Prevalence, incidence and predictors of volunteer community health worker attrition in Kwale County, Kenya.
In underserved populations, the contribution of community health workers (CHWs) is vital to the healthcare systems. Attrition of these workers causes critical breakdowns in the delivery of essential services to these populations. Literature on reasons for attrition is limited, although some have been identified in studies on sustainability of CHW programmes. These factors are, however, likely to be influenced by context. We measured CHW attrition and its predictors in a rural area in Kenya. We conducted a nested case-control study and focus group discussions among CHWs involved in a maternal and child health project. A training register of 1005 CHWs was used to sample and follow CHWs for attrition. Incidence of CHW attrition was calculated using a Poisson model. Separately, we used logistic regression to determine predictors of CHW attrition. Of the 1005 CHWs, 498 (49.6%) had left the project by the time of the study. The incidence of attrition was 46.8/1000 person-years (95% CI 38.7 to 56.5). In the case-control study, lack of interest in peer organisation membership (OR 5.3; 95% CI 1.3 to 20.6) was associated with attrition. Absence of refresher training (OR 4.0; 95% CI 2.2 to 7.1) and receiving no feedback from supervisors (OR 2.0; 95% CI 1.0 to 3.9) were also associated with attrition. Discordance in expectations and perceived heavy workload were also identified as key reasons for attrition in the qualitative study. This study estimates high prevalence and incidence of CHW attrition in Kwale County, Kenya. Ongoing training, feedback and peer support are also important in enhancing retention of CHWs. Additionally, expectations regarding the roles and benefits of involvement in CHW work should be communicated clearly, and workload should be kept reasonable or negotiated with the CHWs.


Defective kinetics of cytochrome c oxidase and alteration of mitochondrial membrane potential in fibroblasts and cytoplasmic hybrid cells with the mutation for myoclonus epilepsy with ragged-red fibres ('MERRF') at position 8344 nt.
We have investigated pathogenic effects of the tRNA(Lys) A8344G mutation associated with the syndrome myoclonus epilepsy with ragged-red fibres (MERRF) by using fibroblasts and fibroblast-derived cytoplasmic hybrid cells harbouring different percentages of mutated mitochondrial DNA (mtDNA). The activity of cytochrome c oxidase (COX) in patient fibroblasts with 89% mutated mtDNA was decreased to 20% of the control levels. COX exhibited altered kinetics, with a decreased V(max) for both the low-affinity and high-affinity phases; however, the K(m) values were not significantly changed. The substrate-dependent synthesis of ATP was decreased to 50% of the control. Analysis of the mitochondrial membrane potential, DeltaPsi, in digitonin-treated cells with tetramethylrhodamine methyl ester (TMRM) with the use of flow cytometry showed a 80% decrease in DeltaPsi at state 4 and an increased sensitivity of DeltaPsi to an uncoupler in fibroblasts from the patient. The investigation of transmitochondrial cytoplasmic hybrid clones derived from the patient's fibroblasts enabled us to characterize the relationship between heteroplasmy of the MERRF mutation, COX activity and DeltaPsi. Within the range of 87-73% mutated mtDNA, COX activity was decreased to 5-35% and DeltaPsi was decreased to 6-78%. These results demonstrate that the MERRF mutation affects COX activity and DeltaPsi in different proportions with regard to mutation heteroplasmy and indicate that the biochemical manifestation of the MERRF mutation exerts a very steep threshold of DeltaPsi inhibition.


USP22 exerts tumor-suppressive functions in colorectal cancer by decreasing mTOR activity.
USP22, the deubiquitinating subunit of the SAGA transcriptional cofactor complex, is a member of an 11-gene "death-from-cancer" signature. USP22 has been considered an attractive therapeutic target since high levels of its expression were associated with distant metastasis, poor survival, and high recurrence rates in a wide variety of solid tumors, including colorectal cancer (CRC). We sought to investigate the role of Usp22 during tumorigenesis in vivo using a mouse model for intestinal carcinogenesis with a tissue-specific Usp22 ablation. In addition, we assessed the effects of USP22 depletion in human CRC cells on tumorigenic potential and identified underlying molecular mechanisms. For the first time, we report that USP22 has an unexpected tumor-suppressive function in vivo. Intriguingly, intestine-specific Usp22 deletion exacerbated the tumor phenotype caused by Apc mutation, resulting in significantly decreased survival and higher intestinal tumor incidence. Accordingly, human CRC cells showed increased tumorigenic properties upon USP22 reduction in vitro and in vivo and induced gene expression signatures associated with an unfavorable outcome in CRC patients. Notably, USP22 loss resulted in increased mTOR activity with the tumorigenic properties elicited by the loss of USP22 being reversible by mTOR inhibitor treatment in vitro and in vivo. Here, we demonstrate that USP22 can exert tumor-suppressive functions in CRC where its loss increases CRC burden by modulating mTOR activity. Importantly, our data uncover a tumor- and context-specific role of USP22, suggesting that USP22 expression could serve as a marker for therapeutic stratification of cancer patients.


Antinociceptive effects of long-acting nalbuphine decanoate after intramuscular administration to Hispaniolan Amazon parrots (Amazona ventralis).
To evaluate the thermal antinociceptive effects and duration of action of nalbuphine decanoate after IM administration to Hispaniolan Amazon parrots (Amazona ventralis). 10 healthy adult Hispaniolan Amazon parrots of unknown sex. Nalbuphine decanoate (33.7 mg/kg) or saline (0.9% NaCl) solution was administered IM in a randomized complete crossover experimental design (periods 1 and 2). Foot withdrawal threshold to a noxious thermal stimulus was used to evaluate responses. Baseline thermal withdrawal threshold was recorded 1 hour before drug or saline solution administration, and thermal foot withdrawal threshold measurements were repeated 1, 2, 3, 6, 12, 24, 48, and 72 hours after drug administration. Nalbuphine decanoate administered IM at a dose of 33.7 mg/kg significantly increased thermal foot withdrawal threshold, compared with results after administration of saline solution during period 2, and also caused a significant change in withdrawal threshold for up to 12 hours, compared with baseline values. Nalbuphine decanoate increased the foot withdrawal threshold to a noxious thermal stimulus in Hispaniolan Amazon parrots for up to 12 hours and provided a longer duration of action than has been reported for other nalbuphine formulations. Further studies with other types of nociceptive stimulation, dosages, and dosing intervals as well as clinical trials are needed to fully evaluate the analgesic effects of nalbuphine decanoate in psittacine birds.


Olfactory memory formation and the influence of reward pathway during appetitive learning by honey bees.
Animals possess the ability to assess food quality via taste and via changes in state that occur after ingestion. Here, we investigate the extent to which a honey bee's ability to assess food quality affected the formation of association with an odor stimulus and the retention of olfactory memories associated with reward. We used three different conditioning protocols in which the unconditioned stimulus (food) was delivered as sucrose stimulation to the proboscis (mouthparts), the antennae or to both proboscis and antennae. All means of delivery of the unconditioned stimulus produced robust associative conditioning with an odor. However, the memory of a conditioned odor decayed at a significantly greater rate for subjects experiencing antennal-only stimulation after either multiple- or single-trial conditioning. Finally, to test whether the act of feeding on a reward containing sucrose during conditioning affected olfactory memory formation, we conditioned honey bees to associate an odor with antennal stimulation with sucrose followed by feeding on a water droplet. We observed that a honey bee's ability to recall the conditioned odor was not significantly different from that of subjects conditioned with an antennal-only sucrose stimulus. Our results show that stimulation of the sensory receptors on the proboscis and/or ingestion of the sucrose reward during appetitive olfactory conditioning are necessary for long-term memory formation.


Involvement of 26-kDa cell-associated TNF-alpha in experimental hepatitis and exacerbation of liver injury with a matrix metalloproteinase inhibitor.
TNF-alpha is a pleiotropic cytokine that exists both as a 26-kDa cell-associated and a 17-kDa soluble form. Recently, a class of matrix metalloproteinase inhibitors has been identified that can prevent the processing by TNF convertase of 26-kDa TNF-alpha to its 17-kDa form and can reduce mortality from normally lethal doses of D-galactosamine plus LPS (D-GalN/LPS). Here we report that a matrix metalloproteinase inhibitor, GM-6001, improves survival but does not protect against liver injury from D-GalN/LPS-induced shock in the mouse. In Con A-induced hepatitis, GM-6001 actually exacerbates hepatocellular necrosis and apoptosis despite greater than 90% reduction in plasma TNF-alpha concentrations. Treatment with GM-6001 also has minimal effect on the concentration of membrane-associated TNF-alpha in the livers of animals with Con A induced hepatitis. In contrast, a TNF binding protein (TNF-bp), which neutralizes both membrane-associated and soluble TNF-alpha, prevents D-GalN/LPS- and Con A-induced hepatitis. Our studies suggest that cell-associated TNF-alpha plays a role in the hepatocellular necrosis and apoptosis that accompany D-GalN/LPS- or Con A-induced hepatitis, and that matrix metalloproteinase inhibitors are ineffective in preventing this hepatic injury.


[Pregnancy after direct oocyte-sperm transfer (DOST)--a simple alternative to in vitro fertilization (IVF)].
We report on the first pregnancies achieved at our clinic following direct oocyte-sperm-transfer (DOST). Immediately after follicular aspiration, oocytes together with spermatozoa prepared by the swim-up-technique are transferred to the uterus in a manner similar to the embryo transfer following IVF. Our study population consists of 9 patients with bilateral tubal occlusion, accompanied by a male factor in three cases. Up to now, we have achieved four pregnancies, including one twin pregnancy, all occurring, when a male factor was absent. The method described appears promising in terms of efficiency, reduction in the costs and technical procedures during treatment of human infertility.


Identification of a Free Non-tryptophan Fluorophore in Water-soluble Fraction of Human Brunescent Cataractous Lens Nucleus.
Purpose: We previously reported that a unique free fluorophore (Fl-Glc), presumably a beta-glucoside, is particularly abundant in human brunescent cataractous lens nuclei. Our preliminary experiments indicated that incubation of low-molecular weight (MW) fraction of non-brunescent lens nuclei causes an increase in a particular fluorophore (Fl-X). This study was undertaken to compare the Fl-Glc with the Fl-X and subsequently to identify the Fl-X.Methods: Experiment (1) The purified Fl-X and its beta-glucosidase digest (aglycon) were compared with the Fl-Glc and its aglycon, respectively, by high-performance liquid chromatography (HPLC). Experiment (2) i) The Fl-X and its aglycon were analysed by liquid chromatography/mass spectrometry (LC/MS). ii) Authentic xanthurenic acid was analysed by HPLC and LC/MS.Results: Experiment (1) The retention times of the Fl-X and the Fl-Glc exactly coincided. The fluorescence peaks of both disappeared after beta-glucosidase treatment. Experiment (2) i) LC/MS results suggested that the MWs of the Fl-X and its aglycon were 367 and 205, respectively. ii) HPLC and LC/MS results for xanthurenic acid (MW = 205) were exactly the same as those for the aglycon of the Fl-X.Conclusions: The Fl-Glc and the Fl-X are identifical, and the Fl-X (= Fl-Glc) is a glucoside of xanthurenic acid.


