How Well Are Hand Hygiene Practices and Promotion Implemented in Sierra Leone? A Cross-Sectional Study in 13 Public Hospitals.
Healthcare-associated infections (HAIs) result in millions of avoidable deaths or prolonged lengths of stay in hospitals and cause huge economic loss to health systems and communities. Primarily, HAIs spread through the hands of healthcare workers, so improving hand hygiene can reduce their spread. We evaluated hand hygiene practices and promotion across 13 public health hospitals (six secondary and seven tertiary hospitals) in the Western Area of Sierra Leone in a cross-sectional study using the WHO hand hygiene self-Assessment framework in May 2021. The mean score for all hospitals was 273 ± 46, indicating an intermediate level of hand hygiene. Nine hospitals achieved an intermediate level and four a basic level. More secondary hospitals 5 (83%) were at the intermediate level, compared to tertiary hospitals 4 (57%). Tertiary hospitals were poorly rated in the reminders in workplace and institutional safety climate domains but excelled in training and education. Lack of budgets to support hand hygiene implementation is a priority gap underlying this poor performance. These gaps hinder hand hygiene practice and promotion, contributing to the continued spread of HAIs. Enhancing the distribution of hand hygiene resources and encouraging an embedded culture of hand hygiene practice in hospitals will reduce HAIs.


Implementing routine communication about costs of cancer treatment: perspectives of providers, patients, and caregivers.
Rising costs in oncology care often impact patients and families directly, making communication about costs and financial impacts of treatment crucial. Cost expenditures could offer opportunities for estimation and prediction, affording personalized conversations about financial impact. We sought to explore providers', patients', and caregivers' preferences towards implementing communication about cost, including when, how, and by whom such information might be provided. We conducted semi-structured phone interviews with a diverse population including 12 oncology providers, 12 patients, and 8 patient caregivers (N = 32). The constant comparative method was used to identify mutually agreed upon themes. Participant groups differed in their concerns surrounding cost communication, namely whether they want to receive this information and how such information might impact provider and patient treatment decisions. All participants agreed that oncology providers should not be leading cost conversations. Patients and caregivers identified social workers or financial advisors as most equipped to communicate about cost. Participants emphasized timely cost conversations, ideally around the time of diagnosis. Participants favored various metrics of financial impact beyond overall costs of care including disability, days lost from work, and out-of-pocket expenses. Cost transparency should be incorporated into usual care; however, there are several challenges to making cost conversations a part of everyday practice. Patients and family members need resources related to cost to aid in decision-making and those delivering cost information should have competency in oncology, financial advisement, and patient-centered care.


Lung cancer signatures in plasma based on proteome profiling of mouse tumor models.
We investigated the potential of in-depth quantitative proteomics to reveal plasma protein signatures that reflect lung tumor biology. We compared plasma protein profiles of four mouse models of lung cancer with profiles of models of pancreatic, ovarian, colon, prostate, and breast cancer and two models of inflammation. A protein signature for Titf1/Nkx2-1, a known lineage-survival oncogene in lung cancer, was found in plasmas of mouse models of lung adenocarcinoma. An EGFR signature was found in plasma of an EGFR mutant model, and a distinct plasma signature related to neuroendocrine development was uncovered in the small-cell lung cancer model. We demonstrate relevance to human lung cancer of the protein signatures identified on the basis of mouse models.


Predictors of poor CD4 and weight recovery in HIV-infected children initiating ART in South Africa.
To identify baseline demographic and clinical risk factors associated with poor CD4 and weight response after initiation of antiretroviral therapy (ART) in a cohort of human immunodeficiency virus (HIV)-infected children in KwaZulu-Natal, South Africa. We performed a retrospective cohort study of 674 children initiating antiretroviral therapy at McCord and St. Mary's hospitals in KwaZulu-Natal, South Africa, from August 2003 to December 2008. We extracted data from paper charts and electronic medical records to assess risk factors associated with CD4 and weight response using logistic regression. From the initial cohort of 901 children <10 years old initiating ART between August 2003 and December 2008, we analyzed 674 children with complete baseline data. Viral suppression rates (<400 copies/ml) were 84% after six months of therapy and 88% after 12 months of therapy. Seventy-three percent of children achieved CD4 recovery after six months and 89% after 12 months. Weight-for-age Z-score (WAZ) improvements were seen in 58% of children after six months of ART and 64% after 12 months. After six months of ART, lower baseline hemoglobin (p = 0.037), presence of chronic diarrhea (p = 0.007), and virologic failure (p = 0.046) were all associated with poor CD4 recovery by multivariate logistic regression. After 12 months of ART, poor CD4 recovery was associated with higher baseline CD4% (p = 0.005), chronic diarrhea (p = 0.02), and virologic failure (p<0.001). Age less than 3 years at ART initiation (p = 0.0003), higher baseline CD4% (p<0.001), and higher baseline WAZ (p<0.001) were all associated with poor WAZ improvements after 6 months by multivariate logistic regression. The presence of chronic diarrhea at baseline, independent of nutritional status and viral response, predicts poor CD4 recovery. Age at initiation of ART is an important factor in early WAZ response to ART, while viral suppression strongly predicts CD4 recovery but not WAZ improvement.


A robust targeted sequencing approach for low input and variable quality DNA from clinical samples.
Next-generation deep sequencing of gene panels is being adopted as a diagnostic test to identify actionable mutations in cancer patient samples. However, clinical samples, such as formalin-fixed, paraffin-embedded specimens, frequently provide low quantities of degraded, poor quality DNA. To overcome these issues, many sequencing assays rely on extensive PCR amplification leading to an accumulation of bias and artifacts. Thus, there is a need for a targeted sequencing assay that performs well with DNA of low quality and quantity without relying on extensive PCR amplification. We evaluate the performance of a targeted sequencing assay based on Oligonucleotide Selective Sequencing, which permits the enrichment of genes and regions of interest and the identification of sequence variants from low amounts of damaged DNA. This assay utilizes a repair process adapted to clinical FFPE samples, followed by adaptor ligation to single stranded DNA and a primer-based capture technique. Our approach generates sequence libraries of high fidelity with reduced reliance on extensive PCR amplification-this facilitates the accurate assessment of copy number alterations in addition to delivering accurate single nucleotide variant and insertion/deletion detection. We apply this method to capture and sequence the exons of a panel of 130 cancer-related genes, from which we obtain high read coverage uniformity across the targeted regions at starting input DNA amounts as low as 10 ng per sample. We demonstrate the performance using a series of reference DNA samples, and by identifying sequence variants in DNA from matched clinical samples originating from different tissue types.


Removal of truncation artefacts in magnetic resonance images by recovering missing spectral data.
Truncation artefacts are often present in many archived clinical magnetic resonance (MR) images due to the need of shortening the acquisition time by sampling a part of their k-space. This artificial information degrades the quality of the image and may hamper clinical diagnosis. In this paper, we propose a novel method to remove the artefacts by recovering the missing k-space or spectral data. The method consists of four steps: (a) estimating the truncated k-space from the images containing truncations artefacts, (b) computing the parameters of the sparse representation of the difference image of an image from the estimated truncated k-space, (c) recovering the missing spectral data using the parameters computed in (b), and (d) obtaining the artefact-removed image through inverse Fourier transform of the estimated and the recovered spectral data. Experiments on both simulated and real MR images have shown that the proposed method effectively removes truncation artefacts while preserving image quality and outperforms both the conventional Hamming window method and the popular TV method.


Macronutrients, diet quality, and frailty in older men.
Frailty, a phenotype of multisystem impairment and expanding vulnerability, is associated with higher risk of adverse health outcomes not entirely explained by advancing age. We investigated associations of macronutrients, dietary fiber, and overall diet quality with frailty status in older community-dwelling men. Participants were 5,925 men aged ≥ 65 years enrolled in the Osteoporotic Fractures in Men (MrOS) study at six U.S. centers. Diet was assessed at baseline with a food frequency questionnaire. We assessed frailty status (robust, intermediate, or frail) at baseline and at a second clinic visit (a mean of 4.6 years later) using a slightly modified Cardiovascular Health Study frailty index. We used multinomial logistic regression to assess associations between macronutrient intake, dietary fiber, and the Diet Quality Index Revised with frailty status at baseline and at the second clinic visit. At baseline, 2,748 (46.4%) participants were robust, 2,681 (45.2%) were intermediate, and 496 (8.4%) were frail. Carbohydrate, fat, protein, and dietary fiber showed no consistent associations with frailty status. Overall diet quality exhibited fairly consistent associations with frailty status. The Diet Quality Index Revised was inversely associated with frail status relative to robust status at the baseline visit (odds ratio for Q5 vs Q1 = 0.44, 95% confidence interval: 0.30, 0.63; p for trend < .0001) and at the second clinic visit (odds ratio for Q5 vs Q1 = 0.18, 95% confidence interval: 0.03, 0.97; p for trend = .0180). Overall diet quality was inversely associated with prevalent and future frailty status in this cohort of older men.


Investigations on the Tobacco Necrosis Virus D p60 replicase protein.
Tobacco Necrosis Virus D (TNV-D), in the genus Betanecrovirus (family Tombusviridae), possesses a single-stranded, positive-sense RNA genome containing six open reading frames (ORFs). Two 5'-proximal ORFs (1 and 2) encode overlapping polypeptides of 22 and 82 kDa (p22 and p82, respectively) which are both required for replication. The p22 auxiliary protein contains no replication motifs but the C-terminal region, downstream of a leaky stop codon, encodes a 60 kDa polypeptide (p60) which contains conserved RNA-dependent RNA polymerase (RdRP) motifs. Here we have expressed and purified recombinant p60 and show that in vitro it binds and efficiently synthesises both TNV-D RNA and Satellite tobacco necrosis virus C RNA. Alanine scanning mutagenesis of conserved amino acids in characteristic motifs in p60 revealed that some mutations significantly reduced RNA synthesis but mutating the second asparagine residue in the conserved GDD box was lethal. The effects of mutating identical amino acids in p60 on virus replication in vivo were examined in Nicotiana benthamiana plants following infection with RNA transcribed from wild type (wt) and mutant constructs. In inoculated leaves the behaviour of the mutants paralleled the in vitro data but systemic infection was precluded in all but one mutant which had reverted to wt. This study is the first to demonstrate the nucleic acid-binding and synthetic capabilities of a betanecrovirus polymerase.


