue.
A new approach is described for imaging mass spectrometry analysis of drugs and metabolites in tissue using matrix-assisted laser desorption ionization-Fourier transform ion cyclotron resonance (MALDI-FTICR). The technique utilizes the high resolving power to produce images from thousands of ions measured during a single mass spectrometry (MS)-mode experiment. Accurate mass measurement provides molecular specificity for the ion images on the basis of elemental composition. Final structural confirmation of the targeted compound is made from accurate mass fragment ions generated in an external quadrupole-collision cell. The ability to image many small molecules in a single measurement with high specificity is a significant improvement over existing MS/MS based technologies. Example images are shown for olanzapine in kidney and liver and imatinib in glioma.


Evaluation of the Ion Torrent Personal Genome Machine for Gene-Targeted Studies Using Amplicons of the Nitrogenase Gene nifH.
The sequencing chips and kits of the Ion Torrent Personal Genome Machine (PGM), which employs semiconductor technology to measure pH changes in polymerization events, have recently been upgraded. The quality of PGM sequences has not been reassessed, and results have not been compared in the context of a gene-targeted microbial ecology study. To address this, we compared sequence profiles across available PGM chips and chemistries and with 454 pyrosequencing data by determining error types and rates and diazotrophic community structures. The PGM was then used to assess differences in nifH-harboring bacterial community structure among four corn-based cropping systems. Using our suggested filters from mock community analyses, the overall error rates were 0.62, 0.36, and 0.39% per base for chips 318 and 314 with the 400-bp kit and chip 318 with the Hi-Q chemistry, respectively. Compared with the 400-bp kit, the Hi-Q kit reduced indel rates by 28 to 59% and produced one to seven times more reads acceptable for downstream analyses. The PGM produced higher frameshift rates than pyrosequencing that were corrected by the RDP FrameBot tool. Significant differences among platforms were identified, although the diversity indices and overall site-based conclusions remained similar. For the cropping system analyses, a total of 6,182 unique NifH operational taxonomic units at 5% amino acid dissimilarity were obtained. The current crop type, as well as the crop rotation history, significantly influenced the composition of the soil diazotrophic community detected.


The experience of children and families with lymphoedema--a journey within a journey.
This paper reports on a study in the UK that explored the experience of children suffering with Lymphoedema and that of their families. Qualitative data was collected from 20 children between the ages of 6 and 18 and their respective parents. Single, semi-structured interviews were used in which children and their parents were asked to share how lymphoedema impacted on their family life. Children were asked about their school experience, their dreams and their aspirations. Three categories emerged. Firstly, the negotiation of the health care system. Themes included correct diagnosis, finding robust information and reaching a knowledgeable expert. The second category explored the complex role of the parents as advocates. Themes within this category included the dilemmas of parenting and the increasing challenges as children reached adolescence. The final category involved the impact on the family unit. The first theme concerned the integration of lymphoedema into daily activities and the intrusion on family time. The second explored the impact on siblings and the final theme the changing dilemmas as children moved through the stages of childhood and faced adulthood.


Factors affecting wear of composite resin denture teeth--24-month results from a clinical study.
The objectives of this study were to measure the occlusal wear of composite resin denture teeth in patients wearing a complete denture and to evaluate factors affecting wear. Fifty participants provided with complete dentures in at least one jaw were included. Gypsum casts were made from preliminary vinyl polysiloxane impressions 4 weeks after insertion, then after 6 (t(1)), 12 (t(2)), and 24 months (t(3)). Three-hundred and three posterior denture teeth were evaluated after 24 months. Wear was measured indirectly, from the casts, by means of a three-dimensional laser scanner device. Sequential images of the occlusal surfaces were digitized and superimposed (occlusal matching). Statistical analysis was performed by the use of mixed regression models, with the patient being a random effect. Mean wear (median, interquartile range; micrometer) of the entire occlusal surface was 8 (19) at t(1), 18 (34) at t(2), and 40 (61) at t(3). Maximum vertical loss (median, interquartile range; micrometer) was 92 (112) at t(1), 146 (148) at t(2), and 226 (184) at t(3). The dental status of the opposing jaw and the nature of the opposing material significantly affected the wear of denture teeth at t (3). Gender, daily wearing time, jaw, and type of tooth had no significant effects on the extent of wear. Clinically relevant vertical loss of composite resin denture teeth occurs after 24 months. Considering the limitations of this study, wear of denture teeth was affected by dental status and opposing material. The results suggest that wear of composite resin denture teeth exceeds that of enamel.


