Nutrient availability and the RAS/cyclic AMP pathway both induce expression of ribosomal protein genes in Saccharomyces cerevisiae but by different mechanisms.
By differential hybridization, we identified a number of genes in Saccharomyces cerevisiae that are activated by addition of cyclic AMP (cAMP) to cAMP-depleted cells. A majority, but not all, of these genes encode ribosomal proteins. While expression of these genes is also induced by addition of the appropriate nutrient to cells starved for a nitrogen source or for a sulfur source, the pathway for nutrient activation of ribosomal protein gene transcription is distinct from that of cAMP activation: (i) cAMP-mediated transcriptional activation was blocked by prior addition of an inhibitor of protein synthesis whereas nutrient-mediated activation was not, and (ii) cAMP-mediated induction of expression occurred through transcriptional activation whereas nutrient-mediated induction was predominantly a posttranscriptional response. Transcriptional activation of the ribosomal protein gene RPL16A by cAMP is mediated through a upstream activation sequence element consisting of a pair of RAP1 binding sites and sequences between them, suggesting that RAP1 participates in the cAMP activation process. Since RAP1 protein decays during starvation for cAMP, regulation of ribosomal protein genes under these conditions may directly relate to RAP1 protein availability. These results define additional critical targets of the cAMP-dependent protein kinase, suggest a mechanism to couple ribosome production to the metabolic activity of the cell, and emphasize that nutrient regulation is independent of the RAS/cAMP pathway.


Retroviral capsid assembly: a role for the CA dimer in initiation.
In maturing retroviral virions, CA protein assembles to form a capsid shell that is essential for infectivity. The structure of the two folded domains [N-terminal domain (NTD) and C-terminal domain (CTD)] of CA is highly conserved among various retroviruses, and the capsid assembly pathway, although poorly understood, is thought to be conserved as well. In vitro assembly reactions with purified CA proteins of the Rous sarcoma virus (RSV) were used to define factors that influence the kinetics of capsid assembly and provide insights into underlying mechanisms. CA multimerization was triggered by multivalent anions providing evidence that in vitro assembly is an electrostatically controlled process. In the case of RSV, in vitro assembly was a well-behaved nucleation-driven process that led to the formation of structures with morphologies similar to those found in virions. Isolated RSV dimers, when mixed with monomeric protein, acted as efficient seeds for assembly, eliminating the lag phase characteristic of a monomer-only reaction. This demonstrates for the first time the purification of an intermediate on the assembly pathway. Differences in the intrinsic tryptophan fluorescence of monomeric protein and the assembly-competent dimer fraction suggest the involvement of the NTD in the formation of the functional dimer. Furthermore, in vitro analysis of well-characterized CTD mutants provides evidence for assembly dependence on the second domain and suggests that the establishment of an NTD-CTD interface is a critical step in capsid assembly initiation. Overall, the data provide clear support for a model whereby capsid assembly within the maturing virion is dependent on the formation of a specific nucleating complex that involves a CA dimer and is directed by additional virion constituents.


Population health assessment project: An innovative strategy for teaching principles of epidemiology.
Public health nurses (PHNs) use principles of epidemiology in their work with communities; however, teaching these concepts at the baccalaureate level can be challenging. To reinforce the epidemiologic concepts taught in the classroom, two nursing faculty in our baccalaureate nursing program developed a population health project that allows students to explore concepts of epidemiology within the context of a population health concern. This article describes how the project provides students with a realistic opportunity to research the health issue. As a part of the project, students collect and analyze data about the health concern at the county, state, and national level. The project also requires students to identify local agencies who provide services for people who are impacted by the health issue and describe their role and function within the community. Furthermore, students discuss the roles and responsibilities of nurses in managing the population health concern under investigation. In doing so, students recognize how epidemiology informs population level nursing care. If nursing students understand epidemiological concepts, they will be better prepared to educate the communities they serve after graduation when a population health crisis, such as the recent COVID-19 pandemic, occurs.


Tissue miR-193b as a Novel Biomarker for Patients with Ovarian Cancer.
BACKGROUND MiRNAs play important roles in regulating many fundamental biological processes. Deregulation of miRNAs is involved in the initiation and progression of cancer. MiR-193b is regarded as tumor suppressor in many types of cancers. However, the role of miR-193b in ovarian cancer is poorly understood. MATERIAL AND METHODS The expression level of miR-193b in ovarian cancer cell lines and ovarian cancer samples was evaluated using quantitative real-time reverse transcription-PCR (qRT-PCR). The ovarian cancer patients were categorized into a high miR-193b expression group and a low miR-193b expression group according to the median miR-193b expression level. The correlation between tissue miR-193b expression and the patients' clinicopathological factors, as well as survival, was also analyzed. RESULTS The results showed that the miR-193b expression was significantly down-regulated in ovarian cancer cell lines and tumor tissues compared with normal controls. In addition, tissue miR-193b expression was positively correlated with FIGO stage (P=0.001), histological grade (P=0.032), ascites (P=0.019), lymph node metastasis (P=0.003), and tumor size (P=0.041). Among 116 patients with ovarian cancer examined, the 5-year overall survival (OS) rates were 62.5% and 22.01% in patients with high and low miR-193b expression, respectively (P=0.003). Multivariate analysis showed that tissue miR-193b is an independent prognostic factor in patients with ovarian cancer (HR=4.219; P=0.015). CONCLUSIONS Reduction of miR-193b was found in ovarian cancer and its lower expression was associated with poorer prognosis. Tissue miR-193b showed potential as novel biomarker for ovarian cancer.


Vitamin K2 supplementation improves hip bone geometry and bone strength indices in postmenopausal women.
Vitamin K mediates the synthesis of proteins regulating bone metabolism. We have tested whether high vitamin K(2) intake promotes bone mineral density and bone strength. Results showed that K(2) improved BMC and femoral neck width, but not DXA-BMD. Hence high vitamin K(2) intake may contribute to preventing postmenopausal bone loss. Vitamin K is involved in the synthesis of several proteins in bone. The importance of K vitamins for optimal bone health has been suggested by population-based studies, but intervention studies with DXA-BMD as a clinical endpoint have shown contradicting results. Unlike BMC, DXA-BMD does not take into account the geometry (size, thickness) of bone, which has an independent contribution to bone strength and fracture risk. Here we have tested whether BMC and femoral neck width are affected by high vitamin K intake. A randomized clinical intervention study among 325 postmenopausal women receiving either placebo or 45 mg/day of vitamin K(2) (MK-4, menatetrenone) during three years. BMC and hip geometry were assessed by DXA. Bone strength indices were calculated from DXA-BMD, femoral neck width (FNW) and hip axis length (HAL). K(2) did not affect the DXA-BMD, but BMC and the FNW had increased relative to placebo. In the K(2)-treated group hip bone strength remained unchanged during the 3-year intervention period, whereas in the placebo group bone strength decreased significantly. Vitamin K(2) helps maintaining bone strength at the site of the femoral neck in postmenopausal women by improving BMC and FNW, whereas it has little effect on DXA-BMD.


Lipid oxidation fuels recovery from exhaustive exercise in white muscle of rainbow trout.
The oxidative utilization of lipid and carbohydrate was examined in white muscle of rainbow trout (Oncorhynchus mykiss) at rest, immediately after exhaustive exercise, and for 32-h recovery. In addition to creatine phosphate and glycolysis fueling exhaustive exercise, near maximal activation of pyruvate dehydrogenase (PDH) at the end of exercise points to oxidative phosphorylation of carbohydrate as an additional source of ATP during exercise. Within 15 min postexercise, PDH activation returned to resting values, thus sparing accumulated lactate from oxidation. Glycogen synthase activity matched the rate of glycogen resynthesis and represented near maximal activation. Decreases in white muscle free carnitine, increases in long-chain fatty acyl carnitine, and sustained elevations of acetyl-CoA and acetyl carnitine indicate a rapid utilization of lipid to supply ATP for recovery. Increases in malonyl-CoA during recovery suggest that malonyl-CoA may not regulate carnitine palmitoyltransferase-1 in trout muscle during recovery, but instead it may act to elongate short-chain fatty acids for mitochondrial oxidation. In addition, decreases in intramuscular triacylglycerol and in plasma nonesterified fatty acids indicate that both endogenous and exogenous lipid fuels may be oxidized during recovery.


Disability as an outcome in MS clinical trials.
Inferences about long-term effects of therapies in multiple sclerosis (MS) have been based on surrogate markers studied in short-term trials. Preventing progressive disability is the key therapeutic goal but there remains no validated definition for its measurement in a trial context. Meanwhile, MS trials continue to shorten and to depend on unvalidated surrogates. Since there have been no treatment claims for improving unremitting disability, worsening of disability in the placebo/control arm must occur for effectiveness on this outcome to be shown. We examined widely-used clinical surrogates of long-term disability progression in individual patients with MS within a unique database from the placebo arms of 31 randomized clinical trials. Detection of treatment effects in secondary progressive MS trials is undermined by noise in disability measurement. Whereas existing measures can be partially validated in secondary progressive MS, this is not the case in relapsing-remitting MS. Here, examination of widely used definitions of treatment failure demonstrated that disability progression was no more likely than similarly defined improvement. Existing definitions of disease progression in short-term intervention trials in relapsing-remitting patients reflect random variation, measurement error, and remitting relapses. Clinical surrogates of unremitting disability used in trials of relapsing-remitting multiple sclerosis cannot be validated. Trials have been too short or degrees of disability change too small to measure the key outcomes. These analyses highlight the difficulty in determining effectiveness of therapy in chronic diseases.