Perfusion of ischemic ulcers of the extremity: a prognostic indicator of healing.
Forty patients with an ischemic ulcer of the lower extremity had peripheral vascular perfusion studies, performed with intra-arterial injections of aggregated technetium Tc 99m serum albumin microspheres (15mu to 30mu in diameter), in an attempt to develop an objective prognostic criterion for healing. The association between ulcer healing and the presence or absence of diabetes mellitus, palpable peripheral pulses, and patent trifurcation vessels on the arteriogram was reviewed and no association was noted. When, however, there was a relative hyperemia of the ulcer bed in comparison to the adjacent tissue of at least 3.5:1, as determined by counting the amount of radioactivity per unit area, 86% of patients went on to heal their ulcers. In those without this degree of hyperemia, only 11% were healed with conservative nonsurgical management. The results have shown that relative hyperemia of the ulcer bed is a clinically useful prognostic indicator in the patient with ischemic ulcer disease.


Growth rate regulation is associated with developmental modification of source efficiency.
Plants modulate their growth rate according to seasonal and environmental cues using a suite of growth repressors known to interact directly with cellular machinery controlling cell division and growth. Mutants lacking growth repressors show increased growth rates1,2, but the mechanism by which these plants ensure source availability for faster growth is unclear. Here, we undertake a comprehensive analysis of the fast-growth phenotype of a quintuple growth-repressor mutant, using a combination of theoretical and experimental approaches to understand the physiological basis of source-sink coordination. Our results show that, in addition to the control of tissue growth rates, growth repressors also affect tissue composition and leaf thickness, modulating the efficiency of production of new photosynthetic capacity. Modelling suggests that increases in growth efficiency underlie growth-rate differences between the wild type and spatula della growth-repressor mutant, with spatula della requiring less carbon to synthesize a comparable photosynthetic capability to the wild type, and fixing more carbon per unit mass. We conclude that through control of leaf development, growth repressors regulate both source availability and sink strength to achieve growth-rate variation without risking a carbon deficit.


Evidence of incomplete neural control of motor unit properties in cat tibialis anterior after self-reinnervation.
1. The mechanical, morphological and biochemical properties of single motor units from the anterior compartment of the tibialis anterior muscle in adult cats were studied six months after the nerve branches to that compartment were cut and resutured in close proximity to the muscle. 2. In these self-reinnervated muscles, the maximum tetanic tensions were lower in slow than fast units, a relationship similar to that observed among motor units from control adult muscles. The maximum tetanic tensions produced by the fast units were larger than those produced by the same motor unit types in control muscles. Direct counts of muscle fibres belonging to a motor unit showed that factors controlling the number of muscle fibres innervated by a motoneurone type persist during the reinnervation process in that fast motoneurones reinnervated more muscle fibres than slow motoneurones. Thus, the number of muscle fibres reinnervated by a motoneurone principally accounted for the difference in the maximum tension outputs among motor unit types, a relationship similar to that observed in control tibialis anterior muscles. 3. Monoclonal antibodies for specific myosin heavy chains were used to differentiate fibre types. By this criterion, motor units from control muscles were found to contain a homogeneous fibre type composition. In contrast, a heterogeneous, yet markedly biased, fibre type composition was observed in each unit analysed from self-reinnervated muscles. 4. Although not all of the muscle fibres of a motor unit developed the same type-associated parameters after reinnervation, the relationships among myosin heavy chain profile, succinate dehydrogenase activity and the fibre size were similar in fibres of control and self-reinnervated muscles. 5. The processes which dictate both motor unit size and the matching between motoneurone and muscle fibre type during the reinnervation process must be interdependent and result from a hierarchy of decisions which reflects their relative importance. The mechanisms responsible for these two processes may be a combination of: (1) selective innervation which may or may not incorporate a pruning process if multiple synaptic connections are initially formed and/or (2) conversion of enough fibres of a motor unit to form a predominant type.


Simulation applied to health services: opportunities for applying the system dynamics approach.
The aim of this essay is to raise awareness and broaden understanding within the health services community of the system dynamics (SD) simulation approach to policy analysis. The application of simulation in health services is reviewed. A comparison is made between the SD and traditional simulation approaches and is illustrated by considering reductions in waiting times for coronary heart disease treatment. Traditionally, simulation studies have tended to focus on the analysis of localized decisions and therefore on problems orientated towards individual patients. Although these methods are extremely powerful and effective, there is scope for an alternative modelling approach which is based on a more holistic perspective; SD is one such approach. It can assist in the design of robust policies by supporting debate on how the underlying structure might influence the evolutionary behaviour of a system. Using this method we can consider the time variation both of tangibles, such as waiting times and health care costs, and intangibles, such as patient anxiety and the effects of various pressures on purchasing decisions. We propose that SD holds great potential in assisting policy formation in health care.


The Influence of Fatigued Core Muscles on Head Acceleration during Headers in Soccer.
The core muscles play a central role in stabilizing the head during headers in soccer. The objective of this study was to examine the influence of a fatigued core musculature on the acceleration of the head during jump headers and run headers. Acceleration of the head was measured in a pre-post-design in 68 soccer players (age: 21.5 ± 3.8 years, height: 180.0 ± 13.9 cm, weight: 76.9 ± 8.1 kg). Data were recorded by means of a telemetric 3D acceleration sensor and with a pendulum header. The treatment encompassed two exercises each for the ventral, lateral, and dorsal muscle chains. The acceleration of the head between pre- and post-test was reduced by 0.3 G (p = 0.011) in jump headers and by 0.2 G (p = 0.067) in run headers. An additional analysis of all pretests showed an increased acceleration in run headers when compared to stand headers (p < 0.001) and jump headers (p < 0.001). No differences were found in the sub-group comparisons: semi-professional vs. recreational players, offensive vs. defensive players. Based on the results, we conclude that the acceleration of the head after fatiguing the core muscles does not increase, which stands in contrast to postulated expectations. More tests with accelerated soccer balls are required for a conclusive statement.


Ectopic expression of epidermal growth factor receptors induces hormone independence in ZR-75-1 human breast cancer cells.
Epidermal growth factor (EGF) receptor is inversely related to expression of estrogen receptor (ER) and progesterone receptor in primary breast tumors and is a negative predictor for response to endocrine therapy. To investigate a possible causal role of EGF receptor expression in breast cancer progression to hormone independence, we have created an experimental cell system. Epidermal growth factor receptor complementary DNA was introduced in estrogen-dependent ZR-75-1 breast cancer cells, and the resulting ZR/HERc cells exhibited a mitogenic response to epidermal growth factor, thus bypassing estrogen dependence. This EGF-induced proliferation could not be inhibited by antiestrogens. In addition, we noted changes in cell morphology and keratin expression of EGF-stimulated ZR/HERc cells, suggestive of an altered differentiation state. Furthermore, intolerance of functional ER and EGF receptor signal transduction pathways in ZR/HERc cells was observed during simultaneous activation, which possibly explains the inverse relationship of ER and EGF receptor expression in primary tumors. In contrast to the parental cells, ZR/HERc cells rapidly progressed to a stable ER-negative phenotype when cultured in the presence of the antiestrogen hydroxy-tamoxifen. These results suggest a possible role for EGF receptor in progression of breast cancer to hormone independence.


Effect of physician specialty on outcomes in diabetic ketoacidosis.
More than 100,000 people are hospitalized annually in the U.S. with diabetic ketoacidosis (DKA). Outcome differences have not been examined for these patients based on whether their primary care provider is a generalist or a diabetes specialist. The objective of this study was to investigate hospital charges and hospital length of stay (LOS) for patients with DKA according to the specialty of their primary care provider. We investigated all patients with a primary diagnosis of DKA during a 3.5-year period (n = 260) in a large urban teaching hospital. Hospital charges and LOS were studied regarding the specialty of the primary care provider. Demographic factors, severity of illness, laboratory data, and readmission rates were compared. Patients cared for by generalists and endocrinologists had a similar case mix and severity of DKA. The age-adjusted mean LOS for patients of generalists was 4.9 days (95% CI 4.5-5.4), and the mean LOS for patients of endocrinologists was 3.3 days (2.6-4.2) (P < 0.0043). Mean hospital charges differed (P < 0.0001) with an age- and sex-adjusted mean for patients of endocrinologists of $5,463 ($4,179-7,141) and a mean for patients of generalists of $10,109 ($9,151-11,166). The additional charges incurred by generalists were due in part to patients undergoing more procedures. No differences in diabetes-related complications occurred during admission, but the endocrinologist-treated group had a lower readmission rate for DKA during the study period than the generalist-treated group (2 vs. 6%, respectively) (P = 0.03). Endocrinologists provide more cost-effective care than generalists do when serving as primary care providers for patients hospitalized with DKA.


Protection against influenza after annually repeated vaccination: a meta-analysis of serologic and field studies.
According to common recommendations, influenza vaccination should be performed annually. It has been suggested that vaccination in previous years reduces vaccine efficacy in the long term. To determine whether the protection of influenza vaccine decreases when vaccination is repeated annually. Articles published between 1966 and 1997 were selected from MEDLINE. The end point for field studies was the influenza-related morbidity or mortality during influenza outbreaks (resulting in field protection rates). The end point for serologic studies was exceeding a protective postvaccination hemagglutination-inhibition titer (serologic protection rates). Protection rate differences between groups with single and multiple vaccinations were subjected to meta-analysis. Seven field studies (including 13 trials) supported the hypothesis that protection in multiple-vaccination groups is at least as good as that in single-vaccination groups. Ten trials with 5117 observations could be subjected to meta-analysis. The pooled protection-rate difference was close to 0 (1.1%; 95% confidence interval, -0.2% to 2.4%), thus detecting no difference between single or multiple vaccination. Twelve serologic studies (including 53 trials) showed heterogeneous results: 9 trials were significantly in favor of single vaccination, and 7 were in favor of multiple vaccination, but in most cases, there was no significant difference between the 2 vaccination groups. The pooled serologic protection-rate difference from 52 trials (12341 observations) was again close to 0 (1.7%; 95% confidence interval, -1.3% to 4.8%). We did not detect any evidence for a decreasing protection with annually repeated influenza vaccination. Annual vaccination should not be discouraged in populations at risk.


