Ten-Minute Protein Purification and Surface Tethering for Continuous-Flow Biocatalysis.
Nature applies enzymatic assembly lines to synthesize bioactive compounds. Inspired by such capabilities, we have developed a facile method for spatially segregating attached enzymes in a continuous-flow, vortex fluidic device (VFD). Fused Hisn -tags at the protein termini allow rapid bioconjugation and consequent purification through complexation with immobilized metal affinity chromatography (IMAC) resin. Six proteins were purified from complex cell lysates to average homogeneities of 76 %. The most challenging to purify, tobacco epi-aristolochene synthase, was purified in only ten minutes from cell lysate to near homogeneity (>90 %). Furthermore, this "reaction-ready" system demonstrated excellent stability during five days of continuous-flow processing. Towards multi-step transformations in continuous flow, proteins were arrayed as ordered zones on the reactor surface allowing segregation of catalysts. Ordering enzymes into zones opens up new opportunities for continuous-flow biosynthesis.


The Oklahoma Thrombotic Thrombocytopenic Purpura-haemolytic Uraemic Syndrome Registry. A model for clinical research, education and patient care.
The Oklahoma Thrombotic Thrombocytopenic Purpura-Haemolytic Uraemic Syndrome (TTP-HUS) Registry has a 24 year record of success for collaborative clinical research, education, and patient care. This article tells the story of how the Registry began and it describes the Registry's structure and function. The Registry provides a model for using a cohort of consecutive patients to investigate a rare disorder. Collaboration between Oklahoma, United States and Bern, Switzerland has been the basis for successful interpretation of Registry data. Registry data have provided new insights into the evaluation and management of TTP. Because recovery from acute episodes of TTP has been assumed to be complete, the increased prevalence of hypertension, diabetes, depression, and death documented by long-term follow-up was unexpected. Registry data have provided opportunities for projects for students and trainees, education of physicians and nurses, and also for patients themselves. During our follow-up, patients have also educated Registry investigators about problems that persist after recovery from an acute episode of TTP. Most important, Registry data have resulted in important improvements for patient care.


Temperature reduces fish dispersal as larvae grow faster to their settlement size.
As species struggle to cope with rising ocean temperatures, temperate marine assemblages are facing major reorganization. Many benthic species have a brief but critical period dispersing through the plankton, when they are particularly susceptible to variations in temperature. Impacts of rising temperatures can thus ripple through the population with community-wide consequences. However, responses are highly species-specific, making it difficult to discern assemblage-wide patterns in the life histories of different fish species. Here, we evaluate the responses to temperature in the early life histories of several fish species using otolith reconstructive techniques. We also assess the consequences of future warming scenarios to this assemblage. We sampled recent settlers of nine common species across a temperature gradient in the Mediterranean Sea and obtained environmental data for each individual. Using otolith microstructure, we measured early life traits including pelagic larval duration (PLD), growth rate, settlement size, hatching and settlement dates. We used a GLM framework to examine how environmental variables influenced early life-history parameters. We show that increasing temperature results in considerable reduction in the dispersal potential of temperate fish. We find a nearly universal, assemblage-wide decline in pelagic larval duration (PLD) of between 10% and 25%. This was because, with increasing temperature, larvae grew quicker to their settlement size. Settlement size itself was less affected by temperature and appears to be an ontogenetically fixed process. Given current estimates of ocean warming, there could be an assemblage-wide reduction in larval dispersal of up to 50 km across the Mediterranean, reducing connectivity and potentially isolating populations as waters warm.


Explaining sex differences in lifespan in terms of optimal energy allocation in the baboon.
We provide a quantitative test of the hypothesis that sex role specialization may account for sex differences in lifespan in baboons if such specialization causes the dependency of fitness upon longevity, and consequently the optimal resolution to an energetic trade-off between somatic maintenance and other physiological functions, to differ between males and females. We present a model in which females provide all offspring care and males compete for access to reproductive females and in which the partitioning of available energy between the competing fitness-enhancing functions of growth, maintenance, and reproduction is modeled as a dynamic behavioral game, with the optimal decision for each individual depending upon his/her state and the behavior of other members of the population. Our model replicates the sexual dimorphism in body size and sex differences in longevity and reproductive scheduling seen in natural populations of baboons. We show that this outcome is generally robust to perturbations in model parameters, an important finding given that the same behavior is seen across multiple populations and species in the wild. This supports the idea that sex differences in longevity result from differences in the value of somatic maintenance relative to other fitness-enhancing functions in keeping with the disposable soma theory.


Cortical and Subcortical Dysmetabolism Are Dynamic Markers of Clinical Disability and Course in Anti-LGI1 Encephalitis.
This [18F]fluorodeoxyglucose (FDG) PET study evaluates the accuracy of semiquantitative measurement of putaminal hypermetabolism in identifying anti-leucine-rich, glioma-inactivated-1 (LGI1) protein autoimmune encephalitis (AE). In addition, the extent of brain dysmetabolism, their association with clinical outcomes, and longitudinal metabolic changes after immunotherapy in LGI1-AE are examined. FDG-PET scans from 49 age-matched and sex-matched subjects (13 in LGI1-AE group, 15 in non-LGI1-AE group, 11 with Alzheimer disease [AD], and 10 negative controls [NCs]) and follow-up scans from 8 patients with LGI1 AE on a median 6 months after immunotherapy were analyzed. Putaminal standardized uptake value ratios (SUVRs) normalized to global brain (P-SUVRg), thalamus (P/Th), and midbrain (P/Mi) were evaluated for diagnostic accuracy. SUVRg was applied for all other analyses. P-SUVRg, P/Th, and P/Mi were higher in LGI1-AE group than in non-LGI1-AE group, AD group, and NCs (all p < 0.05). P/Mi and P-SUVRg differentiated LGI1-AE group robustly from other groups (areas under the curve 0.84-0.99). Mediotemporal lobe (MTL) SUVRg was increased in both LGI1-AE and non-LGI1-AE groups when compared with NCs (both p < 0.05). SUVRg was decreased in several frontoparietal regions and increased in pallidum, caudate, pons, olfactory, and inferior occipital gyrus in LGI1-AE group when compared with that in NCs (all p < 0.05). In LGI1-AE group, both MTL and putaminal hypermetabolism were reduced after immunotherapy. Normalization of regional cortical dysmetabolism associated with clinical improvement at the 6- and 20-month follow-up. Semiquantitative measurement of putaminal hypermetabolism with FDG-PET may be used to distinguish LGI1-AE from other pathologies. Metabolic abnormalities in LGI1-AE extend beyond putamen and MTL into other subcortical and cortical regions. FDG-PET may be used in evaluating disease evolution in LGI1-AE. This study provides Class II evidence that semiquantitative measures of putaminal metabolism on PET can differentiate patients with LGI1-AE from patients without LGI1-AE, patients with AD, or NCs.