Partial posteromedial olecranon resection: a kinematic study.
The posteromedial aspect of the olecranon process is a site of impingement and subsequent osteophyte development in throwing athletes. Treatment with débridement, with resection of osteophytes and varying amounts of normal olecranon bone, is common. We found no reports in the literature concerning the effects of resecting different amounts of normal bone from the posteromedial aspect of the olecranon. We hypothesized that excessive resection would increasingly alter elbow kinematics and that an optimum amount of olecranon resection could be identified. We investigated the kinematic effects of increasing valgus and varus torques and posteromedial olecranon resections, in twelve cadaveric elbows, with use of an electromagnetic tracking device. Two valgus and two varus torques were applied, and three sequential resections were performed in 3-mm steps from 0 mm to 9 mm. Statistical analyses included paired t tests, 95% confidence intervals, a one-factor analysis of variance with repeated measures, and a post hoc test when significance was established. Sequential partial resection of the posteromedial aspect of the olecranon resulted in stepwise increases in valgus angulation with valgus torque. Clear differences were seen at each level of resection. A pattern of increased valgus angulation also was seen in association with increased valgus torque. Increased valgus torque resulted in a trend toward increased axial internal rotation of the ulna, whereas increased osseous resection resulted in a decrease in the absolute degree of internal rotation or, in some specimens, increased external rotation. Although no single critical amount of olecranon resection was identified, valgus angulation of the elbow increased in association with all resections, with a marked increase occurring in association with a 9-mm resection. Our findings challenge the rationale of removing any amount of normal olecranon bone in throwing athletes as doing so may increase strain on the medial collateral ligament. The implications for the professional throwing athlete are important, and we recommend that bone removal from the olecranon be limited to osteophytes, without the removal of normal bone.


Aspects of early arthritis. What determines the evolution of early undifferentiated arthritis and rheumatoid arthritis? An update from the Norfolk Arthritis Register.
Over 3500 patients with recent onset inflammatory polyarthritis (IP) have been recruited by the Norfolk Arthritis Register (NOAR) since 1990. Longitudinal data from this cohort have been used to examine the prevalence and predictors of remission, functional disability, radiological outcome, cardiovascular mortality and co-morbidity and the development of non-Hodgkin's lymphoma. Rheumatoid factor titre, high baseline C-reactive protein and high baseline HAQ score are all predictors of a poor outcome. There is a strong association between possession of the shared epitope and the development of erosions. Patients who satisfy the American College of Rheumatology criteria for rheumatoid arthritis (RA) have a worse prognosis than those who do not. However, it appears that these patients are a poorly defined subset of all those with IP rather than having an entirely separate disease entity. New statistical techniques offer exciting possibilities for using longitudinal datasets such as NOAR to explore the long-term effects of treatment in IP and RA.


Use of community engagement strategies to increase research participation in practice-based research networks (PBRNs).
Practice-based research networks (PBRNs) are increasingly encouraged to use community engagement approaches. The extent to which PBRNs engage clinic and community partners in strategies to recruit and retain participants from their local communities (specifically racial/ethnic communities) is the focus of this study. The design was a cross-sectional survey of PBRN directors in the United States. Survey respondents indicated whether their research network planned for, implemented, and has capacity for activities that engage clinic and community partners in 7 recommended strategies organized into study phases, called the cycle of trust. The objectives of the national survey were to (1) describe the extent to which PBRNs across the United States routinely implement the strategies recommended for recruiting diverse patient groups and (2) identify factors associated with implementing the recommended strategies. The survey response rate was 63%. Activities that build trust often are used more with clinic partners than with community partners. PBRNs that adopt engagement strategies when working with clinic and community partners have less difficulty in recruiting diverse populations. Multivariate analysis showed that the targeting racial/ethnic communities for study recruitment, Clinical and Translational Science Award affiliation, and planning to use community engagement strategies were independent correlates of PBRN implementation of the recommended strategies. PBRNs that successfully engage racial/ethnic communities as research partners use community engagement strategies. New commitments are needed to support PBRN researchers in developing relationships with the communities in which their patients live. Stable PBRN infrastructure funding that appreciates the value of maintaining community engagement between funded studies is critical to the research enterprise that values translating research findings into generalizable care models for patients in the community.


Cognitive-Behavioral and Emotion-Focused Couple Therapy: Similarities and Differences.
Couples and families often seek therapy to deal with relational distress, which is a result of external or internal factors of the relationship. Two approaches are acknowledged to be most effective in dealing with relationship distress or psychological disorders in couples: (a) cognitive behavioral couple therapy with new directions (CBCT) and (b) emotion-focused couple therapy (EFCT). In this article we investigate how much CBCT and EFCT really differ with regard to working with emotions, which is claimed to be a major focus of EFCT, and whether there exist significant differences in efficacy between these two approaches. This article critically reviews the theoretical background, process, techniques and outcomes associated with CBCT and EFCT in an effort to challenge the assumptions noted above. There is no evidence that EFCT is more emotion-focused than CBCT. Both approaches were repeatedly examined with RCT studies with follow-ups. In sum, no significant differences in effect size were found between CBCT and EFCT. CBCT and EFCT are both effective in reducing couples' distress.


Application of forward osmosis membrane in nanofiltration mode to treat reverse osmosis concentrate from wastewater reclamation plants.
Reverse osmosis concentrate (ROC) from wastewater reclamation plants have high concentrations of organic and inorganic compounds, which have to be removed before its disposal. Forward osmosis (FO) and nanofiltration (NF) membranes were tested to treat the ROC for possible water reuse. This research investigated the combined and individual influence of organic and inorganic matter on the fouling of NF and FO membranes. The results revealed that the NF membrane removed most of the organic compounds and some inorganics. The study further highlighted that the FO membrane at NF mode removed the majority of the inorganic compounds and some organics from the ROC. A pretreatment of granulated activated carbon (GAC) adsorption removed 90% of the organic compounds from ROC. In addition, GAC adsorption and acid pretreatment of ROC improved the net water permeate flux by 17% when an FO membrane was used in the NF system. Acid treatment (by bringing the pH down to 5) helped to remove inorganic ions. Therefore, the resultant permeate can be recycled back to the RO water reclamation plant to improve its efficiency.


Guanosine 5'-O-(3-thiotriphosphate) and cations regulate melatonin receptors, and melatonin inhibits cyclic AMP production in the spinal cord.
Effects of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and cations on 2-[125I]iodomelatonin binding were investigated in membrane preparations of the chicken spinal cord. At concentrations of 10 and 50 mumol/l, GTP gamma S dose-dependently increased (p < 0.05) the equilibrium dissociation constant (Kd) and depressed (p < 0.05) the maximum number of binding sites (Bmax). Na+ at a concentration of 125 mmol/l significantly increased (p < 0.05) the Kd and decreased (p < 0.05) the Bmax, and Mg2+ (2.5 mmol/l) significantly increased (p < 0.05) the Bmax without changes in Kd. In addition, Na+ and Mg2+ affected the interactions of GTP gamma S with melatonin receptors. In the spinal cord explants, melatonin (10 nmol/l) attenuated forskolin-stimulated cyclic AMP production by 53.1%, and preincubation with pertussis toxin abolished this effect of melatonin. These results suggest that the melatonin receptors in the chicken spinal cord are linked to its second messenger via a pertussis-toxin-sensitive guanine-nucleotide-binding protein, and that cations modulate these receptors. Our studies further support a previous hypothesis that melatonin exerts a direct action on spinal cord functions.


The antidepressive effects of exercise: a meta-analysis of randomized trials.
Several meta-analyses examining the effects of exercise on depression have been criticized for including studies of poor methodological integrity. More recent meta-analyses addressed the most common criticism by including only randomized control trials; however, these analyses suffer from incomplete literature searches and lack of moderating variable analyses. Using a more extensive search procedure, the current meta-analysis examines the effects of exercise on depressive symptoms in 58 randomized trials (n = 2982). An overall effect size of -0.80 indicates participants in the exercise treatment had significantly lower depression scores than those receiving the control treatment. This frac34; SD advantage represents level 1, Grade A evidence for the effects of exercise upon depression. Analysis of moderating variables examined the influence of population characteristics, exercise characteristics and methodological characteristics. Examination of clinical significance in 16 trials with clinically depressed patients found 9 of 16 exercise treatment groups were classified as 'recovered' at post-treatment, with another three groups classified as 'improved'. Analysis showed dropout rates for the exercise treatment were similar to those found in psychotherapeutic and drug interventions.


Total body potassium in long-term frusemide therapy: is potassium supplementation necessary?
Measurements of total body potassium (T.B.K.) were made by whole-body counting in four groups of patients receiving oral frusemide for one year. Patients in group 1 had essential hypertension and normal renal function and received 40 mg frusemide daily without potassium supplements. Patients in group 2 were similar but received oral potassium supplements for the first four months of treatment. Patients in group 3 had hypertension associated with renal disease and received 120 mg frusemide daily without potassium supplements. Patients in group 4 also had hypertension and renal impairment and in addition to 120 mg frusemide daily they received oral potassium supplements for four months. No evidence of depletion of T.B.K. was found in any of the groups after continuous treatment with frusemide for one year. It is questioned whether potassium supplementation in long term diuretic therapy with frusemide is necessary unless there is evidence of pre-existing potassium depletion or of some other factor such as cardiac failure, cirrhosis of the liver, or the nephrotic syndrome.


Low threshold power density for the generation of frequency up-converted pulses in bismuth glass by two crossing chirped femtosecond pulses.
We investigated the generation of frequency up-converted femtosecond laser pulses by nondegenerate cascaded four-wave mixing (CFWM) in a bismuth-oxide glass (BI glass). Broad-bandwidth light pulses with different propagation directions were simultaneously obtained by using two small-angle crossing femtosecond laser pulses in BI glass. Experimental results show that the threshold power density for the generation of broad-bandwidth femtosecond pulses in BI glass is one order of magnitude lower than that in fused silica.


Theoretical restrictions on longest implicit time scales in Markov state models of biomolecular dynamics.
Markov state models (MSMs) have been widely used to analyze computer simulations of various biomolecular systems. They can capture conformational transitions much slower than an average or maximal length of a single molecular dynamics (MD) trajectory from the set of trajectories used to build the MSM. A rule of thumb claiming that the slowest implicit time scale captured by an MSM should be comparable by the order of magnitude to the aggregate duration of all MD trajectories used to build this MSM has been known in the field. However, this rule has never been formally proved. In this work, we present analytical results for the slowest time scale in several types of MSMs, supporting the above rule. We conclude that the slowest implicit time scale equals the product of the aggregate sampling and four factors that quantify: (1) how much statistics on the conformational transitions corresponding to the longest implicit time scale is available, (2) how good the sampling of the destination Markov state is, (3) the gain in statistics from using a sliding window for counting transitions between Markov states, and (4) a bias in the estimate of the implicit time scale arising from finite sampling of the conformational transitions. We demonstrate that in many practically important cases all these four factors are on the order of unity, and we analyze possible scenarios that could lead to their significant deviation from unity. Overall, we provide for the first time analytical results on the slowest time scales captured by MSMs. These results can guide further practical applications of MSMs to biomolecular dynamics and allow for higher computational efficiency of simulations.


