Cyclooxygenase-derived vasoconstriction restrains hypoxia-mediated cerebral vasodilation in young adults with metabolic syndrome.
Poor cerebrovascular function in metabolic syndrome (MetSyn) likely contributes to elevated risk of cerebrovascular disease in this growing clinical population. Younger MetSyn adults without clinical evidence of cerebrovascular disease exhibit preserved hypercapnic vasodilation yet markedly impaired hypoxic vasodilation, but the mechanisms behind reduced hypoxic vasodilation are unknown. Based on data from rats, we tested the hypothesis that younger adults with MetSyn exhibit reduced cerebral hypoxic vasodilation due to loss of vasodilating prostaglandins. Middle cerebral artery velocity (MCAv) was measured with transcranial Doppler ultrasound in adults with MetSyn (n = 13, 33 ± 3 yr) and healthy controls (n = 15, 31 ± 2 yr). Isocapnic hypoxia was induced by titrating inspired oxygen to lower arterial saturation to 90% and 80% for 5 min each. Separately, hypercapnia was induced by increasing end-tidal CO2 10 mmHg above baseline levels. Cyclooxygenase inhibition (100 mg indomethacin) was conducted in a randomized double-blind, placebo controlled design. MCAv was normalized for group differences in blood pressure (healthy: 89 ± 2 mmHg vs. MetSyn: 102 ± 2 mmHg) as cerebrovascular conductance index (CVCi), and used to assess cerebral vasodilation. Hypoxia increased CVCi in both groups; however, vasodilation was ∼55% lower in MetSyn at SpO2 = 80% (P < 0.05). Indomethacin tended to decrease hypoxic vasodilation in healthy controls, and unexpectedly increased dilation in MetSyn (P < 0.05). In contrast to hypoxia, hypercapnia-mediated vasodilation was similar between groups, as was the decrease in vasodilation with indomethacin. These data indicate increased production of vasoconstrictor prostaglandins restrains hypoxic cerebral vasodilation in MetSyn, preventing them from responding appropriately to this important physiological stressor.


Graphene oxide assisted spontaneous growth of V2O5 nanowires at room temperature.
Graphene-decorated single crystalline V2O5 nanowires (G-VONs) have been synthesized by mixing graphene oxide (GO) and V2O5 suspensions at room temperature. In this process, V2O5 nanowires (VONs) are formed spontaneously from commercial V2O5 particles with the aid of GO. The as-formed one dimensional G-VONs were characterized by using a X-ray diffractometer, a X-ray photoelectron spectrometer, a scanning electron microscope, and a transmission electron microscope. GO plays a vital role in the VON formation with the simultaneous reduction of GO. A single G-VON showed superior electrical conductivity compared with that of the pure VONs obtained from the sol-gel method. This could be ascribed to the insertion of rGO sheets into the V2O5 layered structure, which was further confirmed by electron energy loss spectroscopy.


Does language guide event perception? Evidence from eye movements.
Languages differ in how they encode motion. When describing bounded motion, English speakers typically use verbs that convey information about manner (e.g., slide, skip, walk) rather than path (e.g., approach, ascend), whereas Greek speakers do the opposite. We investigated whether this strong cross-language difference influences how people allocate attention during motion perception. We compared eye movements from Greek and English speakers as they viewed motion events while (a) preparing verbal descriptions or (b) memorizing the events. During the verbal description task, speakers' eyes rapidly focused on the event components typically encoded in their native language, generating significant cross-language differences even during the first second of motion onset. However, when freely inspecting ongoing events, as in the memorization task, people allocated attention similarly regardless of the language they speak. Differences between language groups arose only after the motion stopped, such that participants spontaneously studied those aspects of the scene that their language does not routinely encode in verbs. These findings offer a novel perspective on the relation between language and perceptual/cognitive processes. They indicate that attention allocation during event perception is not affected by the perceiver's native language; effects of language arise only when linguistic forms are recruited to achieve the task, such as when committing facts to memory.


Capitalizing on deliberate, accidental, and GM-driven environmental change caused by crop modification.
The transgenic traits associated with the majority of commercial genetically modified crops are focused on improving herbicide and insecticide management practices. The use of the transgenic technology in these crops and the associated chemistry has been the basis of studies that provide evidence for occasional improvement in environmental benefits due to the use of less residual herbicides, more targeted pesticides, and reduced field traffic. This is nicely exemplified through studies using Environmental Impact Quotient (EIQ) assessments. Whilst EIQ evaluations may sometimes illustrate environmental benefits they have their limitations. EIQ evaluations are not a surrogate for Environmental Risk Assessments and may not reflect real environmental interactions between crops and the environment. Addressing the impact cultivated plants have on the environment generally attracts little public attention and research funding, but the introduction of GM has facilitated an expansion of research to address potential environmental concerns from government, NGOs, industry, consumers, and growers. In this commentary, some evidence from our own research and several key papers that highlight EIQ assessments of the impact crops are having on the environment are presented. This information may be useful as an education tool on the potential benefits of GM and conventional farming. In addition, other deliberate, accidental, and GM-driven benefits derived from the examination of GM cropping systems is briefly discussed.


Expression of connexin36 mRNA in adult rodent brain.
A new member of the connexin gene family, named Connexin36 (Cx36) has, recently, been identified in rodents and shown to be preferentially, if not exclusively, expressed in neurones of the adult CNS. In this study we present a detailed in situ hybridization analysis of the expression pattern of mouse Connexin 36 (mCx36) mRNA in the adult mouse brain, with particular regards to the correlation of mCx36 expression to specific neuronal cell classes and systems. We found that mCx36 was strongly and widely expressed in the brain, including areas where the presence of gap junctions was never detected before. Quantitative analysis of the hybridization signal indicated varying levels of expression in different areas. In particular mCx36 was highly expressed in the neurones at different levels of the motor pathway, the olfactory pathway, the hippocampus, and areas related to the generation of respiratory rhythm. On the contrary, mCx36 was more heterogeneously expressed in nuclei of the sensory pathways. These findings show that mCx36 is the first connexin specifically expressed in neuronal cells in the adult rodent brain. The profiles of expression clearly indicate that mCx36 might play specific roles within different neuronal systems.


Diagnostic loop electrosurgical excisional procedure for discrepancy: do preoperative factors predict presence of significant cervical intraepithelial neoplasia?
Although pathological discrepancy between Pap smear and biopsy is an accepted indication to perform a diagnostic loop electrosurgical excision procedure (LEEP), this procedure is not without complications. Our objective was to determine the incidence of cervical intraepithelial neoplasia (CIN) 2,3 and patient factors that increase the likelihood of detecting CIN 2,3. We performed a retrospective chart review of patients who underwent a diagnostic LEEP for pathological discrepancy at a university-based colposcopy clinic. Pathological discrepancy is defined as a high-grade Pap smear with a colposcopically directed biopsy of CIN 1 or less. Demographic, cytological, and histological information were collected using a computerized database. The patients were divided into 2 groups (CIN 2,3 and CIN 1 or less) based on the pathology from the LEEP specimen. Patient factors were compared with final pathological results using chi(2) test, Student t test, Wilcoxon rank sum test, and multivariate analysis as indicated. A total of 102 patients were identified. Seven patients had normal specimens, 3 had HPV changes, 25 had CIN 1, 29 had CIN 2, and 38 had CIN 3. Thirty-five patients (34%) had CIN 1 or less, whereas 67 patients (66%) had CIN 2,3. The 2 groups were comparable in terms of age (30.4 vs 28.1 years), parity (2.2 vs 1.9), and age of coitarche (16.3 vs 16.4 years). No statistical difference existed between the groups regarding race, smoking status, Pap smear, history of previous cytological abnormality, contraception method, number of previous sexual partners, and HIV status. The majority of patients (75%) had not undergone previous treatment of CIN. The CIN 2,3 group were more likely than the CIN 1 or less group to have had previous treatment or biopsy for CIN (66% vs 34%; p = .004). Univariate (p = .004) and multivariate (p < .001) analysis demonstrated previous treatment of CIN as the only significant factor predicting CIN 2,3. Two thirds of women undergoing a LEEP for pathological discrepancy between Pap smear and cervical biopsy will have CIN 2,3. Women that have had previous treatment of CIN are more likely to have CIN 2,3 detected on their LEEP specimen.


Bacterial Colonization of the Implant-Abutment Interface (IAI) of Dental Implants with a Sloped Marginal Design: An in-vitro Study.
The aim of this study is to utilize an in vitro dynamic loading model to assess the potential risk of bacterial invasion into the Implant Abutment Interface (IAI) microgap of dental implants with sloped marginal design. Forty implants were divided into two groups (n = 20 per group) based on implant marginal design. Group 1 was comprised of implants with Morse-taper connection and conventional marginal design that connected to titanium abutments. Group 2 was comprised of implants with Morse-taper connection and sloped marginal design that connected to titanium abutments. The specimens were immersed in a bacterial solution of E. coli and loaded with 500,000 cycles of 160N using a chewing simulator. Following disconnection of fixtures and abutments, microbial samples were taken from the threaded portion of the abutment, plated and cultured under appropriate conditions. Ten out of twenty implants of Group 1 and eight out of twenty implants of Group 2 had IAI microgaps colonized by E. Coli. There was not a statistically significant difference in the mean number of E. Coli CFU detected between implants of Group 1 (mean 19.2, SD 23.6) and Group 2 (mean 12.5, SD18.9) (p > .05). The present study demonstrated that implants with a sloped marginal design exhibited similar risk for bacterial invasion into the IAI microgap under in vitro dynamic loading conditions compared to implants with conventional marginal design.


Plasmodium Sporozoite Biology.
Plasmodium sporozoite transmission is a critical population bottleneck in parasite life-cycle progression and, hence, a target for prophylactic drugs and vaccines. The recent progress of a candidate antisporozoite subunit vaccine formulation to licensure highlights the importance of sporozoite transmission intervention in the malaria control portfolio. Sporozoites colonize mosquito salivary glands, migrate through the skin, penetrate blood vessels, breach the liver sinusoid, and invade hepatocytes. Understanding the molecular and cellular mechanisms that mediate the remarkable sporozoite journey in the invertebrate vector and the vertebrate host can inform evidence-based next-generation drug development programs and immune intervention strategies.