Direct observation of closed magnetic flux trapped in the high-latitude magnetosphere.
The structure of Earth's magnetosphere is poorly understood when the interplanetary magnetic field is northward. Under this condition, uncharacteristically energetic plasma is observed in the magnetotail lobes, which is not expected in the textbook model of the magnetosphere. Using satellite observations, we show that these lobe plasma signatures occur on high-latitude magnetic field lines that have been closed by the fundamental plasma process of magnetic reconnection. Previously, it has been suggested that closed flux can become trapped in the lobe and that this plasma-trapping process could explain another poorly understood phenomenon: the presence of auroras at extremely high latitudes, called transpolar arcs. Observations of the aurora at the same time as the lobe plasma signatures reveal the presence of a transpolar arc. The excellent correspondence between the transpolar arc and the trapped closed flux at high altitudes provides very strong evidence of the trapping mechanism as the cause of transpolar arcs.


Perineural Dexmedetomidine Is More Effective Than Clonidine When Added to Local Anesthetic for Supraclavicular Brachial Plexus Block: A Systematic Review and Meta-analysis.
Clonidine, an α-2 agonist, has long been used as a local anesthetic adjunct with proven efficacy to prolong peripheral nerve block duration. Dexmedetomidine, a newer α-2 agonist, has a more favorable pharmacodynamic and safety profile; however, data comparing its efficacy as an adjunct to that of clonidine are inconsistent. We sought to compare the clinical efficacy of these 2 α-2 agonists by examining their effects on peripheral nerve block characteristics for upper extremity surgery. A preliminary search found that the overwhelming majority of randomized controlled trials comparing perineural dexmedetomidine to clonidine for upper extremity surgery were in the setting of supraclavicular brachial plexus block (SCB). Therefore, we performed a systematic review and meta-analysis of randomized controlled trials comparing dexmedetomidine with clonidine as perineural adjuncts to single-injection SCB. Sensory and motor block duration and onset, analgesic duration, α-2 agonist side effects, and block complications were analyzed. Sensory block duration was designated as a primary outcome. Data were combined using random-effects modeling, and ratio-of-means was used to analyze the results. A total of 868 patients from 14 clinical studies were included in the analysis. Compared with clonidine, dexmedetomidine prolonged the duration (ratio of means [95% confidence interval {CI}]) of sensory block by an estimate of 1.2 (1.2-1.3; P< .00001). It also prolonged the duration (ratio of means [99% CI]) of motor block by an estimate of 1.2 (1.1-1.3; P < .00001), and analgesia by an estimate of 1.2 (1.1-1.3; P < .00001). It also hastened the onset of sensory block by an estimate of 0.9 (0.8-1.0; P < .00001) and motor block by an estimate of 0.9 (0.9-1.0; P = .002). Dexmedetomidine was associated with an increased odds ratio (99% CI) of transient bradycardia by an estimate of 7.4 (1.3-40.8; P = .003) and postoperative sedation by an estimate of 11.8 (1.9-73.6; P = .0005). There were no differences in other α-2 agonist-related side effects or block-related complications. Compared with clonidine as a local anesthetic adjunct for single-injection SCB, perineural dexmedetomidine enhances sensory, motor, and analgesic block characteristics. These benefits should be weighed against the increased risk of transient bradycardia.


Jugular-infused methionine, lysine and branched-chain amino acids does not improve milk production in Holstein cows experiencing heat stress.
Poor utilization of amino acids contributes to losses of milk protein yield in dairy cows exposed to heat stress (HS). Our objective was to test the effect of essential amino acids on milk production in lactating dairy cows exposed to short-term HS conditions. To achieve this objective, 12 multiparous, lactating Holstein cows were assigned to two environments (thermoneutral (THN) or HS) from days 1 to 14 in a split-plot type cross-over design. All cows received 0 g/day of essential amino acids from days 1 to 7 (negative control (NC)) followed by an intravenous infusion of l-methionine (12 g/day), l-lysine (21 g/day), l-leucine (35 g/day), l-isoleucine (15 g/day) and l-valine (15 g/day, methionine, lysine and branched-chain amino acids (ML+BCAA)) from days 8 to 14. The basal diet was composed of ryegrass silage and hay, and a concentrate mix. This diet supplied 44 g of methionine, 125 g of lysine, 167 g of leucine, 98 g of isoleucine and 109 g of valine per day to the small intestine of THN cows. Temperature-humidity index was maintained below 66 for the THN environment, whereas the index was maintained above 68, peaking at 76, for 14 continuous h/day for the HS environment. Heat stress conditioning increased the udder temperature from 37.0°C to 39.6°C. Cows that received the ML+BCAA treatment had greater p.m. rectal and vaginal temperatures (0.50°C and 0.40°C, respectively), and respiration rate (8 breaths/min) compared with those on the NC treatment and exposed to a HS environment. However, neither NC nor ML+BCAA affected rectal or vaginal temperatures and respiration rates in the THN environment. Compared with THN, the HS environment reduced dry matter intake (1.48 kg/day), milk yield (2.82 kg/day) and milk protein yield (0.11 kg/day). However, compared with NC, the ML+BCAA treatment increased milk protein percent by 0.07 points. For the THN environment, the ML+BCAA treatment increased concentrations of milk urea nitrogen. For the HS environment, the ML+BCAA treatment decreased plasma concentrations of arginine, ornithine and citrulline; however, differences were not observed for the THN environment. In summary, HS elicited expected changes in production; however, infusions of ML+BCAA failed to increase milk protein yield. Lower dry matter intake and greater heat load in response to ML+BCAA contributed to the lack of response in milk production in HS cows. The ML+BCAA treatment may have reduced the breakdown of muscle protein in heat-stressed cows.


Student nurses need more than maths to improve their drug calculating skills.
Nurses need to be able to calculate accurate drug calculations in order to safely administer drugs to their patients (NMC, 2002). Studies have shown however that nurses do not always have the necessary skills to calculate accurate drug dosages and are potentially administering incorrect dosages of drugs to their patients (Hutton, M. 1998. Nursing Mathematics: the importance of application. Nursing Standard 13(11), 35-38; Kapborg, I. 1994. Calculation and administration of drug dosage by Swedish nurses, Student Nurses and Physicians. International Journal for Quality in Health Care 6(4), 389-395; O'Shea, E. 1999. Factors contributing to medication errors: a literature review. Journal of Advanced Nursing 8, 496-504; Wilson, A. 2003. Nurses maths: researching a practical approach. Nursing Standard 17(47), 33-36). The literature indicates that in order to improve drug calculations strategies need to focus on both the mathematical skills and conceptual skills of student nurses so they can interpret clinical data into drug calculations to be solved. A study was undertaken to investigate the effectiveness of implementing several strategies which focussed on developing the mathematical and conceptual skills of student nurses to improve their drug calculation skills. The study found that implementing a range of strategies which addressed these two developmental areas significantly improved the drug calculation skills of nurses. The study also indicates that a range of strategies has the potential ensuring that the skills taught are retained by the student nurses. Although the strategies significantly improved the drug calculation skills of student nurses, the fact that only 2 students were able to achieve 100% in their drug calculation test indicates a need for further research into this area.


Copper-zinc superoxide dismutase of Caulobacter crescentus: cloning, sequencing, and mapping of the gene and periplasmic location of the enzyme.
Although widely found in the cytoplasm of eucaryotes, the copper-zinc form of superoxide dismutase (CuZnSOD) has been identified in only a small number of bacterial species. One species is the freshwater bacterium Caulobacter crescentus, which also contains an SOD with iron as the metal cofactor (FeSOD). To investigate the function of this CuZnSOD and its structural relationship to the eucaryotic CuZnSODs, the gene encoding CuZnSOD (sodC) of C. crescentus CB15 was cloned and sequenced. By hybridization to pulsed-field electrophoresis gels, sodC was mapped near cysE in the C. crescentus chromosome. Through analysis of spheroplasts, the two SODs of C. crescentus were shown to be differently localized, CuZnSOD in the periplasm and FeSOD in the cytoplasm. In its natural habitat, C. crescentus is frequently associated with blue-green algae (cyanobacteria). The oxygen evolved by these photosynthetic algae may create an extracellular oxidative stress against which the periplasmic CuZnSOD may defend more effectively than the cytoplasmic FeSOD. Amino acid sequence alignments of C. crescentus CuZnSOD with eucaryotic CuZnSODs and with CuZnSOD of Photobacterium leiognathi (the only other bacterium from which CuZnSOD has been isolated and sequenced) suggest similar supersecondary structures for bacterial and eucaryotic CuZnSODs but reveal four novel substitutions in C. crescentus CuZnSOD: a phenylalanine critical to intrasubunit hydrophobic bonding replaced by alanine, a histidine ligand of zinc replaced by aspartate, and substitutions of two other previously invariant residues that stabilize zinc or both copper and zinc. These amino acid substitutions in C. crescentus CuZnSOD may have implications for its catalysis and stability.


Nausea and vomiting after surgery under general anesthesia: an evidence-based review concerning risk assessment, prevention, and treatment.
The German-language recommendations for the management of postoperative nausea and vomiting (PONV) have been revised by an expert committee. Major aspects of this revision are presented here in the form of an evidence-based review article. The literature was systematically reviewed with the goal of revising the existing recommendations. New evidence-based recommendations for the management of PONV were developed, approved by consensus, and graded according to the scheme of the Scottish Intercollegiate Guidelines Network (SIGN). The relevant risk factors for PONV include female sex, nonsmoker status, prior history of PONV, motion sickness, use of opioids during and after surgery, use of inhalational anesthetics and nitrous oxide, and the duration of anesthesia. PONV scoring systems provide a rough assessment of risk that can serve as the basis for a risk-adapted approach. Risk-adapted prophylaxis, however, has not been shown to provide any greater benefit than fixed (combination) prophylaxis, and PONV risk scores have inherent limitations; thus, fixed prophylaxis may be advantageous. Whichever of these two approaches to manage PONV is chosen, high-risk patients must be given multimodal prophylaxis, involving both the avoidance of known risk factors and the application of multiple validated and effective antiemetic interventions. PONV should be treated as soon as it arises, to minimize patient discomfort, the risk of medical complications, and the costs involved. PONV lowers patient satisfaction but is treatable. The effective, evidence-based measures of preventing and treating it should be implemented in routine practice.