Electrostatic Control of Polymorphism in Charged Amphiphile Assemblies.
Stimuli-induced structural transformations of molecular assemblies in aqueous solutions are integral to nanotechnological applications and biological processes. In particular, pH responsive amphiphiles as well as proteins with various degrees of ionization can reconfigure in response to pH variations. Here, we use in situ small and wide-angle X-ray scattering (SAXS/WAXS), transmission electron microscopy (TEM), and Monte Carlo simulations to show how charge regulation via pH induces morphological changes in the assembly of a positively charged peptide amphiphile (PA). Monte Carlo simulations and pH titration measurements reveal that ionic correlations in the PA assemblies shift the ionizable amine pK ∼ 8 from pK ∼ 10 in the lysine headgroup. SAXS and TEM show that with increasing pH, the assembly undergoes spherical micelle to cylindrical nanofiber to planar bilayer transitions. SAXS/WAXS reveal that the bilayer leaflets are interdigitated with the tilted PA lipid tails crystallized on a rectangular lattice. The details of the molecular packing in the membrane result from interplay between steric and van der Waals interactions. We speculate that this packing motif is a general feature of bilayers comprised of amphiphilic lipids with large ionic headgroups. Overall, our studies correlate the molecular charge and the morphology for a pH-responsive PA system and provide insights into the Å-scale molecular packing in such assemblies.


Physical activity, but not sedentary time, influences bone strength in late adolescence.
Physical activity is essential for optimal bone strength accrual, but we know little about interactions between physical activity, sedentary time, and bone outcomes in older adolescents. Physical activity (by accelerometer and self-report) positively predicted bone strength and the distal and midshaft tibia in 15-year-old boys and girls. Lean body mass mediated the relationship between physical activity and bone strength in adolescents. To examine the influence of physical activity (PA) and sedentary time on bone strength, structure, and density in older adolescents. We used peripheral quantitative computed tomography to estimate bone strength at the distal tibia (8% site; bone strength index, BSI) and tibial midshaft (50% site; polar strength strain index, SSIp) in adolescent boys (n = 86; 15.3 ± 0.4 years) and girls (n = 106; 15.3 ± 0.4 years). Using accelerometers (GT1M, Actigraph), we measured moderate-to-vigorous PA (MVPAAccel), vigorous PA (VPAAccel), and sedentary time in addition to self-reported MVPA (MVPAPAQ-A) and impact PA (ImpactPAPAQ-A). We examined relations between PA and sedentary time and bone outcomes, adjusting for ethnicity, maturity, tibial length, and total body lean mass. At the distal tibia, MVPAAccel and VPAAccel positively predicted BSI (explained 6-7% of the variance, p < 0.05). After adjusting for lean mass, only VPAAccel explained residual variance in BSI. At the tibial midshaft, MVPAAccel, but not VPAAccel, positively predicted SSIp (explained 3% of the variance, p = 0.01). Lean mass attenuated this association. MVPAPAQ-A and ImpactPAPAQ-A also positively predicted BSI and SSIp (explained 2-4% of the variance, p < 0.05), but only ImpactPAPAQ-A explained residual variance in BSI after accounting for lean mass. Sedentary time did not independently predict bone strength at either site. Greater tibial bone strength in active adolescents is mediated, in part, by lean mass. Despite spending most of their day in sedentary pursuits, adolescents' bone strength was not negatively influenced by sedentary time.


Cartilage oligomeric matrix protein specific antibodies are pathogenic.
Cartilage oligomeric matrix protein (COMP) is a major non-collagenous component of cartilage. Earlier, we developed a new mouse model for rheumatoid arthritis using COMP. This study was undertaken to investigate the epitope specificity and immunopathogenicity of COMP-specific monoclonal antibodies (mAbs). B cell immunodominant regions on the COMP molecule were measured with a novel enzyme-linked immunosorbent assay using mammalian expressed full-length mouse COMP as well as a panel of recombinant mouse COMP fragments. 18 mAbs specific to COMP were generated and the pathogenicity of mAbs was investigated by passive transfer experiments. B cell immunodominant epitopes were localized within 4 antigenic domains of the COMP but with preferential response to the epidermal growth factor (EGF)-like domain. Some of our anti-COMP mAbs showed interactions with the native form of COMP, which is present in cartilage and synovium. Passive transfer of COMP-specific mAbs enhanced arthritis when co-administrated with a sub-arthritogenic dose of a mAb specific to collagen type II. Interestingly, we found that a combination of 5 COMP mAbs was capable of inducing arthritis in naive mice. We have identified the specificities of mAbs to COMP and their contribution to the development of arthritis. These findings will further improve our understanding of the autoantibody mediated immunopathologies occurring widely in rheumatoid arthritis (RA), as well as in other autoimmune disorders.


iDS372, a Phenotypically Reconciled Model for the Metabolism of Streptococcus pneumoniae Strain R6.
A high-quality GSM model for Streptococcus pneumoniae R6 model strain (iDS372), comprising 372 genes and 529 reactions, was developed. The construction of this model involved performing a genome-wide reannotation to identify the metabolic capacity of the bacterium. A reaction representing the abstraction of the biomass composition was reconciled from several studies reported in the literature and previous models, and included in the model. The final model comprises two compartments and manifold automatically generated gene rules. The validation was performed with experimental data from recent studies, regarding the usability of carbon sources, the effect of the presence of oxygen, and the requirement of amino acids for growth. This model can be used to better understand the metabolism of this major pathogen, provide clues regarding new drug targets, and eventually design strategies for fighting infections by these bacteria.


A human factors investigation of medication alerts: barriers to prescriber decision-making and clinical workflow.
Computerized medication alerts (e.g., drug-drug interaction alerts), which are intended to protect patient safety, should also be designed to support prescriber workflow. However, relatively few studies have examined the use of medication alerts during patient care processes. To assess barriers associated with the use of medication alerts, we directly observed medication prescribing during routine patient care. Prescribers (physicians, pharmacists, and nurse practitioners) were recruited from five outpatient primary care clinics at a major Midwestern Veterans Affairs Medical Center (VAMC). A total of 199 alerts were observed across 91 patients and 20 prescribers during normal patient care tasks. Through inductive qualitative analysis, we identified 15 barriers associated with medication alerts; herein, we describe five of the key barriers in detail. Results may be used to create alert redesigns, which have the potential to more fully support clinical workflow, prescriber decision-making, and patient safety.


Hyperbaric Oxygen Therapy: Side Effects Defined and Quantified.
Significance: Hyperbaric oxygen therapy (HBOT) is an important advanced therapy in the treatment of problem wounds, including diabetic foot ulcers and late effect radiation injury. HBOT remains among the safest therapies used today. Nonetheless, there are side effects associated with HBOT. It is important for providers to be able to identify, understand, and quantify these side effects for prevention, management, and informed consent. Recent Advances: The past two decades have seen significant advancements in our understanding of the underlying mechanisms of HBOT. This has led to a better understanding of the underlying reason for clinical benefit. It has also led to a better understanding of its side effects. Moreover, more recent literature allows for better quantification of these side effects. This review will highlight these side effects. Critical Issues: Wound healing in the case of problem nonhealing wounds requires the use of various advanced treatment modalities, including HBOT. HBOT has been shown to significantly improve healing rates in certain problem wounds, including advanced diabetic foot ulcers and late effect radiation injury. It is provided in a variety of clinical settings by providers with varying levels of expertise. It is important for those providing this therapy to understand the potential side effects. Future Directions: Research in HBOT has led to significant advancements in the area of wound healing. At the same time, there remains a variety of treatment protocols used at different institutions. It is important to quantify risk and benefit at different treatment pressures and times to better standardize treatment and improve patient care.


Mechanisms of action of disease-modifying agents and brain volume changes in multiple sclerosis.
Disease-modifying agents (DMAs), including interferon beta (IFNbeta) and glatiramer acetate (GA), are the mainstays of long-term treatment of multiple sclerosis (MS). Other potent anti-inflammatory agents like natalizumab and different types of chemotherapeutics are increasingly being used for treatment of MS, particularly in patients with breakthrough disease activity. Brain volume (BV) loss occurs early in the disease process, accelerates over time, and may be only partially affected by DMA therapy. Low-dose, low frequency IFNbeta administered once weekly and GA appear to partially reduce BV decline over the second and third years of treatment. High dose, high frequency IFNbeta demonstrated no clear effect on BV loss during this time period. Current evidence suggests that changes in BV after immunoablation may not be due entirely to the resolution of edema but may be related to potential chemotoxicity of high dose cyclophosphamide. Natalizumab reduces the development of BV decline in the second and third years of treatment. IV immunoglobulin showed a positive effect on decelerating BV reduction in relapsing and advanced stages of MS. These differences between DMAs may be explained by the extent of their therapeutic effects on inflammation and on the balance between inhibition or promotion of remyelination and neuronal repair in the CNS. We described the mechanisms of action by which DMAs induce accelerated, non-tissue-related BV loss (pseudoatrophy) in the short term but, in the long run, may still potentially lead to permanent BV decline. The effects of corticosteroid therapy on changes in BV in patients with MS help clarify the mechanisms through which potent anti-inflammatory treatments may prevent, stabilize, or induce BV loss.