Effect of lemongrass water extract supplementation on atherogenic index and antioxidant status in rats.
Cymbopogon citratus (DC) Stapf., commonly known as lemongrass, possesses strong antioxidant and cardiotonic properties. Lemongrass water extract contains several polyphenolic compounds including gallic acid, isoquercetin, quercetin, rutin, catechin and tannic acid. Rutin, isoquercetin catechin and quercetin are the flavonoids most abundantly found in the extract. The extract significantly decreased total cholesterol, low-density lipoprotein and atherogenic index in rats after treatment (p < 0.05). Expression of genes and protein of sterol regulatory element binding protein-1c (SREBP1c) and HMG-CoA reductase (HMGR) was also lowered significantly in treated groups (p < 0.05). Moreover, serum antioxidant capacity increased in treated rats in comparison with untreated ones (p < 0.05) and was associated with decreased serum lipid peroxidation.


Proposed criteria for identifying GE crop plants that pose a low or negligible risk to the environment under conditions of low-level presence in seed.
The low-level presence (LLP) of genetically engineered (GE) seeds that have been approved in the country of origin but not the country of import presents challenges for regulators in both seed importing and exporting countries, as well as for the international seed trade and the farmers who rely on it. In addition to legal, financial and regulatory challenges, such LLP situations in seed may also require an environmental risk assessment by the country of import. Such assessments have typically been informed by the national framework established to support decisions related to wide scale cultivation, and frequently do not take into account the low environmental exposure and prior regulatory history of the GE plant. In addition, such assessment processes may not be well suited to the decision-making timeframe that is necessary when dealing with an LLP situation in imported seed. In order to facilitate regulatory decision making, this paper proposes a set of scientific criteria for identifying GE crop plants that are expected to pose a low or negligible risk to the environment under LLP conditions in seed. Regulatory decision makers in some importing countries may decide to use these criteria to assist in risk analysis associated with LLP situations they are experiencing or could experience in the future, and might choose to proactively apply the criteria to identify existing GE plants with regulatory approvals in other countries that would be expected to pose low risk under conditions of LLP in seed.


The native 3D organization of bacterial polysomes.
Recent advances have led to insights into the structure of the bacterial ribosome, but little is known about the 3D organization of ribosomes in the context of translating polysomes. We employed cryoelectron tomography and a template-matching approach to map 70S ribosomes in vitrified bacterial translation extracts and in lysates of active E. coli spheroplasts. In these preparations, polysomal arrangements were observed in which neighboring ribosomes are densely packed and exhibit preferred orientations. Analysis of characteristic examples of polysomes reveals a staggered or pseudohelical organization of ribosomes along the mRNA trace, with the transcript being sequestered on the inside, the tRNA entrance sites being accessible, and the polypeptide exit sites facing the cytosol. Modeling of elongating nascent polypeptide chains suggests that this arrangement maximizes the distance between nascent chains on adjacent ribosomes, thereby reducing the probability of intermolecular interactions that would give rise to aggregation and limit productive folding.


Sensory Pattern Contributions to Developmental Performance in Children With Autism Spectrum Disorder.
Sensory processing differences in preschool-age children with autism spectrum disorder (ASD) affect their engagement in everyday activities, thereby influencing opportunities to practice and develop skills such as social communication and adaptive behavior. The purpose of this study was to investigate the extent to which specific sensory processing patterns relate to aspects of development (i.e., adaptive behavior, expressive and receptive language, fine and gross motor skills, social behavior) in a sample of preschool-age children with ASD (N=400). A retrospective chart review was used to gather clinical data. Results suggest that sensory processing patterns differentially affect children's developmental skills and adaptive behavior. Certain sensory processing patterns predicted children's development of language, motor, and adaptive skills. These findings have clear implications for occupational therapy practice with young children with ASD. Practitioners should consider how sensory processing in ASD both supports and limits children's ability to engage in social communication and learning opportunities.


A Randomized, Double-Blind, Placebo-Controlled Phase 2 Study of Brodalumab in Patients With Moderate-to-Severe Crohn's Disease.
To assess the safety and efficacy of brodalumab, a human anti-interleukin-17 receptor monoclonal antibody, in patients with moderate-to-severe Crohn's disease (CD). Phase 2, randomized, double-blind, placebo-controlled, dose-ranging study in patients with moderate-to-severe CD and evidence of active inflammation. Patients were randomized 1:1:1:1 to receive brodalumab (210, 350, or 700 mg at baseline and week 4) or placebo. The primary end point was proportion of patients achieving Crohn's disease activity index (CDAI) remission (≤150) at week 6. Secondary end points included proportion of patients with CDAI response (reduction from baseline of ≥100) at week 6 and change from baseline in CDAI at week 6. The study was terminated early based on an imbalance in worsening CD in active treatment groups. At the time of termination, 130 patients had been randomized. At week 6, remission rates were 3% (210 mg), 15% (350 mg), 9% (700 mg), and 3% (placebo) and CDAI response occurred in 16% (210 mg), 27% (350 mg), 15% (700 mg), and 13% (placebo) of patients. Mean change in CDAI at week 6 was -8.7 (95.3) (210 mg), -35.4 (105.6) (350 mg), -0.6 (105.9) (700 mg), and -28.2 (86.0) (placebo). Besides worsening of CD, overall incidences of adverse events were similar across treatment groups. Treatment with brodalumab resulted in a disproportionate number of cases of worsening CD in patients with active CD and no evidence of meaningful efficacy. These analyses did not suggest additional safety risks of brodalumab beyond worsening of CD symptoms in patients with active CD.


EEG activation differences in the pre-motor cortex and supplementary motor area between normal individuals with high and low traits of autism.
The human mirror neuron system (hMNS) is believed to provide a basic mechanism for social cognition. Event-related desynchronization (ERD) in alpha (8-12Hz) and low beta band (12-20Hz) over sensori-motor cortex has been suggested to index mirror neurons' activity. We tested whether autistic traits revealed by high and low scores on the Autistic Quotient (AQ) in the normal population are linked to variations in the electroencephalogram (EEG) over motor, pre-motor cortex and supplementary motor area (SMA) during action observation. Results revealed that in the low AQ group, the pre-motor cortex and SMA were more active during hand action than static hand observation whereas in the high AQ group the same areas were active both during static and hand action observation. In fact participants with high traits of autism showed greater low beta ERD while observing the static hand than those with low traits and this low beta ERD was not significantly different when they watched hand actions. Over primary motor cortex, the classical alpha and low beta ERD during hand actions relative to static hand observation was found across all participants. These findings suggest that the observation-execution matching system works differently according to the degree of autism traits in the normal population and that this is differentiated in terms of the EEG according to scalp site and bandwidth.


What's New, What's Hot in Solid Organ Transplantation? Summary of the American Transplant Congress 2011.
Breakthroughs in basic and clinical science in solid organ transplantation were presented at the American Transplant Congress 2011. Key areas of presentation included the pathogenesis of late allograft failure, immune regulation and tolerance, pathways in allograft injury, electing appropriate patients for transplantation, determining the best allocation schemes to maximize effective utilization, organ preservation, monitoring the alloimmune response and immunosuppressive management. In this review, we present highlights of the meeting. These presentations demonstrate the exciting promise in translating from the bench to affect patient care.


Evaluation of a nutrition model in predicting performance of vietnamese cattle.
The objective of this study was to evaluate the predictions of dry matter intake (DM) and average daily gain (ADG) of Vietnamese Yellow (Vang) purebred and crossbred (Vang with Red Sindhi or Brahman) bulls fed under Vietnamese conditions using two levels of solution (1 and 2) of the large ruminant nutrition system (LRNS) model. Animal information and feed chemical characterization were obtained from five studies. The initial mean body weight (BW) of the animals was 186, with standard deviation ±33.2 kg. Animals were fed ad libitum commonly available feedstuffs, including cassava powder, corn grain, Napier grass, rice straw and bran, and minerals and vitamins, for 50 to 80 d. Adequacy of the predictions was assessed with the Model Evaluation System using the root of mean square error of prediction (RMSEP), accuracy (Cb), coefficient of determination (r(2)), and mean bias (MB). When all treatment means were used, both levels of solution predicted DMI similarly with low precision (r(2) of 0.389 and 0.45 for level 1 and 2, respectively) and medium accuracy (Cb of 0.827 and 0.859, respectively). The LRNS clearly over-predicted the intake of one study. When this study was removed from the comparison, the precision and accuracy considerably increased for the level 1 solution. Metabolisable protein was limiting ADG for more than 68% of the treatment averages. Both levels differed regarding precision and accuracy. While level 1 solution had the least MB compared with level 2 (0.058 and 0.159 kg/d, respectively), the precision was greater for level 2 than level 1 (0.89 and 0.70, respectively). The accuracy (Cb) was similar between level 1 and level 2 (p = 0.8997; 0.977 and 0.871, respectively). The RMSEP indicated that both levels were on average under- or over-predicted by about 190 g/d, suggesting that even though the accuracy (Cb) was greater for level 1 compared to level 2, both levels are likely to wrongly predict ADG by the same amount. Our analyses indicated that the level 1 solution can predict DMI reasonably well for this type of animal, but it was not entirely clear if animals consumed at their voluntary intake and/or if the roughness of the diet decreased DMI. A deficit of ruminally-undegradable protein and/or a lack of microbial protein may have limited the performance of these animals. Based on these evaluations, the LRNS level 1 solution may be an alternative to predict animal performance when, under specific circumstances, the fractional degradation rates of the carbohydrate and protein fractions are not known.


Effects of combined dietary chromium(III) propionate complex and thiamine supplementation on insulin sensitivity, blood biochemical indices, and mineral levels in high-fructose-fed rats.
Insulin resistance is the first step in glucose intolerance and the development of type 2 diabetes mellitus, thus effective prevention strategies should also include dietary interventions to enhance insulin sensitivity. Nutrients, such as microelement chromium(III) and thiamine, play regulatory roles in carbohydrate metabolism. The objective of this study was to evaluate the insulin-sensitizing potential of the combined supplementary chromium(III) propionate complex (CrProp) and thiamine in insulin resistance animal model (rats fed a high-fructose diet). The experiment was carried out on 40 nine-week-old male Wistar rats divided into five groups (eight animals each). Animals were fed ad libitum: the control diet (AIN-93 M) and high-fructose diets with and without a combination of two levels of CrProp (0.1 and 1 mg Cr/kg body mass/day) and two levels of thiamine (0.5 and 10 mg/kg body mass/day) for 8 weeks. At the end of the experiment rats were sacrificed to collect blood and internal organs for analyses of blood biochemical and hematologic indices as well as tissular microelement levels that were measured using appropriate methods. It was found that both supplementary CrProp and thiamine (given alone) have significant insulin-sensitizing and moderate blood-lipid-lowering properties, while the combined supplementation with these agents does not give synergistic effects in insulin-resistant rats. CrProp given separately increased kidney Cu and Cr levels, while thiamine alone increased hepatic Cu contents and decreased renal Zn and Cu contents.