Hyperlipidemia is associated with lower risk of poststroke mortality independent of statin use: A population-based study.
Background Although statin therapy is associated with reduced stroke and mortality risk, some studies report that higher lipid levels are associated with improved outcomes following ischemic stroke. Aims We examined the association of hyperlipidemia (HLD) combined with statin therapy on all-cause mortality in stroke patients. Methods All stroke patients in the Greater Cincinnati Northern Kentucky region of ∼1.3 million were identified using ICD-9 discharge codes in 2005 and 2010. Stroke patients with and without HLD were categorized based on their reported statin use at baseline or discharge into three groups: no-HLD/no-statins, HLD/no-statins, and HLD/on-statins. Cox proportional hazards model was used to estimate the risk of mortality at 30 days, 1 year, and 3 years poststroke. Results Overall, 77% (2953) of the 3813 ischemic stroke patients were diagnosed with HLD and 72% ( n = 2123) of those patients were on statin medications. The mean age was 70.0 ± 14.6 years, 56% were women, and 21% were black. In adjusted analyses, the HLD/no-statins group showed 35% (adjusted hazard ratio (aHR) = 0.65, 95% CI: 0.46-0.92), 27% (aHR = 0.73, 95% CI: 0.59-0.90), and 17% (aHR = 0.83, 95% CI: 0.70-0.97) reduced risk of mortality at 30 days, 1 year, and 3 years, respectively, poststroke, compared with no-HLD/no-statins group. The HLD/on-statins group showed an additional 17% significant survival benefit at 3 years poststroke compared with HLD/no-statins group. Conclusions A diagnosis of HLD in ischemic stroke patients is associated with reduced short- and long-term mortality, irrespective of statin use. Statin therapy is associated with significant, additional long-term survival benefit.


What Contributes Most to High Health Care Costs? Health Care Spending in High Resource Patients.
U.S. health care spending nearly doubled in the decade from 2000-2010. Although the pace of increase has moderated recently, the rate of growth of health care costs is expected to be higher than the growth in the economy for the near future. Previous studies have estimated that 5% of patients account for half of all health care costs, while the top 1% of spenders account for over 27% of costs. The distribution of health care expenditures by type of service and the prevalence of particular health conditions for these patients is not clear, and is likely to differ from the overall population. To examine health care spending patterns and what contributes to costs for the top 5% of managed health care users based on total expenditures. This retrospective observational study employed a large administrative claims database analysis of health care claims of managed care enrollees across the full age and care spectrum. Direct health care expenditures were compared during calendar year 2011 by place of service (outpatient, inpatient, and pharmacy), payer type (commercially insured, Medicare Advantage, and Medicaid managed care), and therapy area between the full population and high resource patients (HRP). The mean total expenditure per HRP during calendar year 2011 was $43,104 versus $3,955 per patient for the full population. Treatment of back disorders and osteoarthritis contributed the largest share of expenditures in both HRP and the full study population, while chronic renal failure, heart disease, and some oncology treatments accounted for disproportionately higher expenditures in HRP. The share of overall expenditures attributed to inpatient services was significantly higher for HRP (40.0%) compared with the full population (24.6%), while the share of expenditures attributed to pharmacy (HRP = 18.1%, full = 21.4%) and outpatient services (HRP = 41.9%, full = 54.1%) was reduced. This pattern was observed across payer type. While the use of physician-administered pharmaceuticals was slightly higher in HRP, their use did not alter this spending pattern. Overall, expenditures in the HRP population are more than 10-fold higher compared with the full population. Managed care pharmacy can benefit from understanding what contributes to these higher costs, and managed care directors should consider an appropriately balanced assessment of the share of total spend by service and therapeutic category in HRP when devising drug usage and related cost-management strategies.


How physicians use clinical information in diagnosing pulmonary embolism: an application of conjoint analysis.
To investigate what diagnostic strategies physicians adopt when the literature is unclear about the best use of diagnostic information, the authors examined how physicians weighted eight items of clinical information in diagnosing pulmonary embolism. Thirteen faculty members, 23 house officers, and 19 students estimated the likelihood of pulmonary embolism and predicted whether they would begin treatment in 27 simulated cases. The vignettes were constructed using a fractional factorial design and analyzed using conjoint analysis. Overall the lung scan results (34.7%) and leg examination (19.0%) were weighted most heavily in making the diagnosis, whereas the leg examination (35.7%) was given the greatest weight when deciding about therapy. Weights given by groups at different levels of training were similar, but there was considerable variation within groups. Heterogeneity of diagnostic strategies did not appear to decrease in groups with more training and experience. Multivariate analysis of predictors of pulmonary embolism in 102 actual cases showed that although lung scan results were important in both actual and simulated cases, heart rate accounted for the most variance in the actual cases but was hardly used in the physicians' strategies. There is considerable variation among physicians in how clinical information is used in diagnosing pulmonary embolism, and the variation may not decrease with increased experience.


Late-onset rheumatoid arthritis registry study, LORIS study: study protocol and design.
Although drug treatment strategies for rheumatoid arthritis (RA) are relatively well established, there is a paucity of evidence on the treatment in older patients. The purpose of this study is to build a registry for late-onset RA (LORA), which is expected to increase rapidly worldwide. In addition, we aim to propose optimal treatment strategies according to the patient background including frailty, thereby contributing to improving the quality of treatment and daily living in patients with RA. The LORIS (Late-onset Rheumatoid Arthritis Registry) Study is a prospective nation-wide multicenter observational study of patients with LORA. The inclusion criteria were patients aged ≥ 65 years at onset, meeting 2010 ACR/EULAR classification criteria for RA, and starting either any disease-modifying antirheumatic drugs (DMARDs) in a DMARD-naïve patient or the first biologic/targeted synthetic DMARDs during the study period. Enrollment was started on 11 January, 2022 and will be closed on 31 December, 2023. Patients will undergo a comprehensive baseline assessment including clinical data, medication, cognitive and physical function, psychosocial factors, and frailty. Data will be collected at baseline, Month 3, 6, 12, 18, 24, 36, and summarized descriptively. The factors associated with adverse events and achieving remission will be determined. A multi-disciplinary panel including patients, rheumatologists, and geriatric specialists will discuss the results and build a consensus regarding the treatment goals of LORA. We expect to provide a broad range of information for evidence-based shared decision making in the treatment of LORA. Registered at the UMIN registry (UMIN000046086) on 1 January 2022.


The effect of platelet-rich plasma on posterolateral lumbar fusion in a rat model.
Our purpose was to examine platelet-rich plasma (PRP) for its effect on bone formation and to follow the immunohistochemical changes in calcitonin gene-related peptide (CGRP) in dorsal root ganglion neurons innervating the discs as a possible index of nociceptive nerve transmission in a rat model. A total of seventy rats were used. Ten constituted a non-punctured disc sham group, while another ten rats constituted a group that underwent puncture of the L4-L5 discs. Forty rats were in experimental groups in which the L4-L5 discs were punctured; posterolateral lumbar arthrodesis was performed with PRP (the PRP group) or without the use of PRP (the normal arthrodesis group), with twenty rats in each group. The remaining ten rats were used as blood donors. Four and eight weeks after surgery, microcomputed tomography examinations were done to evaluate the amount of bone and the L4-L5 spines were harvested to evaluate bone union, followed by resection of dorsal root ganglion neurons. The percentages of Fluoro-Gold-labeled and CGRP-immunoreactive neurons were calculated. The platelet count and the concentration of growth factors in PRP were higher than those in blood (p < 0.05). The bone volumes observed in the PRP group were significantly greater than those of the normal arthrodesis group at four and eight weeks (p < 0.05). Three (30%; 95% confidence interval [CI], 6% to 65%) of ten rats in the normal arthrodesis group and eight (80%; 95% CI, 44% to 98%) of ten rats in the PRP group were considered to have bone fusion four weeks after surgery (p < 0.05). At eight weeks, seven (70%; 95% CI, 34% to 94%) of ten rats in the normal arthrodesis group and nine (90%; 95% CI, 55% to 99%) of ten rats in the PRP group were considered to have bone fusion (p = 0.27). The proportion of CGRP-immunoreactive neurons was significantly greater in the punctured group than in the other groups. There were no significant differences between the normal arthrodesis group and the PRP group. PRP appears to promote bone formation in rats and has a tendency to shorten the period of bone union in this rat model of posterolateral lumbar arthrodesis, but it did not influence the proportion of CGRP-immunoreactive neurons, a likely indicator of inflammatory pain originating from the degenerative intervertebral disc. The ability of PRP in this model suggests that it may be able to shorten the period of union and lead to an early return to social activities after treatment.


Amino acid metabolic signaling influences Aedes aegypti midgut microbiome variability.
The mosquito midgut microbiota has been shown to influence vector competence for multiple human pathogens. The microbiota is highly variable in the field, and the sources of this variability are not well understood, which limits our ability to understand or predict its effects on pathogen transmission. In this work, we report significant variation in female adult midgut bacterial load between strains of A. aegypti which vary in their susceptibility to dengue virus. Composition of the midgut microbiome was similar overall between the strains, with 81-92% of reads coming from the same five bacterial families, though we did detect differences in the presence of some bacterial families including Flavobacteriaceae and Entobacteriaceae. We conducted transcriptomic analysis on the two mosquito strains that showed the greatest difference in bacterial load, and found that they differ in transcript abundance of many genes implicated in amino acid metabolism, in particular the branched chain amino acid degradation pathway. We then silenced this pathway by targeting multiple genes using RNA interference, which resulted in strain-specific bacterial proliferation, thereby eliminating the difference in midgut bacterial load between the strains. This suggests that the branched chain amino acid (BCAA) degradation pathway controls midgut bacterial load, though the mechanism underlying this remains unclear. Overall, our results indicate that amino acid metabolism can act to influence the midgut microbiota. Moreover, they suggest that genetic or physiological variation in BCAA degradation pathway activity may in part explain midgut microbiota variation in the field.