Micro-habitat distribution drives patch quality for sub-tropical rocky plateau amphibians in the northern Western Ghats, India.
The importance of patch quality for amphibians is frequently overlooked in distribution models. Here we demonstrate that it is highly important for the persistence of endemic and endangered amphibians found in the threatened and fragile ecosystems that are the rocky plateaus in Western Maharashtra, India. These plateaus are ferricretes of laterite and characterise the northern section of the Western Ghats/Sri Lanka Biodiversity Hotspot, the eighth most important global hotspot and one of the three most threatened by population growth. We present statistically supported habitat associations for endangered and data-deficient Indian amphibians, demonstrating significant relationships between individual species and their microhabitats. Data were collected during early monsoon across two seasons. Twenty-one amphibian taxa were identified from 14 lateritic plateaus between 67 and 1179m above sea level. Twelve of the study taxa had significant associations with microhabitats using a stepwise analysis of the AICc subroutine (distLM, Primer-e, v7). Generalist taxa were associated with increased numbers of microhabitat types. Non-significant associations are reported for the remaining 9 taxa. Microhabitat distribution was spatially structured and driven by climate and human activity. Woody plants were associated with 44% of high-elevation taxa. Of the 8 low-elevation taxa 63% related to water bodies and 60% of those were associated with pools. Rock size and abundance were important for 33% of high elevation specialists. Three of the 4 caecilians were associated with rocks in addition to soil and stream presence. We conclude the plateaus are individualistic patches whose habitat quality is defined by their microhabitats within climatic zones.


Adults with severe reading and learning difficulties: a challenge for the family physician.
An estimated 40 to 44 million adults living in the United States have severe difficulty reading, writing, spelling, and doing arithmetic. These deficiencies interfere with their receiving adequate health care. Many of these adults have reading or other learning disabilities that further compromise their ability to understand their medical conditions and to participate fully in their own care. The literature on the cognitive and effective characteristics of adults with reading and learning disabilities was searched using the MEDLINE, PsychLIT, and ERIC databases. This literature is reviewed with an emphasis placed on how these characteristics might challenge a family physician's ability to provide optimal patient care, and what can be done to meet these challenges. Illustrative case vignettes of adults with these disabilities are described. The cognitive and affective characteristics of this patient population make it difficult for the family physician to provide optimal medical services. Suggestions are given to make medical care more accessible and appropriate for these patients.


Patient outcomes and length of hospital stay after radical prostatectomy for prostate cancer: analysis of hospital episodes statistics for England.
To investigate the morbidity and mortality after radical prostatectomy (RP) in relation to the numbers of RPs carried out at individual hospitals, as recent studies of complex surgery report worse outcomes in low-volume hospitals, and there has been a large increase in RPs for localized prostate cancer. We analysed hospital episode statistics data for all 18 027 RPs in English National Health Service hospitals between 1997 and 2004. In multivariate analysis, there was a U-shaped association of hospital volume with mortality (P for nonlinear trend, 0.004), but this finding was based on only 59 (0.3%) deaths. The mean length of stay was 6 days and decreased by 2.96% (95% confidence interval, CI, 1.98-3.92; P < 0.001) per quintile increase in hospital volume. In all, 16.1% of men had 30-day in-hospital complications; 20.3% were readmitted with complications within a year. The odds of 30-day in-hospital wound/bleeding complications decreased by 6% (95% CI 1-11; P = 0.02), and miscellaneous medical complications decreased by 10% (0-19; P = 0.04) per increase in hospital volume quintile. For re-admissions within a year, the hazard of vascular complications decreased by 15% (6-22; P = 0.001), wound/bleeding complications decreased by 8% (2-13; P = 0.01) and genitourinary complications decreased by 5% (2-8; P = 0.002), per increase in hospital volume quintile. In men undergoing RP the length of hospital stay and rates of some short- and long-term postoperative complications afterward are lower in high-volume hospitals. The magnitudes of these effects on the outcomes studied may be too small and inconsistent to indicate a policy of selective referral to high-volume hospitals. Quality of life and oncological outcomes, however, could not be investigated in this dataset.


Perceptual learning as a possible new approach for remediation and prevention of developmental dyslexia.
Learning to read is extremely difficult for about 10% of children across cultures because they are affected by developmental dyslexia (DD). According to the dominant view, DD is considered an auditory-phonological processing deficit. However, accumulating evidence from developmental and clinical vision science, suggests that the basic cross-modal letter-to-speech sound integration deficit in DD might arise from a mild atypical development of the magnocellular-dorsal pathway which also contains the main fronto-parietal attentional network. Letters have to be precisely selected from irrelevant and cluttering letters by rapid orienting of visual attention before the correct letter-to-speech sound integration applies. Our aim is to review the literature supporting a possible role of perceptual learning (PL) in helping to solve the puzzle called DD. PL is defined as improvement of perceptual skills with practice. Based on the previous literature showing how PL is able to selectively change visual abilities, we here propose to use PL to improve the impaired visual functions characterizing DD and, in particular, the visual deficits that could be developmentally related to an early magnocellular-dorsal pathway and selective attention dysfunction. The crucial visual attention deficits that are causally linked to DD could be, indeed, strongly reduced by training the magnocellular-dorsal pathway with the PL, and learning to read for children with DD would not be anymore such a difficult task. This new remediation approach - not involving any phonological or orthographic training - could be also used to develop new prevention programs for pre-reading children at DD risk.


Cancer-Related Cognitive Impairment in Breast Cancer Patients: Influences of Psychological Variables.
Approximately 3.1 million women in the US are living with breast cancer and up to 75% of these women experience cancer-related cognitive impairment (CRCI). CRCI is described as impairments in memory, verbal fluency, thought processes, and attention span. Despite the high prevalence of breast cancer, only a few studies have been published on CRCI and most of these studies primarily focused on its pathophysiological mechanism. However, recent evidence has demonstrated that breast cancer patients with CRCI are more likely to have high level of psychologic distress, suggesting a possible relationship between CRCI and psychologic distress. This review aims to examine existing literature that describes CRCI in relation to psychological distress among breast cancer patients. One thousand four hundred and ninety-eight articles were searched using PubMed, CINAHL, and PsycINFO. Thirteen studies met inclusion criteria, and one article was additionally pulled from article reference lists. Of these19 studies, psychologic distress has been operationalized in varied ways such as anxiety (n = 3), depression (n = 2), both anxiety and depression (n = 4), stress (n = 4), worry (n = 2), mental fatigue (n = 1), and undefined psychological distress (n = 2). Except for six studies designed as a longitudinal study, the rest of studies used a cross-sectional design. Twelve studies used both subjective and objective measures to assess cognitive function. We found that the patients with high psychological distress displayed lower performance on cognitive function tests. Our finding indicates that psychological variables contributed to CRCI that breast cancer patients experienced. Areas for further investigation are proposed that will advance the care of breast cancer patients with CRCI.


Transport activity of MCT1 expressed in Xenopus oocytes is increased by interaction with carbonic anhydrase.
Injection of carbonic anhydrase isoform II (CA) into Xenopus frog oocytes increased the rate of H+ flux via the rat monocarboxylate transporter isoform 1 (MCT1) expressed in the oocytes. MCT1 activity was assessed by changes of intracellular H+ concentration measured by pH-selective microelectrodes during application of lactate. CA-induced augmentation of the rate of H+ flux mediated by MCT1 was not inhibited by ethoxyzolamide (10 microM) and did not depend on the presence of added CO2/HCO3- but was suppressed by injection of an antibody against CA. Deleting the C terminus of the MCT1 greatly reduced its transport rate and removed transport facilitation by CA. Injected CA accelerated the CO2/HCO3(-)-induced acidification severalfold, which was blocked by ethoxyzolamide and was independent of MCT1 expression. Mass spectrometry confirmed activity of CA as injected into the frog oocytes. With pulldown assays we demonstrated a specific binding of CA to MCT1 that was not attributed to the C terminus of MCT1. Our results suggest that CA enhances MCT1 transport activity, independent of its enzymatic reaction center, presumably by binding to MCT1.


Iron mineralogy and uranium-binding environment in the rhizosphere of a wetland soil.
Wetlands mitigate the migration of groundwater contaminants through a series of biogeochemical gradients that enhance multiple contaminant-binding processes. The hypothesis of this study was that wetland plant roots contribute organic carbon and release O2 within the rhizosphere (plant-impact soil zone) that promote the formation of Fe(III)-(oxyhydr)oxides. In turn, these Fe(III)-(oxyhydr)oxides stabilize organic matter that together contribute to contaminant immobilization. Mineralogy and U binding environments of the rhizosphere were evaluated in samples collected from contaminated and non-contaminated areas of a wetland on the Savannah River Site in South Carolina. Based on Mössbauer spectroscopy, rhizosphere soil was greatly enriched with nanogoethite, ferrihydrite-like nanoparticulates, and hematite, with negligible Fe(II) present. X-ray computed tomography and various microscopy techniques showed that root plaques were tens-of-microns thick and consisted of highly oriented Fe-nanoparticles, suggesting that the roots were involved in creating the biogeochemical conditions conducive to the nanoparticle formation. XAS showed that a majority of the U in the bulk wetland soil was in the +6 oxidation state and was not well correlated spatially to Fe concentrations. SEM/EDS confirm that U was enriched on root plaques, where it was always found in association with P. Together these findings support our hypothesis and suggest that plants can alter mineralogical conditions that may be conducive to contaminant immobilization in wetlands.


Frequency of nonspecific clinical signs in dogs with separation anxiety, thunderstorm phobia, and noise phobia, alone or in combination.
To determine the frequency of nonspecific clinical signs in dogs with separation anxiety, thunderstorm phobia, noise phobia, or any combination of these conditions and determine whether these conditions are associated in dogs. Case series. 141 dogs. Diagnoses were established using specific criteria. Owners of dogs completed a questionnaire on how frequently their dogs exhibited destructive behavior, urination, defecation, vocalization, and salivation when the owners were absent and the types and frequency of reactions to thunderstorms, fireworks, and other noises. Associations of the 3 conditions and of various nonspecific clinical signs within and between diagnoses were nonrandom. The probability that a dog would have separation anxiety given that it had noise phobia was high (0.88) and approximately the same as the probability it would have separation anxiety given that it had thunderstorm phobia (0.86). However, the probability that a dog would have noise phobia given that it had separation anxiety (0.63) was higher than the probability that it would have thunderstorm phobia given that it had separation anxiety (0.52). The probability that a dog would have noise phobia given that it had thunderstorm phobia (0.90) was not equivalent to the converse (0.76). Results suggested that dogs with any of these conditions should be screened for the others. Interactions among these conditions are important in the assessment and treatment of dogs with > 1 of these conditions. Responses to noise were different from those to thunderstorms, possibly because of the unpredictability and uncertainty of thunderstorms.