Reduced Mortality Associated with Acute Kidney Injury Requiring Dialysis in the United States.
Dialysis-requiring acute kidney injury (AKI-D) is a documented complication of hospitalization and procedures. Temporal incidence of AKI-D and related hospital mortality in the US population has not been recently characterized. We describe the epidemiology of AKI-D as well as associated in-hospital mortality in the US. Retrospective cohort of a national discharge data (n = 86,949,550) from the Healthcare Cost and Utilization Project's National Inpatient Sample, 2001-2011 of patients' hospitalization with AKI-D. Primary outcomes were AKI-D and in-hospital mortality. We determined the annual incidence rate of AKI-D in the US from 2001 to 2011. We estimated ORs for AKI-D and in-hospital mortality for each successive year compared to 2001 using multiple logistic regression models, adjusted for patient and hospital characteristics, and stratified the analyses by sex and age. We also calculated population-attributable risk of in-hospital mortality associated with AKI-D. The adjusted odds of AKI-D increased by a factor of 1.03 (95% CI 1.02-1.04) each year. The number of AKI-D-related (19,886-34,195) in-hospital deaths increased almost 2-fold, although in-hospital mortality associated with AKI-D (28.0-19.7%) declined significantly from 2001 to 2011. Over the same period, the adjusted odds of mortality for AKI-D patients were 0.60 (95% CI 0.56-0.67). Population-attributable risk of mortality associated with AKI-D increased (2.1-4.2%) over the study period. The incidence rate of AKI-D has increased considerably in the US since 2001. However, in-hospital mortality associated with AKI-D hospital admissions has decreased significantly.


HIV-1 vaccine immunogen design strategies.
An effective human immunodeficiency virus type 1 (HIV-1) vaccine is expected to have the greatest impact on HIV-1 spread and remains a global scientific priority. Only one candidate vaccine has significantly reduced HIV-1 acquisition, yet at a limited efficacy of 31%, and none have delayed disease progression in vaccinated individuals. Thus, the challenge remains to develop HIV-1 immunogens that will elicit protective immunity. A combination of two independent approaches - namely the elicitation of broadly neutralising antibodies (bNAb) to prevent or reduce acquisition of infection and stimulation of effective cytotoxic T lymphocyte (CTL) responses to slow disease progression in breakthrough infections (recent evidence suggests that CTLs could also block HIV-1 from establishing persistent infection) - is the current ideal. The purpose of this review is to summarise strategies and progress in the design and testing of HIV-1 immunogens to elicit bNAb and protective CTL immune responses. Recent advances in mimicking the functional native envelope trimer structure and in designing structurally-stabilised bNAb epitope forms to drive development of germline precursors to mature bNAb are highlighted. Systematic or computational approaches to T cell immunogen design aimed at covering viral diversity, increasing the breadth of immune responses and/or reducing viable viral escape are discussed. We also discuss a recent novel vaccine vector approach shown to induce extremely broad and persistent T cell responses that could clear highly pathogenic simian immunodeficiency virus (SIV) early after infection in the monkey model. While in vitro and animal model data are promising, Phase II and III human clinical trials are ultimately needed to determine the efficacy of immunogen design approaches.


Recent advances in N-heterocyclic carbene catalyzed achiral synthesis.
N-Heterocyclic carbenes (NHCs) have emerged as powerful and elegant organocatalysts in a variety of newly developed and unprecedented enantioselective transformations due to their unique umpolung capacity. As a supplement to conventional enantioselective organocatalysis, NHC-induced non-asymmetric catalysis has gradually attracted much interest in recent years. Herein, this review aims to reveal the recent developments in NHC-promoted non-asymmetric umpolung transformations resulting in the expeditious construction of versatile achiral natural heterocycles, carbocycles and acylated products.


Mapping two functional domains of clathrin light chains with monoclonal antibodies.
The two forms of clathrin light chains (LCA and LCB) or clathrin-associated proteins (CAP1 and CAP2) have presented an immunochemical paradox. Biochemically similar, both possess two known functional parameters: binding the clathrin heavy chain and mediating the action of an uncoating ATPase. All previously reported anti-CAP mAbs, however, react specifically with only CAP1 (Brodsky, F. M., 1985, J. Cell Biol., 101:2047-2054; Kirchhausen, T., S. C. Harrison, P. Parham, and F. M. Brodsky, 1983, Proc. Natl. Acad. Sci. USA, 80:2481-2485). Four new anti-CAP mAbs are reported here: two, C-7H12 and C-6C1, react with both forms; two others, C-10B2 and C-4E5, react only with the lower form. Sandwich ELISAs indicated that C-10B2, C-4E5, C-6C1, and C-7H12 react with distinct epitopes. Monoclonal antibodies C-10B2 and C-4E5 immunoprecipitate clathrin-coated vesicles (CCVs) and react with CAP2 epitopes accessible to chymotrypsin on the vesicle. These mAbs inhibit phosphorylation of CAP2 by endogenous CCV casein kinase II. In contrast, C-6C1 and C-7H12 react with epitopes that are relatively insensitive to chymotrypsin. CAP peptide fragments containing these epitopes remain bound to reassembled cages or CCVs after digestion. Immunoprecipitation and ELISAs demonstrate that C-7H12 and C-6C1 react with unbound CAPs but not with CAPs bound to triskelions or CCVs. The data indicate that the CAPs consist of at least two discernible structural domains: a nonconserved, accessible domain that is relevant to the phosphorylation of CAP2 and a conserved, inaccessible domain that mediates the binding of CAPs to CCVs.


Sensory testing associates with pain quality descriptors during acute dental pain.
Pain descriptors capture the multidimensional nature of pain and can elucidate underlying pathophysiological mechanisms. This study determined whether the pain descriptors chosen by subjects experiencing acute dental pain associate with the outcomes of two commonly performed dental sensory tests. The goal of the study is to clarify whether pain descriptors are useful in discriminating the underlying biological processes contributing to dental pain. Participants (n = 228) presenting with acute toothache underwent standardized clinical dental sensory testing and described their pain in reference to 22 pain quality descriptors. Univariate and two-way ANOVA determined the relationship between groups defined by cold detection (positive or negative) and percussion hypersensitivity (painful or not) on the affected tooth, and pain descriptor reporting. Subjects experiencing painful toothache most frequently reported evoked pain to temperature and chewing, and pain descriptors such as "throbbing" and "aching." They also reported neuropathic pain descriptors such as "tingling" and "electric shock." Subjects who detected a cold stimulus (thermal) on the affected tooth, frequently reported high intensity paroxysmal shooting pain compared to those that did not detect cold. By contrast, patients with percussion (mechanical) hypersensitivity on the affected tooth, reported higher levels of global pain intensity at rest and in function, and reported significantly higher intensity "radiating" and "throbbing" pain, than subjects with non-painful percussion. The reporting of neuropathic pain descriptors by subjects experiencing acute toothache was more frequent than expected, suggesting that neuropathic mechanisms could contribute to typical toothache pain. Subjects experiencing toothache with mechanical hypersensitivity experience more intense pain overall. In subjects experiencing acute toothache, specific pain descriptors associate with the responses to routine clinical sensory tests performed on the injured tooth. The frequent reporting of neuropathic pain descriptors suggests that neuropathic mechanisms could create a diagnostic challenge to differentiate toothache from intraoral neuropathic conditions. Persons experiencing toothache with mechanical hypersensitivity experience more intense pain overall, suggesting patients with this clinical feature will have distinct clinical pain management needs.


Epigenetic modulation of secondary metabolite profiles in Aspergillus calidoustus and Aspergillus westerdijkiae through histone deacetylase (HDAC) inhibition by vorinostat.
The effects of epigenetic modulation on secondary metabolite biosynthesis were investigated with five Aspergillus species cultured in the presence of either the DNA methyltransferase inhibitor 5-azacitidine or the histone deacetylase inhibitor vorinostat. With Aspergillus calidoustus and Aspergillus westerdijkiae, fermentation in the presence of vorinostat (100 μM) induced significant changes in secondary metabolite profile with examples of both induction and repression. We identified putative biosynthetic gene clusters for emericellamide in A. calidoustus and ochratoxin in A. westerdijkiae. A substantial induction in production levels was observed for two secondary metabolites: the diketopiperazine alkaloid phenylahistin in A. calidoustus and the polyketide penicillic acid in A. westerdijkiae, indicating the potential of epigenetic regulation for the activation of silent fungal biosynthetic pathways.


Yellow laser acupuncture - A new option for prevention and early intervention of lifestyle-related diseases: A randomized, placebo-controlled trial in volunteers.
The yellow laser constitutes a totally new option in the field of laser acupuncture, in addition to the already existing red, near infrared, green and violet lasers. Especially for so called lifestyle-related diseases, this could open up new methods of integrative therapy. The goal of the present study was to investigate among other parameters blood pressure (BP), heart rate (HR), heart rate variability (HRV), and temperature effects before, during, and after stimulation of different acupoints with yellow laser. We recruited 26 healthy volunteers (13 female, 13 male; mean age ± SD 24.1 ± 3.3 years) at the Medical University of Graz. The acupoints Baihui, Neiguan, Taichong and a placebo point were stimulated with a 589 nm (50 mW, 500 µm; 5 min) yellow laser. Blood pressure was measured noninvasively at the wrist; for the registration of the electrocardiogram a medilog AR12 HRV system was used. Effects on temperature were measured with a Flir i7 infrared camera. There were significant decreases after yellow laser acupuncture in the systolic BP, diastolic BP also decreased (n.s.). HRV in both (men and women) increased. The temperature during the yellow laser stimulation decreased significantly in all measured points. After the stimulation it increased again significantly. Based on a questionnaire volunteers reported a significantly decreased level of stress after yellow laser stimulation. Significant positive effects on BP and well-being were found after yellow laser stimulation. The results are very promising and can be very important especially for the treatment of lifestyle related diseases.