DnmA and FisA Mediate Mitochondria and Peroxisome Fission, and Regulate Mitochondrial Function, ROS Production and Development in Aspergillus nidulans.
The dynamin-like protein Drp1 and its receptor Fis-1 are required for mitochondria and peroxisome fission in animal and yeast cells. Here, we show that in the fungus Aspergillus nidulans the lack of Drp1 and Fis-1 homologs DnmA and FisA has strong developmental defects, leading to a notable decrease in hyphal growth and asexual and sexual sporulation, with some of these defects being aggravated or partially remediated by different carbon sources. Although both DnmA and FisA, are essential for mitochondrial fission, participate in peroxisomal division and are fully required for H2O2-induced mitochondrial division, they also appear to play differential functions. Despite their lack of mitochondrial division, ΔdnmA and ΔfisA mutants segregate mitochondria to conidiogenic cells and produce viable conidia that inherit a single mitochondrion. During sexual differentiation, ΔdnmA and ΔfisA mutants develop fruiting bodies (cleistothecia) that differentiate excessive ascogenous tissue and a reduced number of viable ascospores. ΔdnmA and ΔfisA mutants show decreased respiration and notably high levels of mitochondrial reactive oxygen species (ROS), which likely correspond to superoxide. Regardless of this, ΔdnmA mutants can respond to an external H2O2 challenge by re-localizing the MAP kinase-activated protein kinase (MAPKAP) SrkA from the cytoplasm to the nuclei. Our results show that ROS levels regulate mitochondrial dynamics while a lack of mitochondrial fission results in lower respiration, increased mitochondrial ROS and developmental defects, indicating that ROS, mitochondrial division and development are critically interrelated processes.


Language Evolution in Swarm Robotics: A Perspective.
While direct local communication is very important for the organization of robot swarms, so far it has mostly been used for relatively simple tasks such as signaling robots preferences or states. Inspired by the emergence of meaning found in natural languages, more complex communication skills could allow robot swarms to tackle novel situations in ways that may not be a priori obvious to the experimenter. This would pave the way for the design of robot swarms with higher autonomy and adaptivity. The state of the art regarding the emergence of communication for robot swarms has mostly focused on offline evolutionary approaches, which showed that signaling and communication can emerge spontaneously even when not explicitly promoted. However, these approaches do not lead to complex, language-like communication skills, and signals are tightly linked to environmental and/or sensory-motor states that are specific to the task for which communication was evolved. To move beyond current practice, we advocate an approach to emergent communication in robot swarms based on language games. Thanks to language games, previous studies showed that cultural self-organization-rather than biological evolution-can be responsible for the complexity and expressive power of language. We suggest that swarm robotics can be an ideal test-bed to advance research on the emergence of language-like communication. The latter can be key to provide robot swarms with additional skills to support self-organization and adaptivity, enabling the design of more complex collective behaviors.


Far-red light photoacclimation (FaRLiP) in Synechococcus sp. PCC 7335. II.Characterization of phycobiliproteins produced during acclimation to far-red light.
Phycobilisomes (PBS) are antenna complexes that harvest light for photosystem (PS) I and PS II in cyanobacteria and some algae. A process known as far-red light photoacclimation (FaRLiP) occurs when some cyanobacteria are grown in far-red light (FRL). They synthesize chlorophylls d and f and remodel PS I, PS II, and PBS using subunits paralogous to those produced in white light. The FaRLiP strain, Leptolyngbya sp. JSC-1, replaces hemidiscoidal PBS with pentacylindrical cores, which are produced when cells are grown in red or white light, with PBS with bicylindrical cores when cells are grown in FRL. This study shows that the PBS of another FaRLiP strain, Synechococcus sp. PCC 7335, are not remodeled in cells grown in FRL. Instead, cells grown in FRL produce bicylindrical cores that uniquely contain the paralogous allophycocyanin subunits encoded in the FaRLiP cluster, and these bicylindrical cores coexist with red-light-type PBS with tricylindrical cores. The bicylindrical cores have absorption maxima at 650 and 711 nm and a low-temperature fluorescence emission maximum at 730 nm. They contain ApcE2:ApcF:ApcD3:ApcD2:ApcD5:ApcB2 in the approximate ratio 2:2:4:6:12:22, and a structural model is proposed. Time course experiments showed that bicylindrical cores were detectable about 48 h after cells were transferred from RL to FRL and that synthesis of red-light-type PBS continued throughout a 21-day growth period. When considered in comparison with results for other FaRLiP cyanobacteria, the results here show that acclimation responses to FRL can differ considerably among FaRLiP cyanobacteria.


The challenges of innovation in the organization of home enteral tube feeding.
The number of patients discharged from hospital who need home enteral tube feeding has been increasing steadily in the UK. Arrangements for support of these patients is extremely variable. The unsatisfactory arrangements for home enteral tube feeding which existed in Avon in 1996 prompted an innovative reorganization. On the basis of that experience, this review examines the key issues involved and the questions to be considered, which may be of benefit to other trusts faced with similar challenges.


Phenobarbital protection against methyl mercury neophrotoxicity.
Phenobarbital injections to rats given a low oral dose level of methl mercury for 2 or 4 wk decreased methyl mercury-induced ultrastructural alterations in kidney proximal tubule cells, increased urinaryexcretion of inorganic mercury and increased blood concentrations of methylmercury. These effects were not seen after 2 wk of treatment but were highly significant after 4 wk.


Polymorphisms of alcohol dehydrogenase-1B and aldehyde dehydrogenase-2 and the blood and salivary ethanol and acetaldehyde concentrations of Japanese alcoholic men.
The effects of genetic polymorphism of aldehyde dehydrogenase-2 (ALDH2) on alcohol metabolism are striking in nonalcoholics, and the effects of genetic polymorphism of alcohol dehydrogenase-1B (ADH1B) are modest at most, whereas genetic polymorphisms of both strongly affect the susceptibility to alcoholism and upper aerodigestive tract (UADT) cancer of drinkers. We evaluated associations between ADH1B/ADH1C/ALDH2 genotypes and the blood and salivary ethanol and acetaldehyde levels of 168 Japanese alcoholic men who came to our hospital for the first time in the morning and had been drinking until the day before. The ethanol levels in their blood and saliva were similar, but the acetaldehyde levels in their saliva were much higher than in their blood, probably because of acetaldehyde production by oral bacteria. Blood and salivary ethanol and acetaldehyde levels were both significantly higher in the subjects with the less active ADH1B*1/*1 genotype than in the ADH1B*2 carriers, but none of the levels differed according to ALDH2 genotype. Significant linkage disequilibrium was detected between the ADH1B and ADH1C genotypes, but ADH1C genotype did not affect the blood or salivary ethanol or acetaldehyde levels. High blood acetaldehyde levels were found even in the active ALDH2*1/*1 alcoholics, which were comparable with the levels of the inactive heterozygous ALDH2*1/*2 alcoholics with less active ADH1B*1/*1. The slope of the increase in blood acetaldehyde level as the blood ethanol level increased was significantly steeper in alcoholics with inactive heterozygous ALDH2*1/*2 plus ADH1B*2 allele than with any other genotype combinations, but the slopes of the increase in salivary acetaldehyde level as the salivary ethanol level increased did not differ between the groups of subjects with any combinations of ALDH2 and ADH1B genotypes. The ADH1B/ALDH2 genotype affected the blood and salivary ethanol and acetaldehyde levels of nonabstinent alcoholics in a different manner from nonalcoholics, and clear effects of ADH1B genotype and less clear effects of ALDH2 were observed in the alcoholics. Alterations in alcohol metabolism as a result of alcoholism may modify the gene effects, and these findings provide some clues in regard to associations between the genotypes and the risks of alcoholism and UADT cancer.