Modulation of extracellular matrix metabolism in rabbit articular chondrocytes and human rheumatoid synovial cells by the non-steroidal anti-inflammatory drug etodolac. II: Glycosaminoglycan synthesis.
The effect of Etodolac on glycosaminoglycan (GAG) synthesis by human synovial cells and rabbit articular chondrocytes in culture was studied at doses ranging from 0.01 to 10 micrograms/ml. In chondrocyte cultures, short-term exposure to Etodolac decreased the total amount of GAGs (mainly chondroitin sulfate) affecting essentially the cell layer-associated fraction. When cells were incubated with Interleukin-1 (IL-1), Etodolac still exerted its inhibiting effect on GAG synthesis. Long-term exposure of chondrocytes to Etodolac caused a diminution of the total GAG synthesis for the highest concentration studied (1 micrograms/ml). IL-1 alone or in combination with Etodolac decreased the synthesis of GAGs suggesting that pretreatment with Etodolac cannot prevent the action of IL-1. In synoviocyte cultures, the drug caused an inhibition of the GAG biosynthesis (mainly hyaluronic acid). In presence of IL-1, Etodolac at the highest concentration partially diminished the stimulatory effect of IL-1. A 8-day exposure of the cells to Etodolac led to a decrease of GAG synthesis. In the same experiment, IL-1 alone caused a slight increase of GAG production that was not abolished by Etodolac treatment.


Chitosan-Grafted Halloysite Nanotubes-Fe3O4 Composite for Laccase-Immobilization and Sulfamethoxazole-Degradation.
A surface-engineered nano-support for enzyme laccase-immobilization was designed by grafting the surface of halloysite nanotubes (HNTs) with Fe3O4 nanoparticles and chitosan. Herein, HNTs were magnetized (HNTs-M) by a cost-effective reduction-precipitation method. The synthesized HNTs-M were grafted with 0.25%, 0.5%, 1%, and 2% chitosan (HNTs-M-chitosan), respectively. Synthesized HNTs-M-chitosan (0.25%), HNTs-M-chitosan (0.5%), HNTs-M-chitosan (1%) and HNTs-M-chitosan (2%) were linked with glutaraldehyde (GTA) for laccase immobilization. Among these formulations, HNTs-M-chitosan (1%) exhibited the highest laccase immobilization with 95.13% activity recovery and 100.12 mg/g of laccase loading. The optimized material was characterized thoroughly by scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HR-TEM), X-ray powder diffraction (XRD), thermal gravimetric analysis (TGA), and vibrating sample magnetometer (VSM) analysis. The immobilized laccase (HNTs-M-chitosan (1%)-GTA-Laccase) exhibited higher pH, temperature, and storage stabilities. The HNTs-M-chitosan (1%)-GTA-Laccase possesses excellent reusability capabilities. At the end of 10 cycles of the reusability experiment, HNTs-M-chitosan (1%)-GTA-Laccase retained 59.88% of its initial activity. The immobilized laccase was utilized for redox-mediated degradation of sulfamethoxazole (SMX), resulting in 41%, 59%, and 62% degradation of SMX in the presence of 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), guaiacol (GUA), and syringaldehyde (SA), respectively. Repeated SMX degradation (57.10% after the sixth cycle) confirmed the potential of HNTs-M-chitosan (1%)-GTA-Laccase for environmental pollutant degradation. Thus, we successfully designed chitosan-based, rapidly separable super-magnetic nanotubes for efficacious enhancement of laccase biocatalysis, which can be applied as nano-supports for other enzymes.


Identification of a common sphingolipid-binding domain in Alzheimer, prion, and HIV-1 proteins.
The V3 loop of the human immunodeficiency virus (HIV)-1 surface envelope glycoprotein gp120 is a sphingolipid-binding domain mediating the attachment of HIV-1 to plasma membrane microdomains (rafts). Sphingolipid-induced conformational changes in gp120 are required for HIV-1 fusion. Galactosylceramide and sphingomyelin have been detected in highly purified preparations of prion rods, suggesting that the prion protein (PrP) may interact with selected sphingolipids. Moreover, a major conformational transition of the Alzheimer beta-amyloid peptide has been observed upon interaction with sphingolipid-containing membranes. Structure similarity searches with the combinatorial extension method revealed the presence of a V3-like domain in the human prion protein PrP and in the Alzheimer beta-amyloid peptide. In each case, synthetic peptides derived from the predicted V3-like domain were found to interact with monomolecular films of galactosylceramide and sphingomyelin at the air-water interface. The V3-like domain of PrP is a disulfide-linked loop (Cys(179)-Cys(214)) that includes the E200K mutation site associated with familial Creutzfeldt-Jakob disease. This mutation abrogated sphingomyelin recognition. The identification of a common sphingolipid-binding motif in gp120, PrP, and beta-amyloid peptide underscores the role of lipid rafts in the pathogenesis of HIV-1, Alzheimer, and prion diseases and may provide new therapeutic strategies.


Deriving ethnic-specific BMI cutoff points for assessing diabetes risk.
The definition of obesity (BMI ≥ 30 kg/m(2)), a key risk factor of diabetes, is widely used in white populations; however, its appropriateness in nonwhite populations has been questioned. We compared the incidence rates of diabetes across white, South Asian, Chinese, and black populations and identified equivalent ethnic-specific BMI cutoff values for assessing diabetes risk. We conducted a multiethnic cohort study of 59,824 nondiabetic adults aged ≥ 30 years living in Ontario, Canada. Subjects were identified from Statistics Canada's population health surveys and followed for up to 12.8 years for diabetes incidence using record linkages to multiple health administrative databases. The median duration of follow-up was 6 years. After adjusting for age, sex, sociodemographic characteristics, and BMI, the risk of diabetes was significantly higher among South Asian (hazard ratio 3.40, P < 0.001), black (1.99, P < 0.001), and Chinese (1.87, P = 0.002) subjects than among white subjects. The median age at diagnosis was lowest among South Asian (aged 49 years) subjects, followed by Chinese (aged 55 years), black (aged 57 years), and white (aged 58 years) subjects. For the equivalent incidence rate of diabetes at a BMI of 30 kg/m(2) in white subjects, the BMI cutoff value was 24 kg/m(2) in South Asian, 25 kg/m(2) in Chinese, and 26 kg/m(2) in black subjects. South Asian, Chinese, and black subjects developed diabetes at a higher rate, at an earlier age, and at lower ranges of BMI than their white counterparts. Our findings highlight the need for designing ethnically tailored prevention strategies and for lowering current targets for ideal body weight for nonwhite populations.


Happiness is best kept stable: positive emotion variability is associated with poorer psychological health.
Positive emotion has been shown to be associated with adaptive outcomes in a number of domains, including psychological health. However, research has largely focused on overall levels of positive emotion with less attention paid to how variable versus stable it is across time. We thus examined the psychological health correlates of positive emotion variability versus stability across 2 distinct studies, populations, and scientifically validated approaches for quantifying variability in emotion across time. Study 1 used a daily experience approach in a U.S. community sample (N = 244) to examine positive emotion variability across 2 weeks (macrolevel). Study 2 adopted a daily reconstruction method in a French adult sample (N = 2,391) to examine variability within 1 day (microlevel). Greater macro- and microlevel variability in positive emotion was associated with worse psychological health, including lower well-being and life satisfaction and greater depression and anxiety (Study 1), and lower daily satisfaction, life satisfaction, and happiness (Study 2). Taken together, these findings support the notion that positive emotion variability plays an important and incremental role in psychological health above and beyond overall levels of happiness, and that too much variability might be maladaptive.


Microinjection of leptin into the ventromedial hypothalamus increases glucose uptake in peripheral tissues in rats.
We studied the effects of microinjection of leptin into the ventromedial hypothalamus (VMH) and lateral hypothalamus (LH) on glucose uptake in peripheral tissues in unanesthetized rats. The rate of glucose uptake was assessed in vivo by 2-[3H]deoxyglucose incorporation. Single injection of leptin into VMH increased glucose uptake in brown adipose tissue (BAT), heart, skeletal muscles, and spleen but not in white adipose tissue or skin. On the other hand, microinjection of leptin into LH had little effect on glucose uptake in those tissues. The plasma concentrations of glucose and insulin were unaltered by intrahypothalamic injection of leptin into either VMH or LH. Among skeletal muscles, the increase in glucose uptake induced by intrahypothalamic injection of leptin was greater in the soleus than in the extensor digitorum longus. Likewise, the increased glucose uptake in the gastrocnemius in response to leptin was more prominent in the red part than in the white part of the tissue. When surgical sympathetic denervation of the interscapular BAT was performed, the enhanced glucose uptake by BAT in response to intrahypothalamic leptin was completely suppressed. These findings suggest that intrahypothalamic injection of leptin preferentially increases glucose uptake by some peripheral tissues through activation of the VMH-sympathetic (or its neighboring medial hypothalamus-sympathetic) nervous system, thereby contributing to the maintenance of energy balance.