Miocene mammal reveals a Mesozoic ghost lineage on insular New Zealand, southwest Pacific.
New Zealand (NZ) has long been upheld as the archetypical example of a land where the biota evolved without nonvolant terrestrial mammals. Their absence before human arrival is mysterious, because NZ was still attached to East Antarctica in the Early Cretaceous when a variety of terrestrial mammals occupied the adjacent Australian portion of Gondwana. Here we report discovery of a nonvolant mammal from Miocene (19-16 Ma) sediments of the Manuherikia Group near St Bathans (SB) in Central Otago, South Island, NZ. A partial relatively plesiomorphic femur and two autapomorphically specialized partial mandibles represent at least one mouse-sized mammal of unknown relationships. The material implies the existence of one or more ghost lineages, at least one of which (based on the relatively plesiomorphic partial femur) spanned the Middle Miocene to at least the Early Cretaceous, probably before the time of divergence of marsupials and placentals > 125 Ma. Its presence in NZ in the Middle Miocene and apparent absence from Australia and other adjacent landmasses at this time appear to reflect a Gondwanan vicariant event and imply persistence of emergent land during the Oligocene marine transgression of NZ. Nonvolant terrestrial mammals disappeared from NZ some time since the Middle Miocene, possibly because of late Neogene climatic cooling.


Type 1 protein phosphatase is required for maintenance of cell wall integrity, morphogenesis and cell cycle progression in Saccharomyces cerevisiae.
GLC7 encodes the catalytic subunit of type 1 protein serine/threonine phosphatase (PP1) in the yeast Saccharomyces cerevisiae. Here we have characterized the temperature-sensitive glc7-10 allele, which displays aberrant bud morphology and an abnormal actin cytoskeleton at the restrictive temperature. At 37 degrees C glc7-10 strains accumulated a high proportion of budded cells with an unmigrated nucleus, duplicated spindle pole bodies, a short spindle, delocalized cortical actin and 2C DNA content, indicating a cell cycle block prior to the metaphase to anaphase transition. glc7-10 was suppressed by growth on high osmolarity medium and exhibited temperature-sensitive cell lysis upon hypo-osmotic stress. Pkc1p, the yeast protein kinase C homolog which is thought to regulate the Mpk1p MAP kinase pathway involved in cell wall remodelling and polarized cell growth, was found to act as a dosage suppressor of glc7-10. Although neither activation of BCK1 (MEKK) by the dominant BCK1-20 mutation nor increased dosage of MKK1 (MEK) or MPK1 (MAP kinase) mimicked PKC1 as a glc7-10 dosage suppressor, extra copies of genes encoding upstream components of the Pkc1p pathway such as ROM2, RHO2, HCS77/WSC1/SLG1 and MID2 also suppressed glc7-10 effectively. Conversely, mpk1delta glc7-10 and bck1delta glc7-10 double mutants displayed a synthetic cell lysis defect compared with each single mutant and glc7-10 was hypersensitive to reduced PKC1 function, displaying highly aberrant morphologies and inviability even at the normally permissive temperature of 26 degrees C. Dephosphorylation by PP1 therefore functions positively to promote cell integrity, bud morphology and polarization of the actin cytoskeleton and glc7-10 cells require higher levels of Pkc1p activity to sustain these functions.


Ubiquitin-positive intranuclear inclusions in neuronal and glial cells in a mouse model of the fragile X premutation.
Fragile X-associated tremor/ataxia syndrome (FXTAS) is an adult-onset neurodegenerative disorder caused by CGG trinucleotide repeat expansions in the fragile X mental retardation 1 (FMR1) gene. The neuropathological hallmark of the disease is the presence of ubiquitin-positive intranuclear inclusions in neurons and in astrocytes. Ubiquitin-positive intranuclear inclusions have also been found in the neurons of transgenic mice model carrying an expanded CGG((98)) trinucleotide repeat of human origin but have not previously been described in glial cells. Therefore, we used immunocytochemical methods to determine the pathological features of nuclear and/or cytoplasmic inclusions in astrocytes, Bergmann glia, and neurons, as well as relationships between inclusion patterns, age, and repeat length in CGG knock-in (KI) mice in comparison with wild-type mice. In CGG KI mice, ubiquitin-positive intranuclear inclusions were found in neurons (e.g., pyramidal cells, GABAergic neurons) throughout the brain in cortical and subcortical brain regions; these inclusions increased in number and size with advanced age. Ubiquitin-positive intranuclear inclusions were also present in protoplasmic astrocytes, including Bergmann glia in the cerebellum. The morphology of intranuclear inclusions in CGG KI mice was compared to that of typical inclusions in human neurons and astrocytes in postmortem FXTAS brain tissue. This new finding of previously unreported pathology in astrocytes of CGG KI mice now provides an important mouse model to study astrocyte pathology in human FXTAS.


Autosomal genes of autosomal/X-linked duplicated gene pairs and germ-line proliferation in Caenorhabditis elegans.
We report molecular genetic studies of three genes involved in early germ-line proliferation in Caenorhabditis elegans that lend unexpected insight into a germ-line/soma functional separation of autosomal/X-linked duplicated gene pairs. In a genetic screen for germ-line proliferation-defective mutants, we identified mutations in rpl-11.1 (L11 protein of the large ribosomal subunit), pab-1 [a poly(A)-binding protein], and glp-3/eft-3 (an elongation factor 1-alpha homolog). All three are members of autosome/X gene pairs. Consistent with a germ-line-restricted function of rpl-11.1 and pab-1, mutations in these genes extend life span and cause gigantism. We further examined the RNAi phenotypes of the three sets of rpl genes (rpl-11, rpl-24, and rpl-25) and found that for the two rpl genes with autosomal/X-linked pairs (rpl-11 and rpl-25), zygotic germ-line function is carried by the autosomal copy. Available RNAi results for highly conserved autosomal/X-linked gene pairs suggest that other duplicated genes may follow a similar trend. The three rpl and the pab-1/2 duplications predate the divergence between C. elegans and C. briggsae, while the eft-3/4 duplication appears to have occurred in the lineage to C. elegans after it diverged from C. briggsae. The duplicated C. briggsae orthologs of the three C. elegans autosomal/X-linked gene pairs also display functional differences between paralogs. We present hypotheses for evolutionary mechanisms that may underlie germ-line/soma subfunctionalization of duplicated genes, taking into account the role of X chromosome silencing in the germ line and analogous mammalian phenomena.


Sagittal alignment and complications following lumbar 3-column osteotomy: does the level of resection matter?
OBJECTIVE Three-column osteotomy (3CO) is a demanding technique that is performed to correct sagittal spinal malalignment. However, the impact of the 3CO level on pelvic or truncal sagittal correction remains unclear. In this study, the authors assessed the impact of 3CO level and postoperative apex of lumbar lordosis on sagittal alignment correction, complications, and revisions. METHODS In this retrospective study of a multicenter spinal deformity database, radiographic data were analyzed at baseline and at 1- and 2-year follow-up to quantify spinopelvic alignment, apex of lordosis, and resection angle. The impact of 3CO level and apex level of lumbar lordosis on the sagittal correction was assessed. Logistic regression analyses were performed, controlling for cofounders, to investigate the effects of 3CO level and apex level on intraoperative and postoperative complications as well as on the need for subsequent revision surgery. RESULTS A total of 468 patients were included (mean age 60.8 years, mean body mass index 28.1 kg/m2); 70% of patients were female. The average 3CO resection angle was 25.1° and did not significantly differ with regard to 3CO level. There were no significant correlations between the 3CO level and amount of sagittal vertical axis or pelvic tilt correction. The postoperative apex level significantly correlated with greater correction of pelvic tilt (2° per more caudal level, R = -0.2, p = 0.006). Lower-level 3CO significantly correlated with revisions for pseudarthrosis (OR = 3.88, p = 0.001) and postoperative motor deficits (OR = 2.02, p = 0.026). CONCLUSIONS In this study, a more caudal lumbar 3CO level did not lead to greater sagittal vertical axis correction. The postoperative apex of lumbar lordosis significantly impacted pelvic tilt. 3CO levels that were more caudal were associated with more postoperative motor deficits and revisions.


Encapsulation of active cytoskeletal protein networks in cell-sized liposomes.
We demonstrate that cytoskeletal actin-myosin networks can be encapsulated with high efficiency in giant liposomes by hydration of lipids in an agarose hydrogel. The liposomes have cell-sized diameters of 10-20 μm and a uniform actin content. We show by measurements of membrane fluorescence intensity and bending rigidity that the majority of liposomes are unilamellar. We further demonstrate that the actin network can be specifically anchored to the membrane by biotin-streptavidin linkages. These protein-filled liposomes are useful model systems for quantitative studies of the physical mechanisms by which the cytoskeleton actively controls cell shape and mechanics. In a broader context, this new preparation method should be widely applicable to encapsulation of proteins and polymers, for instance, to create polymer-reinforced liposomes for drug delivery.


MicroRNA 146 (Mir146) modulates spermatogonial differentiation by retinoic acid in mice.
Impaired biogenesis of microRNAs disrupts spermatogenesis and leads to infertility in male mice. Spermatogonial differentiation is a key step in spermatogenesis, yet the mechanisms that control this event remain poorly defined. In this study, we discovered microRNA 146 (Mir146) to be highly regulated during spermatogonial differentiation, a process dependent on retinoic acid (RA) signaling. Mir146 transcript levels were diminished nearly 180-fold in differentiating spermatogonia when compared with undifferentiated spermatogonia. Luciferase assays revealed the direct binding of Mir146 to the 3' untranslated region of the mediator complex subunit 1 (Med1), a coregulator of retinoid receptors (RARs and RXRs). Overexpression of Mir146 in cultured undifferentiated spermatogonia reduced Med1 transcript levels, as well as those of differentiation marker kit oncogene (Kit). MED1 protein was also diminished. Conversely, inhibition of Mir146 increased the levels of Kit. When undifferentiated spermatogonia were exposed to RA, Mir146 was downregulated along with a marker for undifferentiated germ cells, zinc finger and BTB domain containing 16 (Zbtb16; Plzf); Kit was upregulated. Overexpression of Mir146 in RA-treated spermatogonia inhibited the upregulation of Kit, stimulated by retinoic acid gene 8 (Stra8), and spermatogenesis- and oogenesis-specific basic helix-loop-helix 2 (Sohlh2). Inhibition of Mir146 in RA-treated spermatogonia greatly enhanced the upregulation of these genes. We conclude that Mir146 modulates the effects of RA on spermatogonial differentiation.