Modulation of the proteolytic activity of the complement protease C1s by polyanions: implications for polyanion-mediated acceleration of interaction between C1s and SERPING1.
The complement system plays crucial roles in the immune system, but incorrect regulation causes inflammation and targeting of self-tissue, leading to diseases such as systemic lupus erythematosus, rheumatoid arthritis and age-related macular degeneration. In vivo, the initiating complexes of the classical complement and lectin pathways are controlled by SERPING1 [(C1 inhibitor) serpin peptidase inhibitor, clade G, member 1], which inactivates the components C1s and MASP-2 (mannan-binding lectin serine peptidase 2). GAGs (glycosaminoglycan) and DXS (dextran sulfate) are able to significantly accelerate SERPING1-mediated inactivation of C1s, the key effector enzyme of the classical C1 complex, although the mechanism is poorly understood. In the present study we have shown that C1s can bind to DXS and heparin and that these polyanions enhanced C1s proteolytic activity at low concentrations and inhibited it at higher concentrations. The recent determination of the crystal structure of SERPING1 has given rise to the hypothesis that both the serpin (serine protease inhibitor)-polyanion and protease-polyanion interactions might be required to accelerate the association rate of SERPING1 and C1s. To determine what proportion of the acceleration was due to protease-polyanion interactions, a chimaeric mutant of alpha1-antitrypsin containing the P4-P1 residues from the SERPING1 RCL (reactive-centre loop) was produced. Like SERPING1, this molecule is able to effectively inhibit C1s, but is unable to bind polyanions. DXS exerted a biphasic effect on the association rate of C1s which correlated strongly with the effect of DXS on C1s proteolytic activity. Thus, whereas polyanions are able to bind C1s and modulate its activity, polyanion interactions with SERPING1 must also play a vital role in the mechanism by which these cofactors accelerate the C1s-SERPING1 reaction.


Evidence based postoperative treatment of distal radius fractures following internal locking plate fixation.
Originally, the treatment method of choice for distal radial fractures (DRF) has been a non-operative approach with six to eight weeks of plaster casting. The introduction of volar locking plate systems at the beginning of the 21 st century has pushed trends towards open reduction and internal fixation (ORIF). While the introduction of fixed angle locking plates together with the increasing knowledge on wrist function and related variable outcomes has led to consensus that operative fixation in instable DRF is the treatment method of choice, there is no agreement on a postoperative care of these injuries. The authors will discuss the available evidence for current concepts of postoperative treatment of DRFs following fixed angle fixation under socioeconomical, biomechanical and burden of disease aspects. Further, relevant randomized controlled trials are evaluated with regard to applied postoperative treatment regimes and related risks for complications.


Pregnancy and melanoma.
Malignant melanoma is the most common malignancy during pregnancy, and is diagnosed during childbearing age in approximately one-third of women diagnosed with melanoma. The impact of hormonal changes during pregnancy and from iatrogenic hormones on melanoma is controversial. Women undergo immunologic changes during pregnancy that may decrease tumor surveillance. In addition, hormone receptors are found on some melanomas. In spite of these observations, the preponderance of evidence does not support a poorer prognosis for pregnancy-associated melanomas. There is also a lack of evidence that oral contraceptives or hormone replacement therapy worsens melanoma prognosis.


Comparison of the irritation potentials of Boswellia serrata gum resin and of acetyl-11-keto-beta-boswellic acid by in vitro cytotoxicity tests on human skin-derived cell lines.
Indian frankincense is a gum resin from Boswellia serrata of Burseraceae used in Ayurveda and Western medicine for the antinflammatory effects of boswellic acids, particularly 3-O-acetyl-11-keto-beta-boswellic acid (AKBA). We evaluated in vitro cytotoxicities of B. serrata extract and AKBA on differentiated and undifferentiated keratinocytes (HaCaT and NCTC 2544), and foetal dermal fibroblasts (HFFF2), using neutral red uptake (NRU), MTT, and DNA assays. Comparison between NRU and MTT, and between the extract and AKBA, suggested a relatively higher toxicity of both substances on lysosomes respect to mitochondria. Extract cytotoxicity on lysosomes was higher in NCTC and HFFF2 than on the more differentiated HaCaT. DNA assay showed low extract inhibition on HFFF2 proliferation, possibly due to lower growth rate, and a stronger effect on NCTC than on HaCaT, possibly related to higher proapoptotic effect on the less differentiated NCTC, as also suggested by higher AKBA toxicity on NCTC than on HaCaT. In general, gum resin and AKBA toxicities were slightly lower or higher than that of the reference compound SDS. Our in vitro model allowed to compare the sensitivities of different human skin cells to B. serrata, and indicated that the gum resin and AKBA exert moderate to low toxicity on the skin.


Opportunities and challenges of pluripotent stem cell neurodegenerative disease models.
Human neurodegenerative disorders are among the most difficult to study. In particular, the inability to readily obtain the faulty cell types most relevant to these diseases has impeded progress for decades. Recent advances in pluripotent stem cell technology now grant access to substantial quantities of disease-pertinent neurons both with and without predisposing mutations. While this suite of technologies has revolutionized the field of 'in vitro disease modeling', great care must be taken in their deployment if robust, durable discoveries are to be made. Here we review what we perceive to be several of the stumbling blocks in the use of stem cells for the study of neurological disease and offer strategies to overcome them.


The involvement of protein kinase C in exocytosis in mast cells.
Diacylglycerol generated from inositolphospholipid hydrolysis and tumor-promoting phorbol esters stimulate protein kinase C. The synthetic diacylglycerol 1-oleoyl-2-acetyl-rac-glycerol and the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) have been used in pure rat peritoneal mast cells. Both caused histamine release associated with exocytosis. The release by the stimulation of protein kinase C alone in the absence of secretagogues was slow although up to 50% of the histamine content was released by TPA in 120 min. Remarkable potentiation of histamine release was observed when the mast cells were preincubated with TPA before exposure to the calcium ionophore A23187. The potentiation of histamine release corresponded with an intensification of exocytosis. The potentiation is consistent with a participation of protein kinase C in the secretory process. An inhibitory effect due to protein kinase C activity was also demonstrated using TPA and mast cells from sensitized rats. When sensitized mast cells preincubated with 50 nM TPA for 5 min were exposed to the antigen, the histamine release was substantially reduced compared to the sum of the release by the antigen and TPA or by the antigen alone. There was a corresponding decrease in exocytosis. The inhibition of exocytosis and histamine release seems to reflect a regulatory function of protein kinase C for the termination of the response, as demonstrated in other types of cells apparently acting through an inhibition of inositolphospholipid hydrolysis.