Heterogeneity of cyclic nucleotide phosphodiesterases in liver endoplasmic reticulum.
A microsomal fraction from rat liver was subfractionated into three rough endoplasmic reticulum fractions RIII, RII and RI, together with a smooth endoplasmic reticulum plus Golgi fraction. Cyclic nucleotide phosphodiesterase activity was found in all fractions. Subsequently it was shown that Golgi fractions were essentially devoid of cyclic AMP phosphodiesterase activity and the activity resided in the smooth endoplasmic reticulum fraction. The activity of the endoplasmic reticulum constituted some 20% of the homogenate activity, with the major fraction of this being associated with the RII fraction and the least with the RI fraction. With the exception of the activity of the RI fraction, which was a peripheral enzyme, all of the other enzyme activities were integral, requiring detergent or repeated freeze-thawing to effect solubilization. All of the activities appeared to be exposed at the external surface of the endoplasmic reticulum, as they were inactivated by trypsin under conditions where glucose 6-phosphatase was not. All of these activities displayed distinct sensitivities to both thermal and trypsin inactivation, yielding activity decays consistent with a single enzyme species being present in each case. The freeze-thaw-solubilized enzymes yielded single symmetrical peaks on sucrose-density-gradient centrifugation and polyacrylamide-gel electrophoresis. The sedimentation coefficients for the enzymes in the smooth-endoplasmic-reticulum-plus-Golgi, RIII, RII and RI fractions were 3.2S, 4.2S, 4.5S and 4.5S respectively. Whereas the activity in the smooth-endoplasmic-reticulum-plus-Golgi fraction exhibited normal Michaelis kinetics, those in the other fractions yielded kinetics indicative of apparent negative co-operativity. All of the enzymes exhibited low Km values towards cyclic AMP. The enzymes did not appear to be regulated by Ca2+ or calmodulin. ZnCl2 was found to be a potent non-competitive inhibitor of the enzyme in all fractions. NaF was a weak non-competitive inhibitor. The bilayer fluidizing agent benzyl alcohol exerted dissimilar effects on the enzyme activities. It is concluded that the endoplasmic reticulum displays lateral heterogeneity, with single, rather distinct, cyclic AMP phosphodiesterases being found in the different fractions.


Surface alignment disorder and thermal Casimir forces in smectic-A liquid crystalline films.
Disorder components in surface anchoring orientation of a smectic-A liquid crystalline film (occurring, e.g., due to surface contamination sources) modify the thermal pseudo-Casimir interaction mediated between the bounding surfaces of the film. By considering a plane-parallel slab with bounding surfaces positioned normal to the layering field of the film, we study the anchoring disorder effects by assuming that the disorder source is present on one of the two substrates, producing a Gaussian-weighted distribution for the preferred molecular anchoring orientation (easy axis) on that substrate, with a finite mean and variance or, more generally, with a homogeneous in-plane, two-point correlation function. We show that the presence of disorder, either of quenched or annealed type, leads to a significant reduction in the magnitude of the net pseudo-Casimir force between the confining substrates of the film. This force can be attractive or repulsive depending on the boundary conditions. In the quenched case, the interaction force reduction is a direct consequence of an additive free energy term dependent on the variance of the disorder, while in the annealed case, the suppression of the interaction force can be understood based on a disorder-renormalized, effective anchoring strength. We predict a regime of behavior, exhibiting non-monotonic dependence for the interaction pressure as a function of separation. We also show that, by increasing the disorder variance, the interaction pressure changes sign and becomes a monotonically decreasing function of the separation between the substrates.


Impact of epididymal maturation, ejaculation and in vitro capacitation on tyrosine phosphorylation patterns exhibited of boar (Sus domesticus) spermatozoa.
Mammalian spermatozoa acquire functionality during epididymal maturation and ability to penetrate and fertilize the oocyte during capacitation. The aim of this study was to investigate the impact of epididymal maturation, ejaculation and capacitation on phosphotyrosine content of sperm proteins. Western blot, immunocytochemical and flow cytometry analyses demonstrated that epididymal maturation in vivo is associated with a progressive loss of phosphotyrosine residues of the sperm head followed by a subtle increase after in vitro capacitation. As cells pass from caput to cauda epididymis, tyrosine phosphorylation becomes confined to a triangular band over the posterior part of midacrosome region, whereas in vitro capacitation causes a spread labeling over the whole head. Different bands with phosphotyrosine residues were detected during epididymal maturation and after in vitro capacitation: 1) 93, 66 and 45 kDa bands with specific phosphotyrosine expression in immature spermatozoa; 2) 76, 23 and 12 kDa bands with specific phosphotyrosine expression in mature spermatozoa, being significantly increased in their expression after in vitro capacitation; 3) 49, 40, 37, 30, 26 and 25 kDa constitutive bands that increased their phosphotyrosine expression after maturation and/or in vitro capacitation; and 4) 28 and 20 kDa bands with a specific phosphotyrosine expression in in vitro capacitated spermatozoa. These results provided integral novel data of expression and location of phosphotyrosine residues during epididymal maturation, ejaculation and in vitro capacitation of boar spermatozoa. Two new constitutive proteins bands of 26 and 25 kDa with phosphotyrosine residues were also identified.


Quantifying operational lifetimes for coal power plants under the Paris goals.
A rapid transition away from unabated coal use is essential to fulfilling the Paris climate goals. However, many countries are actively building and operating coal power plants. Here we use plant-level data to specify alternative trajectories for coal technologies in an integrated assessment model. We then quantify cost-effective retirement pathways for global and country-level coal fleets to limit long-term temperature change. We present our results using a decision-relevant metric: the operational lifetime limit. Even if no new plants are built, the lifetimes of existing units are reduced to approximately 35 years in a well-below 2 °C scenario or 20 years in a 1.5 °C scenario. The risk of continued coal expansion, including the near-term growth permitted in some Nationally Determined Contributions (NDCs), is large. The lifetime limits for both 2 °C and 1.5 °C are reduced by 5 years if plants under construction come online and 10 years if all proposed projects are built.


Circulating Phylloquinone Concentrations and Risk of Type 2 Diabetes: A Mendelian Randomization Study.
This study investigated the causal relation between circulating phylloquinone (vitamin K1) concentrations and type 2 diabetes by using a Mendelian randomization (MR) approach. We used data from three studies: the European Prospective Investigation into Cancer and Nutrition (EPIC)-InterAct case-cohort study, Diabetes Genetics Replication and Meta-analysis (DIAGRAM), and the UK Biobank, resulting in 69,647 subjects with type 2 diabetes. We calculated a weighted genetic risk score including four genetic variants previously found to be associated with circulating phylloquinone concentrations. Inverse-variance weighted analysis was used to obtain a risk ratio (RR) for the causal relation between circulating phylloquinone concentrations and risk of type 2 diabetes. Presence of pleiotropy and the robustness of the results were assessed using MR-Egger and weighted-median analyses. Genetically predicted concentrations of circulating phylloquinone were associated with lower risk of type 2 diabetes with an RR of 0.93 (95% CI 0.89; 0.97) per every natural logarithm (Ln)-nmol/L-unit increase in circulating phylloquinone. The MR-Egger and weighted median analyses showed RRs of 0.94 (0.86; 1.02) and 0.93 (0.88; 0.98), respectively, indicating no pleiotropy. In conclusion, our study supports that higher circulating phylloquinone may be causally related with lower risk of type 2 diabetes, highlighting the importance of sufficient phylloquinone in the human diet.


Ascorbic acid as an adjunctive therapy in critically ill patients with COVID-19: a propensity score matched study.
Ascorbic acid represents an appealing option for clinicians to utilize in the context of the global COVID-19 pandemic due to its proposed clinical efficacy, relative safety, and low cost. The aim of this study was to evaluate the efficacy and safety of using ascorbic acid in supplemental doses as adjunctive therapy for patients critically ill with COVID-19. This was a two-center, non-interventional, retrospective cohort study. All critically ill adult patients admitted to ICU with a confirmed COVID-19 diagnosis between March 1st and December 31st, 2020, were included in the final analysis. The study was conducted at two large governmental tertiary hospitals in Saudi Arabia. The purpose was to investigate the clinical outcomes of low-dose ascorbic acid as adjunctive therapy in COVID-19 after propensity score matching using baseline severity scores, systematic use of corticosteroids, and study centers. A number of 739 patients were included in this study, among whom 296 patients were included after propensity score matching. There was no association between the administration of ascorbic acid and in-hospital mortality or the 30-day mortality [OR (95% CI) 0.77 (0.47, 1.23), p value = 0.27 and OR (95% CI) 0.73 (0.43, 1.20), p value = 0.21, respectively]. Using ascorbic acid was associated with a lower incidence of thrombosis compared with the non-ascorbic-acid group [6.1% vs. 13% respectively; OR (95% CI) 0.42 (0.184, 0.937), p value = 0.03]. Low dose of ascorbic acid as an adjunctive therapy in COVID-19 critically ill patients was not associated with mortality benefits, but it was associated with a lower incidence of thrombosis. Further studies are required to confirm these findings.


Analysis of the brain palmitoyl-proteome using both acyl-biotin exchange and acyl-resin-assisted capture methods.
Palmitoylation is a reversible post-translational protein modification in which palmitic acid is added to cysteine residues, allowing association with different cellular membranes and subdomains. Recently, techniques have been developed to identify palmitoylation on a proteome-wide scale in order to reveal the full cellular complement of palmitoylated proteins. However, in the studies reported to date, there is considerable variation between the sets of identified palmitoyl-proteins and so there remains some uncertainty over what constitutes the definitive complement of palmitoylated proteins even in well-studied tissues such as brain. To address this issue, we used both acyl-biotin exchange and acyl-resin-assisted capture approaches using rat brain as a common protein source. The palmitoyl-proteins identified from each method by mass spectrometry were then compared with each other and previously published studies. There was generally good agreement between the two methods, although many identifications were unique to one method, indicating that at least some of the variability in published palmitoyl proteomes is due to methodological differences. By combining our new data with previous publications using mammalian cells/tissues, we propose a high confidence set of bona fide palmitoylated proteins in brain and provide a resource to help researchers prioritise candidate palmitoyl-proteins for investigation.