Further validation of the Satisfaction with Life Scale: evidence for the cross-method convergence of well-being measures.
The structure of subjective well-being has been conceptualized as consisting of two major components: the emotional or affective component and the judgmental or cognitive component (Diener, 1984; Veenhoven, 1984). The judgmental component has also been conceptualized as life satisfaction (Andrews & Withey, 1976). Although the affective component of subjective well-being has received considerable attention from researchers, the judgmental component has been relatively neglected. The Satisfaction With Life Scale (SWLS; Diener, Emmons, Larsen, & Griffin, 1985) was developed as a measure of the judgmental component of subjective well-being (SWB). Two studied designed to validate further the SWLS are reported. Peer reports, a memory measure, and clinical ratings are used as external criteria for validation. Evidence for the reliability and predictive validity of the SWLS is presented, and its performance is compared to other related scales. The SWLS is shown to be a valid and reliable measure of life satisfaction, suited for use with a wide range of age groups and applications, which makes possible the savings of interview time and resources compared to many measures of life satisfaction. In addition, the high convergence of self- and peer-reported measures of subjective well-being and life satisfaction provide strong evidence that subjective well-being is a relatively global and stable phenomenon, not simply a momentary judgment based on fleeting influences.


Measuring timbre discrimination with cross-faded synthetic tones.
The identification and discrimination of timbre are essential features of music perception. As timbre differences appear as multidimensional cues, the spectral shape, the spectral fluctuation, and the rise time are the most dominating parameters of timbre in normal-hearing listeners. We developed a psychoacoustical test to determine the timbre discrimination abilities using only the spectral difference as a cue. Therefore, a synthetically generated tone continuum was used in an adaptive alternative forced choice paradigm. The spectral difference was modified by cross-fading the tones adaptively, depending on the listeners' response which allows very precise determinations of the just noticeable difference (JND). We measured the JND for the spectral difference with 18 normal-hearing listeners. The results confirm the applicability of the test to measure timbre discrimination with the spectral difference as solely cue. Further, the portability of the test to further dimensions of timbre is discussed.


High-performance liquid chromatography of complex mixtures using monodisperse dual-chemistry polymer beads prepared by a pore-size-specific functionalization process. A single column combination of hydrophobic interaction and reversed-phase chromatography.
A novel separation medium for HPLC combining hydrophobic interaction and reversed-phase separation modes in a single column has been prepared from monodisperse 10-microns poly-(glycidyl methacrylate-co-ethylene dimethacrylate) beads using a pore-size-specific functionalization process. In this approach, the large pores of each bead were provided with phenyl groups interspersed among hydrophilic functionalities while a much higher surface concentration of hydrophobic phenyl groups was introduced into the small pores. Due to the size-specific character of the modification process, no protein interaction with any highly hydrophobic surface was observed during chromatography. The beads were used for the separation of samples containing both proteins and small hydrocarbon or drug molecules. A plot of log k' against salt concentration in the mobile phase clearly documents the clean hydrophobic interaction mechanism of protein separation and the absence of charged groups while the linear plot of log k' against acetonitrile concentration for numerous compounds demonstrates the reversed-phase separation ability. No decrease of the efficiency of the test column (23,000 plates/m) was observed in long-term experiments during which more than 1000 injections and many changes between the modes were performed.


Akt-activated endothelium constitutes the niche for residual disease and resistance to bevacizumab in ovarian cancer.
Ovarian cancer is the second leading cause of cancer-related death in women worldwide. Despite optimal cytoreduction and adequate adjuvant therapies, initial tumor response is often followed by relapse suggesting the existence of a tumor niche. Targeted therapies have been evaluated in ovarian cancer to overcome resistant disease. Among them, antiangiogenic therapies inhibit new blood vessel growth, induce endothelial cell apoptosis, and block the incorporation of hematopoietic and endothelial progenitor cells into new blood vessels. Despite in vitro and in vivo successes, antivascular therapy with bevacizumab targeting VEGF-A has limited efficacy in ovarian cancer. The precise molecular mechanisms underlying clinical resistance to anti-VEGF therapies are not yet well understood. Among them, tumor and stromal heterogeneity might determine the treatment outcomes. The present study investigates whether abnormalities in the tumor endothelium may contribute to treatment resistance to bevacizumab and promote a residual microscopic disease. Here, we showed that ovarian cancer cells activate Akt phosphorylation in endothelial cells inducing resistance to bevacizumab leading to an autocrine loop based on FGF2 secretion. Altogether, our results point out the role of an activated endothelium in the resistance to bevacizumab and in the constitution of a niche for a residual disease.


Rapid production of specialized animal handling devices using computer-aided design and solid freeform fabrication.
To develop a process for rapidly and inexpensively producing customized animal handling devices for small animal imaging. To meet the specific needs of a particular imaging experiment, measurements are taken from imaging data and the animal handling devices are designed using 3D computer-aided design (CAD) software. Parts are produced in a few days using solid freeform fabrication (SFF, a.k.a. rapid prototyping). This process is illustrated with the production of an animal handling system for stereotaxically prescribed therapeutic ultrasound and MRI of the mouse brain. The device provides integrated head-fixation, anesthesia delivery, and physiological monitoring in a modular system. Design and production took approximately 1 week and the cost was a small fraction of a traditional machine shop. Commercial animal handling products typically have limited functionality and are not integrated with other laboratory infrastructure. However, using CAD and SFF, sophisticated animal handling devices can be produced to meet the specific experimental needs. This process is typically faster and less expensive than using a traditional machine shop, and the products are more robust than typical homemade devices. Using high-quality purpose-built devices permits experiments to be executed with greater consistency and higher throughput.


Association of Acute Increases in Plasma Creatinine after Renin-Angiotensin Blockade with Subsequent Outcomes.
Data from observational and interventional studies provide discordant results regarding the relationship between creatinine increase after renin-angiotensin system inhibition (RASi) and adverse outcomes. We compared health outcomes among patients with different categories of increase in creatinine upon initiation of RASi in a large population-based cohort. We performed a retrospective analysis of the Stockholm CREAtinine Measurements database, which contains complete information on diagnoses, medication dispensation claims, and laboratory test results for all Stockholm citizens accessing health care. Included were 31,951 adults initiating RASi during 2007-2011 with available pre- and postinitiation creatinine monitoring. Multivariable Cox regression was used to compare mortality, cardiovascular and ESKD events among individuals with different ranges of creatinine increases within 2 months after starting treatment. In a median follow-up of 3.5 years, acute increases in creatinine were associated with mortality (3202 events) in a graded manner: compared with creatinine increases <10%, a 10%-19% increase showed an adjusted hazard ratio (HR) of 1.15 (95% confidence interval [95% CI], 1.05 to 1.27); HR 1.22 (95% CI, 1.07 to 1.40) for 20%-29%; HR 1.55 (95% CI, 1.36 to 1.77) for ≥30%. Similar graded associations were present for heart failure (2275 events, P<0.001) and ESKD (52 events; P<0.001), and, less consistently, myocardial infarction (842 events, P=0.25). Results were robust across subgroups, among continuing users, when patients with decreases in creatinine were excluded from the reference group, and after accounting for death as a competing risk. Among real-world monitored adults, increases in creatinine (>10%) after initiation of RASi are associated with worse health outcomes. These results do not address the issue of discontinuation of RASi when plasma creatinine increases but do suggest that patients with increases in creatinine have higher subsequent risk of cardiovascular and kidney outcomes.


Food Insecurity and Health Care Expenditures in the United States, 2011-2013.
To determine whether food insecurity, limited or uncertain food access owing to cost, is associated with greater health care expenditures. Nationally representative sample of the civilian noninstitutionalized population of the United States (2011 National Health Interview Survey [NHIS] linked to 2012-2013 Medication Expenditure Panel Survey [MEPS]). Longitudinal retrospective cohort. A total of 16,663 individuals underwent assessment of food insecurity, using the 10-item adult 30-day food security module, in the 2011 NHIS. Their total health care expenditures in 2012 and 2013 were recorded in MEPS. Expenditure data were analyzed using zero-inflated negative binomial regression and adjusted for age, gender, race/ethnicity, education, income, insurance, and residence area. Fourteen percent of individuals reported food insecurity, representing 41,616,255 Americans. Mean annualized total expenditures were $4,113 (standard error $115); 9.2 percent of all individuals had no health care expenditures. In multivariable analyses, those with food insecurity had significantly greater estimated mean annualized health care expenditures ($6,072 vs. $4,208, p < .0001), an extra $1,863 in health care expenditure per year, or $77.5 billion in additional health care expenditure annually. Food insecurity was associated with greater subsequent health care expenditures. Future studies should determine whether food insecurity interventions can improve health and reduce health care costs.


Intermolecular interactions and solvation effects of dimethylsulfoxide on type III deep eutectic solvents.
Reline, a typical type III Deep Eutectic Solvent (DES), composed of urea and choline chloride, is widely used and highly investigated for its thermo-physical properties and intermolecular interactions. The formation of hydrogen bonds between urea and chloride ions and a strong suppression in the hydrogen bonds between the urea molecules lead to a significantly low melting point of reline. Considering commercial applications, aqueous solutions of reline have been extensively analyzed; however, water, being a protic solvent, strongly affects the hydrogen bond network in reline and disrupts its structure. A mixture of reline with aprotic solvent has not been systematically investigated in the literature. To this end, we herein explore the intermolecular interactions between reline and dimethylsulfoxide (DMSO), a commonly used industrial solvent, which unlike water, does not have labile hydrogen. Our experiments revealed that a decrease in the density of the mixture with increasing DMSO amount is less significant as compared with water. Furthermore, molecular dynamics simulation results, as analyzed in terms of the hydrogen bonds between the components and interaction energies, suggest a very limited effect on urea-urea, urea-chloride, and urea-choline interactions in the presence of DMSO. Moreover, DMSO preferentially interacts with urea in the mixture; whereas in the case with water, the chloride ion was preferentially hydrated. Radial distribution analysis indicates that reline, while getting diluted by DMSO, maintains its structure up to 0.7-mole fraction of DMSO. Taken together the results reveal that DMSO reserves intermolecular interactions of reline better than water.