Salt-taste polymorphism TRPV1-rs8065080 is associated with increased likelihood of depression in an elderly cohort.
Despite the prevalence of depression and anxiety worldwide, their aetiologies remain unclear, and they can be difficult to diagnose and treat. Changes in salt-taste perception have been found in both conditions. Single nucleotide polymorphisms (SNPs) in the salt-taste-related gene TRPV1, have been associated with alterations to salt-taste perception, preference, and sodium consumption. Diet quality is a known modifier of depression and anxiety and recently, sodium intake has been studied in mental health. However, the relationships between salt-taste genetics, depression, anxiety, and these dietary factors are yet to be elucidated. Data from the well-characterised cross-sectional Retirement Health and Lifestyle Study (n = 536, ≥65y) were used to explore the relationships between the salt-taste SNP TRPV1-rs8065080, levels of depression and anxiety (Hospital Anxiety and Depression Scale, HADS), estimated sodium intake and diet quality in this secondary analysis. Standard least squares regression and nominal logistic regression modelling were used to compare continuous and categorical variables respectively, with analyses stratified by sex. Presence of the TRPV1-rs8065080 variant allele (C) was found to increase the likelihood of having depression (HADS) in the total population, and in males. The associations remained significant after adjusting for sodium intake, three diet quality indices and demographic variables, suggesting that TRPV1-rs8065080 genotype is driving the association with depression. Future studies should explore extra-oral functions of the SNP and salt-taste receptors in the brain, and the roles of neurotransmitters common to both depression and salt-taste to improve the management of this increasingly prevalent and difficult to treat condition.


Virtual reality hypnosis pain control in the treatment of multiple fractures: a case series.
This case series evaluated the use of virtual reality hypnosis (VRH) for the treatment of pain associated with multiple fractures from traumatic injuries. VRH treatment was administered on 2 consecutive days, and pain and anxiety were assessed each day before and after VRH treatment as well as on Day 3, which was 24 hours after the second treatment session. Pain reduction from baseline to Day 3 was from 70% to 30%, despite opioid analgesic use remaining stable. The subjective pain reduction reported by patients was encouraging, and the results of this case series suggest the importance of further study of VRH with larger samples using randomized controlled trials.


ETS transcription factors Etv2 and Fli1b are required for tumor angiogenesis.
ETS transcription factor ETV2/Etsrp functions as a key regulator of embryonic vascular development in multiple vertebrates. However, its role in pathological vascular development has not been previously investigated. To analyze its role in tumor angiogenesis, we utilized a zebrafish xenotransplantation model. Using a photoconvertible kdrl:NLS-KikGR line, we demonstrated that all tumor vessels originate from the existing embryonic vasculature by the mechanism of angiogenesis. Xenotransplantation of mouse B16 melanoma cells resulted in a significant increase in expression of the ETS transcription factors etv2 and fli1b expression throughout the embryonic vasculature. etv2 null mutants which undergo significant recovery of embryonic angiogenesis during later developmental stages displayed a strong inhibition of tumor angiogenesis. We utilized highly specific and fully validated photoactivatable morpholinos to inhibit Etv2 function after embryonic vasculogenesis has completed. Inducible inhibition of Etv2 function resulted in a significant reduction of tumor angiogenesis and inhibition of tumor growth. Furthermore, inducible inhibition of Etv2 function in fli1b mutant embryos resulted in even stronger reduction in tumor angiogenesis and growth, demonstrating that Etv2 and Fli1b have a partially redundant requirement during tumor angiogenesis. These results demonstrate the requirement for Etv2 and Fli1b in tumor angiogenesis and suggest that inhibition of these ETS factors may present a novel strategy to inhibit tumor angiogenesis and reduce tumor growth.


Population genomics of domestic and wild yeasts.
Since the completion of the genome sequence of Saccharomyces cerevisiae in 1996 (refs 1, 2), there has been a large increase in complete genome sequences, accompanied by great advances in our understanding of genome evolution. Although little is known about the natural and life histories of yeasts in the wild, there are an increasing number of studies looking at ecological and geographic distributions, population structure and sexual versus asexual reproduction. Less well understood at the whole genome level are the evolutionary processes acting within populations and species that lead to adaptation to different environments, phenotypic differences and reproductive isolation. Here we present one- to fourfold or more coverage of the genome sequences of over seventy isolates of the baker's yeast S. cerevisiae and its closest relative, Saccharomyces paradoxus. We examine variation in gene content, single nucleotide polymorphisms, nucleotide insertions and deletions, copy numbers and transposable elements. We find that phenotypic variation broadly correlates with global genome-wide phylogenetic relationships. S. paradoxus populations are well delineated along geographic boundaries, whereas the variation among worldwide S. cerevisiae isolates shows less differentiation and is comparable to a single S. paradoxus population. Rather than one or two domestication events leading to the extant baker's yeasts, the population structure of S. cerevisiae consists of a few well-defined, geographically isolated lineages and many different mosaics of these lineages, supporting the idea that human influence provided the opportunity for cross-breeding and production of new combinations of pre-existing variations.


Hic-5 is required for fetal gene expression and cytoskeletal organization of neonatal cardiac myocytes.
Cardiac costameres link the extracellular matrix to the sarcomere at the z-disc and contain proteins such as integrins and other signaling molecules implicated in the regulation of pathological hypertrophy. Paxillin family members, hic-5 and paxillin, are scaffolding proteins associated with the integrin complex that have been shown to mediate numerous protein interactions in other cell types. While paxillin has been described in postnatal heart, hic-5 has not been identified. Our results provide evidence of hic-5 in neonatal cardiac myocytes co-localized with paxillin and alpha-actinin at the z-discs and the ends of actin filaments. Treatment with the hypertrophic agonist phenylephrine resulted in increased hic-5 expression while having no effect on paxillin levels. To see if increased hic-5 expression was sufficient to induce changes in cytoskeletal organization, hic-5 was overexpressed in myocytes by adenoviral infection. Hic-5 overexpression significantly increased the number of cells with organized cytoskeleton. Using siRNA mediated knockdown, we examined the requirement of hic-5 and paxillin in regulation of phenylephrine induced gene expression and cytoskeletal organization. Our results indicate that hic-5, not paxillin is required for upregulation of ANF and alpha-skeletal actin genes as well as in cytoskeletal reorganization. Finally, we demonstrated that hic-5 upregulation occurs downstream of MEK1/2-ERK1/2 signaling as inhibition of MEK1/2 using U0126 inhibitor completely inhibited hic-5 upregulation by PE. In a complimentary study, we showed that hic-5 knockdown had no effect on PE induced ERK1/2 phosphorylation. These findings demonstrate a novel role for hic-5 in the regulation of actin cytoskeleton and fetal gene expression.


Guggulsterone inhibits NF-kappaB and IkappaBalpha kinase activation, suppresses expression of anti-apoptotic gene products, and enhances apoptosis.
Guggulsterone, derived from Commiphora mukul and used to treat obesity, diabetes, hyperlipidemia, atherosclerosis, and osteoarthritis, has been recently shown to antagonize the farnesoid X receptor and decrease the expression of bile acid-activated genes. Because activation of NF-kappaB has been closely linked with inflammatory diseases affected by guggulsterone, we postulated that it must modulate NF-kappaB activation. In the present study, we tested this hypothesis by investigating the effect of this steroid on the activation of NF-kappaB induced by inflammatory agents and carcinogens. Guggulsterone suppressed DNA binding of NF-kappaB induced by tumor necrosis factor (TNF), phorbol ester, okadaic acid, cigarette smoke condensate, hydrogen peroxide, and interleukin-1. NF-kappaB activation was not cell type-specific, because both epithelial and leukemia cells were inhibited. Guggulsterone also suppressed constitutive NF-kappaB activation expressed in most tumor cells. Through inhibition of IkappaB kinase activation, this steroid blocked IkappaBalpha phosphorylation and degradation, thus suppressing p65 phosphorylation and nuclear translocation. NF-kappaB-dependent reporter gene transcription induced by TNF, TNFR1, TRADD, TRAF2, NIK, and IKK was also blocked by guggulsterone but without affecting p65-mediated gene transcription. In addition, guggulsterone decreased the expression of gene products involved in anti-apoptosis (IAP1, xIAP, Bfl-1/A1, Bcl-2, cFLIP, and survivin), proliferation (cyclin D1 and c-Myc), and metastasis (MMP-9, COX-2, and VEGF); this correlated with enhancement of apoptosis induced by TNF and chemotherapeutic agents. Overall, our results indicate that guggulsterone suppresses NF-kappaB and NF-kappaB-regulated gene products, which may explain its anti-inflammatory activities.


Dealing with the difficult older patient.
Physicians are often asked to intervene when elderly patients in acute or long-term care facilities are disruptive, demanding or abusive. Though medication may be the immediate response, it often fails to solve the problem, which, being situational and behavioural, calls for a situational and behavioural solution. An organized approach to solving such problems is presented that takes into account the interactions among the patient, family, staff, setting and circumstances. The role of the physician as leader in the process is also discussed.


Photosensitization by coumarin derivatives.
Coumarin and several of its derivatives were investigated for their photosensitizing properties. With a few exceptions, the coumarins are potentially strong photocontact sensitizers but do not evoke phototoxic reactions. Substitution at the 6 or 7 positions of the benzopyrone ring confers photoallergenic capability, whereas hydrogenation abolishes activity. Photo-cross-reactions among closely related derivatives can develop. The action spectrum for contact photodermatitis extends from 360 nm down to at least 300 nm. Data are presented to show that the photocontact allergic response is related to the ultraviolet A dose and to the concentration of the offending chemical.


Modified rice bran hemicellulose inhibits vascular endothelial growth factor-induced angiogenesis in vitro via VEGFR2 and its downstream signaling pathways.
Angiogenesis is implicated in diverse pathological conditions such as cancer, rheumatoid arthritis, psoriasis, atherosclerosis, and retinal neovascularization. In the present study, we investigated the effects of modified rice bran hemicellulose (MRBH), a water-soluble hemicellulose preparation from rice bran treated with shiitake enzymes, on vascular endothelial growth factor (VEGF)-induced angiogenesis in vitro and its mechanism. We found that MRBH significantly inhibited VEGF-induced tube formation in human umbilical vein endothelial cells (HUVECs) co-cultured with human dermal fibroblasts. We also observed that MRBH dose-dependently suppressed the VEGF-induced proliferation and migration of HUVECs. Furthermore, examination of the anti-angiogenic mechanism indicated that MRBH reduced not only VEGF-induced activation of VEGF receptor 2 but also of the downstream signaling proteins Akt, extracellular signal-regulated protein kinase 1/2, and p38 mitogen-activated protein kinase. These findings suggest that MRBH has in vitro anti-angiogenic effects that are partially mediated through the inhibition of VEGF signaling.