Breast reconstruction following subcutaneous mastectomy for cancer: a critical appraisal of the nipple-areola complex.
Subcutaneous mastectomy for women with advanced breast cancer has been historically controversial because of the increased risk for tumor recurrence. Despite this, some women remain interested in this method of treatment as a means of preserving the appearance of the breast and nipple-areola complex. Several studies have evaluated the feasibility of subcutaneous mastectomy; however, there has been no study that has critically analyzed the aesthetic outcome of the nipple-areola complex following this approach. Over a 14-month interval, 12 women had subcutaneous mastectomy with preservation of the nipple-areola complex. The reconstruction was unilateral in 10 women and bilateral in two women, totaling 14 breasts. The mastectomy was for cancer in 11 and for prophylaxis in three breasts. Outcomes were assessed based on the sensation, appearance, and secondary procedures of the nipple-areola complex, and tumor recurrence and patient satisfaction. Of the five parameters, sensation was present in six breasts (42.9 percent), delayed healing was noted in four breasts (28.6 percent), symmetry with the contralateral breast was achieved in five of 10 women (50 percent) following unilateral reconstruction, tumor recurrence was noted in three of 11 breasts (27.3 percent), and secondary procedures related to the nipple-areola complex were necessary in five of the 14 breasts (35.7 percent). Outcome was graded as excellent in three, good in eight, and poor in three breasts. This study has demonstrated that aesthetic outcome of the nipple-areola complex is variable following subcutaneous mastectomy and immediate breast reconstruction. However, patient satisfaction was graded as good to excellent in 11 of 14 breasts (78.6 percent). Subcutaneous mastectomy with flap reconstruction results in fewer secondary procedures and improved aesthetic outcome when compared with implant reconstruction.


Estimation of a method detection limit for an in vivo XRF arsenic detection system.
An x-ray fluorescence measurement system has been developed with an 125I source to detect arsenic in superficial layers of phantoms and tissue. Based on in vivo measurements, in conjunction with Monte Carlo simulations, the detection limit for arsenic in skin ranges between 2.6+/-0.5 and 5.7+/-1.1 microg g(-1), depending on skin thickness and assuming that arsenic is uniformly distributed in the skin. The effect of skin arsenic distribution was also examined.


Bridging the humanities and health care with theatre: Theory and outcomes of a theatre-based model for enhancing psychiatric care via stigma reduction.
This article describes the rational, methods, implementation, and effectiveness of Identity Development Evolution and Sharing (IDEAS), an evidence-supported, narrative theater-based training that reduces stigma among health care providers to increase health care equity in psychiatric rehabilitation. The IDEAS model has been used to reduce provider bias toward patients. From May 2017 to January 2020, we interviewed people from three patient groups who have been harmed by stigma, including Black women, transgender, and gender-diverse people, and people with substance use disorders. These interviews informed the creation of three theatrical scripts that were performed by professional actors for audiences of health care providers from January 2020 to May 2022. The performances aimed to raise conscious awareness of implicit provider biases and to provide a reflective opportunity to ameliorate these biases. The purpose of IDEAS is to improve experiences in health care settings such as psychiatric rehabilitation of patients from groups who have been harmed by stigma. We used paired-samples t tests to compare pre/postprovider stigma, measured via the Acceptance and Action Questionnaire-Stigma (AAQ-S). Sociodemographic factors for providers who viewed IDEAS were similar across all three performances. IDEAS significantly decreased AAQ-S scores (t = 11.32, df = 50, M = 13.65, 95% confidence limit: [11.32, 15.97], p < .0001). IDEAS reduces provider stigma to support positive clinical encounters with diverse patient populations. These findings are relevant for psychiatric rehabilitation settings, which seek to establish positive rapport between providers and patients. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Optimization of mold wheat bread fortified with soy flour, pea flour and whey protein concentrate.
The objective of this work was to study the effect of replacing a selected wheat flour for defatted soy flour, pea flour and whey protein concentrate on both dough rheological characteristics and the performance and nutritional quality of bread. A mixture design was used to analyze the combination of the ingredients. The optimization process suggested that a mixture containing 88.8% of wheat flour, 8.2% of defatted soy flour, 0.0% of pea flour and 3.0% of whey protein concentrate could be a good combination to achieve the best fortified-bread nutritional quality. The fortified bread resulted in high protein concentration, with an increase in dietary fiber content and higher calcium levels compared with those of control (wheat flour 100%). Regarding protein quality, available lysine content was significantly higher, thus contributing with the essential amino acid requirement.


Safety and antitumor activity of the anti-PD-1 antibody pembrolizumab in patients with recurrent carcinoma of the anal canal.
Safety and efficacy of pembrolizumab, a humanized programmed death 1 monoclonal antibody, was assessed in KEYNOTE-028, a multicohort, phase Ib trial for patients with programmed death ligand 1 (PD-L1)-positive advanced solid tumors. We report results for the cohort of patients with advanced anal carcinoma. Patients with PD-L1-positive tumors (≥1%) received intravenous pembrolizumab 10 mg/kg once every 2 weeks for up to 2 years or until confirmed progression or unacceptable toxicity. Response was assessed every 8 weeks for the first 6 months and every 12 weeks thereafter per Response Evaluation Criteria In Solid Tumors, version 1.1. Primary endpoints were safety and overall response rate per investigator review. Secondary endpoints included progression-free survival, overall survival, and response duration. Data cutoff date was 1 July 2015. Of the 43 patients with advanced anal carcinoma evaluable for PD-L1 expression, 32 (74%) had PD-L1-positive tumors as assessed with the 22C3 prototype assay, of whom 25 were enrolled between April and September 2014. Sixteen patients (64%) experienced treatment-related adverse events; the most common ones were diarrhea and fatigue in four patients (16%) each and nausea in three patients (12%). There were no treatment-related deaths or discontinuations as of the data cutoff date. Among the 24 patients with squamous cell carcinoma histology, four had confirmed partial response, for an overall response rate of 17% [95% confidence interval (CI), 5%-37%) and 10 (42%) had confirmed stable disease, for a disease control rate of 58%. One additional patient with non-squamous histology had confirmed stable disease. In this population of patients with PD-L1-positive advanced squamous cell anal carcinoma, pembrolizumab demonstrated a manageable safety profile and encouraging antitumor activity. These data support further study of pembrolizumab for this patient population. NCT02054806.


Resilience of informal settlements to climate change in the mountainous areas of Konso, Ethiopia and QwaQwa, South Africa.
Managing change is essential for human survival; thus, the importance of adapting to climate change has been increasingly recognised by researchers and governments alike. This is reflected in the growing literature on climate change and the imperative for action including building resilience in our socio-ecological systems. Despite the large body of research that now exists, few studies have considered the resilience of informal rural or peri-urban settlements in mountainous regions. This article considered the resilience of two rural settlements in mountainous areas, namely Konso, Ethiopia, and QwaQwa, South Africa, to the influences of climate change based on the assets available to them. The authors obtained the local communities' perception of their risks throsugh interviews with community leaders and a survey of 384 residents, divided equally between each settlement. Furthermore, the resilience of each community was assessed on the basis of their environmental, social, economic, human, institutional and physical capitals using a climate change resilience indicator. The findings showed that both communities faced major challenges because of climate change, particularly from drought and poverty. We found that both communities retained some forms of indigenous knowledge, but its greater application in Konso appeared to improve resilience to a greater extent than QwaQwa, where it played a lesser role and the community was more dependent on the government. However, indigenous knowledge alone is not sufficient to support these communities in the long term, given the growing aridity of the regions, and other approaches are also necessary, including government support, to enhance and grow their capitals.