Motor and non-motor symptoms in Turkish patients with Parkinson's disease affecting family caregiver burden and quality of life.
Parkinson's disease (PD) decreases the quality of life (QoL) of both caregivers and patients by increasing stress and burden, and it has a negative impact on their psychological state. In the present study, the authors examined the impact of PD-patient motor and non-motor clinical symptoms on the psychological health, burden, and QoL of Turkish caregivers. The study included 50 patients with PD and their caregivers. Patients' disease severity and disability, motor and non-motor symptoms, disease complications, sleep disturbances, anxiety, and depressive symptoms were assessed with various scales. Data were analyzed by multiple linear-regression models to identify variables associated with caregiver burden, psychological status, and QoL. Anxiety and depressive symptoms exhibited by the patients significantly affected the psychological state of the caregivers. Caregiver burden was increased by disease severity; the patient's degree of disability, anxiety, and depressive symptoms; and excessive daytime sleepiness. Caregiver QoL was significantly affected by the presence of dyskinesia, sialorrhea, and anxiety symptoms in patients with PD. These findings indicate that caregiving for patients with PD, particularly those in later stages, with psychiatric symptoms, affects caregiver psychological status, QoL, and caregiver burden. These results can be used to develop treatment approaches to improve caregivers' psychological status and QoL and decrease caregiver burden.


Combination of letrozole, metronomic cyclophosphamide and sorafenib is well-tolerated and shows activity in patients with primary breast cancer.
To assess whether the combination of letrozole, metronomic cyclophosphamide and sorafenib (LCS) is well tolerated and shows activity in primary breast cancer (BC). Thirteen oestrogen receptor-positive, postmenopausal, T2-4, N0-1 BC patients received the LCS combination for 6 months. In these patients we examined the pharmacokinetics of sorafenib and cyclophosphamide, toxicity of the regimen, the clinical response to therapy and changes in the levels of biologically relevant biomarkers. Adequate plasma concentrations of sorafenib were achieved in patients when it was dosed in combination with L+C. The mean plasma concentrations of C were consistently lower following administration of LCS, compared with administration of L+C only. The most common drug-related grade 3/4 adverse events were skin rash (69.3%), hand-foot skin reaction (69.3%) and diarrhoea (46.1%). According to RECIST Criteria, a clinical complete response was observed in 6 of 13 patients. A significant reduction in tumour size, evaluated with MRI, was also observed between baseline and 14 days of treatment in all 13 patients (P=0.005). A significant reduction in SUV uptake, measured by (18)FDG-PET/CT, was observed in all patients between baseline and 30 days of treatment (P=0.015) and between baseline and definitive surgery (P=0.0002). Using modified CT Criteria, a response was demonstrated in 8 out of 10 evaluable patients at 30 days and in 11 out of 13 evaluable patients at the definitive surgery. A significant reduction in Ki67 expression was observed in all patients at day 14 compared with baseline (P<0.00001) and in 9 out of 13 patients at the definitive surgery compared with baseline (P<0.03). There was also a significant suppression of CD31 and VEGF-A expression in response to treatment (P=0.01 and P=0.007, respectively). The LCS combination is feasible and tolerable. The tumour response and target biomarker modulation indicate that the combination is clinically and biologically active.


Role of nitric oxide signaling in endothelial differentiation of embryonic stem cells.
Signaling pathways that govern embryonic stem cell (ESCs) differentiation are not well characterized. Nitric oxide (NO) is a potent vasodilator that modulates other signaling pathways in part by activating soluble guanylyl cyclase (sGC) to produce cyclic guanosine monophosphate (cGMP). Because of its importance in endothelial cell (EC) growth in the adult, we hypothesized that NO may play a critical role in EC development. Accordingly, we assessed the role of NO in ESC differentiation into ECs. Murine ESCs differentiated in the presence of NO synthase (NOS) inhibitor NG-nitroarginine methyl ester (L-NAME) for up to 11 days were not significantly different from vehicle-treated cells in EC markers. However, by 14 days, L-NAME-treated cells manifested modest reduction in EC markers CD144, FLK1, and endothelial NOS. ESC-derived ECs generated in the presence of L-NAME exhibited reduced tube-like formation in Matrigel. To understand the discrepancy between early and late effects of L-NAME, we assessed the NOS machinery and observed low mRNA expression of NOS and sGC subunits in ESCs, compared to differentiating cells after 14 days. In response to NO donors or activation of NOS or sGC, cellular cGMP levels were undetectable in undifferentiated ESCs, at low levels on day 7, and robustly increased in day 14 cells. Production of cGMP upon NOS activation at day 14 was inhibited by L-NAME, confirming endogenous NO dependence. Our data suggest that NOS elements are present in ESCs but inactive until later stages of differentiation, during which period NOS inhibition reduces expression of EC markers and impairs angiogenic function.


Temperature dependence and bidirectional cation fluxes in red blood cells from spontaneously hypertensive rats.
The net passive influx of Na+ and efflux of K+ (orthodirection) through the red blood cell membranes from spontaneously hypertensive rats (SHR) were observed to be significantly higher (p less than 0.05) than those of three strains of normotensive rats when the measurements were made at 4 degrees C. Similar comparative studies, carried out at 37 degrees C, in the absence or presence of ouabain, showed no difference in these fluxes through this membrane from SHR compared to those from Wistar-Kyoto (WKY) rats, one of the normotensive strains. A study was undertaken to determine the temperature at which the greater cation fluxes in SHR red blood cells occurred. The net fluxes of Na+ and K+ decreased as the temperature was reduced from 37 degrees to 15 degrees C, but a paradoxical increase in the fluxes was observed as the temperature was decreased from 15 degrees to 4 degrees C. Only with this temperature shift (15 degrees to 4 degrees C) was the increase in flux significantly greater in SHR than in WKY cells. Subsequent studies were designed to determine whether the difference in the transport systems of red blood cells of SHR and WKY could be observed in fluxes of these cations in either direction across the membrane. For "reverse direction" flux studies, red blood cells were loaded with Na+ (to 130 mEq/liter cell water) and depleted of K+ (to 30 mEq/liter cell water) by incubation with the ionophore monensin.(ABSTRACT TRUNCATED AT 250 WORDS)


Importation of exotic ticks and tick-borne spotted fever group rickettsiae into the United States by migrating songbirds.
Birds are capable of carrying ticks and, consequently, tick-transmitted microorganisms over long distances and across geographical barriers such as oceans and deserts. Ticks are hosts for several species of spotted fever group rickettsiae (SFGR), which can be transmitted to vertebrates during blood meals. In this study, the prevalence of this group of rickettsiae was examined in ticks infesting migratory songbirds by using polymerase chain reaction (PCR). During the 2009 and 2010 spring migration season, 2064 northward-migrating passerine songbirds were examined for ticks at Johnson Bayou, Louisiana. A total of 91 ticks was removed from 35 individual songbirds for tick species identification and spotted fever group rickettsia detection. Ticks were identified as Haemaphysalis juxtakochi (n=38, 42%), Amblyomma longirostre (n=22, 24%), Amblyomma nodosum (n=17, 19%), Amblyomma calcaratum (n=11, 12%), Amblyomma maculatum (n=2, 2%), and Haemaphysalis leporispalustris (n=1, 1%) by comparing their 12S rDNA gene sequence to homologous sequences in GenBank. Most of the identified ticks were exotic species originating outside of the United States. The phylogenetic analysis of the 71 ompA gene sequences of the rickettsial strains detected in the ticks revealed the occurrence of 6 distinct rickettsial genotypes. Two genotypes (corresponding to a total of 28 samples) were included in the Candidatus Rickettsia amblyommii clade (less than 1% divergence), 2 of them (corresponding to a total of 14 samples) clustered with Rickettsia sp. "Argentina" with less than 0.2% sequence divergence, and 2 of them (corresponding to a total of 27 samples), although closely related to the R. parkeri-R. africae lineage (2.50-3.41% divergence), exhibited sufficient genetic divergence from its members to possibly constitute a new rickettsial genotype. Overall, there does not seem to be a specific relationship between exotic tick species, the rickettsiae they harbor, or the reservoir competence of the corresponding bird species.


Carry-over effects of the social environment on future divorce probability in a wild bird population.
Initial mate choice and re-mating strategies (infidelity and divorce) influence individual fitness. Both of these should be influenced by the social environment, which determines the number and availability of potential partners. While most studies looking at this relationship take a population-level approach, individual-level responses to variation in the social environment remain largely unstudied. Here, we explore carry-over effects on future mating decisions of the social environment in which the initial mating decision occurred. Using detailed data on the winter social networks of great tits, we tested whether the probability of subsequent divorce, a year later, could be predicted by measures of the social environment at the time of pairing. We found that males that had a lower proportion of female associates, and whose partner ranked lower among these, as well as inexperienced breeders, were more likely to divorce after breeding. We found no evidence that a female's social environment influenced the probability of divorce. Our findings highlight the importance of the social environment that individuals experience during initial pair formation on later pairing outcomes, and demonstrate that such effects can be delayed. Exploring these extended effects of the social environment can yield valuable insights into processes and selective pressures acting upon the mating strategies that individuals adopt.


High-sensitivity multispeckle diffuse correlation spectroscopy.
Significance: Cerebral blood flow is an important biomarker of brain health and function as it regulates the delivery of oxygen and substrates to tissue and the removal of metabolic waste products. Moreover, blood flow changes in specific areas of the brain are correlated with neuronal activity in those areas. Diffuse correlation spectroscopy (DCS) is a promising noninvasive optical technique for monitoring cerebral blood flow and for measuring cortex functional activation tasks. However, the current state-of-the-art DCS adoption is hindered by a trade-off between sensitivity to the cortex and signal-to-noise ratio (SNR). Aim: We aim to develop a scalable method that increases the sensitivity of DCS instruments. Approach: We report on a multispeckle DCS (mDCS) approach that is based on a 1024-pixel single-photon avalanche diode (SPAD) camera. Our approach is scalable to > 100,000 independent speckle measurements since large-pixel-count SPAD cameras are becoming available, owing to the investments in LiDAR technology for automotive and augmented reality applications. Results: We demonstrated a 32-fold increase in SNR with respect to traditional single-speckle DCS. Conclusion: A mDCS system that is based on a SPAD camera serves as a scalable method toward high-sensitivity DCS measurements, thus enabling both high sensitivity to the cortex and high SNR.