Basal and touch-evoked fos-like immunoreactivity during experimental inflammation in the rat.
Fos-immunoreactivity can readily be induced in spinal cord neurones by noxious, but to a much more limited extent, by innocuous peripheral stimuli. The present study has investigated whether low intensity stimuli and electrical stimulation of A beta afferents elicit greater c-fos expression during the behavioural sensory hypersensitivity generated by experimental peripheral inflammation. We have examined the time-course of c-fos expression after inflammation produced by either an intra-plantar injection of the irritant turpentine oil or of complete Freund's adjuvant (CFA). In the former case, a significant initial expression in all dorsal horn laminae was followed by a gradual decrease, whereas after CFA injection, an initial expression limited to the superficial laminae subsequently extended into the deep laminae, with a decrease at 24 h and an increase in labelling at later times. Low intensity touch stimuli repeated for 10 min, when applied at 24 h and 48 h after CFA injection, elicited a significant increase in the number of Fos-immunoreactive neurons in both the superficial and deep laminae of the dorsal horn compared to non-inflamed animals. Electrical stimulation of the sciatic nerve 24 h post-CFA injection, at a strength sufficient only to activate A beta-afferents fibres (100 microA, 50 microseconds, 10 min), also elicited a significant increase in labelling relative to the same stimuli applied in control animals, especially in laminae V-VI. The present results demonstrate that low intensity cutaneous stimuli elicit a significantly greater increase in c-fos expression in dorsal horn neurons during peripheral inflammation and that A beta-afferent input contributes to this, a finding that may relate to the allodynia experienced during inflammation.


Enteric sensory neurons communicate with interstitial cells of Cajal to affect pacemaker activity in the small intestine.
Enteric sensory neurons (the AH neurons) play a role in control of gastrointestinal motor activity; AH neuron activation has been proposed to change propulsion into segmentation. We sought to find a mechanism underlying this phenomenon. We formulated the hypothesis that AH neurons increase local ICC-MP (interstitial cells of Cajal associated with the myenteric plexus) pacemaker frequency to disrupt peristalsis and promote absorption. To that end, we sought structural and physiological evidence for communication between ICC-MP and AH neurons. We designed experiments that allowed us to simultaneously activate AH neurons and observe changes in ICC calcium transients that underlie its pacemaker activity. Neurobiotin injection in AH neurons together with ICC immunohistochemistry proved the presence of multiple contacts between AH neuron varicosities and the cell bodies and processes of ICC-MP. Generating action potential activity in AH neurons led to increase in the frequency and amplitude of calcium transients underlying pacemaker activity in ICC. When no rhythmicity was seen, rhythmic calcium transients were evoked in ICC. As a control, we stimulated nitrergic S neurons, which led to reduction in ICC calcium transients. Hence, we report here the first demonstration of communication between AH neurons and ICC. The following hypothesis can now be formulated: AH neuron activation can disrupt peristalsis directed by ICC-MP slow wave activity, through initiation of a local pacemaker by increasing ICC pacemaker frequency through increasing the frequency of ICC calcium transients. Evoking new pacemakers distal to the proximal lead pacemaker will initiate both retrograde and antegrade propulsion causing back and forth movements that may disrupt peristalsis.


Comorbidities in the Elderly and Their Possible Influence on Vaccine Response.
The following chapter is focused on the impact of comorbidities on the effectiveness of vaccination in older persons. Relevant comorbidities are cardiovascular diseases like hypertension, coronary artery disease or congestive heart failure, which lead to reduction of vaccine immunogenicity; or chronic obstructive pulmonary disease with a decline in lung function and a higher risk for pneumonia or infections due to influenza. End-stage renal disease has a high impact on developing infections and causes immune dysfunction over all parts of the immune system. Depression and dementia as well as psychological stress are associated with poor antibody response and a higher range of inflammation markers. Chronic inflammatory processes like rheumatoid arthritis also alter the immune system. In addition, geriatric syndromes and lowered functional status have implications for the vaccination response. Malnutrition is characterized by depletion of structural and functional proteins. This leads to a low antibody response. Negative immunomodulatory effects are also observed in vitamin D insufficiency. Frailty as well is associated with immunological changes and lowered performance in the activities of daily living, but moderate physical activity improves immune function.


Dynamic interaction of cAMP with the Rap guanine-nucleotide exchange factor Epac1.
Epac1 is a Rap-specific guanine-nucleotide exchange factor (GEF) which is activated by the binding of cAMP to a cyclic nucleotide monophosphate (cNMP)-binding domain. We investigated the equilibrium and dynamics of the interaction of cAMP and Epac1 using a newly designed fluorescence analogue of cAMP, 8-MABA-cAMP. We observed that the interaction of cAMP, measured by competition with 8-MABA-cAMP, with an isolated cNMP binding domain of Epac1 has an overall equilibrium constant (Kd) of 4 microM and that the kinetics of the interaction are highly dynamic. The binding properties of cAMP are apparently not affected when the catalytic domain is present, despite the fact that binding of cAMP results in activation of Epac1. This indicates that for the activation process, no appreciable binding energy is required. However, when bound to Rap1b, the apparent Kd of Epac to cAMP was about fivefold lower, suggesting that substrate interaction stabilizes cAMP binding. Since the fluorescent analogues used here were either less able or unable to induce activation of Epac1, we concluded that the binding of nucleotide to Epac and the activation of GEF activity are uncoupled processes and that thus appropriate cAMP analogues can be used as inhibitors of the Epac1-mediated signal transduction pathway of Rap.


Residential Proximity to Major Roadways at Birth, DNA Methylation at Birth and Midchildhood, and Childhood Cognitive Test Scores: Project Viva(Massachusetts, USA).
Epigenetic variability is hypothesized as a regulatory pathway through which prenatal exposures may influence child development and health. We sought to examine the associations of residential proximity to roadways at birth and epigenome-wide DNA methylation. We also assessed associations of differential methylation with child cognitive outcomes. We estimated residential proximity to roadways at birth using a geographic information system (GIS) and cord blood methylation using Illumina's HumanMethylation450-array in 482 mother-child pairs in Project Viva. We identified individual CpGs associated with residential-proximity-to-roadways at birth using robust linear regression [[Formula: see text]]. We also estimated association between proximity-to-roadways at birth and methylation of the same sites in blood samples collected at age 7-11 y ([Formula: see text]). We ran the same analyses in the Generation R Study for replication ([Formula: see text]). In Project Viva, we investigated associations of differential methylation at birth with midchildhood cognition using linear regression. Living closer to major roadways at birth was associated with higher cord blood (and-more weakly-midchildhood blood) methylation of four sites in LAMB2. For each halving of residential-proximity-to-major-roadways, we observed a 0.82% increase in DNA methylation at cg05654765 [95% confidence interval (CI): (0.54%, 1.10%)], 0.88% at cg14099457 [95% CI: (0.56%, 1.19%)], 0.19% at cg03732535 [95% CI: (0.11%, 0.28)], and 1.08% at cg02954987 [95% CI: (0.65%, 1.51%)]. Higher cord blood methylation of these sites was associated with lower midchildhood nonverbal cognitive scores. Our results did not replicate in the Generation R Study. Our discovery results must be interpreted with caution, given that they were not replicated in a separate cohort. However, living close to major roadways at birth was associated with cord blood methylation of sites in LAMB2-a gene known to be linked to axonal development-in our U.S. cohort. Higher methylation of these sites associated with lower nonverbal cognitive scores at age 7-11 y in the same children. https://doi.org/10.1289/EHP2034.


Chemical composition and antibacterial activity of essential oils from different parts of Leonurus japonicus Houtt.
The herb and fruits of Leonurus japonicus Houtt., named "Yimucao" and "Chongweizi", respectively, in Chinese, have been widely used in China as gynecological medicines. The components of the essential oils obtained by hydrodistillation were investigated by GC-MS. The antibacterial activity of the essential oils was determined by micro-dilution assay. The results showed large variations in the chemical composition and antibacterial activity of the oils. The oil of "Yimucao" showed antibacterial activity against various Gram-positive bacteria and consisted mainly of sesquiterpenes and diterpenes, with phytone, phytol, caryophyllene oxide and β-caryophyllene being the most significant constituents, whereas the oil of "Chongweizi", mainly made up of bornyl acetate and aliphatic hydrocarbons, was inactive in the antibacterial assay. Further study of the main compounds in "Yimucao oil" showed that β-caryophyllene had wide-spectrum activity against Gram-positive bacteria.


Integrating play in neurodevelopmental treatment.
This article presents one perspective on the integration of play activities within a neurodevelopmental frame of reference. Based on the premise that activities are characteristic of occupational therapy intervention, issues related to combining play activities with neurodevelopmental principles are discussed. Clinical examples are also provided to illustrate the value of integrating play activities within the occupational therapy treatment of the child with cerebral palsy.


Public understanding of the purpose of cancer screening: A population-based survey.
Objectives In examining informed choice in cancer screening, we investigated public awareness that some screening programmes aim to prevent cancer, while others seek to detect cancer at an early stage. Methods A population-based survey of adults aged 50-70 in England (n = 1433), including data on demographic characteristics and screening experience. Participants were asked to select the main purpose of cervical, breast, and colorectal cancer screening (both faecal occult blood testing and flexible sigmoidoscopy). Results Across all four screening programmes, most people thought the main aim was to catch cancer early (71-78%). Only 18 and 14% knew that cervical screening and flexible sigmoidoscopy, respectively, are primarily preventive. Knowledge of the preventive aspect of these two programmes was low across the board, with few demographic patterns. By contrast, 78 and 73% of the sample were aware that breast screening and the faecal occult blood test, respectively, predominantly aim to detect cancer early. For these programmes, accurate knowledge was socially graded, lower in ethnic minority groups, and positively associated with previous participation in the programmes. Conclusions Our findings suggest that although awareness of the purpose of early detection screening is high, awareness that screening can prevent cancer is low across all demographic groups. Understanding the purpose of screening is a key aspect of informed choice but despite current communication strategies highlighting these differences, people do not seem to have a nuanced understanding of these differing aims. Our findings may be indicative of a broader public scepticism about the preventability of cancer.