HMG-coenzyme A reductase inhibition, type 2 diabetes, and bodyweight: evidence from genetic analysis and randomised trials.
Statins increase the risk of new-onset type 2 diabetes mellitus. We aimed to assess whether this increase in risk is a consequence of inhibition of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), the intended drug target. We used single nucleotide polymorphisms in the HMGCR gene, rs17238484 (for the main analysis) and rs12916 (for a subsidiary analysis) as proxies for HMGCR inhibition by statins. We examined associations of these variants with plasma lipid, glucose, and insulin concentrations; bodyweight; waist circumference; and prevalent and incident type 2 diabetes. Study-specific effect estimates per copy of each LDL-lowering allele were pooled by meta-analysis. These findings were compared with a meta-analysis of new-onset type 2 diabetes and bodyweight change data from randomised trials of statin drugs. The effects of statins in each randomised trial were assessed using meta-analysis. Data were available for up to 223 463 individuals from 43 genetic studies. Each additional rs17238484-G allele was associated with a mean 0·06 mmol/L (95% CI 0·05-0·07) lower LDL cholesterol and higher body weight (0·30 kg, 0·18-0·43), waist circumference (0·32 cm, 0·16-0·47), plasma insulin concentration (1·62%, 0·53-2·72), and plasma glucose concentration (0·23%, 0·02-0·44). The rs12916 SNP had similar effects on LDL cholesterol, bodyweight, and waist circumference. The rs17238484-G allele seemed to be associated with higher risk of type 2 diabetes (odds ratio [OR] per allele 1·02, 95% CI 1·00-1·05); the rs12916-T allele association was consistent (1·06, 1·03-1·09). In 129 170 individuals in randomised trials, statins lowered LDL cholesterol by 0·92 mmol/L (95% CI 0·18-1·67) at 1-year of follow-up, increased bodyweight by 0·24 kg (95% CI 0·10-0·38 in all trials; 0·33 kg, 95% CI 0·24-0·42 in placebo or standard care controlled trials and -0·15 kg, 95% CI -0·39 to 0·08 in intensive-dose vs moderate-dose trials) at a mean of 4·2 years (range 1·9-6·7) of follow-up, and increased the odds of new-onset type 2 diabetes (OR 1·12, 95% CI 1·06-1·18 in all trials; 1·11, 95% CI 1·03-1·20 in placebo or standard care controlled trials and 1·12, 95% CI 1·04-1·22 in intensive-dose vs moderate dose trials). The increased risk of type 2 diabetes noted with statins is at least partially explained by HMGCR inhibition. The funding sources are cited at the end of the paper.


Reliability of N95 respirators for respiratory protection before, during, and after nursing procedures.
The adequate fit of an N95 respirator is important for health care workers to reduce the transmission of airborne infectious diseases in the clinical setting. This study aimed to evaluate whether adequately sealed N95 respirators may provide consistent protection for the wearer while performing nursing procedures. Participants were a group of nursing students (N = 120). The best fitting respirator for these participants was identified from the 3 common models, 1860, 1860S, and 1870+ (3M), using the quantitative fit test (QNFT) method. Participants performed nursing procedures for 10-minute periods while wearing a backpack containing the portable aerosol spectrometers throughout the assessment to detect air particles inside the respirator. The average fit factor of the best fitting respirator worn by the participants dropped significantly after nursing procedures (184.85 vs 134.71) as detected by the QNFT. In addition, significant differences in particle concentration of different sizes (>0.3, >0.4, >1.0, and >4.0 µm) inside the respirator were detected by the portable aerosol spectrometers before, during, and after nursing procedures. Body movements during nursing procedures may increase the risk of face seal leakage. Further research, including the development of prototype devices for better respirator fit, is necessary to improve respiratory protection of users.


Change in Caco-2 cells following treatment with various lavender essential oils.
Lavender is an aromatic evergreen shrub diffused in the Mediterranean basin appreciated since antiquity. The genus Lavandula is part of Lamiaceae family and includes more than 20 species, among which true lavender (L. vera D.C. or L. angustifolia Miller.) and spike lavender (L. latifolia Medikus); there are also numerous hybrids known as lavandins (L. hybrida Rev.). L. vera, spike lavender and several hybrids are the most intensely used breeding species for the production of essential oils. Lavender and lavandin essential oils have been applied in food, pharmaceutical and other agro industries as biological products. In their chemical composition, terpenes linalool and linalyl acetate along with terpenoids such as 1,8-cineole are mostly responsible for biological and therapeutic activities. This study evaluates cytotoxic activity of essential oils derived from four lavender species on human epithelial colorectal adenocarcinoma cells. Analysis of pre- and post-treatment cell morphology has been performed using scanning electron microscope.


The contribution of latent factors of executive functioning to mind wandering: an experience sampling study.
Accumulating evidence suggests that individuals with greater executive resources spend less time mind wandering. Independent strands of research further suggest that this association depends on concentration and a guilty-dysphoric daydreaming style. However, it remains unclear whether this association is specific to particular features of executive functioning or certain operationalizations of mind wandering, including task-unrelated thoughts (TUTs, comprising external distractions and mind wandering) and stimulus-independent and task-unrelated thoughts (SITUTs, comprising mind wandering only). This study sought to clarify these associations by using confirmatory factor analysis to compute latent scores for distinct executive functions based on nine cognitive tasks and relating them to experience sampling reports of mind wandering. We expected that individuals with greater executive control (specifically updating) would show a stronger reduction in SITUTs as momentary concentration and guilty-dysphoric style increase. A bifactor model of the cognitive battery indicated a general factor (common executive functioning) and ancillary factors (updating and shifting). A significant interaction between updating and concentration on mind wandering was observed with mind wandering defined as TUTs, but not as SITUTs (N = 187). A post hoc analysis clarified this discrepancy by showing that as concentration increases, both external distractions and mind wandering decrease more strongly among people with greater updating. Moreover, common executive functioning predicted a more negative slope of guilty-dysphoric style on SITUTs, whereas updating and shifting predicted more positive slopes. The opposite slopes of these executive functions on daily life mind wandering may reflect a stability-flexibility trade-off between goal maintenance and goal replacement abilities.


Lymphomas of the mediastinum.
The purpose of this clinicopathologic overview is to describe the types of lymphomas that present in the mediastinum. A comparison of the frequency of the different subtypes of lymphoma that are found in children and adults is provided. In general, immunohistochemistry and immunophenotyping studies are essential to the laboratory workup of neoplasms presenting in the mediastinum. An assessment of proliferative index in lymphoma is most helpful to determine tumor aggressiveness and patient prognosis. Electron microscopy is most helpful in the differential diagnosis of mediastinal neoplasms, where lymphomas may be distinguished from nonlymphomatous neoplasms using key ultrastructural features. The role of electron microscopy in the subclassification of lymphomas is mostly academic, with a few exceptions. The varied ultrastructural appearance of Hodgkin's cells and of different subtypes of non-Hodgkin's lymphoma is illustrated, using cases from our patient files. An ultrastructural study of lacunar cells in Hodgkin's disease provides evidence that the formation of lacunae may have a structural and/or physiologic basis. Mummified cells showing some of the features of a physiologic form of cell death, called apoptosis, are also described.