Types of Myocardial Infarction Among Human Immunodeficiency Virus-Infected Individuals in the United States.
The Second Universal Definition of Myocardial Infarction (MI) divides MIs into different types. Type 1 MIs result spontaneously from instability of atherosclerotic plaque, whereas type 2 MIs occur in the setting of a mismatch between oxygen demand and supply, as with severe hypotension. Type 2 MIs are uncommon in the general population, but their frequency in human immunodeficiency virus (HIV)-infected individuals is unknown. To characterize MIs, including type; identify causes of type 2 MIs; and compare demographic and clinical characteristics among HIV-infected individuals with type 1 vs type 2 MIs. This longitudinal study identified potential MIs among patients with HIV receiving clinical care at 6 US sites from January 1, 1996, to March 1, 2014, using diagnoses and cardiac biomarkers recorded in the centralized data repository. Sites assembled deidentified packets, including physician notes and electrocardiograms, procedures, and clinical laboratory tests. Two physician experts adjudicated each event, categorizing each definite or probable MI as type 1 or type 2 and identifying the causes of type 2 MI. The number and proportion of type 1 vs type 2 MIs, demographic and clinical characteristics among those with type 1 vs type 2 MIs, and the causes of type 2 MIs. Among 571 patients (median age, 49 years [interquartile range, 43-55 years]; 430 men and 141 women) with definite or probable MIs, 288 MIs (50.4%) were type 2 and 283 (49.6%) were type 1. In analyses of type 1 MIs, 79 patients who underwent cardiac interventions, such as coronary artery bypass graft surgery, were also included, totaling 362 patients. Sepsis or bacteremia (100 [34.7%]) and recent use of cocaine or other illicit drugs (39 [13.5%]) were the most common causes of type 2 MIs. A higher proportion of patients with type 2 MIs were younger than 40 years (47 of 288 [16.3%] vs 32 of 362 [8.8%]) and had lower current CD4 cell counts (median, 230 vs 383 cells/µL), lipid levels (mean [SD] total cholesterol level, 167 [63] vs 190 [54] mg/dL, and mean (SD) Framingham risk scores (8% [7%] vs 10% [8%]) than those with type 1 MIs or who underwent cardiac interventions. Approximately half of all MIs among HIV-infected individuals were type 2 MIs caused by heterogeneous clinical conditions, including sepsis or bacteremia and recent use of cocaine or other illicit drugs. Demographic characteristics and cardiovascular risk factors among those with type 1 and type 2 MIs differed, suggesting the need to specifically consider type among HIV-infected individuals to further understand MI outcomes and to guide prevention and treatment.


Novel application of multi-stimuli network inference to synovial fibroblasts of rheumatoid arthritis patients.
Network inference of gene expression data is an important challenge in systems biology. Novel algorithms may provide more detailed gene regulatory networks (GRN) for complex, chronic inflammatory diseases such as rheumatoid arthritis (RA), in which activated synovial fibroblasts (SFBs) play a major role. Since the detailed mechanisms underlying this activation are still unclear, simultaneous investigation of multi-stimuli activation of SFBs offers the possibility to elucidate the regulatory effects of multiple mediators and to gain new insights into disease pathogenesis. A GRN was therefore inferred from RA-SFBs treated with 4 different stimuli (IL-1 β, TNF- α, TGF- β, and PDGF-D). Data from time series microarray experiments (0, 1, 2, 4, 12 h; Affymetrix HG-U133 Plus 2.0) were batch-corrected applying 'ComBat', analyzed for differentially expressed genes over time with 'Limma', and used for the inference of a robust GRN with NetGenerator V2.0, a heuristic ordinary differential equation-based method with soft integration of prior knowledge. Using all genes differentially expressed over time in RA-SFBs for any stimulus, and selecting the genes belonging to the most significant gene ontology (GO) term, i.e., 'cartilage development', a dynamic, robust, moderately complex multi-stimuli GRN was generated with 24 genes and 57 edges in total, 31 of which were gene-to-gene edges. Prior literature-based knowledge derived from Pathway Studio or manual searches was reflected in the final network by 25/57 confirmed edges (44%). The model contained known network motifs crucial for dynamic cellular behavior, e.g., cross-talk among pathways, positive feed-back loops, and positive feed-forward motifs (including suppression of the transcriptional repressor OSR2 by all 4 stimuli. A multi-stimuli GRN highly concordant with literature data was successfully generated by network inference from the gene expression of stimulated RA-SFBs. The GRN showed high reliability, since 10 predicted edges were independently validated by literature findings post network inference. The selected GO term 'cartilage development' contained a number of differentiation markers, growth factors, and transcription factors with potential relevance for RA. Finally, the model provided new insight into the response of RA-SFBs to multiple stimuli implicated in the pathogenesis of RA, in particular to the 'novel' potent growth factor PDGF-D.


Nanoscale DNA tetrahedra improve biomolecular recognition on patterned surfaces.
The bottom-up approach of DNA nano-biotechnology can create biomaterials with defined properties relevant for a wide range of applications. This report describes nanoscale DNA tetrahedra that are beneficial to the field of biosensing and the targeted immobilization of biochemical receptors on substrate surfaces. The DNA nanostructures act as immobilization agents that are able to present individual molecules at a defined nanoscale distance to the solvent thereby improving biomolecular recognition of analytes. The tetrahedral display devices are self-assembled from four oligonucleotides. Three of the four tetrahedron vertices are equipped with disulfide groups to enable oriented binding to gold surfaces. The fourth vertex at the top of the bound tetrahedron presents the biomolecular receptor to the solvent. In assays testing the molecular accessibility via DNA hybridization and protein capturing, tetrahedron-tethered receptors outperformed conventional immobilization approaches with regard to specificity and amount of captured polypeptide by a factor of up to seven. The bottom-up strategy of creating DNA tetrahedrons is also compatible with the top-down route of nanopatterning of inorganic substrates, as demonstrated by the specific coating of micro- to nanoscale gold squares amid surrounding blank or poly(ethylene glycol)-passivated glass surfaces. DNA tetrahedra can create biofunctionalized surfaces of rationally designed properties that are of relevance in analytical chemistry, cell biology, and single-molecule biophysics.


Directional biases in phylogenetic structure quantification: a Mediterranean case study.
Recent years have seen an increasing effort to incorporate phylogenetic hypotheses to the study of community assembly processes. The incorporation of such evolutionary information has been eased by the emergence of specialized software for the automatic estimation of partially resolved supertrees based on published phylogenies. Despite this growing interest in the use of phylogenies in ecological research, very few studies have attempted to quantify the potential biases related to the use of partially resolved phylogenies and to branch length accuracy, and no work has examined how tree shape may affect inference of community phylogenetic metrics. In this study, using a large plant community and elevational dataset, we tested the influence of phylogenetic resolution and branch length information on the quantification of phylogenetic structure; and also explored the impact of tree shape (stemminess) on the loss of accuracy in phylogenetic structure quantification due to phylogenetic resolution. For this purpose, we used 9 sets of phylogenetic hypotheses of varying resolution and branch lengths to calculate three indices of phylogenetic structure: the mean phylogenetic distance (NRI), the mean nearest taxon distance (NTI) and phylogenetic diversity (stdPD) metrics. The NRI metric was the less sensitive to phylogenetic resolution, stdPD showed an intermediate sensitivity, and NTI was the most sensitive one; NRI was also less sensitive to branch length accuracy than NTI and stdPD, the degree of sensitivity being strongly dependent on the dating method and the sample size. Directional biases were generally towards type II errors. Interestingly, we detected that tree shape influenced the accuracy loss derived from the lack of phylogenetic resolution, particularly for NRI and stdPD. We conclude that well-resolved molecular phylogenies with accurate branch length information are needed to identify the underlying phylogenetic structure of communities, and also that sensitivity of phylogenetic structure measures to low phylogenetic resolution can strongly differ depending on phylogenetic tree shape.


Peripheral clonal elimination of functional T cells.
A major mechanism for generating tolerance in developing T cells is the intrathymic clonal deletion of T cells that have receptors for those self antigens that are presented on hematopoietic cells. The mechanisms of tolerance induction to antigens not expressed in the thymus remain unclear. Tolerance to self antigens can be generated extrathymically through the induction of clonal nonresponsiveness in T cells with self-reactive receptors. A second mechanism of extrathymic tolerance was identified: clonal elimination of mature T cells with self-reactive receptors that had previously displayed functional reactivity.


Does neuromuscular electrical stimulation strengthen the quadriceps femoris? A systematic review of randomised controlled trials.
Devices for neuromuscular electrical stimulation (NMES) are increasingly used by individuals without specific injuries and are standard equipment in most physical therapy practices. The most often stimulated muscle group is the quadriceps femoris. We designed a systematic review and meta-analysis of randomised controlled trials to determine whether NMES is an effective modality for strength augmentation of the quadriceps femoris. A full content search for randomised controlled trials was performed in Medline, Embase, Cinahl, the Cochrane Controlled Trials Register and the Physical Therapy Evidence Database. Maximum volitional isometric or isokinetic muscle torque in Nm was used as main outcome measure.Thirty-five trials were included and evaluated. A fundamental distinction was made between the trials using subjects with unimpaired quadriceps femoris muscles and the trials using post-injury or post-operative subjects. In the unimpaired quadriceps subgroup, meta-analyses were performed for the comparisons 'NMES versus no exercises' and 'NMES versus volitional exercises'. All other comparisons were evaluated descriptively. The included trials were generally of poor quality and meta-analytic data indicate that publication bias may be present. The evaluated data suggest that, both for the unimpaired and impaired quadriceps, NMES makes sense compared with doing no exercises but volitional exercises appear to be more effective in most situations.Based on the available evidence, NMES may only be preferred over volitional training for within-cast muscle training and perhaps in specific situations where volitional training does not receive sufficient patient compliance. Further research should be directed toward identifying the clinical impact at activity and participation levels and the optimal stimulation parameters of this modality.