Mortality rates and causes of death among all HIV-positive individuals with hemophilia in Canada over 21 years of follow-up.
Many individuals with hemophilia were infected with human immunodeficiency virus (HIV) in the early 1980s through contaminated blood products. Most also were co-infected with hepatitis C virus (HCV). Deaths among the entire cohort of HIV-positive hemophiliacs in Canada up to 2003 are described. Using registry data, we analyzed Kaplan-Meier survival curves, determined the effect of age at HIV seroconversion on mortality, and described cause-specific proportional mortality patterns over time. Of 2427 Canadians with hemophilia, 660 (27.2%) were HIV-positive, of whom 406 (61.5%) died. In contrast, 114 (6.5%) deaths occurred in HIV-negative controls. Median age at HIV seroconversion was 20 (range, < 1-67 years), and median survival was 15.0 years (95% confidence interval, 13.6-16.4 years). Younger age at HIV seroconversion was associated with improved survival; however, this finding was not explained by differences in causes of death across age groups. Following the introduction of highly active antiretroviral therapy, the proportion of deaths due to acquired immune deficiency syndrome has decreased, while the proportion of deaths due to liver disease has increased. There were 1134 HCV-positive individuals, of whom only 444 (39.2%) were also HIV-positive. Liver disease is a growing health concern among many hemophiliacs, not only those who are HIV-positive.


[Treatment of malnutrition in elderly patients].
Although often undiagnosed, undernutrition is prevalent among elderly patients in hospitals. The effect of undernutrition is reduced mental and pulmonary function, increased prevalence of infection, and increased morbidity and mortality. This article presents an overview of the diagnosis and treatment of undernutrition in elderly patients with nonmalignant disease, based on relevant literature. There are no generally accepted definitions of undernutrition. Undernutrition should be suspected clinically and screened for, using anthropometric methods (body weight, body mass index, triceps skinfold, arm muscle circumference) or functional tests (hand dynamometry, laboratory parameters). Results from several studies indicate that nutritional intervention could increase body weight and physical capacity of patients and reduce length of hospitalisation and mortality. Undernutrition is common in clinical practice. Hospitalised patients should be screened for risk of undernutrition. The screening should be combined with a nutrition plan, including strategies for nutrition therapy.


Dysregulation of the immune response in severe sepsis.
Sepsis is systemic expression of a generalized activation of the host's innate immunity as a result of varied types of insults. This expression involves a cellular inflammatory response that has both proinflammatory and antiinflammatory components, the primary trigger for which is an intracellular oxidative stress, induced by receptor-mediated transmembrane signal transduction or direct noxious injury. Sepsis reflects the interaction between pro- and anti-inflammatory intracellular mechanisms, the uncontrolled activation of which leads to cell exhaustion, organ dysfunction, and death. Successful clinical trials of novel treatments for the management of severe sepsis share a common ability to down-regulate this overall response, restoring normal proinflammatory responsiveness and mitochondrial energetic function.


Doubled single-frequency Nd:YLF ring laser coupled to a passive nonresonant cavity.
We demonstrate an original solution to obtain a single-frequency ring laser coupled to an external passive nonresonant ring cavity, which plays the role of an optical diode. This system provides more output power than systems with an intracavity unidirectional device. To the best of our knowledge, this work marks the first demonstration of a unidirectional planar ring laser at 1.3 microm. Using 12 W at 797 nm to pump a Nd:YLF laser, combined with intracavity second-harmonic generation, we achieve yields of 440 mW at 661.3 nm and 340 mW at 656.0 nm.


Evaluating the performance of the National Early Warning Score in different diagnostic groups.
The National Early Warning Score (NEWS) is used in hospitals across the UK to detect deterioration of patients within care pathways. It is used for most patients, but there are relatively few studies validating its performance in groups of patients with specific conditions. The performance of NEWS was evaluated against 36 other Early Warning Scores, in 123 patient groups, through use of the area under the receiver operating characteristic (AUROC) curve technique, to compare the abilities of each Early Warning Score to discriminate an outcome within 24hrs of vital sign recording. Outcomes evaluated were death, ICU admission, or a combined outcome of either death or ICU admission within 24 hours of an observation set. The National Early Warning Score 2 performs either best or joint best within 120 of the 123 patient groups evaluated and is only outperformed in prediction of unanticipated ICU admission. When outperformed by other Early Warning Scores in the remaining 3 patient groups, the performance difference was marginal. Consistently high performance indicates that NEWS is a suitable early warning score to use for all diagnostic groups considered by this analysis, and patients are not disadvantaged through use of NEWS in comparison to any of the other evaluated Early Warning Scores.


Intrinsically low thermal conductivity of bismuth oxychalcogenides originating from interlayer coupling.
The anharmonicity of phonons in a solid is ultimately rooted in the chemical bonding. However, the direct connection between phonon anharmoncity and chemical bonding is difficult to make experimentally or theoretically, mainly due to their complicated lattice structures. Here, with the help of first-principles calculations, we show that the intrinsically low lattice thermal conductivity (κ) of Bi2O2X (X = S, Se, Te) shows a strong connection to the electrostatic inter-layer coupling. We explain our results by the strong anharmonic chemical bonding between Bi and chalcogen atoms. Additionally, due to the strong anharmonicity, a large portion of phonon modes has a mean free path shorter than the average atomic distance. We employ a recently proposed two-channel model to take into account their contribution to κ.


Disease burden in four populations of dog and cat breeds compared to mixed-breed dogs and European shorthair cats.
Current public and professional opinion is that many dog breeds suffer from health issues related to inherited diseases or extreme phenotypes. The aim of this historical comparative observational study was to evaluate the breed-related disease burden in three purebred dog populations (Chihuahua, French bulldog, Labrador retriever) and one purebred cat breed (Persian cats) in the Netherlands by comparison to a control population of mixed-breed dogs and European Shorthair cats. A qualitative query was performed, consisting of a literature review and collecting the expert opinions of University veterinary specialists, to gather insight into potential diseases of the study population. Next, a referral clinic case control study of the patients referred to specific medical disciplines in the University Clinic was performed. The odds ratio (OR) was calculated to determine the likelihood of a patient referred to a particular medical discipline being a certain breed. Together, the qualitative query and the case control study resulted in a list of potentially relevant diseases limited to five organ systems per breed. These were analysed in data from primary practices. Patient files from ten primary practices over a period of two years were manually extracted and examined. Four-hundred individual patient records per breed as well as 1000 non-breed records were randomly selected from the 10 practices, weighted per practice size. Records were then examined and the presence or absence of certain diseases was identified. To evaluate the disease burden per breed, proportional difference (PD) was estimated, as well as the animal's age at presentation in months. The results of the referral clinic case control study showed an overrepresentation (Odds Ratio>1.5) of the selected breeds in several medical specialties, while median age at presentation was in some cases significantly lower than in the non-breed animals. Results of the practice-based extended cross-sectional study showed that only a few of the selected diseases contribute to the disease burden in these purebred populations, which was different from the expectations derived from the literature or expert opinion. Additional results included age difference at presentation, which may be interpreted as age of onset, and could indicate a higher disease burden for the individual animal. Also, only a small percentage of purebred dogs was registered with the national kennel club. Our final recommendation is that population-based data mining is needed to evaluate country-specific companion animal health and welfare.


Intestinal antiviral signaling is controlled by autophagy gene Epg5 independent of the microbiota.
Mutations in the macroautophagy/autophagy gene EPG5 are responsible for Vici syndrome, a human genetic disease characterized by combined immunodeficiency. Previously, we found that epg5-/- mice exhibit hyperinflammation in the lungs mediated by IL1B/IL-1β and TNF/TNFα, resulting in resistance to influenza. Here, we find that disruption of Epg5 results in protection against multiple enteric viruses including norovirus and rotavirus. Gene expression analysis reveals IFNL/IFN-λ responsive genes as a key alteration. Further, mice lacking Epg5 exhibit substantial alterations of the intestinal microbiota. Surprisingly, germ-free mouse studies indicate Epg5-associated inflammation of both the intestine and lung is microbiota-independent. Genetic studies support IFNL signaling as the primary mediator of resistance to enteric viruses, but not of microbial dysbiosis, in epg5-/- mice. This study unveils an important role, unexpectedly independent of the microbiota, for autophagy gene Epg5 in host organism protection by modulating intestinal IFNL responses.Abbreviations: CTNNB1: catenin (cadherin associated protein), beta 1; DAPI: 4',6-diamidino-2-phenylindole; EPG5: ectopic P-granules autophagy protein 5 homolog (C. elegans); FT: fecal transplant; IFI44: interferon-induced protein 44; IFIT1: interferon-induced protein with tetratricopeptide repeats 1; IFNG/IFN-γ: interferon gamma; IFNL/IFN-λ: interferon lambda; IFNLR1: interferon lambda receptor 1; IL1B/IL-1β: interleukin 1 beta; ISG: interferon stimulated gene; GF: germ-free; LEfSe: linear discriminant analysis effect size; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MNoV: murine norovirus; MX2: MX dynamin-like GTPase 2; OAS1A: 2'-5' oligoadenylate synthetase 1A; RV: rotavirus; SPF: specific-pathogen free; SQSTM1/p62: sequestosome 1; STAT1: signal transducer and activator of transcription 1; STING1: stimulator of interferon response cGAMP interactor 1; TBK1: TANK-binding kinase 1; TNF/TNFα: tumor necrosis factor.


Dynamic nuclear polarization enhanced NMR spectroscopy for pharmaceutical formulations.
Dynamic nuclear polarization (DNP) enhanced solid-state NMR spectroscopy at 9.4 T is demonstrated for the detailed atomic-level characterization of commercial pharmaceutical formulations. To enable DNP experiments without major modifications of the formulations, the gently ground tablets are impregnated with solutions of biradical polarizing agents. The organic liquid used for impregnation (here 1,1,2,2-tetrachloroethane) is chosen so that the active pharmaceutical ingredient (API) is minimally perturbed. DNP enhancements (ε) of between 40 and 90 at 105 K were obtained for the microparticulate API within four different commercial formulations of the over-the-counter antihistamine drug cetirizine dihydrochloride. The different formulations contain between 4.8 and 8.7 wt % API. DNP enables the rapid acquisition with natural isotopic abundances of one- and two-dimensional (13)C and (15)N solid-state NMR spectra of the formulations while preserving the microstructure of the API particles. Here this allowed immediate identification of the amorphous form of the API in the tablet. API-excipient interactions were observed in high-sensitivity (1)H-(15)N correlation spectra, revealing direct contacts between povidone and the API. The API domain sizes within the formulations were determined by measuring the variation of ε as a function of the polarization time and numerically modeling nuclear spin diffusion. Here we measure an API particle radius of 0.3 μm with a single particle model, while modeling with a Weibull distribution of particle sizes suggests most particles possess radii of around 0.07 μm.