Widely Disseminated Cryptococcosis Manifesting in a Previously Undiagnosed Human Immunodeficiency Virus (HIV)-Positive 18-Year-Old.
BACKGROUND After initial infection with HIV, loss of CD4+ T cells progresses along a predictable timeline. The clinical latency stage lasts an average of 10 years, until the CD4+ T cell count falls below 200 cells/uL or the patient develops an AIDS-defining opportunistic infection/cancer. This report describes an unusual opportunistic infection in a young patient with no prior clinical evidence of HIV infection. CASE REPORT An 18-year-old man presented with fever, abdominal pain, and dyspnea for the previous 2 weeks and was symptomatically treated for gastroenteritis. He presented 2 weeks later with extreme fatigue, and a CT scan revealed diffuse lymphadenopathy. He was transferred to a regional hospital, but upon arrival and prior to detailed investigative work-up, he developed cardiac arrest. Despite maximal resuscitative efforts, he died approximately 8 h after admission. At autopsy, diffuse lymphadenopathy, splenomegaly, and pulmonary congestion were noted. Disseminated cryptococcal infection involving almost every organ system was identified at autopsy. A postmortem HIV-1 antibody test was positive. The cause of death was severe immunodeficiency as a result of advanced HIV infection resulting in disseminated cryptococcal infection, with cerebral edema, herniation, and respiratory failure. CONCLUSIONS This patient's non-specific symptoms in conjunction with his rapid decline made arriving at a correct diagnosis challenging. Only during autopsy was the disseminated fungal infection discovered, leading to suspicion of HIV infection. HIV autopsies are not uncommon, but the clinical history is usually known beforehand. This case report highlights the importance of considering HIV-related conditions in patients presenting with this array of symptoms, as well as to alert healthcare providers and staff to the need for increased biosafety precautions.


Cyclic adenosine 5'-diphosphate ribose analogs without a "southern" ribose inhibit ADP-ribosyl cyclase-hydrolase CD38.
Cyclic adenosine 5'-diphosphate ribose (cADPR) analogs based on the cyclic inosine 5'-diphosphate ribose (cIDPR) template were synthesized by recently developed stereo- and regioselective N1-ribosylation. Replacing the base N9-ribose with a butyl chain generates inhibitors of cADPR hydrolysis by the human ADP-ribosyl cyclase CD38 catalytic domain (shCD38), illustrating the nonessential nature of the "southern" ribose for binding. Butyl substitution generally improves potency relative to the parent cIDPRs, and 8-amino-N9-butyl-cIDPR is comparable to the best noncovalent CD38 inhibitors to date (IC50 = 3.3 μM). Crystallographic analysis of the shCD38:8-amino-N9-butyl-cIDPR complex to a 2.05 Å resolution unexpectedly reveals an N1-hydrolyzed ligand in the active site, suggesting that it is the N6-imino form of cADPR that is hydrolyzed by CD38. While HPLC studies confirm ligand cleavage at very high protein concentrations, they indicate that hydrolysis does not occur under physiological concentrations. Taken together, these analogs confirm that the "northern" ribose is critical for CD38 activity and inhibition, provide new insight into the mechanism of cADPR hydrolysis by CD38, and may aid future inhibitor design.


Extrachromosomal DNA in the Apicomplexa.
Malaria and related apicomplexan parasites have two highly conserved organellar genomes: one is of plastid (pl) origin, and the other is mitochondrial (mt). The organization of both organellar DNA molecules from the human malaria parasite Plasmodium falciparum has been determined, and they have been shown to be tightly packed with genes. The 35-kb circular DNA is the smallest known vestigial plastid genome and is presumed to be functional. All but two of its recognized genes are involved with genetic expression: one of the two encodes a member of the clp family of molecular chaperones, and the other encodes a conserved protein of unknown function found both in algal plastids and in eubacterial genomes. The possible evolutionary source and intracellular location of the plDNA are discussed. The 6-kb tandemly repeated mt genome is the smallest known and codes for only three proteins (cytochrome b and two subunits of cytochrome oxidase) as well as two bizarrely fragmented rRNAs. The organization of the mt genome differs somewhat among genera. The mtDNA sequence provides information not otherwise available about the structure of apicomplexan cytochrome b as well as the unusually fragmented rRNAs. The malarial mtDNA has a phage-like replication mechanism and undergoes extensive recombination like the mtDNA of some other lower eukaryotes.


chopBAI: BAM index reduction solves I/O bottlenecks in the joint analysis of large sequencing cohorts.
Advances in sequencing capacity have led to the generation of unprecedented amounts of genomic data. The processing of this data frequently leads to I/O bottlenecks, e. g. when analyzing a small genomic region across a large number of samples. The largest I/O burden is, however, often not imposed by the amount of data needed for the analysis but rather by index files that help retrieving this data. We have developed chopBAI, a program that can chop a BAM index (BAI) file into small pieces. The program outputs a list of BAI files each indexing a specified genomic interval. The output files are much smaller in size but maintain compatibility with existing software tools. We show how preprocessing BAI files with chopBAI can lead to a reduction of I/O by more than 95% during the analysis of 10 kb genomic regions, eventually enabling the joint analysis of more than 10 000 individuals. The software is implemented in C ++, GPL licensed and available at http://github.com/DecodeGenetics/chopBAIContact:firstname.lastname@example.com.


Youth and parents' experiences of a new inter-agency transition model for spina bifida compared to youth who did not take part in the model.
Young adults with spina bifida are underserved in health care and are at risk as they transition to adult health care. A pediatric rehabilitation hospital and an adult community health center partnered to help address this gap. Although some research has explored general transition experiences of youth with disabilities, little is known about their experiences in a transition model that involves an inter-agency partnership, continuous, age-appropriate, and client-centered care. To explore youth and parent experiences of a new transition model for youth with spina bifida, compared to the experiences of young adults with spina bifida who did not participate in the model. Using a descriptive, qualitative design involving a thematic analysis we conducted semi-structured interviews with 32 participants (9 youth, 11 parents, 12 young adults). Most youth and parents in our sample who took part in the new model felt supported by pediatric providers and benefitted from gradually transferring responsibility from parents to youth. They also reported experiencing challenges, including lack of support from primary care providers and lack of clarity about the new model. Many young adults who did not take part in the model reported receiving some transition-related thought support from pediatric specialists, parents, and in some cases, primary care providers. However, they also reported experiencing gaps in their continuity of care and needed more support with employment, relationships, finances, and housing. Our findings show the new transition model for youth with spina bifida can help enhance participants' transition experiences and preparation for adulthood. However, the model needs further development to address the varied abilities and support needs of youth with spina bifida.


Strengthening health information systems to address health equity challenges.
Special studies and isolated initiatives over the past several decades in low-, middle- and high-income countries have consistently shown inequalities in health among socioeconomic groups and by gender, race or ethnicity, geographical area and other measures associated with social advantage. Significant health inequalities linked to social (dis)advantage rather than to inherent biological differences are generally considered unfair or inequitable. Such health inequities are the main object of health development efforts, including global targets such as the Millennium Development Goals, which require monitoring to evaluate progress. However, most national health information systems (HIS) lack key information needed to assess and address health inequities, namely, reliable, longitudinal and representative data linking measures of health with measures of social status or advantage at the individual or small-area level. Without empirical documentation and monitoring of such inequities, as well as country-level capacity to use this information for effective planning and monitoring of progress in response to interventions, movement towards equity is unlikely to occur. This paper reviews core information requirements and potential databases and proposes short-term and longer term strategies for strengthening the capabilities of HIS for the analysis of health equity and discusses HIS-related entry points for supporting a culture of equity-oriented decision-making and policy development.


Running combined simple and vertical mattress suture: a rapid skin-everting stitch.
Vertical mattress sutures are used in skin surgery to produce eversion of the wound edges, which produces a better, cosmetically acceptable scar. The disadvantages of this closure include the risk of wound hypereversion if tied too tightly, longer time to perform the stitch, and difficulties with grasping the externalized loops during suture removal. To examine and discuss the new modification of running vertical mattress sutures, which can overcome the disadvantages. Running combined simple and vertical mattress suture techniques are presented in a series of schematic diagrams that follow. The running combined simple and vertical mattress is easier and quicker to close the wound than the classic interrupted or running vertical mattress suture. Whereas the mattress suture everts the wound edges, the simple suture coapts the sides, preventing undue inversion or hypereversion of the wound. Suture removal is less uncomfortable for the patient and is performed faster by the medical staff owing to a reduction in the number of externalized loops by half compared with the classic vertical mattress stitch. Alternation of the vertical mattress and simple running suture saves the surgeon's time without lessening the advantages of good wound eversion and hence the formation of a less apparent, cosmetically more appealing scar after wound contraction.


Biomechanical and leaf-climate relationships: a comparison of ferns and seed plants.
Relationships of leaf size and shape (physiognomy) with climate have been well characterized for woody non-monocotyledonous angiosperms (dicots), allowing the development of models for estimating paleoclimate from fossil leaves. More recently, petiole width of seed plants has been shown to scale closely with leaf mass. By measuring petiole width and leaf area in fossils, leaf mass per area (MA) can be estimated and an approximate leaf life span inferred. However, little is known about these relationships in ferns, a clade with a deep fossil record and with the potential to greatly expand the applicability of these proxies. We measured the petiole width, MA, and leaf physiognomic characters of 179 fern species from 188 locations across six continents. We applied biomechanical models and assessed the relationship between leaf physiognomy and climate using correlational approaches. The scaling relationship between area-normalized petiole width and MA differs between fern fronds and pinnae. The scaling relationship is best modeled as an end-loaded cantilevered beam, which is different from the best-fit biomechanical model for seed plants. Fern leaf physiognomy is not influenced by climatic conditions. The cantilever beam model can be applied to fossil ferns. The lack of sensitivity of leaf physiognomy to climate in ferns argues against their use to reconstruct paleoclimate. Differences in climate sensitivity and biomechanical relationships between ferns and seed plants may be driven by differences in their hydraulic conductivity and/or their differing evolutionary histories of vein architecture and leaf morphology.


Cardiac exposures in breast cancer radiotherapy: 1950s-1990s.
To estimate the doses to the heart and coronary arteries from common breast cancer radiotherapy (RT) regimens used worldwide from the 1950s to the 1990s. Virtual simulation and computed tomography planning were used to reconstruct the megavoltage and electron regimens. Manual planning was used for the orthovoltage and brachytherapy regimens. Several sources of variability associated with the dose estimates were assessed. Breast or chest wall RT resulted in whole heart doses of 0.9-14 Gy for left-sided and of 0.4-6 Gy for right-sided irradiation. Internal mammary chain RT delivered heart doses of 3-17 Gy and 2-10 Gy for left- and right-sided irradiation, respectively. For most regimens, the dose to the left anterior descending coronary artery was greater than the heart dose. Scar boost, supraclavicular fossa, and axillary RT delivered mean cardiac doses of <or=3 Gy. The greatest source of variability in estimating dose from a given regimen was patient anatomy. For most techniques, the greatest radiation doses were received by the anterior part of the heart and the left anterior descending coronary artery, a common site of atherosclerosis causing myocardial infarction. Irradiation of these structures might have contributed to the excess risk of death from heart disease seen after some past breast cancer RT regimens.