The waist-to-body mass index ratio as an anthropometric predictor for cardiovascular outcome in subjects with established atherosclerotic cardiovascular disease.
Obesity is an independent risk factor for atherosclerotic cardiovascular disease (ASCVD). However, 'obesity paradox' is observed in patients with coronary artery disease while defining obesity by body mass index (BMI). The purpose of this study is to identify a better anthropometric parameter to predict cardiovascular events in patients with ASCVD. The study was conducted using the Taiwanese Secondary Prevention for patients with AtheRosCLErotic disease (T-SPARCLE) Registry. A total of 6,920 adult patients with stable ASCVD, enrolled from January 2010 to November 2014, were included, with a mean age of 65.9 years, 73.9% males, and a mean BMI of 26.3 kg/m2 at baseline. These patients were followed up for a median of 2.5 years. The study endpoint was the composite major adverse cardiovascular event (MACE), defined as cardiovascular death, nonfatal myocardial infarction or stroke, or cardiac arrest with resuscitation. Multivariable Cox proportional hazards regression showed a significant positive association between waist-to-BMI ratio and MACE (adjusted hazard ratio 1.69 per cm‧m2/kg increase in waist-to-BMI ratio, 95% CI 1.12-2.49, p = 0.01) after adjusting for potential risk factors and confounders. Traditional anthropometric parameters, such as BMI, weight, waist and waist-hip ratio, or newer waist-based indices, such as body roundness index and a body shape index, did not show any significant linear associations (p = 0.09, 0.30, 0.89, 0.54, 0.79 and 0.06, respectively). In the restricted cubic spline regression analysis, the positive dose-response association between waist-to-BMI ratio and MACE persisted across all the range of waist-to-BMI ratio. The positive dose-response association was non-linear with a much steeper increase in the risk of MACE for waist-to-BMI ratio > 3.6 cm‧m2/kg. In conclusion, waist-to-BMI ratio may function as a positive predictor for the risk of MACE in established ASCVD patients.


Promoting electronic health record adoption among small independent primary care practices.
To describe the benefits and limitations of incentive arrangements used to engage small primary care practices to adopt electronic health records (EHRs). Retrospective review of program outreach efforts and EHR enrollment rates. Comparison of benefits and limitations of various promotional strategies in addition to a city subsidy. Measures of enrollment progress include the mean number of outreach attempts, the mean number of days from first attempt to enrollment, and the enrollment yield. Selected practice demographics were collected for comparison purposes. Of 890 providers representing 217 practices who were eligible for the city subsidy, 37.7% enrolled, with a mean of 96.6 days from first attempt to enrollment and a mean of 10 outreach attempts. The offer for full payment of technical assistance fees and hardware yielded an additional 100 providers representing 43 practices, a 14.1% enrollment rate. This group also had the highest mean number of days from first attempt to enrollment (236.0 days) and the highest mean number of outreach attempts (22 attempts). The offer for a partial rebate had the lowest yield (a 6.2% enrollment rate), a mean of 169.3 days from first attempt to enrollment, and a mean number of 19 outreach attempts. Because of diverse needs and levels of awareness in the EHR adoption process, a flexible milestone-based process is needed to engage primary care providers. In particular, community influence and additional funding were necessary for increasing enrollment among providers in medically underserved neighborhoods. These providers also required persistent and numerous follow-up attempts. Because billions of dollars in federal aid are being offered, effective local recruitment strategies are needed to facilitate provider engagement to increase EHR adoption rates.


Competing Interactions in Hierarchical Porphyrin Self-Assembly Introduce Robustness in Pathway Complexity.
Pathway complexity in supramolecular polymerization has recently sparked interest as a method to generate complex material behavior. The response of these systems relies on the existence of a metastable, kinetically trapped state. In this work, we show that strong switch-like behavior in supramolecular polymers can also be achieved through the introduction of competing aggregation pathways. This behavior is illustrated with the supramolecular polymerization of a porphyrin-based monomer at various concentrations, solvent compositions, and temperatures. It is found that the monomers aggregate via an isodesmic mechanism in weakly coupled J-type aggregates at intermediate solvent quality and temperature, followed by nucleated H-aggregates at lower solvent qualities and temperatures. At further increased thermodynamic driving forces, such as high concentration and low temperature, the H-aggregates can form hierarchical superhelices. Our mathematical models show that, contrary to a single-pathway polymerization, the existence of the isodesmic aggregation pathway buffers the free monomer pool and renders the nucleation of the H-aggregates insensitive to concentration changes in the limit of high concentrations. We also show that, at a given temperature or solvent quality, the thermodynamically stable aggregate morphology can be selected by controlling the remaining free external parameter. As a result, the judicious application of pathway complexity allows us to synthesize a diverse set of materials from only a single monomer. We envision that the engineering of competing pathways can increase the robustness in a wide variety of supramolecular polymer materials and lead to increasingly versatile applications.


Uncertainties influencing health-based prioritization of ozone abatement options.
The primary goal of air quality management is protection of human health. Therefore, formulation of ground-level ozone mitigation policies could be informed by considering not just attainment of regulatory standards but also how control measures benefit public health. However, evaluation of health impacts is complicated by uncertainties associated with photochemical modeling and epidemiological studies. This study demonstrates methods to characterize uncertainties influencing health-benefits estimation of ozone reduction (averted premature mortalities due to short-term exposure) in the Dallas-Fort Worth (DFW) region. Uncertainty in photochemical modeling and the selection of temporal metric (duration of ozone exposure) for concentration-response relationships can each affect the health-based prioritization of ozone control options. For example, deterministic results (neglecting uncertainties) based on 8-h daily maximum ozone reduction shows DFW anthropogenic NO(x) controls to yield 9.23 times as much benefit per ton as VOC controls. However, the rankings reverse under 5.7% of the cases (including 2.8% cases that exhibit incremental mortalities due to NO(X) control) when uncertainties in the photochemical model are considered. Evaluated ozone exposure on a 24-h rather than an 8-h basis also reverses the rankings.


Measuring Perceived Barriers to Physical Activity in Adolescents.
Perceived barriers to moderate-to-vigorous physical activity (PA) may contribute to the low rates of moderate-to-vigorous PA in adolescents. We examined the psychometric properties of scores from the perceived barriers to moderate-to-vigorous PA scale (PB-MVPA) by examining composite reliability and validity evidence based on the internal structure of the PB-MVPA and relations with other variables. This study was a cross-sectional analysis of data collected in 2013 from adolescents (N = 507; Mage = 12.40, SD = .62) via self-report scales. Using exploratory and confirmatory factor analyses, we found that perceived barriers were best represented as two factors representing internal (e.g., "I am not interested in physical activity") and external (e.g., "I need equipment I don't have") dimensions. Composite reliability was over .80. Using multiple regression to examine the relationship between perceived barriers and moderate-to-vigorous PA, we found that perceived internal barriers were inversely related to moderate-to-vigorous PA (β = -.32, p < .05). Based on results of the analysis of variances, there were no known-group sex differences for perceived internal and external barriers (p > .26). The PB-MVPA scale demonstrated evidence of score reliability and validity. To improve the understanding of the impact of perceived barriers on moderate-to- vigorous PA in adolescents, researchers should examine internal and external barriers separately.


Phorbol esters increase MLC phosphorylation and actin remodeling in bovine lung endothelium without increased contraction.
Direct protein kinase C (PKC) activation with phorbol myristate acetate (PMA) results in the loss of endothelial monolayer integrity in bovine lung endothelial cells (EC) but produces barrier enhancement in human lung endothelium. To extend these findings, we studied EC contractile events and observed a 40% increase in myosin light chain (MLC) phosphorylation in bovine endothelium following PMA challenge. The increase in PMA-mediated MLC phosphorylation occurred at sites distinct from Ser19/Thr18, sites catalyzed by MLC kinase (MLCK), and immunoblotting with antibodies specific to phosphorylated Ser19/Thr18 demonstrated profound time-dependent Ser19/Thr18 dephosphorylation. These events occurred in conjunction with rearrangement of stress fibers into a grid-like network, but without an increase in cellular contraction as measured by silicone membrane wrinkling assay. The PMA-induced MLC dephosphorylation was not due to kinase inhibition but, rather, correlated with rapid increases in myosin-associated phosphatase 1 (PPase 1) activity. These data suggest that PMA-mediated EC barrier regulation may involve dual mechanisms that alter MLC phosphorylation. The increase in bovine MLC phosphorylation likely occurs via direct PKC-dependent MLC phosphorylation in conjunction with decreases in Ser19/Thr18 phosphorylation catalyzed by MLCK due to PMA-induced increases in PPase 1 activity. Together, these events result in stress fiber destabilization and profound actin rearrangement in bovine endothelium, which may result in the physiological alterations observed in these models.


The Microbial Quality Aspects and Decontamination Approaches for the Herbal Medicinal Plants and Products: An in-Depth Review.
The present review article provides an overview of the published literature concerning microbial quality of medicinal plants and products and their decontamination methods. It is important to analyze different aspects regarding the cultivation, growing, harvesting, storage, manufacturing, and decontamination of medicinal plant products. Herbal medicinal plants bear a massive microbial load leading to contamination and mycotoxin, which needs to be considered, and properly controlled using suitable sterilization and decontamination methods. The main focus of this review is on the definition, advantages, disadvantages and applications of decontamination methods, particularly to show that one must consider the characteristics of the initial sample to be decontaminated. The effects of various methods (ozone, plasma, irradiation) on medicinal herbs and products treated for microbiological decontamination are dependent on factors related to microbial load (i.e., nature and amount of initial contamination), herb/product matrix (i.e., complexity of chemical composition, physical state - solid or liquid) and treatment conditions (i.e., time, irradiation dose, absence or presence of oxygen). In addition, it is important to accept some loss of the chemical compounds, while decreasing microbial load to acceptable limits according to official herbal pharmacopoeias and literature, thus ensuring a final product with quality, safety and therapeutic efficacy. The conclusion, which comes from this contribution, is that herbal medicine has more contaminants than a chemically welldefined drug, thus, good manufacturing practices should be followed.


Effects of a single exposure to carbon disulphide on the rate of urea production and on plasma free fatty acid and glucose concentrations in the rat.
The concentration of plasma free fatty acids in rats was significantly increased after a short period of exposure to inhalation of carbon disulphide (4 h, 2 mg/1). In contrast, after a longer period of exposure (15 h overnight, 2 mg/1) the concentration of plasma free fatty acid was significantly decreased despite a small hypoglycaemia. At the same time plasma urea concentration was significantly higher in CS2-treated rats. The total esterified fatty acid content of plasma was lower after exposure, but there was no change in plasma glycerol. Following an intragastric water load, no differences were observed in urine flow rate nor in renal clearances of urea and inulin between control and treated rats. It is concluded that the rate of urea production is significantly increased during acute CS2-intoxication, and it is suggested that two factors contribute to this effect: first, an increased breakdown of proteins with which CS2 or its metabolic products have reacted; and secondly an increased rate of utilization of plasma glucose associated with increased gluconeogenesis from amino acid precursors. It is further suggested that the stress effects of CS2 dominate in the short term before being overcome by a diminished sympathetic response. When rats were exposed to CS2 overnight without free access to water, the great vessel haematocrit was significantly lower than in corresponding controls. This was shown to be accounted for by differences in plasma volume. No such difference was observed when rats had free access to water during exposure. These effects probably reflect differing rates of water loss under mildly dehydrating conditions, but a direct effect of CS2 on the cardiovascular system is not excluded.