Mineral status of beef cows and sheep on spring pasture fertilized with kieserite.
The utilization of minerals in orchardgrass (Dactylis glomerata L.) fertilized with kieserite (2,240 kg/ha) was examined in spring with lactating beef cows and wether lambs. Kieserite increased the concentration of Mg in vegetative herbage from .22 to .29% ( P less than .01) and of S from .20 to .29% (P less than .01), with little effect on other minerals. Fertilization also increased (P less than .01) apparent absorptions of Mg and S by lambs fed cut orchardgrass, from 24.5 to 36.5% and from 58.6 to 66.9%, respectively, and increased (P less than .01) apparent Mg retention from +.16 to +.33 g/day. Fertilization did not affect dry matter digestibility or apparent absorption of N, Ca or P by lambs. composition of blood and saliva of beef cows grazing fertilized and unfertilized pastures was determined during a 2-week period in early spring. Cows grazing kieserite-treated pastures showed a small increase (P greater than .05) in serum Mg, ultrafiltrable Mg and red blood cell (RBC) Mg, and fewer cows on the fertilized pastures showed severe hypomagnesemia (less than 1.0 mg/100 ml) after the first day of grazing. The decline in blood Mg concentrations was not accompanied by hypocalcemia or by changes in serum Na or K concentrations, but serum inorganic P decreased (P less than .01) markedly during the first 10 days of grazing. Ultrafiltrable Ca in blood was affected by fertilizer and period, but it is questionable whether either ultrafiltrable Mg or Ca would provide a more sensitive index of mineral status than serum concentrations. Concentrations of K, NA and P in saliva were more susceptible to fertilizer and period effects than were concentrations in blood, although all values fell within a normal range. There was no evidence of an inverse relationship between serum Mg and concentration of K in saliva of cows after a change from a winter-type diet to spring herbage containing high levels of N and K.


LR_Gapcloser: a tiling path-based gap closer that uses long reads to complete genome assembly.
Completing a genome is an important goal of genome assembly. However, many assemblies, including reference assemblies, are unfinished and have a number of gaps. Long reads obtained from third-generation sequencing (TGS) platforms can help close these gaps and improve assembly contiguity. However, current gap-closure approaches using long reads require extensive runtime and high memory usage. Thus, a fast and memory-efficient approach using long reads is needed to obtain complete genomes. We developed LR_Gapcloser to rapidly and efficiently close the gaps in genome assembly. This tool utilizes long reads generated from TGS sequencing platforms. Tested on de novo assembled gaps, repeat-derived gaps, and real gaps, LR_Gapcloser closed a higher number of gaps faster and with a lower error rate and a much lower memory usage than two existing, state-of-the art tools. This tool utilized raw reads to fill more gaps than when using error-corrected reads. It is applicable to gaps in the assemblies by different approaches and from large and complex genomes. After performing gap-closure using this tool, the contig N50 size of the human CHM1 genome was improved from 143 kb to 19 Mb, a 132-fold increase. We also closed the gaps in the Triticum urartu genome, a large genome rich in repeats; the contig N50 size was increased by 40%. Further, we evaluated the contiguity and correctness of six hybrid assembly strategies by combining the optimal TGS-based and next-generation sequencing-based assemblers with LR_Gapcloser. A proposed and optimal hybrid strategy generated a new human CHM1 genome assembly with marked contiguity. The contig N50 value was greater than 28 Mb, which is larger than previous non-reference assemblies of the diploid human genome. LR_Gapcloser is a fast and efficient tool that can be used to close gaps and improve the contiguity of genome assemblies. A proposed hybrid assembly including this tool promises reference-grade assemblies. The software is available at http://www.fishbrowser.org/software/LR_Gapcloser/.


Mutational landscape and clonal architecture in grade II and III gliomas.
Grade II and III gliomas are generally slowly progressing brain cancers, many of which eventually transform into more aggressive tumors. Despite recent findings of frequent mutations in IDH1 and other genes, knowledge about their pathogenesis is still incomplete. Here, combining two large sets of high-throughput sequencing data, we delineate the entire picture of genetic alterations and affected pathways in these glioma types, with sensitive detection of driver genes. Grade II and III gliomas comprise three distinct subtypes characterized by discrete sets of mutations and distinct clinical behaviors. Mutations showed significant positive and negative correlations and a chronological hierarchy, as inferred from different allelic burdens among coexisting mutations, suggesting that there is functional interplay between the mutations that drive clonal selection. Extensive serial and multi-regional sampling analyses further supported this finding and also identified a high degree of temporal and spatial heterogeneity generated during tumor expansion and relapse, which is likely shaped by the complex but ordered processes of multiple clonal selection and evolutionary events.


Do patient-related blood donors represent a threat to the safety of the blood supply?
Patient-related blood donors contribute to a significant proportion of the blood units collected in hospital-based blood banks. However, there is some concern on the safety of this kind of donation because of the possible existence of incentives for the donor to conceal deferrable risk factors, thus increasing the risk of donation within the window-period of transfusion-transmitted infections. We tested the hypothesis that if patient-related blood donors are less safe than community ones, the former would display both a higher prevalence of viral markers and a predominance of undisclosed risk-factors with low social acceptability. Comparison of virus reactivity rates against hepatitis C virus (HCV), hepatitis B virus (HBV) and human immunodeficiency virus (HIV), and the associated risk-factors, between patient-related and community donors who donated whole blood in our center during a five-year period. During the period under study 72,226 donors gave 149,944 whole blood units, of which 22,888 (15.3%) were provided by patient-related donors. There were 273 confirmed virus-reactive donations (15 anti-HIV, 148 anti-HCV and 110 HBsAg). The adjusted prevalence of virus reactivity was 19 (95% CI: 11-35) times higher in first-time donors than in repeat donors, 3.5 (95% CI: 2.3-4.1) times higher in donors > or = 30 years old than in younger ones, and 2.5 (95% CI: 1.9-3.2) times higher in patient-related donors than in community donors. With regard to deferrable risk-factors not disclosed at the time of donation, there was no significant difference between patient-related and community donors in the frequency of people who denied any risk-factor or who admitted intravenous drug use or high-risk sex. Past household contact with individuals having liver disease was significantly more frequent in patient-related donors than in community ones. Our results do not support the hypothesis that patient-related donors represent an increased risk of window-period donation because they conceal deferrable risk factors more frequently than community donors.


Xanthogranulomatous pyelonephritis: critical analysis of 30 patients.
Xanthogranulomatous pyelonephritis (XGP) is a chronic inflammatory condition of the kidneys. Nevertheless, disparities between the pre-operative and pathological diagnoses are frequently encountered. We reviewed all patients with XGP over a 17-year period to identify and characterize the pre-operative and pathological characteristics of the disease in our patients. A comprehensive review of all nephrectomy patients with a pre-operative diagnosis of pyelonephritis at National Taiwan University Hospital from 1991 to 2008 with the pathological diagnosis of XGP was conducted to demonstrate the clinical and radiological characteristics of XGP. XGP was diagnosed in 30 (18.6%) of the 160 nephrectomies performed for pyelonephritis. Of the 30 patients with XGP, 25 were women (83.3%) and 20 (66.7%) were overweight (body mass index >23). Their mean age was 55.17 years. The average serum creatinine level was 1.68 mg/dL. The image findings included renal calculi (56.7%), staghorn stones (26.7%), and spread of the disease to the retroperitoneum and psoas muscle (33.3%). Escherichia coli (36.7%) was the most prevalent pathogen. The mortality of the two-stage surgical treatment was zero, and morbidity developed in only 1 patient (4.8%). The key to accurate pre-operative diagnosis is to keep risk factors in mind such as age, sex, and renal calculi. Clinicians should maintain a high suspicion of XGP for early recognition and be aware of the care of chronic kidney disease. Finally, the association between XGP and central obesity warrants further research.


Why Health Insurance Matters-and Why Research Evidence Should Too.
In the current debate over the future of the Affordable Care Act (ACA), research evidence on the impact of the law and the effects of health insurance coverage in general is critical. Studies of health insurance expansion over the past decade have demonstrated that coverage expansions can produce significant reductions in mortality-particularly among minorities, those living in poorer areas, and those with chronic conditions potentially treatable with timely medical care. More recent studies of the ACA in particular demonstrate that the law has produced historically large reductions in the uninsured rate, with resulting improvements in access to care, perceived quality of care, and self-reported health. Yet much of the general public and many policy makers remain unaware of this evidence. Researchers and clinicians in academic medicine have a role to play in ensuring that critically important health policy decisions are made using rigorous evidence to best protect the interests of our patients.


Masking of diabetic phenotype on a low-energy diet despite persistence of impaired insulin response.
Type 2 diabetes has been postulated to be a "thrifty genotype" that aids fuel conservation; to investigate this hypothesis, type 2 diabetic and nondiabetic subjects, normal weight and obese, were studied after both 6 days on a 700-calorie diet and 6 days on a 2,800-calorie diet. On 2,800 calories per day, the median basal plasma glucose was raised in diabetics compared with nondiabetic subjects (median, 5.5 and 4.5 mmol/L, respectively; P = .003), but on the low-energy diet there was no difference (4.1 and 3.7 mmol/L, respectively; P greater than .05). Basal plasma insulin levels were similar in the diabetic and nondiabetic subjects and were twofold to threefold higher on the high- than low-energy diet in both diabetic (median, 7.8 and 3.1 mU/L on high- and low-energy diets, respectively; P = .007) and nondiabetic subjects (6.8 and 2.6 mU/L, respectively; P = .005). Similarly, the postprandial insulin concentrations increased to the same degree on the high- compared with the low-energy diet in diabetic (median postprandial increment, 14.9 and 3.4 mU/L; P = .005) and nondiabetic patients (9.5 and 3.0 mU/L; P = .005) and the beta cells in type 2 diabetes appeared to modulate normally to different calorie intakes. On both diets, the diabetic subjects had an impaired first-phase insulin response to an intravenous glucose load compared with nondiabetic subjects (high-energy diet median first-phase increment, 4.3 and 21.2 mU/L in diabetic and nondiabetic subjects, P = .0007; low-energy diet, 4.0 and 20.4 mU/L, respectively, P = .003).(ABSTRACT TRUNCATED AT 250 WORDS)


A principal component analysis approach to correcting the knee flexion axis during gait.
Accurate and precise knee flexion axis identification is critical for prescribing and assessing tibial and femoral derotation osteotomies, but is highly prone to marker misplacement-induced error. The purpose of this study was to develop an efficient algorithm for post-hoc correction of the knee flexion axis and test its efficacy relative to other established algorithms. Gait data were collected on twelve healthy subjects using standard marker placement as well as intentionally misplaced lateral knee markers. The efficacy of the algorithm was assessed by quantifying the reduction in knee angle errors. Crosstalk error was quantified from the coefficient of determination (r(2)) between knee flexion and adduction angles. Mean rotation offset error (αo) was quantified from the knee and hip rotation kinematics across the gait cycle. The principal component analysis (PCA)-based algorithm significantly reduced r(2) (p<0.001) and caused αo,knee to converge toward 11.9±8.0° of external rotation, demonstrating improved certainty of the knee kinematics. The within-subject standard deviation of αo,hip between marker placements was reduced from 13.5±1.5° to 0.7±0.2° (p<0.001), demonstrating improved precision of the knee kinematics. The PCA-based algorithm performed at levels comparable to a knee abduction-adduction minimization algorithm (Baker et al., 1999) and better than a null space algorithm (Schwartz and Rozumalski, 2005) for this healthy subject population.