Early-Life Antibiotic Exposure, Gut Microbiota Development, and Predisposition to Obesity.
Antibiotics are often prescribed inappropriately to infants and young children, with potentially adverse effects on the developing gut microbiota and related metabolic processes. We review evidence from 17 epidemiologic studies suggesting that antibiotic exposure during critical periods of early development may influence weight gain and the development of obesity. Complementary research in both humans and rodents indicates that gut microbiota play a key role in this process, although further research is needed to confirm and characterize the causal mechanisms involved. Obesity is a complex and multifactorial condition; thus, a multipronged prevention strategy will be required to curb the current obesity epidemic. Evidence to date suggests this strategy should include the judicious use of antibiotics, especially in early life when the developing gut microbiota is particularly susceptible to perturbations with long-lasting implications for metabolic programming and obesity risk.


Isolation and Partial Nucleic Acid Characterization of a New Isolate of Potato virus V with Distinct Biological and Serological Properties.
In vitro cultures of potato (Solanum tuberosum cv. Papa Amarillo) were imported into the United States and were subjected to routine pathogen screening for newly introduced varieties. These cultures indexed positive for several of the six viruses tested and thus were treated using standard methods for eliminating viruses in in vitro cultures. Although the therapy apparently freed the potato of the tested viruses, some of the cultures continued to exhibit mosaic symptoms when planted in the greenhouse. When examined by electron microscopy, these plants contained flexuous rods. Tested samples did not index positive with any potato virus-specific polyclonal antisera but reacted with the "universal" potyvirus monoclonal antibody. These results indicated that these samples were likely infected with an as yet undescribed potyvirus. Further investigation indicated that this virus is a new strain of Potato virus V (PVV) that is serologically distinct from the common strain. This is the first description of a distinct strain of PVV. This particular strain has diverged sufficiently from other isolates of PVV to no longer be detectable by some commonly used antisera, and therefore is a concern for the future restriction of PVV spread into regions of the world where it is not currently present.


Oligonucleotide delivery with cell surface binding and cell penetrating Peptide amphiphile nanospheres.
A drug delivery system designed specifically for oligonucleotide therapeutics can ameliorate the problems associated with the in vivo delivery of these molecules. The internalization of free oligonucleotides is challenging, and cytotoxicity is the main obstacle for current transfection vehicles. To develop nontoxic delivery vehicles for efficient transfection of oligonucleotides, we designed a self-assembling peptide amphiphile (PA) nanosphere delivery system decorated with cell penetrating peptides (CPPs) containing multiple arginine residues (R4 and R8), and a cell surface binding peptide (KRSR), and report the efficiency of this system in delivering G-3129, a Bcl-2 antisense oligonucleotide (AON). PA/AON (peptide amphiphile/antisense oligonucleotide) complexes were characterized with regards to their size and secondary structure, and their cellular internalization efficiencies were evaluated. The effect of the number of arginine residues on the cellular internalization was investigated by both flow cytometry and confocal imaging, and the results revealed that uptake efficiency improved as the number of arginines in the sequence increased. The combined effect of cell penetration and surface binding property on the cellular internalization and its uptake mechanism was also evaluated by mixing R8-PA and KRSR-PA. R8 and R8/KRSR decorated PAs were found to drastically increase the internalization of AONs compared to nonbioactive PA control. Overall, the KRSR-decorated self-assembled PA nanospheres were demonstrated to be noncytotoxic delivery vectors with high transfection rates and may serve as a promising delivery system for AONs.


Functional studies on hemoglobin opossum. Conclusions drawn regarding the role of the distal histidine.
In the hemoglobin of the opossum, the alpha chains have different residues at positions E7 and E11 than do most other mammalian hemoglobins. In the opossum, the usual histidine at alpha E7 is replaced by glutamine, the valine at alpha E11 by isoleucine, and the hemoglobin is known to have a low oxygen affinity and a low Hill coefficient. Comparison of kinetic studies of opossum hemoglobin with normal human hemoglobin shows that alpha chains in Hb opossum, despite the lack of distal histidine, do not differ significantly in CO-combination rates in either the T or R states. These rates are much slower than the rates reported for Hb Zurich, the hemoglobin from Chironomus thumi thumi, or the monomeric components of glycera hemoglobin, all of which also have a different residue at E7. As compared with Hb A, the changes in ligand affinities in the T and R states are small and cannot account for the unusually high values of p50 for Hb opossum. The equilibrium and kinetic data indicate that the L = (T)/(R) is about 100 times higher for Hb opossum than for Hb A; CCO = KR/KT approximately equal to 0.014. The kinetic data on l'4 and l also indicate that the R leads to T equilibrium for Hb4(CO)4 and Hb4(CO)2 can be shifted in either direction by adding inositol hexaphosphate or by changing the pH.


Association Between Medicaid Expansion and Rates of Opioid-Related Hospital Use.
The rate of opioid-related emergency department (ED) visits and inpatient hospitalizations has increased rapidly in recent years. Medicaid expansions have the potential to reduce overall opioid-related hospital events by improving access to outpatient treatment for opioid use disorder. To examine the association between Medicaid expansions and rates of opioid-related ED visits and inpatient hospitalizations. A difference-in-differences observational design was used to compare changes in opioid-related hospital events in US nonfederal, nonrehabilitation hospitals in states that implemented Medicaid expansions between the first quarter of 2005 and the last quarter of 2017 with changes in nonexpansion states. All-payer ED and hospital discharges from 45 states in the Healthcare Cost and Utilization Project FastStats were included. State implementation of Medicaid expansions between 2005 and 2017. Rates of all opioid-related ED visits and inpatient hospitalizations, measured as the quarterly numbers of treat-and-release ED discharges and hospital discharges related to opioid abuse, dependence, and overdose, per 100 000 state population. In the 46 states and District of Columbia included in the study, 1524 observations of emergency department data and 2219 observations of opioid-related inpatient hospitalizations were analyzed. The post-2014 Medicaid expansions were associated with a 9.74% (95% CI, -18.83% to -0.65%) reduction in the rate of opioid-related inpatient hospitalizations. There appeared to be no association between the pre-2014 or post-2014 Medicaid expansions and the rate of opioid-related ED visits (post-2014 Medicaid expansions, -3.98%; 95% CI, -14.69% to 6.72%; and pre-2014 Medicaid expansions, 1.02%; 95% CI, -5.25% to 7.28%). Medicaid expansion appears to be associated with meaningful reductions in opioid-related hospital use, possibly attributable to improved care for opioid use disorder in other settings.


C. elegans EIF-3.K promotes programmed cell death through CED-3 caspase.
Programmed cell death (apoptosis) is essential for the development and homeostasis of metazoans. The central step in the execution of programmed cell death is the activation of caspases. In C. elegans, the core cell death regulators EGL-1(a BH3 domain-containing protein), CED-9 (Bcl-2), and CED-4 (Apaf-1) act in an inhibitory cascade to activate the CED-3 caspase. Here we have identified an additional component eif-3.K (eukaryotic translation initiation factor 3 subunit k) that acts upstream of ced-3 to promote programmed cell death. The loss of eif-3.K reduced cell deaths in both somatic and germ cells, whereas the overexpression of eif-3.K resulted in a slight but significant increase in cell death. Using a cell-specific promoter, we show that eif-3.K promotes cell death in a cell-autonomous manner. In addition, the loss of eif-3.K significantly suppressed cell death-induced through the overexpression of ced-4, but not ced-3, indicating a distinct requirement for eif-3.K in apoptosis. Reciprocally, a loss of ced-3 suppressed cell death induced by the overexpression of eif-3.K. These results indicate that eif-3.K requires ced-3 to promote programmed cell death and that eif-3.K acts upstream of ced-3 to promote this process. The EIF-3.K protein is ubiquitously expressed in embryos and larvae and localizes to the cytoplasm. A structure-function analysis revealed that the 61 amino acid long WH domain of EIF-3.K, potentially involved in protein-DNA/RNA interactions, is both necessary and sufficient for the cell death-promoting activity of EIF-3.K. Because human eIF3k was able to partially substitute for C. elegans eif-3.K in the promotion of cell death, this WH domain-dependent EIF-3.K-mediated cell death process has potentially been conserved throughout evolution.


Study of the mechanisms of crocetin-induced differentiation and apoptosis in human acute promyelocytic leukemia cells.
Crocetin, the major carotenoid in saffron, exhibits potent anticancer effects. However, the antileukemic effects of crocetin are still unclear, especially in primary acute promyelocytic leukemia (APL) cells. In the current study, the potential antipromyelocytic leukemia activity of crocetin and the underlying molecular mechanisms were investigated. Crocetin (100 µM), like standard anti-APL drugs, all-trans retinoic acid (ATRA, 10 µM) and As2 O 3 (arsenic trioxide, 50 µM), significantly inhibited proliferation and induced apoptosis in primary APL cells, as well as NB4 and HL60 cells. The effect was associated with the decreased expressions of prosurvival genes Akt and BCL2, the multidrug resistance (MDR) proteins, ABCB1 and ABCC1 and the inhibition of tyrosyl-DNA phosphodiesterase 1 (TDP1), while the expressions of proapoptotic genes CASP3, CASP9, and BAX/BCL2 ratio were significantly increased. In contrast, crocetin at relatively low concentration (10 µM), like ATRA (1 µM) and As 2 O 3 (0.5 µM), induced differentiation of leukemic cells toward granulocytic pattern, and increased the number of differentiated cells expressing CD11b and CD14, while the number of the immature cells expressing CD34 or CD33 was decreased. Furthermore, crocetin suppressed the expression of clinical marker promyelocytic leukemia/retinoic acid receptor-α ( PML/RARα) in NB4 and primary APL cells, and reduced the expression of histone deacetylase 1 ( HDAC1) in all leukemic cells. The results suggested that crocetin can be considered as a candidate for future preclinical and clinical trials of complementary APL treatment.