A Mechanistic Model of Annual Sulfate Concentrations in the United States.
Understanding how individual pollution sources contribute to ambient sulfate pollution is critical for assessing past and future air quality regulations. Since attribution to specific sources is typically not encoded in spatial air pollution data, we develop a mechanistic model which we use to estimate, with uncertainty, the contribution of ambient sulfate concentrations attributable specifically to sulfur dioxide (SO2) emissions from individual coal-fired power plants in the central United States. We propose a multivariate Ornstein-Uhlenbeck (OU) process approximation to the dynamics of the underlying space-time chemical transport process, and its distributional properties are leveraged to specify novel probability models for spatial data that are viewed as either a snapshot or time-averaged observation of the OU process. Using US EPA SO2 emissions data from 193 power plants and state-of-the-art estimates of ground-level annual mean sulfate concentrations, we estimate that in 2011 - a time of active power plant regulatory action - existing flue-gas desulfurization (FGD) technologies at 66 power plants reduced population-weighted exposure to ambient sulfate by 1.97 μg/m3 (95% CI: 1.80 - 2.15). Furthermore, we anticipate future regulatory benefits by estimating that installing FGD technologies at the five largest SO2-emitting facilities would reduce human exposure to ambient sulfate by an additional 0.45 μg/m3 (95% CI: 0.33 - 0.54).


Scientific validation of the antimicrobial and antiproliferative potential of Berberis aristata DC root bark, its phytoconstituents and their biosafety.
Berberis aristata is an important part of traditional healing system from more than 2500 years. The aqueous extract of Berberis aristata root bark displayed broad spectrum activity against 13 test pathogens, ranging from 12 to 25 mm. In classical optimization, 15% concentration prepared at 40 °C for 40 min was optimal and thermostable. Statistical optimization enhanced the activity by 1.13-1.30-folds. Ethyl acetate was the best organic solvent to elute out the potential compound responsible for antimicrobial activity. Diterpenes were the most abundant phytoconstituent (15.3%) and showed broad spectrum antimicrobial activity ranging from 16.66 to 42.66 mm. Ethyl acetate extract displayed the lowest minimum inhibitory concentration (0.05-1 mg/mL), followed by diterpenes (0.05-5 mg/mL) and flavonoids (0.05-10 mg/mL). The test extracts were microbicidal in nature and showed a prolonged post antibiotic effect ranging from 2 to 8 h. They were found to be biosafe as per Ames and MTT assay. The in vitro cytotoxicity evaluation of diterpenes against L20B, RD and Hep 2 cell lines revealed its IC50 ranging from 245 to 473 µg/mL. Acute oral toxicity of diterpenes on Swiss albino mice did not show any changes in behavioral pattern, body weight, biochemical parameters as well as organs' architecture. The study thus indicates B. aristata could be a potential candidate for development of potent drug owing to its antimicrobial potential and biosafe profile.


Selected observations on rickettsiae and their host cells.
The study of the interaction between rickettsiae and their host cells is in its infancy. Members of the genera Rickettsia, Coxiella and Rochalimaea show considerable diversity in host cell range (in vivo vs. in vitro), kind of association with host cell (pericellular, intracellular), mode of entry, interactions with various host cell membranes, intracellular localization (intraphagosomal, free in cytoplasm, intranuclear), adaptation to preferred microhabitat (e.g., optimal pH for enzymes), details of growth cycle, mechanisms of host cell damage. Quantitative in vitro methods exist for the study of infection cycles. Knowledge of nutritional requirements is almost non-existent. Host factors, (e.g., antibody, lymphokines, immune interferon) influence intracellular rickettsiae. Rickettsia-host cell interactions remain a fertile field for discovery.


Differentiation between Candida albicans and Candida dubliniensis using hypertonic Sabouraud broth and tobacco agar.
Opportunistic fungal infections in immunocompromised hosts are caused by Candida species, and the majority of such infections are due to Candida albicans. However, the emerging pathogen Candida dubliniensis demonstrates several phenotypic characteristics in common with C. albicans, such as production of germ tubes and chlamydospores, calling attention to the development of stable resistance to fluconazole in vitro. The aim of this study was to evaluate the performance of biochemistry identification in the differentiating between C. albicans and C. dubliniensis, by phenotyping of yeast identified as C. albicans. Seventy-nine isolates identified as C. albicans by the API system ID 32C were grown on Sabouraud dextrose agar at 30°C for 24-48h and then inoculated on hypertonic Sabouraud broth and tobacco agar. Our results showed that 17 (21.5%) isolates were growth-inhibited on hypertonic Sabouraud broth, a phenotypic trait inconsistent with C. albicans in this medium. However, the results observed on tobacco agar showed that only 9 (11.4%) of the growth-inhibited isolates produced characteristic colonies of C. dubliniensis (rough colonies, yellowish-brown with abundant fragments of hyphae and chlamydospores). The results suggest that this method is a simple tool for screening C. albicans and non-albicans yeast and for verification of automated identification.


Psychological characteristics of Italian gender dysphoric adolescents: a case-control study.
Gender dysphoria (GD) is associated with clinically significant distress and impairment in social, scholastic, and other important areas of functioning, especially when early onset is reported. The aim of the present study is to assess the psychopathological features associated with GD in adolescence, comparing a group of gender dysphoric adolescents (GDs) with a group of non-referred adolescents (NRs), in terms of body uneasiness, suicide risk, psychological functioning, and intensity of GD. A sample of 46 adolescents with GD and 46 age-matched NRs was evaluated (mean ± SD age = 16.00 ± 1.49 and 16.59 ± 1.11 respectively, p > 0.05). Subjects were asked to complete the Body Uneasiness Test (BUT) to explore body uneasiness, the Youth Self Report (YSR) to measure psychological functioning, the Multi-Attitude Suicide Tendency Scale (MAST) for suicide risk, and the Gender Identity/Gender Dysphoria Questionnaire for Adolescents and Adults (GIDYQ-AA) for GD assessment. Adolescents with GD reported significantly higher levels of body uneasiness (BUT-GSI, F = 380.13, p < 0.0001), as well as a worse psychological functioning (YSR, F = 13.06 and p < 0.0001 for "total problem scale" and F = 12.53, p = 0.001 for "internalizing" scale) as compared to NRs. When YSR subscales were considered, GDs showed significantly higher scores in the "withdrawal/depression", "anxiety/depression", and "social problems" (all p < 0.0001). In addition, GDs showed significantly higher levels in the "attraction to death" and "repulsion by life" scales and lower scores in the "attraction to life" scale (all p < 0.0001). Finally, GIDYQ-AA score was significantly lower (meaning a higher level of gender dysphoria symptoms) in GDs vs. NRs (p < 0.0001). GD adolescents reported significantly higher body dissatisfaction and suicidal risk compared to NRs. In addition, results confirmed a significant impairment in social psychological functioning in adolescents with GD.