A regimen containing bedaquiline and delamanid compared to bedaquiline in patients with drug-resistant tuberculosis.
There are limited data on combining delamanid and bedaquiline in drug-resistant tuberculosis (DR-TB) regimens. Prospective long-term outcome data, including in HIV-infected persons, are unavailable.We prospectively followed up 122 South African patients (52.5% HIV-infected) with DR-TB and poor prognostic features between 2014 and 2018. We examined outcomes and safety in those who received a bedaquiline-based regimen (n=82) compared to those who received a bedaquiline-delamanid combination regimen (n=40).There was no significant difference in 6-month culture conversion (92.5% versus 81.8%; p=0.26) and 18-month favourable outcome rate (63.4% versus 67.5%; p=0.66) in the bedaquiline versus the bedaquiline-delamanid combination group, despite the latter having more advanced drug resistance (3.7% versus 22.5% resistant to at least five drugs; p=0.001) and higher pre-treatment failure rates (12.2% versus 52.5% with pre-treatment multidrug-resistant TB therapy failure; p<0.001). Although the proportion of prolongation of the QT interval corrected using Fridericia's formula was higher in the combination group (>60 ms from baseline (p=0.001) or >450 ms during treatment (p=0.001)), there were no symptomatic cases or drug withdrawals in either group. Results were similar in HIV-infected patients.A bedaquiline-delamanid combination regimen showed comparable long-term safety compared to a bedaquiline-based regimen in patients with DR-TB, irrespective of HIV status. These data inform regimen selection in patients with DR-TB from TB-endemic settings.


Bilateral reflex vasodilation in the palatal mucosa evoked by unilateral tooth-pulp stimulation in the cat.
In a previous study in cats, we found that electrical stimulation of the tooth pulp caused blood flow increases at various sites in the ipsilateral oral mucosa (upper and lower gingivae, lower lip, buccal mucosa and tongue, and, notably, bilaterally in the palatal mucosa). Tooth-pulp stimulation is well-known to induce severe pain and to evoke autonomic reflex responses in other organs and tissues. The purpose of this study was: to confirm that tooth-pulp stimulation may indeed induce autonomically mediated vasodilator responses in the feline oral mucosa away from the stimulated tooth, and to test our hypothesis that the reflex pathway involves parasympathetic vasodilator fibers as efferents. Dynamic changes in palatal mucosal blood flow (PMBF), with lower lip blood flow (LBF) as a control, were investigated in anesthetized, cervically sympathectomized cats by means of Laser Doppler Flowmetry. Unilateral electrical stimulation of the maxillary canine tooth pulp produced hexamethonium-sensitive bilateral increases in PMBF in a stimulus-intensity-dependent manner, without an increase in systemic blood pressure; LBF increased only ipsilaterally. Bilateral section of the glossopharyngeal nerve roots had no effect on the vasodilator responses, while unilateral section of the facial nerve root or lesion of the pterygopalatine ganglion (PPG) abolished the response on that side. Intracranial electrical stimulation of the peripheral cut ends of the facial or glossopharyngeal nerve roots caused an increase in ipsilateral PMBF. These results indicate that unilateral tooth-pulp stimulation induces a bilateral reflex vasodilator response in the palatal mucosa mediated via parasympathetic vasodilator fibers that emerge from the brain stem with the facial nerve and reach the blood vessels via PPG. Although there is a dual innervation of the cat palatal mucosa by parasympathetic vasodilator fibers running via the facial and glossopharyngeal nerve roots, the latter are not involved in the bilateral reflex responses.


Scaffold of Selenium Nanovectors and Honey Phytochemicals for Inhibition of Pseudomonas aeruginosa Quorum Sensing and Biofilm Formation.
Honey is an excellent source of polyphenolic compounds that are effective in attenuating quorum sensing (QS), a chemical process of cell-to-cell communication system used by the opportunistic pathogen Pseudomonas aeruginosa to regulate virulence and biofilm formation. However, lower water solubility and inadequate bioavailability remains major concerns of these therapeutic polyphenols. Its therapeutic index can be improved by using nano-carrier systems to target QS signaling potently. In the present study, we fabricated a unique drug delivery system comprising selenium nanoparticles (SeNPs; non-viral vectors) and polyphenols of honey (HP) for enhancement of anti-QS activity of HP against P. aeruginosa PAO1. The developed selenium nano-scaffold showed superior anti-QS activity, anti-biofilm efficacy, and anti-virulence potential in both in-vitro and in-vivo over its individual components, SeNPs and HP. LasR is inhibited by selenium nano-scaffold in-vitro. Using computational molecular docking studies, we have also demonstrated that the anti-virulence activity of selenium nano-scaffold is reliant on molecular binding that occurs between HP and the QS receptor LasR through hydrogen bonding and hydrophobic interactions. Our preliminary investigations with selenium-based nano-carriers hold significant promise to improve anti-virulence effectiveness of phytochemicals by enhancing effective intracellular delivery.


Radiation Dose Reduction in Digital Breast Tomosynthesis by MTANN with Multi-scale Kernels.
Digital breast tomosynthesis (DBT) is an advanced three-dimensional screening modality for the early detection of breast cancer. DBT is able to reduce the problem of tissue overlap in standard two-dimensional mammograms, thus improving the sensitivity and specificity of cancer detection. Although DBT can improve diagnostic accuracy, it leads to higher radiation dose to patients compared to two-dimensional mammography. In this paper, we propose a novel radiation dose reduction technique that introduces multi-scale kernels to our original massive-training artificial neural network (MTANN) to reduce radiation dose substantially, while maintaining high image quality in DBT. After training our new MTANN with low-dose (LD) images and the corresponding "teaching" high-dose (HD) images, we can convert new LD images to "virtual" high-dose (VHD) images where noise and artifact in the LD images are significantly reduced. In VHD images, it is critical to preserve subtle structures and tiny patterns such as microcalcifications (MCs) which are essential for breast cancer diagnosis. We developed anatomical MTANN experts including an MC-specific expert with multi-scale kernels, which are combined by gating layers to generate whole VHD images. Our MTANN scheme was able to achieve a 79% dose reduction while preserving details of MCs. Experimental results demonstrated that our method achieved the highest performance among the best-known noise-reduction techniques and state-of-the-art deep-learning techniques.Clinical Relevance- Our method can decrease the dose radiation dose in DBT and maintain the image quality.


Targeting of the A2A adenosine receptor counteracts immunosuppression in vivo in a mouse model of chronic lymphocytic leukemia.
Tumor immunosuppression is a major cause for treatment failure and disease relapse, both in solid tumors and leukemia. Local hypoxia is among the conditions that cause immunosuppression, acting at least in part through the upregulation of extracellular adenosine levels, which potently suppress T cell responses and skew macrophages towards an M2 phenotype. Hence, there is intense investigation to identify drugs that target this axis. By using the TCL1 adoptive transfer CLL mouse model, we show that adenosine production and signaling are upregulated in the hypoxic lymphoid niches, where intense colonization of leukemic cells occurs. This leads to a progressive remodeling of the immune system towards tolerance, with expansion of T regulatory cells (Tregs), loss of CD8+ T cell cytotoxicity and differentiation of murine macrophages towards the patrolling (M2-like) subset. In vivo administration of SCH58261, an inhibitor the A2A adenosine receptor, re-awakens T cell responses, while limiting Tregs expansion, and re-polarizes monocytes towards the inflammatory (M1-like) phenotype. These results show for the first time the in vivo contribution of adenosine signaling to immune tolerance in CLL, and the translational implication of drugs interrupting this pathway. Although the effects of SCH58261 on leukemic cells are limited, interfering with adenosine signaling may represent an appealing strategy for combination-based therapeutic approaches.


On the generalized Wertheim law in integrated photoelasticity.
Two-dimensional photoelasticity is based on the classical Wertheim law. The integral Wertheim law can be used in integrated photoelasticity only in the case when the directions of the secondary principal stresses are constant on the light beam. We generalize the integral Wertheim law for the case when a slight rotation of the secondary principal directions takes place.


Ultrathin Nonlinear Metasurface for Optical Image Encoding.
Security of optical information is of great importance in modern society. Many cryptography techniques based on classical and quantum optics have been widely explored in the linear optical regime. Nonlinear optical encryption in which encoding and decoding involve nonlinear frequency conversions represents a new strategy for securing optical information. Here, we demonstrate that an ultrathin nonlinear photonic metasurface, consisting of meta-atoms with 3-fold rotational symmetry, can be used to hide optical images under illumination with a fundamental wave. However, the hidden image can be read out from second harmonic generation (SHG) waves. This is achieved by controlling the destructive and constructive interferences of SHG waves from two neighboring meta-atoms. In addition, we apply this concept to obtain gray scale SHG imaging. Nonlinear metasurfaces based on space variant optical interference open new avenues for multilevel image encryption, anticounterfeiting, and background free image reconstruction.