Time to implement 9-month infant HIV testing in South Africa.
South Africa (SA) is likely to meet the National Strategic Plan target of <2% mother-to-child HIV transmission at 6 weeks of age in 2015. Children infected with HIV after 6 weeks often remain undiagnosed because of poor implementation of post-weaning and final outcome 18-month HIV testing. The World Health Organization recommends a screening HIV rapid test (HRT) in HIV-exposed infants at the 9-month immunisation visit to exclude postnatal infection, with a confirmatory HIV polymerase chain reaction (PCR) test if the HRT is positive. To evaluate the impact of substituting this recommendation for the post-weaning HIV testing recommended by SA guidelines. Rates of seroreversion and probability of infection at 9 months of age were applied to a theoretical population of 100 HIV-exposed infants, uninfected at birth and breastfed for 1 year with antiretroviral prophylaxis. Nine scenarios were developed and the number of HIV PCRs saved compared with current guidelines was calculated. Nine-month testing using the HRT reduced the number of follow-up PCR tests done in all scenarios by >50%, with differences ranging from 51% to 59% and 81% to 89% for low and high seroreversion rates, respectively. Nine-month testing using HRT would increase identification and early treatment of HIV-infected infants, improve monitoring of postnatal transmission rates, and reduce the number of HIV PCR tests done with resultant cost saving. Training of healthcare workers implementing HRT would be required. Ongoing efforts to improve implementation and monitoring of testing at 9 and 18 months will be essential.


Pedestrian Dead Reckoning-Assisted Visual Inertial Odometry Integrity Monitoring.
Visual inertial odometers (VIOs) have received increasing attention in the area of indoor positioning due to the universality and convenience of the camera. However, the visual observation of VIO is more susceptible to the environment, and the error of observation affects the final positioning accuracy. To address this issue, we analyzed the causes of visual observation error that occur under different scenarios and their impact on positioning accuracy. We propose a new method of using the short-time reliability of pedestrian dead reckoning (PDR) to aid in visual integrity monitoring and to reduce positioning error. The proposed method selects optimized positioning by automatically switching between outputs from VIO and PDR. Experiments were carried out to test and evaluate the proposed PDR-assisted visual integrity monitoring. The sensor suite of experiments consisted of a stereo camera and an inertial measurement unit (IMU). Results were analyzed in detailed and indicated that the proposed system performs better for indoor positioning within an environment that contains low illumination, little background texture information, or few moving objects.


Recodable surfaces based on switchable hydrogen bonds.
We introduce recodable surfaces solely based on reversible artificial hydrogen bonding interactions. We show that a symmetrical oligoamide (SOA) attached to poly(methyl methacrylate) (PMMA) can be repeatedly immobilized and cleaved off spatially defined surface domains photochemically functionalized with asymmetric oligoamides (AOAs). The spatially resolved recodability is imaged and quantified via ToF-SIMS.


Effect of density on the thermal decomposition mechanism of ε-CL-20: a ReaxFF reactive molecular dynamics simulation study.
The explosive detonation reaction occurs when explosives are compressed by different shock strengths, and the degree of compression affects the chemical reaction of the detonation process. The thermal decomposition mechanism of explosives under different compression densities has thus attracted significant research interest, and a better understanding of this mechanism would be helpful for determining the mechanism of the detonation reaction of explosives. In this study, a ε-CL-20 supercell was constructed, and the thermal decomposition was calculated at different compression densities and temperatures using molecular dynamics simulations based on the ReaxFF-lg reactive force field. We analyzed the effect of density on the main elementary reaction, which consists of the initial reaction and the formation of final products. In addition, we studied the effect of density on the generation of clusters and the reaction kinetics of the thermal decomposition. The results indicate that the initial reaction pathway of the CL-20 molecule is the cleavage of the N-NO2 bond at different densities and that the frequency of N-NO2 bond breakage decreases at high density. As the density increases, clusters easily form and are resistant to decomposition at the later stage of thermal decomposition, which eventually leads to a decrease in the number of final products. Increasing the initial density of CL-20 significantly increases the reaction rate of the initial decomposition but hardly changes the activation energy of the decomposition.


Sexual conflict and the tragedy of the commons.
It is widely understood that the costs and benefits of mating can affect the fecundity and survival of individuals. Sexual conflict may have profound consequences for populations as a result of the negative effects it causes males and females to have on one another's fitness. Here we present a model describing the evolution of sexual conflict, in which males inflict a direct cost on female fitness. We show that these costs can drive the entire population to extinction. To males, females are an essential but finite resource over which they have to compete. Population extinction owing to sexual conflict can therefore be seen as an evolutionary tragedy of the commons. Our model shows that a positive feedback between harassment and the operational sex ratio is responsible for the demise of females and, thus, for population extinction. We further show that the evolution of female resistance to counter harassment can prevent a tragedy of the commons. Our findings not only demonstrate that sexual conflict can drive a population to extinction but also highlight how simple mechanisms, such as harassment costs to males and females and the coevolution between harassment and resistance, can help avert a tragedy of the commons caused by sexual conflict.


Engineering signal peptides for enhanced protein secretion from Lactococcus lactis.
Lactococcus lactis is an attractive vehicle for biotechnological production of proteins and clinical delivery of therapeutics. In many such applications using this host, it is desirable to maximize secretion of recombinant proteins into the extracellular space, which is typically achieved by using the native signal peptide from a major secreted lactococcal protein, Usp45. In order to further increase protein secretion from L. lactis, inherent limitations of the Usp45 signal peptide (Usp45sp) must be elucidated. Here, we performed extensive mutagenesis on Usp45sp to probe the effects of both the mRNA sequence (silent mutations) and the peptide sequence (amino acid substitutions) on secretion. We screened signal peptides based on their resulting secretion levels of Staphylococcus aureus nuclease and further evaluated them for secretion of Bacillus subtilis α-amylase. Silent mutations alone gave an increase of up to 16% in the secretion of α-amylase through a mechanism consistent with relaxed mRNA folding around the ribosome binding site and enhanced translation. Targeted amino acid mutagenesis in Usp45sp, combined with additional silent mutations from the best clone in the initial screen, yielded an increase of up to 51% in maximum secretion of α-amylase while maintaining secretion at lower induction levels. The best sequence from our screen preserves the tripartite structure of the native signal peptide but increases the positive charge of the n-region. Our study presents the first example of an engineered L. lactis signal peptide with a higher secretion yield than Usp45sp and, more generally, provides strategies for further enhancing protein secretion in bacterial hosts.


Telaprevir for the treatment of chronic hepatitis C infection.
Telaprevir is an NS3/4A protease inhibitor that has recently received US FDA approval for the treatment of chronic HCV infection. Telaprevir is given in combination with peg-IFN-α and ribavirin and is indicated for both treatment-naive and treatment-experienced patients with genotype 1 infection. Along with the other first generation NS3/4A protease inhibitor boceprevir, these combination regimens have immediately become the standard of care for genotype 1 patients. The adverse event profile for the combination regimen remains dominated by peg-IFN-α and ribavirin, but there is additional anemia and rash with telaprevir. Owing to telaprevir's metabolism by the CYP3/4A pathway, drug-drug interactions could lead to toxicity from other medications or decreased efficacy of telaprevir. Viral resistance can develop during treatment with telaprevir, and patients will need to be educated on their role in adherence to minimize the risk of resistance and improve their chances of cure of HCV infection.


Role of contact-angle hysteresis for fluid transport in wet granular matter.
The stability of sand castles is determined by the structure of wet granulates. Experimental data on the size distribution of fluid pockets are ambiguous with regard to their origin. We discovered that contact-angle hysteresis plays a fundamental role in the equilibrium distribution of bridge volumes, and not geometrical disorder as commonly conjectured. This has substantial consequences on the mechanical properties of wet granular beds, including a history-dependent rheology and lowered strength. Our findings are obtained using a model in which the Laplace pressures, bridge volumes, and contact angles are dynamical variables associated with the contact points. While accounting for contact line pinning, we track the temporal evolution of each bridge. We observe a crossover to a power-law decay of the variance of capillary pressures at late times and a saturation of the variance of bridge volumes to a finite value connected to contact line pinning. Large-scale simulations of liquid transport in the bridge network reveal that the equilibration dynamics at early times is well described by a mean-field model. The spread of final bridge volumes can be directly related to the magnitude of contact-angle hysteresis.


Reciprocal backcross mice confirm major loci linked to hyperoxic acute lung injury survival time.
Morbidity and mortality associated with acute lung injury (ALI) and acute respiratory distress syndrome remain substantial. Although many candidate genes have been tested, a clear understanding of the pathogenesis is lacking, as is our ability to predict individual outcome. Because ALI is a complex disease, single gene approaches cannot easily identify effectors that must be treated concurrently. We employed a strategy to help identify critical genes and gene combinations involved in ALI mortality. Using hyperoxia to induce ALI, a mouse model for genetic analyses of ALI survival time was identified: C57BL/6J (B) mice are sensitive (i.e., die early), whereas 129X1/SvJ (S) mice are significantly more resistant, but with low penetrance. Segregation analysis of reciprocal F(2) mice generated from B and S strains revealed significant sex, cross, and parent of origin effects. Quantitative trait locus (QTL) analysis identified five chromosomal regions significantly linked to hyperoxic ALI survival time (named Shali1-Shali5). Further analyses demonstrated that both parental strains contribute resistance alleles to their offspring and that the phenotype demonstrated parent of origin effects. To validate earlier findings, we generated and tested mice from all eight possible B-S-derived backcrosses. Results from segregation and QTL analyses of 935 backcrosses, alone and combined with the previous 840 B-S-derived F(2) population, further supported the highly significant QTLs on chromosomes 1 (Shali1) and 4 (Shali2) and confirmed that the sex, cross, and parent of origin all contribute to survival time with hyperoxic ALI.