[Hard x-ray therapy with special reference to postoperative irradiation of breast carcinoma].
Five-year-experiences have shown that a considerably intensified tolerance of the skin and an absorption equability of tissues and bones and muscles are obtained with the hard ray roentgen irradiator, RT 305, for half-value depths from 20 mm to 70 mm H-2O and half-value thickness to about 7 mm Cu. By means of a rod anode tube of 300 kV with a super-normal filtration to 3.5 mm Pb, a hard and nearly monochromatic radiation, which already possesses the above mentioned advantages of the ultra-hard ray therapy with an effective energy about 0.2 MeV, appears. The application of this hard ray therapy is given preferably for disease foci in the environment of bone tissues, i.e. in the head and throat, the vertebral column, the osseous pelvic ring and the extremities and as the conventional roentgen therapy ist not able to give it. The hard ray therapy seems to be particularly adapted for the postoperative irradiation of the cancer of the breast with the ribs lying in the region of the irradiation fields, with the sternum and the bones of the shoulder girdle. With regard to these radiotherapeutic possibilities, corresponding cases are demonstrated.


Prospective associations of insomnia markers and symptoms with depression.
Whether insomnia, a known correlate of depression, predicts depression longitudinally warrants elucidation. The authors examined 555 Wisconsin Sleep Cohort Study participants aged 33-71 years without baseline depression or antidepressant use who completed baseline and follow-up overnight polysomnography and had complete questionnaire-based data on insomnia and depression for 1998-2006. Using Poisson regression, they estimated relative risks for depression (Zung scale score > or =50) at 4-year (average) follow-up according to baseline insomnia symptoms and polysomnographic markers. Twenty-six participants (4.7%) developed depression by follow-up. Having 3-4 insomnia symptoms versus none predicted depression risk (age-, sex-, and comorbidity-adjusted relative risk (RR) = 3.2, 95% confidence interval: 1.1, 9.6). After multiple adjustments, frequent difficulty falling asleep (RR = 5.3, 95% confidence interval: 1.1, 27.9) and polysomnographically assessed (upper or lower quartiles) sleep latency, continuity, and duration (RRs = 2.2-4.7; P's < or = 0.05) predicted depression. Graded trends (P-trend < or = 0.05) were observed with increasing number of symptoms, difficulty falling asleep, and difficulty returning to sleep. Given the small number of events using Zung > or =50 (depression cutpoint), a limitation that may bias multivariable estimates, continuous depression scores were analyzed; mean values were largely consistent with dichotomous findings. Insomnia symptoms or markers increased depression risk 2.2- to 5.3-fold. These results support prior findings based on self-reported insomnia and may extend similar conclusions to objective markers. Heightened recognition and treatment of insomnia may prevent subsequent depression.


CRISPR genome engineering and viral gene delivery: a case of mutual attraction.
The adaptation of the CRISPR/Cas9 DNA engineering machinery for mammalian cells has revolutionized our approaches to low- or high-throughput genome annotation and paved the way for conceptually novel therapeutic strategies. A large part of the attraction of CRISPR stems from the small size of its two core components--Cas9 and gRNA--and hence its compatibility with virtually any available viral vector delivery system. As a result, over the past two years, four major classes of viral vectors have already been engineered and applied as CRISPR delivery tools--retroviruses, lentiviruses, adenoviruses, and adeno-associated viruses (AAVs). The juxtaposition of these two technologies reflects a case of tremendous mutual attraction and holds unprecedented promises for biology and medicine. Here, we provide an overview of the state-of-the-art of this rapidly emerging field, from a comparative description of the principal vector designs, to a synopsis of some of the most exciting applications that were reported to date, including the use of viral CRISPR vectors for genome-wide loss-of-function screens, multiplexed gene editing or disease modeling in animals. Once specificity and safety have been improved further, viral vector-mediated in vitro/in vivo CRISPR delivery and expression promise to radically transform basic and applied biomedical research.


Systems biology markup language: Level 2 and beyond.
The SBML (systems biology markup language) is a standard exchange format for computational models of biochemical networks. We continue developing SBML collaboratively with the modelling community to meet their evolving needs. The recently introduced SBML Level 2 includes several enhancements to the original Level 1, and features under development for SBML Level 3 include model composition, multistate chemical species and diagrams.


Circulating folic acid in plasma: relation to folic acid fortification.
The implementation of folic acid fortification in the United States has resulted in unprecedented amounts of this synthetic form of folate in the American diet. Folic acid in circulation may be a useful measure of physiologic exposure to synthetic folic acid, and there is a potential for elevated concentrations after fortification and the possibility of adverse effects. We assessed the effect of folic acid fortification on circulating concentrations of folic acid and 5-methyltetrahydrofolate in the Framingham Offspring Cohort. This is a cross-sectional study that used plasma samples from fasting subjects before and after fortification. Samples were measured for folate distribution with the use of an affinity-HPLC method with electrochemical detection. Among nonsupplement users, the median concentration of folic acid in plasma increased from 0.25 to 0.50 nmol/L (P < 0.001) after fortification, and among supplement users the median increased from 0.54 to 0.68 nmol/L (P = 0.001). Among nonsupplement users, the prevalence of high circulating folic acid (>/=85th percentile) increased from 9.4% to 19.1% (P = 0.002) after fortification. Among supplement users, the prevalence of high circulating folic acid increased from 15.9% to 24.3% (P = 0.02). Folic acid intake and total plasma folate were positively and significantly related to high circulating folic acid after adjustment for potential confounding factors (P for trend < 0.001). Folic acid fortification has resulted in increased exposure to circulating folic acid. The biochemical and physiologic consequences of this are unknown, but these findings highlight the need to understand the effects of chronic exposure to circulating folic acid.


A framework for whole-cell mathematical modeling.
The default framework for modeling biochemical processes is that of a constant-volume reactor operating under steady-state conditions. This is satisfactory for many applications, but not for modeling growth and division of cells. In this study, a whole-cell modeling framework is developed that assumes expanding volumes and a cell-division cycle. A spherical newborn cell is designed to grow in volume during the growth phase of the cycle. After 80% of the cycle period, the cell begins to divide by constricting about its equator, ultimately affording two spherical cells with total volume equal to twice that of the original. The cell is partitioned into two regions or volumes, namely the cytoplasm (Vcyt) and membrane (Vmem), with molecular components present in each. Both volumes change during the cell cycle; Vcyt changes in response to osmotic pressure changes as nutrients enter the cell from the environment, while Vmem changes in response to this osmotic pressure effect such that membrane thickness remains invariant. The two volumes change at different rates; in most cases, this imposes periodic or oscillatory behavior on all components within the cell. Since the framework itself rather than a particular set of reactions and components is responsible for this behavior, it should be possible to model various biochemical processes within it, affording stable periodic solutions without requiring that the biochemical process itself generates oscillations as an inherent feature. Given that these processes naturally occur in growing and dividing cells, it is reasonable to conclude that the dynamics of component concentrations will be more realistic than when modeled within constant-volume and/or steady-state frameworks. This approach is illustrated using a symbolic whole cell model.