The names Hungateiclostridium Zhang et al. 2018, Hungateiclostridium thermocellum (Viljoen et al. 1926) Zhang et al. 2018, Hungateiclostridium cellulolyticum (Patel et al. 1980) Zhang et al. 2018, Hungateiclostridium aldrichii (Yang et al. 1990) Zhang et al. 2018, Hungateiclostridium alkalicellulosi (Zhilina et al. 2006) Zhang et al. 2018, Hungateiclostridium clariflavum (Shiratori et al. 2009) Zhang et al. 2018, Hungateiclostridium straminisolvens (Kato et al. 2004) Zhang et al. 2018 and Hungateiclostridium saccincola (Koeck et al. 2016) Zhang et al. 2018 contravene Rule 51b of the International Code of Nomenclature of Prokaryotes and require replacement names in the genus Acetivibrio Patel et al. 1980.
A recent publication has created the genus name HungateiclostridiumZhang et al. 2018 and the new combinations Hungateiclostridium cellulolyticum (Patel et al. 1980) Zhang et al. 2018, Hungateiclostridium aldrichii (Yang et al. 1990.) Zhang et al. 2018, Hungateiclostridium alkalicellulosi (Zhilina et al. 2006) Zhang et al. 2018, Hungateiclostridium clariflavum (Shiratori et al. 2009) Zhang et al. 2018, Hungateiclostridium straminisolvens (Kato et al. 2004) Zhang et al. 2018 and Hungateiclostridium saccincola (Koeck et al. 2016) Zhang et al. 2018 for names at the rank of species that were previously either included in the genus ClostridiumPrazmowski 1880, Acetivibrio Patel et al. 1980 or HerbivoraxKoeck et al. 2016. Rules 23a, 38, 39b, 41a, 42 and 44 have not been followed and an illegitimate name at the rank of genus or illegitimate combinations at the rank of species as defined in Rule 51b(1) and (2) have been created. Another aspect is recognising the fact that an instance of heterotypic synonym has been created between Acetivibrio Patel et al. 1980, HerbivoraxKoeck et al. 2016 and HungateiclostridiumZhang et al. 2018, where the earliest validly published genus name is Acetivibrio Patel et al. 1980, of which the nomenclatural type is Acetivibrio cellulolyticus Patel et al. 1980. It follows from Rules 23a, 38, 39a, 39b, 41a, 42 and 44 that the genus name to be used is Acetivibrio Patel et al. 1980, with new combinations in that genus replacing those published in the genus HungateiclostridiumZhang et al. 2018, which together with the genus name are illegitimate according to Rule 51b of the International Code of Nomenclature of Prokaryotes. Additional issues are also addressed with regards to the names Pseudoclostridium thermosuccinogenes (Drent et al. 1995) Zhang et al. 2018, PseudoclostridiumZhang et al. 2018 OscillospiraceaePeshkoff 1940 (Approved Lists 1980), RuminococcaceaeRainey 2010, Eubacteriales Buchanan 1917 (Approved Lists 1980) and ClostridialesPrévot 1953 (Approved Lists 1980).


Cholesterogenesis: derepression in extrahepatic tissues with 4-aminopyrazolo (3,4-d) pyrimidine.
Administration of 4-aminopyrazolo [3,4-d] pyrimidine decreased serum cholesterol levels in the rat to less than 5 milligrams per deciliter. Coincident with this change, there was a 2.1 - to 16.0-fold increase in the rate of sterol synthesis in seven extrahepatic tissues. This suggests that cholesterol carried in serum lipoproteins plays a major role in regulating sterol synthesis in many extrahepatic tissues.


Normative PROMIS Scores in Healthy Collegiate Athletes: Establishing a Target for Return to Function in the Young Adult Athlete.
The Patient-Reported Outcomes Measurement Information System (PROMIS) computer-adaptive testing (CAT) has been shown to be a valid and reliable means of assessing patient-reported outcomes. However, normal scores and distributions for a subset of a healthy young athletic population have not been established. To establish normative PROMIS scores for the domains of Physical Function (PF-CAT), Mobility (M-CAT), Upper Extremity Function (UE-CAT), and Pain Interference (PI-CAT) and determine the frequency of floor and ceiling effects in a population of healthy collegiate athletes. Cross-sectional study; Level of evidence, 3. Healthy collegiate athletes (18-23 years of age) were prospectively enrolled to complete the 4 PROMIS CAT domains. Additionally, the athletes provided information regarding their age, sex, and sport(s). Mean scores (±SD) and identification of ceiling or floor effects were calculated. Ceiling and floor effects were considered significant if >15% of the participants obtained the highest or lowest possible score on a domain. A total of 194 healthy athletes (mean age, 19.1 years) were included in the study: 118 (60.8%) men and 76 (39.2%) women. Mean scores were 62.9 ± 6.7 for PF-CAT, 58.2 ± 4.1 for M-CAT, 57.4 ± 5.8 for UE-CAT, and 43.2 ± 6.2 for PI-CAT. Distributions of scores for M-CAT and UE-CAT indicated strong ceiling effects by 77.3% and 66.0% of the participants, respectively. In healthy athletes, the PF-CAT differed most from the expected population-based mean score (50), with the mean being >1 SD above (62.9), without a ceiling effect observed. There were no significant sex- or age-based differences on any of the PROMIS domain scores. Healthy collegiate athletes scored nearly 1 SD from population-based means for all of the domains tested. M-CAT and UE-CAT demonstrated ceiling effects in more than two-thirds of healthy athletes, which may limit their utility in this population. The PF-CAT did not demonstrate floor or ceiling effects and demonstrated differences in a young adult athletic population from the population mean. The mean PF-CAT score of 62.9 can represent a target for return of function in injured athletes.