The role of aluminum sensing and signaling in plant aluminum resistance.
As researchers have gained a better understanding in recent years into the physiological, molecular, and genetic basis of how plants deal with aluminum (Al) toxicity in acid soils prevalent in the tropics and sub-tropics, it has become clear that an important component of these responses is the triggering and regulation of cellular pathways and processes by Al. In this review of plant Al signaling, we begin by summarizing the understanding of physiological mechanisms of Al resistance, which first led researchers to realize that Al stress induces gene expression and modifies protein function during the activation of Al resistance responses. Subsequently, an overview of Al resistance genes and their function provides verification that Al induction of gene expression plays a major role in Al resistance in many plant species. More recent research into the mechanistic basis for Al-induced transcriptional activation of resistance genes has led to the identification of several transcription factors as well as cis-elements in the promoters of Al resistance genes that play a role in greater Al-induced gene expression as well as higher constitutive expression of resistance genes in some plant species. Finally, the post-transcriptional and translational regulation of Al resistance proteins is addressed, where recent research has shown that Al can both directly bind to and alter activity of certain organic acid transporters, and also influence Al resistance proteins indirectly, via protein phosphorylation.


Quantitative inferences on the locomotor behaviour of extinct species applied to Simocyon batalleri (Ailuridae, Late Miocene, Spain).
Inferences of function and ecology in extinct taxa have long been a subject of interest because it is fundamental to understand the evolutionary history of species. In this study, we use a quantitative approach to investigate the locomotor behaviour of Simocyon batalleri, a key taxon related to the ailurid family. To do so, we use 3D surface geometric morphometric approaches on the three long bones of the forelimb of an extant reference sample. Next, we test the locomotor strategy of S. batalleri using a leave-one-out cross-validated linear discriminant analysis. Our results show that S. batalleri is included in the morphospace of the living species of musteloids. However, each bone of the forelimb appears to show a different functional signal suggesting that inferring the lifestyle or locomotor behaviour of fossils can be difficult and dependent on the bone investigated. This highlights the importance of studying, where possible, a maximum of skeletal elements to be able to make robust inferences on the lifestyle of extinct species. Finally, our results suggest that S. batalleri may be more arboreal than previously suggested.


Within- and between-city contrasts in nitrogen dioxide and mortality in 10 Canadian cities; a subset of the Canadian Census Health and Environment Cohort (CanCHEC).
The independent and joint effects of within- and between-city contrasts in air pollution on mortality have been investigated rarely. To examine the differential effects of between- versus within-city contrasts in pollution exposure, we used both ambient measurements and land use regression models to assess associations with mortality and exposure to nitrogen dioxide (NO2) among ~735,600 adults in 10 of the largest Canadian cities. We estimated exposure contrasts partitioned into within- and between-city contrasts, and the sum of these as overall exposures, for every year from 1984 to 2006. Residential histories allowed us to follow subjects annually during the study period. We calculated hazard ratios (HRs) adjusted for many personal and contextual variables. In fully-adjusted, random-effects models, we found positive associations between overall NO2 exposures and mortality from non-accidental causes (HR per 5 p.p.b.: 1.05; 95% confidence interval (CI): 1.03-1.07), cardiovascular disease (HR per 5 p.p.b.: 1.04; 95% CI: 1.01-1.06), ischaemic heart disease (HR per 5 p.p.b.: 1.05; 95% CI: 1.02-1.08) and respiratory disease (HR per 5 p.p.b.: 1.04; 95% CI: 0.99-1.08), but not from cerebrovascular disease (HR per 5 p.p.b.: 1.01; 95% CI: 0.96-1.06). We found that most of these associations were determined by within-city contrasts, as opposed to by between-city contrasts in NO2. Our results suggest that variation in NO2 concentrations within a city may represent a more toxic mixture of pollution than variation between cities.


Experimental systems implementation of a hybrid optical-digital correlator.
A high-speed hybrid optical-digital correlator system was designed, constructed, modeled, and demonstrated experimentally. This correlator is capable of operation at approximately 3000 correlations/s. The input scene is digitized at a resolution of 512 x 512 pixels and the phase information of the two-dimensional fast Fourier transform calculated and displayed in the correlator filter plane at normal video frame rates. High-fidelity reference template images are stored in a phase-conjugating optical memory placed at the nominal input plane of the correlator and reconstructed with a high-speed acousto-optic scanner; this allows for cross correlation of the entire reference data set with the input scene within one frame period. A high-speed CCD camera is used to capture the correlation-plane image, and rapid correlation-plane processing is achieved with a parallel processing architecture.


Adherence to disease-specific drug treatment among patients with pulmonary arterial hypertension or chronic thromboembolic pulmonary hypertension.
Pulmonary arterial hypertension (PAH) and chronic thromboembolic pulmonary hypertension (CTEPH) require lifelong treatment. The aim of the present study was to investigate adherence to disease-specific treatment in patients with PAH or CTEPH. The study comprised an adult population diagnosed with PAH (n=384) or CTEPH (n=187) alive in 2016-2017. The study utilised three registries: the Swedish PAH registry, the National Board of Health and Welfare, and Statistics Sweden. Withdrawals from pharmacies of disease-specific oral treatments were studied. Adherence was assessed as: 1) Number of days covered defined as the difference between the total number of daily dosages dispensed and the total number of days covered; and 2) Manual assessment by two persons that independently reviewed each patient's prescription fill history to detect anomalies or patterns of deteriorating or improving adherence over time. The mean age was 61±16 years, 61% were female and mean time since diagnosis was 4.6 years. Adherence was 62% using the Number of days covered method and 66% by the Manual assessment method. Drug-specific adherence varied from 91% for riociguat to 60% for sildenafil. Good adherence was associated with shorter time since diagnosis in patients with PAH and with lower number of concomitant other chronic treatments in patients with CTEPH. Age, sex, socioeconomic status or number of pulmonary hypertension (PH) treatments were not associated with adherence. Adherence to oral disease-specific treatment was 60-66% and associated with time since diagnosis and number of concomitant chronic treatments. Sex, age or socioeconomic factors did not affect adherence.


Substrate specificity of human liver neutral alpha-mannosidase.
The digestion of radiolabelled natural oligosaccharide substrates by human liver neutral alpha-mannosidase has been studied by h.p.l.c. and h.p.t.l.c. The high-mannose oligosaccharides Man9GlcNAc and Man8GlcNAc are hydrolysed by the enzyme by two distinct non-random routes to a common product of composition Man6GlcNAc, which is then slowly converted into a unique Man5GlcNAc oligosaccharide, Man alpha(1----2)Man alpha(1----2)Man alpha(1----3)[Man alpha (1----6)] Man beta(1----4)GlcNAc. These pathways are different from the processing and lysosomal catabolic pathways for these structures. In particular, the alpha(1----2)-linked mannose residues attached to the core alpha(1----3)-linked mannose residue are resistant to hydrolysis. The key processing intermediate, Man alpha(1----3)[Man alpha(1----6)]Man alpha(1----6)[Man alpha(1----3)] Man beta(1----4)GlcNAc, is not produced in the digestion of high-mannose glycans by the neutral alpha-mannosidase, but it is hydrolysed by the enzyme by a non-random route to Man beta(1----4)GlcNAc via the core structure Man alpha(1----3)[Man alpha(1----6)]Man beta(1----4)GlcNAc. In contrast with its ready hydrolysis by lysosomal alpha-mannosidase, the core alpha(1----3)-mannosidic linkage is quite resistant to hydrolysis by neutral alpha-mannosidase. The precise specificity of neutral alpha-mannosidase towards high-mannose oligosaccharides suggests that it has a role in the modification of such structures in the cytosol.


Genome Data Provides High Support for Generic Boundaries in Burkholderia Sensu Lato.
Although the taxonomy of Burkholderia has been extensively scrutinized, significant uncertainty remains regarding the generic boundaries and composition of this large and heterogeneous taxon. Here we used the amino acid and nucleotide sequences of 106 conserved proteins from 92 species to infer robust maximum likelihood phylogenies with which to investigate the generic structure of Burkholderia sensu lato. These data unambiguously supported five distinct lineages, of which four correspond to Burkholderia sensu stricto and the newly introduced genera Paraburkholderia, Caballeronia, and Robbsia. The fifth lineage was represented by P. rhizoxinica. Based on these findings, we propose 13 new combinations for those species previously described as members of Burkholderia but that form part of Caballeronia. These findings also suggest revision of the taxonomic status of P. rhizoxinica as it is does not form part of any of the genera currently recognized in Burkholderia sensu lato. From a phylogenetic point of view, Burkholderia sensu stricto has a sister relationship with the Caballeronia+Paraburkholderia clade. Also, the lineages represented by P. rhizoxinica and R. andropogonis, respectively, emerged prior to the radiation of the Burkholderia sensu stricto+Caballeronia+Paraburkholderia clade. Our findings therefore constitute a solid framework, not only for supporting current and future taxonomic decisions, but also for studying the evolution of this assemblage of medically, industrially and agriculturally important species.


Optoelectronic reconfigurable microchannels.
This paper reports a new optoelectronic reconfigurable microchannel (OERM) technology which uses low-power light to create and reconfigure microchannels within seconds. This technology thereby removes the need for complicated, on-chip fluidic interconnects and enables real-time modification of microchannel networks. In essence, OERM technology is based on the local thawing of a frozen solution using an optoelectronic heating effect powered by an external voltage supplier; a light pattern of microchannels is projected on an OERM chip containing a thin layer of frozen liquid, which produces microchannels corresponding to the light pattern via melting. When the pattern is modified, specific regions freeze and others melt according to the new light pattern, which causes the microchannel network to reconfigure. The versatility of this platform is illustrated by performing several experiments: single-phase flows are used to draw a smiley-face pattern, and multi-phase flow operations, such as generating and merging droplets and bubbles are demonstrated. Due to its low optical power requirement and its fast reconfiguration speed, OERM can dynamically alter microfluidic circuits while benefiting from the flexibility of light patterns and thereby offers a myriad of possible microfluidic applications.