Global mismatch between fishing dependency and larval supply from marine reserves.
Marine reserves are viewed as flagship tools to protect exploited species and to contribute to the effective management of coastal fisheries. Yet, the extent to which marine reserves are globally interconnected and able to effectively seed areas, where fisheries are most critical for food and livelihood security is largely unknown. Using a hydrodynamic model of larval dispersal, we predict that most marine reserves are not interconnected by currents and that their potential benefits to fishing areas are presently limited, since countries with high dependency on coastal fisheries receive very little larval supply from marine reserves. This global mismatch could be reversed, however, by placing new marine reserves in areas sufficiently remote to minimize social and economic costs but sufficiently connected through sea currents to seed the most exploited fisheries and endangered ecosystems.


Treatment with depleting CD4 monoclonal antibody results in a preferential loss of circulating naive T cells but does not affect IFN-gamma secreting TH1 cells in humans.
CD4(pos) TH1 T cells are considered to play a central role in a number of human autoimmune diseases such as rheumatoid arthritis (RA) and multiple sclerosis. Experimental treatment protocols aimed at selectively eliminating CD4(pos) T cells thus far have yielded disappointing clinical results. Here we analyzed phenotype and function of circulating T cells in multiple sclerosis patients treated with the chimeric CD4 mAb cM-T412 in a randomized, double-blind, placebo-controlled, magnetic resonance imaging-monitored phase II trial. Treatment resulted in a long-lasting depletion of CD4(pos) T cells but did not affect CD8(pos) T cell numbers. Analysis of CD4(pos) subpopulations showed that unprimed, CD45RA(pos)/R0(neg) lymphocytes were approximately three times more sensitive to the mAb than primed, CD45RA(neg)/R0(pos) T cells. Notably, within the CD45RA(pos) subset, T cells with phenotypic evidence of prior activation, i.e., expressing Fas, were relatively insensitive to cM-T412, compared with Fas(neg) cells. Remarkably, while a decrease in the number of IL-4-producing T helper 2 (TH2)-type cells in the anti-CD4 treated group was observed, numbers of IFN-gamma-producing T helper 1 (TH1)-type cells remained stable, resulting in a significant increase in the TH1/TH2 ratio. Our data show that treatment with depleting CD4 mAb does not eliminate the cells most strongly involved in the disease process, i.e., primed, IFN-gamma-producing TH1-type cells, and may therefore give an explanation for the lack of beneficial clinical effects of depleting CD4 mAb in human chronic autoimmune disease.


Taking the trophic bypass: aquatic-terrestrial linkage reduces methylmercury in a terrestrial food web.
Ecosystems can be linked by the movement of matter and nutrients across habitat boundaries via aquatic insect emergence. Aquatic organisms tend to have higher concentrations of certain toxic contaminants such as methylmercury (MeHg) compared to their terrestrial counterparts. If aquatic organisms come to land, terrestrial organisms that consume them are expected to have elevated MeHg concentrations. But emergent aquatic insects could have other impacts as well, such as altering consumer trophic position or increasing ecosystem productivity as a result of nutrient inputs from insect carcasses. We measure MeHg in terrestrial arthropods at two lakes in northeastern Iceland and use carbon and nitrogen stable isotopes to quantify aquatic reliance and trophic position. Across all terrestrial focal arthropod taxa (Lycosidae, Linyphiidae, Acari, Opiliones), aquatic reliance had significant direct and indirect (via changes in trophic position) effects on terrestrial consumer MeHg. However, contrary to our expectations, terrestrial consumers that consumed aquatic prey had lower MeHg concentrations than consumers that ate mostly terrestrial prey. We hypothesize that this is due to the lower trophic position of consumers feeding directly on midges relative to those that fed mostly on terrestrial prey and that had, on average, higher trophic positions. Thus, direct consumption of aquatic inputs results in a trophic bypass that creates a shorter terrestrial food web and reduced biomagnification of MeHg across the food web. Our finding that MeHg was lower at terrestrial sites with aquatic inputs runs counter to the conventional wisdom that aquatic systems are a source of MeHg contamination to surrounding terrestrial ecosystems.


Suprathreshold compound action potential amplitude as a measure of auditory function in cochlear implant users.
Electrically evoked compound action potential (eCAP) amplitudes elicited at suprathreshold levels were assessed as a measure of the effectiveness of cochlear implant (CI) stimulation. Twenty-one individuals participated; one was excluded due to facial stimulation during eCAP testing. For each participant, eCAPs were elicited with stimulation from seven electrodes near the upper limit of the individual's electrical dynamic range. A reduced-channel CI program was created using those same seven electrodes, and participants performed a vowel discrimination task. Consistent with previous reports, eCAP amplitudes varied across tested electrodes; the profiles were unique to each individual. In 6 subjects (30%), eCAP amplitude variability was partially explained by the impedance of the recording electrode. The remaining amplitude variability within subjects, and the variability observed across subjects could not be explained by recording electrode impedance. This implies that other underlying factors, such as variations in neural status across the array, are responsible. Across-site mean eCAP amplitude was significantly correlated with vowel discrimination scores (r2 = 0.56). A single eCAP amplitude measured from the middle of the array was also significantly correlated with vowel discrimination, but the correlation was weaker (r2 = 0.37), though not statistically different from the across-site mean. Normalizing each eCAP amplitude by its associated recording electrode impedance did not improve the correlation with vowel discrimination (r2 = 0.52). Further work is needed to assess whether combining eCAP amplitude with other measures of the electrode-neural interface and/or with more central measures of auditory function provides a more complete picture of auditory function in CI recipients.


Automatic classification of focal liver lesions based on MRI and risk factors.
Accurate classification of focal liver lesions is an important part of liver disease diagnostics. In clinical practice, the lesion type is often determined from the abdominal MR examination, which includes T2-weighted and dynamic contrast enhanced (DCE) MR images. To date, only T2-weighted images are exploited for automatic classification of focal liver lesions. In this study additional MR sequences and risk factors are used for automatic classification to improve the results and to make a step forward to a clinically useful aid for radiologists. Clinical MRI data sets of 95 patients with in total 125 benign lesions (40 adenomas, 29 cysts and 56 hemangiomas) and 88 malignant lesions (30 hepatocellular carcinomas (HCC) and 58 metastases) were included in this study. Contrast curve, gray level histogram, and gray level co-occurrence matrix texture features were extracted from the DCE-MR and T2-weighted images. In addition, risk factors including the presence of steatosis, cirrhosis, and a known primary tumor were used as features. Fifty features with the highest ANOVA F-score were selected and fed to an extremely randomized trees classifier. The classifier evaluation was performed using the leave-one-out principle and receiver operating characteristic (ROC) curve analysis. The overall accuracy for the classification of the five major focal liver lesion types is 0.77. The sensitivity/specificity is 0.80/0.78, 0.93/0.93, 0.84/0.82, 0.73/0.56, and 0.62/0.77 for adenoma, cyst, hemangioma, HCC, and metastasis, respectively. The proposed classification system using features derived from clinical DCE-MR and T2-weighted images, with additional risk factors is able to differentiate five common types of lesions and is a step forward to a clinically useful aid for focal liver lesion diagnosis.


Phenotyping and genetic studies of 357 consecutive patients presenting with premature ovarian failure.
Premature ovarian failure (POF) encompasses a heterogeneous spectrum of conditions, with phenotypic variability among patients. The etiology of POF remains unknown in most cases. We performed a global phenotyping of POF women with the aim of better orienting attempts at an etiological diagnosis. We performed a mixed retrospective and prospective study of clinical, biological, histological, morphological, and genetic data relating to 357 consecutive POF patients between 1997 and 2008. The study was conducted at a reproductive endocrinology referral center. Seventy-six percent of the patients presented with normal puberty and secondary amenorrhea. Family history was present in 14% of the patients, clinical and/or biological autoimmunity in 14.3%. Fifty-six women had a fluctuating form of POF. The presence of follicles was suggested at ultrasonography in 50% of the patients, and observed in 29% at histology; the negative predictive value of the presence of follicles at ultrasonography was 77%. Bone mineral density alterations were found in 58% of the women. Eight patients had X chromosomal abnormalities other than Turner's syndrome, eight other patients evidenced FMR1 pre-mutation. Two other patients had autoimmune polyendocrine syndrome type 2 and 1. A genetic cause of POF was identified in 25 patients, i.e. 7% of the whole cohort. POF etiology remains most often undiscovered. Novel strategies of POF phenotyping are in such content mandatory to improve the rate of POF patients for whom etiology is identified.


Glycosylation affects rat Kv1.1 potassium channel gating by a combined surface potential and cooperative subunit interaction mechanism.
The effect of glycosylation on Kv1.l potassium channel function was investigated in mammalian cells stably transfected with Kv1.l or Kv1.1N207Q. Macroscopic current analysis showed that both channels were expressed but Kv1.1N207Q, which was not glycosylated, displayed functional differences compared with wild-type, including slowed activation kinetics, a positively shifted V 1/2, a shallower slope for the conductance versus voltage relationship, slowed C-type inactivation kinetics, and a reduced extent of and recovery from C-type inactivation. Kv1. 1N207Q activation properties were also less sensitive to divalent cations compared with those of Kv1.l. These effects were largely due to the lack of trans-Golgi added sugars, such as galactose and sialic acid, to the N207 carbohydrate tree. No apparent change in ionic current deactivation kinetics was detected inKv1.1N207Q compared with wild-type. Our data, coupled with modelling, suggested that removal of the N207 carbohydrate tree had two major effects. The first effect slowed the concerted channel transition from the last dosed state to the open state without changing the voltage dependence of its kinetics. This effect contributed to the G-V curve depolarization shift and together with the lower sensitivity to divalent cations suggested that the carbohydrate tree and its negatively charged sialic acids affected the negative surface charge density on the channel's extracellular face that was sensed by the activation gating machinery. The second effect reduced a cooperativity factor that slowed the transition from the open state to the dosed state without changing its voltage dependence. This effect accounted for the shallower G-V slope, and contributed to the depolarized G-V shift, and together with the inactivation changes it suggested that the carbohydrate tree also affected channel conformations. Thus N-glycosylation, and particularly terminal sialylation, affected Kv1.l gating properties both by altering the surface potential sensed by the channel's activation gating machinery and by modifying conformational changes regulating cooperative subunit interactions during activation and inactivation. Differences in glycosylation pattern among closely related channels may contribute to their functional differences and affect their physiological roles.