The floodplain food web mosaic: a study of its importance to salmon and steelhead with implications for their recovery.
Although numerous studies have attempted to place species of interest within the context of food webs, such efforts have generally occurred at small scales or disregard potentially important spatial heterogeneity. If food web approaches are to be employed to manage species, studies are needed that evaluate the multiple habitats and associated webs of interactions in which these species participate. Here, we quantify the food webs that sustain rearing salmon and steelhead within a floodplain landscape of the Methow River, Washington, USA, a location where restoration has been proposed to restore side channels in an attempt to recover anadromous fishes. We combined year-long measures of production, food demand, and diet composition for the fish assemblage with estimates of invertebrate prey productivity to quantify food webs within the main channel and five different, intact, side channels; ranging from channels that remained connected to the main channel at low flow to those reduced to floodplain ponds. Although we found that habitats within the floodplain had similar invertebrate prey production, these habitats hosted different local food webs. In the main channel, 95% of total prey consumption flowed to fishes that are not the target of proposed restoration. These fishes consumed 64% and 47% of the prey resources that were found to be important to fueling chinook and steelhead production in the main channel, respectively. Conversely, in side channels, a greater proportion of prey was consumed by anadromous salmonids. As a result, carrying capacity estimates based on food were 251% higher, on average, for anadromous salmonids in side channels than the main channel. However, salmon and steelhead production was generally well below estimated capacity in both the main and side channels, suggesting these habitats are under-seeded with respect to food, and that much larger populations could be supported. Overall, this study demonstrates that floodplain heterogeneity is associated with the occurrence of a mosaic of food webs, all of which were utilized by anadromous salmonids, and all of which may be important to their recovery and persistence. In the long term, these and other fishes would'likely benefit from restoring the processes that maintain floodplain complexity.


Stimulation of human leukocytes by protein II+ gonococci is mediated by lectin-like gonococcal components.
We investigated the ability of carbohydrates, glycosidases, and concanavalin A to inhibit the stimulation of the human leukocyte oxidative burst by gonococci in the absence of serum. The gonococci used in this study contained protein II (P.II) outer membrane proteins, and neutrophil oxidative burst was measured by luminol-enhanced chemiluminescence (CL). The following carbohydrates inhibited CL induced by nonpiliated P.II+ gonococci: beta-D-glucosamine greater than N-acetylneuraminic acid (sialic acid) greater than mannose greater than alpha-methylmannoside greater than N-acetyl-beta-D-glucosamine greater than or equal to glucose greater than or equal to lactose. Fucose, galactose, or beta-D-galactosamine (all 100 mM) did not inhibit or slightly increased CL, indicating a specificity for the observed effects. Mannose and alpha-methylmannoside also inhibited induction of monocyte CL by P.II+ gonococci. Incubation of neutrophils with concanavalin A inhibited subsequent gonococcus-induced CL but not phorbol myristate acetate-induced CL. Treatment of neutrophils with alpha-mannosidase reduced subsequent gonococcus-induced CL greater than 99%, whereas such treatment of gonococci had no effect on their ability to induce neutrophil CL. Incubation of a P.IIb-containing variant of Neisseria gonorrhoeae FA1090 with anti-P.IIb monoclonal antibody inhibited subsequent stimulation of neutrophil CL in a dose-responsive manner, indicating a specific role for P.IIb in the stimulatory process. The data suggest that one or more lectin-like components on the surface of P.II+ gonococci mediate their ability to stimulate the oxidative burst of human phagocytes.


A differential risk assessment and decision model for Transarterial chemoembolization in hepatocellular carcinoma based on hepatic function.
The decision of transarterial chemoembolization (TACE) initiation and/or repetition remains challenging in patients with unresectable hepatocellular carcinoma (HCC). The aim was to develop a prognostic scoring system to guide TACE initiation/repetition. A total of 597 consecutive patients who underwent TACE as their initial treatment for unresectable HCC were included. We derived a prediction model using independent risk factors for overall survival (OS), which was externally validated in an independent cohort (n = 739). Independent risk factors of OS included Albumin-bilirubin (ALBI) grade, maximal tumor size, alpha-fetoprotein, and tumor response to initial TACE, which were used to develop a scoring system ("ASAR"). C-index values for OS were 0.733 (95% confidence interval [CI] = 0.570-0.871) in the derivation, 0.700 (95% CI = 0.445-0.905) in the internal validation, and 0.680 (95% CI = 0.652-0.707) in the external validation, respectively. Patients with ASAR< 4 showed significantly longer OS than patients with ASAR≥4 in all three datasets (all P < 0.001). Among Child-Pugh class B patients, a modified model without TACE response, i.e., "ASA(R)", discriminated OS with a c-index of 0.788 (95% CI, 0.703-0.876) in the derivation, and 0.745 (95% CI, 0.646-0.862) in the internal validation, and 0.670 (95% CI, 0.605-0.725) in the external validation, respectively. Child-Pugh B patients with ASA(R) < 4 showed significantly longer OS than patients with ASA(R) ≥ 4 in all three datasets (all P < 0.001). ASAR provides refined prognostication for repetition of TACE in patients with unresectable HCC. For Child-Pugh class B patients, a modified model with baseline factors might guide TACE initiation.


Independent clinical validation of a Canadian FRAX tool: fracture prediction and model calibration.
A FRAX model for Canada was constructed for prediction of osteoporotic and hip fracture risk using national hip fracture data with and without the use of femoral neck bone mineral density (BMD). Performance of this system was assessed independently in a large clinical cohort of 36,730 women and 2873 men from the Manitoba Bone Density Program database that tracks all clinical dual-energy X-ray absorptiometry (DXA) test results for the Province of Manitoba, Canada. Linkage with other provincial health databases allowed for the direct comparison of fracture risk estimates from the Canadian FRAX model with observed fracture rates to 10 years (549 individuals with incident hip fractures and 2543 with incident osteoporotic fractures). The 10-year Kaplan-Meier estimate for hip fractures in women was 2.7% [95% confidence interval (CI) 2.1-3.4%] with a predicted value of 2.8% for FRAX with BMD, and in men the observed risk was 3.5% (95% CI 0.8-6.2%) with predicted value of 2.9%. The 10-year estimate of osteoporotic fracture risk for all women was 12.0% (95% CI 10.8-13.4%) with a predicted value of 11.1% for FRAX with BMD, and in men, the observed risk was 10.7% (95% CI 6.6-14.9%) with a predicted value of 8.4%. Discrepancies were observed within some subgroups but generally were small. Fracture discrimination based on receiver operating characteristic curve analysis was comparable with published meta-analyses with area under the curve for osteoporotic fracture prediction of 0.694 (95% CI 0.684-0.705) for FRAX with BMD and for hip fractures 0.830 (95% CI 0.815-0.846), both of which were better than FRAX without BMD or BMD alone. Individual risk factors considered by FRAX made significant independent contributions to fracture prediction in one or more of the models. In conclusion, a Canadian FRAX tool calibrated on national hip fracture data generates fracture risk predictions that generally are consistent with observed fracture rates across a wide range of risk categories.


[The oculocardiac reflex in blepharoplasties].
The oculocardiac reflex (OCR) is a well-known phenomenon in ophthalmic surgery, but is rarely described in aesthetic blepharoplasty surgery. It was first mentioned in 1908 by Ascher and Dagnini. Since then, ophthalmologists and anaesthesiologists have regarded the onset of the oculocardiac reflex as a significant intraoperative problem, which is undermined by several case reports that describe dysrhythmias which have haved caused morbidity and death. Per definition the OCR is caused by ocular manipulation and involves intraoperative bradycardia by a change of 20 beats/minute compared to the preoperative heart rate or any dysrhythmia during the manipulation via a trigeminal-vagal-mediated reflex arc. Having operated on a 48-year-old, healthy woman in our clinic, who underwent a cardiac arrest during the blepharoplasty procedure, followed by a successful resuscitation, we investigated the onset of the OCR in our blepharoplasty patients within the last 3 years. The onset of the OCR was noted in 22 of 110 (20 %) blepharoplasty patients, mainly affecting younger, low-weighted patients operated under local anaesthesia. Awareness and treatment of this potentially life-threatening oculocardiac reflex are necessary. In most cases the onset of the reflex may be avoided by a gentle operation technique and by refraining from severe traction to the muscle or fat pad. The best treatment of a profound bradycardia caused by the OCR is to release tension to the muscle or fat pad in order to permit the heart rate to return to normal. Intraoperative monitoring is of utmost importance.


Preoperative skin preparation of the foot and ankle: bristles and alcohol are better.
The most efficient way to prepare the skin for foot and ankle surgery is unknown. In recent studies, >70% of aerobic bacterial cultures of specimens taken from the nail folds following skin preparation with povidone-iodine were positive. The goal of the current study was to determine the effect of isopropyl alcohol on the eradication of bacteria from the nails and skin of the normal foot and ankle. In addition, the effect of using a bristled brush rather than sponges to scrub the foot was investigated. Four skin-preparation techniques were studied in two sets of twenty-five volunteers. In phase I of the study, the right foot and ankle of each member of the first set of volunteers was prepared with method 1, which consisted of a two-stage povidone-iodine scrub and paint with use of soft sponges. The left foot and ankle was prepared with method 2, which consisted of method 1 as well as an additional pre-wash with 70% isopropyl alcohol. In phase II, the right foot and ankle of each member of the second set of volunteers was prepared with method 3, which consisted of a povidone-iodine scrub and paint with use of a bristled brush to scrub the foot. The left side was prepared with method 4, which consisted of an alcohol scrub and paint with use of a bristled brush to scrub the foot. At the end of the preparation process, specimens for aerobic bacterial cultures were obtained from the hallucal nail fold, interdigital web spaces, and anterior aspect of the ankle. Cultures were interpreted as positive or negative, and the results were also assessed quantitatively. The rates of positive cultures of the nail-fold specimens were 76% and 80% after methods 1 and 2 (soft sponges) and 76% and 12% after methods 3 and 4 (bristled brush). The reduction in the percentage of positive cultures with method 4 was highly significant (p < 0.001). Cultures of the specimens from the web spaces showed a significant difference in the rates of positive results between methods 1 and 2 (36% and 8%, p < 0.05) but no significant difference between methods 3 and 4 (12% and 0%, p = 0.25). The rates of positive cultures of specimens from the anterior aspect of the ankle were consistently low (< or = 4% for all methods). Quantitative analysis of positive cultures demonstrated significant reductions (p < 0.01) in heavy growth when bristled brushes had been used, both with povidone-iodine and isopropyl alcohol. The use of isopropyl alcohol and the use of a bristled brush both have beneficial effects on the skin-preparation process before foot and ankle surgery. In the current investigation, the most effective technique was the use of isopropyl alcohol in conjunction with scrubbing with a bristled brush. Merely washing the foot with alcohol-soaked sponges provided limited benefit to the web spaces only.