Time-varying spectral analysis revealing differential effects of sevoflurane anaesthesia: non-rhythmic-to-rhythmic ratio.
Heart rate variability (HRV) may reflect various physiological dynamics. In particular, variation of R-R peak interval (RRI) of electrocardiography appears regularly oscillatory in deeper levels of anaesthesia and less regular in lighter levels of anaesthesia. We proposed a new index, non-rhythmic-to-rhythmic ratio (NRR), to quantify this feature and investigated its potential to estimate depth of anaesthesia. Thirty-one female patients were enrolled in this prospective study. The oscillatory pattern transition of RRI was visualised by the time-varying power spectrum and quantified by NRR. The prediction of anaesthetic events, including skin incision, first reaction of motor movement during emergence period, loss of consciousness (LOC) and return of consciousness (ROC) by NRR were evaluated by serial prediction probability (PK ) analysis; the ability to predict the decrease of effect-site sevoflurane concentration was also evaluated. The results were compared with Bispectral Index (BIS). NRR well-predicted first reaction (PK > 0.90) 30 s ahead, earlier than BIS and significantly better than HRV indices. NRR well-correlated with sevoflurane concentration, although its correlation was inferior to BIS, while HRV indices had no such correlation. BIS indicated LOC and ROC best. Our findings suggest that NRR provides complementary information to BIS regarding the differential effects of anaesthetics on the brain, especially the subcortical motor activity.


Does Future Diabetes Risk Impair Current Quality of Life? A Cross-Sectional Study of Health-Related Quality of Life in Relation to the Finnish Diabetes Risk Score (FINDRISC).
Present study examines the relationship between the estimated risk of developing type 2 diabetes (T2D) and health-related quality of life (HRQoL). We quantify the association between Finnish Diabetes Risk Score (FINDRISC) and HRQoL, and examine the potential use of FINDRISC as tool to evaluate HRQoL indirectly. We conducted a cross-sectional study comprising 707 Finnish people without a diagnosis of T2D between the ages of 51 and 75 years. The risk of developing T2D was assessed using the validated and widely used FINDRISC (range 0-26 points), and quality of life was measured using two preference-based HRQoL instruments (15D and SF-6D) and one health profile instrument (SF-36). Effects of the individual FINDRISC items and demographic and clinical characteristics, such as co-morbidities, on HRQoL were studied using multivariable Tobit regression models. Low HRQoL was significantly and directly associated with the estimated risk of developing T2D. An approximate 4-5 point change in FINDRISC score was observed to be associated with clinically noticeable changes in the preference-based instrument HRQoL index scores. The association between HRQoL and the risk of developing T2D was also observed for most dimensions of HRQoL in all applied HRQoL instruments. Overall, old age, lack of physical activity, obesity, and history of high blood glucose were the FINDRISC factors most prominently associated with lower HRQoL. The findings may help the health care professionals to substantiate the possible improvement in glucose metabolism and HRQoL potentially achieved by lifestyle changes, and better convince people at high risk of T2D to take action towards healthier lifestyle habits. FINDRISC may also provide an accurate proxy for HRQoL, and thus by estimating the risk of T2D with the FINDRISC, information about patients' HRQoL may also be obtained indirectly, when it is not feasible to use HRQoL instruments.


Is pseudarthrosis after spinal instrumentation caused by a chronic infection?
To assess whether a chronic bacterial infection is present in a subset of patients with pseudarthrosis after instrumented spinal fusion. This was a prospective diagnostic study including adult patients with previous instrumented spinal fusion. Patients underwent revision surgery for either pseudarthrosis or other causes (e.g. implant removal, curve progression or junctional kyphosis) (control group). Five separate biopsies were randomly collected, intraoperatively, from the pseudarthrosis site and cultivated under both aerobic (5 days) and anaerobic (14 days) conditions. If cultivation was positive in at least 2/5 tissue samples, the biopsy was sectioned and stained using peptide nucleic acid fluorescence in situ hybridization (PNA-FISH). Confocal laser scanning microscopy was used to examine the sections and visualize bacterial aggregates. The study included 32 pseudarthrosis and 32 control patients. Cultivation yielded bacteria in at least 1/5 biopsies in 52% of patients with no difference between the groups (p = 1.0). Bacteria of the same species was found in at least 2/5 samples in seven pseudarthrosis patients and four controls (p = 0.509). Propionibacterium acnes was found in 8 of these 11 samples. Microscopy demonstrated tissue-embedded bacterial aggregates in two of these patients but with no inflammatory cells indicating an active infection. The presence of bacteria was not associated with the number of previous spinal procedures or the pre-revision fusion length (p ≥ 0.503). Pseudarthrosis after instrumented spinal surgery was not significantly associated with the presence of bacteria at the pseudarthrosis site. Positive cultivation results are common after spinal instrumentation, but our results indicate that they rarely represent an organized infection. These slides can be retrieved under Electronic Supplementary Material.


CD226 opposes TIGIT to disrupt Tregs in melanoma.
CD4+ Tregs impede T cell responses to tumors. They express multiple inhibitory receptors that support their suppressive functions, including T cell Ig and ITIM domain (TIGIT). In melanoma patients, we show that Tregs exhibit increased TIGIT expression and decreased expression of its competing costimulatory receptor CD226 as compared with CD4+ effector T cells, resulting in an increased TIGIT/CD226 ratio. Tregs failed to upregulate CD226 upon T cell activation. TIGIT+ Tregs are highly suppressive, stable, and enriched in tumors. TIGIT and CD226 oppose each other to augment or disrupt, respectively, Treg suppression and stability. A high TIGIT/CD226 ratio in Tregs correlates with increased Treg frequencies in tumors and poor clinical outcome upon immune checkpoint blockade. Altogether, our findings show that a high TIGIT/CD226 ratio in Tregs regulates their suppressive function and stability in melanoma. They provide the rationale for novel immunotherapies to activate CD226 in Tregs together with TIGIT blockade to counteract Treg suppression in cancer patients.


Hunter-gatherers in southeast Asia: from prehistory to the present.
Anatomically modern hunter-gatherers expanded from Africa into Southeast Asia at least 50,000 years ago, where they probably encountered and interacted with populations of Homo erectus and Homo floresiensis and the recently discovered Denisovans. Simulation studies suggest that these hunter-gatherers may well have followed a coastal route that ultimately led to the settlement of Sahul, while archaeology confirms that they also crossed significant seas and explored well into the interior. They also adapted to marked environmental changes that alternated between relatively cool and dry conditions and warmer, wetter interludes. During the former, the sea fell by up to 120 m below its present level, which opened up a vast low-lying area known as Sundaland. Three principal alignments can be identified: the first involved the occupation of rock shelters in upland regions, the second has identified settlement on broad riverine floodplains, and the last concentrated on the raised beaches formed from about five millennia ago when the sea level was elevated above its present position. This cultural sequence was dislocated about 4 kya when rice and millet farmers infiltrated the lowlands of Southeast Asia ultimately from the Yangtze River valley. It is suggested that this led to two forms of interaction. In the first, the indigenous hunter-gatherers integrated with intrusive Neolithic communities and, while losing their cultural identity, contributed their genes to the present population of Southeast Asia. In the second, hunter-gatherers withdrew to rainforest refugia and, through selective pressures inherent in such an environment, survived as the small-bodied, dark-skinned humans found to this day in the Philippines, Peninsular Malaysia and Thailand, and the Andaman Islands. Beyond the impact of expansive rice farmers in Melanesia and Australia, hunter-gatherers continued to dominate until they encountered European settlement.


Three-dimensional dynamic geometry of the normal canine mitral annulus and papillary muscles.
Despite an incomplete knowledge of the geometry and dynamics of the mitral annulus (MA), papillary muscle (PM), and the chordae tendineac, chordal-sparing MVR is popular. The systolic reduction in three-dimensional distance between each PM tip and eight MA sites (DT-A) was measured in nine normal closed-chest dogs by use of surgically implanted radiopaque markers. Three loci (tip, junction, and base) on each PM were also projected onto the MA plane at end diastole and end systole to assess PM dynamics. The anterior PM tip showed significant shortening of DT-A toward the opposite side of the MA or the midanterior MA region (P < .005 or P < .05, respectively, versus same MA side [MANOVA]); conversely, the posterior PM tip DT-A shortened toward the opposite side of the MA near the anterior commissure or the area between the anterior commissure and midposterior MA (P < .005 versus same MA side). Annular projection revealed three-dimensional motion (relative to the MA) of the anterior PM tip, junction, and base toward the right trigone, while posterior PM motion was oriented toward the opposite side of the MA. Both PMs in normal canine hearts demonstrated systolic relative motion in a direction compatible with the "oblique" chordal configuration, ie, from the anterior PM to the anterior MA near the right trigone and from the posterior PM to the opposite side of the posterior MA. These observations warrant further investigation of three-dimensional PM-MA dynamics with various methods of chorda preservation during MVR to assess their impact on left ventricular systolic and diastolic function.


Innate immune responses of a lepidopteran insect, Manduca sexta.
Many innate immune mechanisms are conserved throughout the animal kingdom. Manduca sexta, a widely used model for insect biochemical research, employs these mechanisms to defend against invading pathogens and parasites. We have isolated from M. sexta hemolymph a group of proteins (hemolin, peptidoglycan recognition proteins, beta-1,3-glucan recognition proteins, and C-type lectins), which serve as a surveillance mechanism by binding to microbial surface molecules (e.g. peptidoglycan, lipopolysaccharide, lipoteichoic acid, and beta-1,3-glucan). The binding triggers diverse responses such as phagocytosis, nodule formation, encapsulation, melanization, and synthesis of anti-microbial peptides/proteins. Some of these responses are mediated and coordinated by serine proteinase cascades, analogous to the complement system in mammals. Our current research is focused on the proteolytic activation of prophenoloxidase (proPO)--a reaction implicated in melanotic encapsulation, wound healing, and protein cross-linking. We have isolated three proPO-activating proteinases, each of which requires serine proteinase homologs as a cofactor for generating active phenoloxidase. The proteinases and proteinase-like molecules, containing one to two clip domains at their amino-terminus, are acute-phase proteins induced upon an immune challenge. Inhibitory regulation of the proteinases by serpins and association of the proteinase homologs with a bacteria-binding lectin are important for ensuring a localized defense response. Additional serine proteinases expressed in M. sexta hemocytes and fat body have been discovered. Future research efforts will be aimed at elucidating the proteinase cascade for proPO activation and investigating the roles of proteinases in other immune responses such as processing of plasmatocyte-spreading peptide.