Hepatitis B virus quantification and detection of YMDD mutants in a single reaction by real-time PCR and annealing curve analysis.
Long-term antiviral therapy, although effective for treatment of hepatitis B, might select the emergence of drug-resistant hepatitis B virus (HBV) mutants. Detection of HBV mutants and determination of viral titre are two crucial parameters for monitoring treatment response and occurrence of mutants. In this study, we take lamivudine resistance as an example to develop a method that can determine both parameters in a single-tube PCR reaction. The method contained two consecutive steps: in the first step, real-time PCR was used for quantification; in the second step, a novel annealing curve analysis was used for detecting YMDD mutants. For accurate quantification, PCR primers and hybridization probes were chosen from highly conserved regions to ensure the equivalent amplification of all HBV genotypes. Within the sensor probe, there were signature nucleotide polymorphisms that could effectively differentiate YMDD mutants from wild type by distinct melting temperatures (Tm) values. The clinical applicability of the assay was tested in serial samples from 90 patients receiving lamivudine treatment. This assay could readily differentiate YMDD, YIDD and YVDD mutants by their distinct Tm values. The quantification results showed great consistency in a linear range from 10(3) to 10(11) copies/ml. Moreover, this assay could detect YMDD mutants accounting for < or = 10% of the total viral population. Its clinical feasibility has been verified in primary specimens. The newly designed YMDD detection method is simple, sensitive, cost-effective, time-saving and provides a useful tool for follow-up of patients treated with lamivudine or other antiviral drugs.


Expression and localisation of aquaporin water channels in human urothelium in situ and in vitro.
Urothelium is generally considered to be impermeable to water and constituents of urine. The possibility that human urothelium expresses aquaporin (AQP) water channels as the basis for water and solute transport has not previously been investigated. To investigate the expression of AQP water channels by human urothelium in situ, in proliferating urothelial cell cultures and in differentiated tissue constructs. AQP expression by human urothelium in situ and cultured urothelial cells was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunolabelling. Expression screening was carried out on samples of freshly isolated urothelia from multiple surgical (bladder and ureteric) specimens and on proliferating and differentiated normal human urothelial (NHU) cells in culture. Urothelial tissue constructs were established and investigated for expression of urothelial differentiation markers and AQPs. Qualitative study. Transcripts for AQP3, AQP4, AQP7, AQP9, and AQP11 were expressed consistently by freshly isolated urothelia as well as by cultured NHU cells. AQP0, AQP1, AQP2, AQP5, AQP6, AQP8, AQP10, and AQP12 were not expressed. Immunochemistry confirmed expression of AQP3, AQP4, AQP7, and AQP9 at the protein level. AQP3 was shown to be intensely expressed at cell borders in the basal and intermediate layers in both urothelium in situ and differentiated tissue constructs in vitro. This is the first study to demonstrate that AQPs are expressed by human urothelium, suggesting a potential role in transurothelial water and solute transport. Our findings challenge the traditional concept of the urinary tract as an impermeable transit and storage unit and provide a versatile platform for further investigations into the biological and clinical relevance of AQPs in human urothelium.


Prognostic factors for return to work after a first compensated episode of back pain.
To determine which factors measured at baseline and during the course of treatment influence time to return to work after a first compensated episode of back pain. The design is a treatment inception cohort including 305 compensated workers out of 402 eligible ones presenting at two rehabilitation centres for conventional treatment. Crude and adjusted rate ratios (RRs) along with 95% confidence intervals (95% CIs) were estimated with the Cox's proportional hazards regression. 50% of workers had not returned to work after 112 days of follow up, and 11.3% still had not after 270 days. At the end of the study period (maximum follow up time was 1228 days), 230 workers (75.4%) had returned to work, 6.5% had not, and a similar percentage had retired, gone into vocational training, or returned to school. In the final model stratified for radiating pain during treatment, which was an important prognostic variable, workers between 21 to 30 years of age had a greater chance of returning to work (RR (95% CI) 1.43 (1.04 to 1.98) than those > or = 30. The other factors associated with a greater chance of returning to work were: a diagnosis of sprain or pain upsilon a diagnosis of intervertebral disc disorder (2.20 (1.23-3.91)), < 30 days of waiting between the accident and the beginning of treatment (1.30 (0.96 to 1.77)), a good flexion at baseline (1.52 (1.04 to 2.23)), absence of neurological symptoms during treatment (1.40 (0.98-2.00)), > 24 months of employment in the industry (1.49 (1.10 to 2.03)), working for a public industry upsilon a private one (1.63 (1.21 to 2.19)), and the ability to take unscheduled breaks (1.45 (1.06 to 1.97)). Even with a first episode of back pain, time to return to work is long and the proportion not returning is high. Return to work as expected is influenced by disease and host characteristics but also by social and work factors. Reinstatement programmes should account for all these factors.


Category deficits and paradoxical dissociations in Alzheimer's disease and Herpes Simplex Encephalitis.
Most studies examining category specificity are single-case studies of patients with living or nonliving deficits. Nevertheless, no explicit or agreed criteria exist for establishing category-specific deficits in single cases regarding the type of analyses, whether to compare with healthy controls, the number of tasks, or the type of tasks. We examined two groups of patients with neurological pathology frequently accompanied by impaired semantic memory (19 patients with Alzheimer's disease and 15 with Herpes Simplex Encephalitis). Category knowledge was examined using three tasks (picture naming, naming-to-description, and feature verification). Both patient groups were compared with age- and education-matched healthy controls. The profile in each patient was examined for consistency across tasks and across different analyses; however, both proved to be inconsistent. One striking finding was the presence of paradoxical dissociations (i.e., patients who were impaired for living things on one task and nonliving things on another task). The findings have significant implications for how we determine category effects and, more generally, for the methods used to document double dissociations across individual cases in this literature.


Acute Respiratory Distress Syndrome Prediction Score: Derivation and Validation.
Despite advances in treatment strategies, acute respiratory distress syndrome (ARDS) after cardiac surgery remains associated with high morbidity and mortality. A method of screening patients for risk of ARDS after cardiac surgery is needed. To develop and validate an ARDS prediction score designed to identify patients at high risk of ARDS after cardiac or aortic surgery. An ARDS prediction score was derived from a retrospective derivation cohort and validated in a prospective cohort. Discrimination and calibration of the score were assessed with area under the receiver operating characteristic curve and the Hosmer-Lemeshow goodness-of-fit test, respectively. A sensitivity analysis was conducted to assess model performance at different cutoff points. The retrospective derivation cohort consisted of 201 patients with and 602 patients without ARDS who had undergone cardiac or aortic surgery. Nine routinely available clinical variables were included in the ARDS prediction score. In the derivation cohort, the score distinguished patients with versus without ARDS with area under the curve of 0.84 (95% CI, 0.81-0.88; Hosmer-Lemeshow P = .55). In the validation cohort, 46 of 1834 patients (2.5%) had ARDS develop within 7 days after cardiac or aortic surgery. Area under the curve was 0.78 (95% CI, 0.71-0.85), and the score was well calibrated (Hosmer-Lemeshow P = .53). The ARDS prediction score can be used to identify high-risk patients from the first day after cardiac or aortic surgery. Patients with a score of 3 or greater should be closely monitored. The score requires external validation before clinical use.


Aspects of cancer immunotherapy.
Cancer immunotherapy has traditionally undergone a 'revolution' every decade, from the use of Bacille Calmette-Guérin by scarification in the 1970s, to interleukin-2 therapies in the 1980s, and monoclonal antibody treatments in the early 1990s. Usually the early reports on the use of such agents were encouraging, but when more patients were studied in multiple centres, the initial promising results could not be confirmed. Now in a new century, we have more reagents and methods available than ever before - indeed, with such a plethora of reagents it is difficult to envisage them being fully and appropriately tested within the next decade, by which time there will be even more reagents to test. However, there have been three major advances which should lead to substantial progress in cancer immunotherapy: (1) the widespread use of genetic engineering, enabling identification of candidate vaccine proteins and manipulation of their sequences; (2) the production of antigens, antibodies and cytokines in large amounts by recombinant technologies, and (3) an understanding of the mode of presentation of peptides by major histocompatibility complex Class I and Class II molecules and their recognition by T cells. Despite these advances, there are major problems facing cancer immunotherapy, such as the ability of tumours to mutate and evade the immune system and the difficulty of precisely defining the interactions of effector cells in mediating 'rejection' or destruction of a tumour. There are clearly immunological similarities with diseases such as malaria and schistosomiasis, where the invading foreign organisms can use a variety of strategies to resist an elicited immune response. The failure to find a suitable vaccine for these diseases must lead to some pessimism for the development of immunotherapy for an autologous tumour. However, there are promising studies now in progress which should give an indication of the most important directions to follow. This review provides a commentary on aspects of cancer immunotherapy and in particular will deal with: (1) the selection of antigens as vaccine components; (2) the modes of presentation of antigens, particularly by major histocompatibility complex Class I molecules; and (3) new modes of delivery of vaccine immunogens.


Slow electron velocity-map imaging photoelectron spectra of the methoxide anion.
High resolution anion photodetachment spectra are presented for the methoxide anion and its fully deuterated counterpart. The spectra were obtained with slow electron velocity-map imaging. Improved electron affinities are determined for CH3O as 1.5690+/-0.0019 eV and for CD3O as 1.5546+/-0.0019 eV. The spectra resolve many features associated with spin-orbit and vibronic coupling that were not seen in previous photodetachment studies. Photoelectron angular distributions taken as a function of detachment wavelength for the ground vibronic state transitions are recorded and are consistent with the removal of a nonbonding, p-type electron localized on the oxygen atom. Several hot bands and sequence bands are observed for the first time, providing insight into the vibrational structure of the methoxide anion. The results are compared to recent calculations of the anion photoelectron spectra that incorporate bilinear coupling terms among the methoxy vibrational modes and are found to be in reasonable agreement.