Experience sampling method and the everyday experiences of adults with intellectual disability: A feasibility study.
Experiences of people with intellectual disability are often reported by proxy, excluding the person's own perception. To assist people with intellectual disabilities ability to communicate their own experiences, the current study explored the feasibility, reliability and validity of experience sampling methods (ESMs) for people with intellectual disability. After a training session, 19 participants carried a mobile device for 7 consecutive days, answering a survey when prompted 7 times daily. Participants were interviewed at the end of data collection. Excluding incomplete entries, the response rate was 33.8%, varying by living arrangement and employment. Split-half reliability and correlations among logically linked internal experiences demonstrated strong reliability and validity. Illustration of the context of responses supported face validity. Technological and content difficulties were discussed in interviews. Experience sampling methods is feasible for some people with intellectual disability, providing valid and reliable information. Future research is needed to further improve feasibility.


Molecular investigation of mixed malaria infections in southwest Saudi Arabia.
To investigate the incidence of mixed-species (MS) malaria infection, and compare the results with microscopically confirmed cases of malaria. During 2010, blood spots collected from 371 clinically suspected cases of malaria were microscopically examined in a cross-sectional study. The DNA was extracted from the samples, and a nested polymerase chain reaction (PCR) was performed. The results obtained by the 2 methods were compared. From the microscopic analysis it was determined that 369 samples (99.5%) were positive for Plasmodium falciparum (P. falciparum) and 2 were Plasmodium vivax (P. vivax) mono-infections. There were no mixed malaria infections. The PCR analysis, however, showed that in 7 cases (1.9%) the infection was caused by MS malaria comprising of P. falciparum and P. vivax, 2 of these representing the cases that were microscopically diagnosed as P. vivax mono-infections. All cases were negative for Plasmodium malariae, Plasmodium ovale, and Plasmodium knowlesi. Mixed malaria infections are currently overlooked when using microscopy. The PCR assays are essential complementary techniques that should be used with microscopic examination of blood smears.


Dietary fat, cooking fat, and breast cancer risk in a multiethnic population.
Our objective was to examine the association between dietary fat intake, cooking fat usage, and breast cancer risk in a population-based, multiethnic, case-control study conducted in the San Francisco Bay area. Intake of total fat and types of fat were assessed with a food frequency questionnaire among 1,703 breast cancer cases diagnosed between 1995 and 1999 and 2,045 controls. In addition, preferred use of fat for cooking was assessed. Unconditional logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). High fat intake was associated with increased risk of breast cancer (highest vs. lowest quartile, adjusted OR = 1.35, 95% CI = 1.10-1.65, P(trend) < 0.01). A positive association was found for oleic acid (OR = 1.55, 95% CI = 1.14-2.10, P(trend) < 0.01) but not for linoleic acid or saturated fat. Risk was increased for women cooking with hydrogenated fats (OR = 1.58, 95% CI = 1.20-2.10) or vegetable/corn oil (rich in linoleic acid; OR = 1.30, 95% CI = 1.06-1.58) compared to women using olive/canola oil (rich in oleic acid). Our results suggest that a low-fat diet may play a role in breast cancer prevention. We speculate that monounsaturated trans fats may have driven the discrepant associations between types of fat and breast cancer.


ZAP-70 promotes the infiltration of malignant B-lymphocytes into the bone marrow by enhancing signaling and migration after CXCR4 stimulation.
ZAP-70 in chronic lymphocytic leukemia (CLL) is associated with enhanced response to microenvironmental stimuli. We analyzed the functional consequences of ZAP-70 ectopic expression in malignant B-cells in a xenograft mouse model of disseminated B-cell leukemia. Mice injected with B-cells expressing ZAP-70 showed a prominently higher infiltration of the bone marrow. In vitro analysis of the response of malignant B-cells to CXCL12, the main attracting chemokine regulating trafficking of lymphocytes to the bone marrow, or to bone marrow stromal cells, revealed that ZAP-70 induces an increased response in terms of signaling and migration. These effects are probably mediated by direct participation of ZAP-70 in CXCL12-CXCR4 signaling since CXCR4 stimulation led to activation of ZAP-70 and downstream signaling pathways, such as MAPK and Akt, whereas ZAP-70 did not alter the expression of the CXCR4 receptor. In addition, subclones of primary CLL cells with high expression of ZAP-70 also showed increased migrative capacity toward CXCL12. Neutralization of CXCR4 with a monoclonal antibody resulted in impaired in vitro responses to CXCL12 and bone marrow stromal cells. We conclude that ZAP-70 enhances the migration of malignant B-cells into the supportive microenvironment found in the bone marrow mainly by enhancing signaling and migration after CXCR4 stimulation.


Expression of HIV-1 Vpu leads to loss of the viral restriction factor CD317/Tetherin from lipid rafts and its enhanced lysosomal degradation.
CD317/tetherin (aka BST2 or HM1.24 antigen) is an interferon inducible membrane protein present in regions of the lipid bilayer enriched in sphingolipids and cholesterol (often termed lipid rafts). It has been implicated in an eclectic mix of cellular processes including, most notably, the retention of fully formed viral particles at the surface of cells infected with HIV and other enveloped viruses. Expression of the HIV viral accessory protein Vpu has been shown to lead to intracellular sequestration and degradation of tetherin, thereby counteracting the inhibition of viral release. There is evidence that tetherin interacts directly with Vpu, but it remains unclear where in the cell this interaction occurs or if Vpu expression affects the lipid raft localisation of tetherin. We have addressed these points using biochemical and cell imaging approaches focused on endogenous rather than ectopically over-expressed tetherin. We find i) no evidence for an interaction between Vpu and endogenous tetherin at the cell surface, ii) the vast majority of endogenous tetherin that is at the cell surface in control cells is in lipid rafts, iii) internalised tetherin is present in non-raft fractions, iv) expression of Vpu in cells expressing endogenous tetherin leads to the loss of tetherin from lipid rafts, v) internalised tetherin enters early endosomes, and late endosomes, in both control cells and cells expressing Vpu, but the proportion of tetherin molecules destined for degradation rather than recycling is increased in cells expressing Vpu vi) lysosomes are the primary site for degradation of endogenous tetherin in cells expressing Vpu. Our studies underlie the importance of studying endogenous tetherin and let us propose a model in which Vpu intercepts newly internalised tetherin and diverts it for lysosomal destruction rather than recycling to the cell surface.


Plant regeneration and Agrobacterium-mediated genetic transformation systems in liliaceous ornamental plants.
The family Liliaceae (Cronquist system) contains various important ornamental plants. We have been examining for about 20 years the establishment of plant regeneration and genetic transformation systems in liliaceous ornamental plants for their biotechnological breeding and elucidation of the molecular mechanisms determining ornamental traits. In this review, studies on in vitro plant regeneration in 7 genera and on Agrobacterium-mediated production of transgenic plants in 4 genera are described. Plant regeneration was achieved via callus cultures in Agapanthus, Hemerocallis, Hosta, Lilium, Muscari and Tricyrtis. Auxins (2,4-dichrolophenoxyacetic acid, α-naphthaleneacetic acid and/or picloram) were effective for inducing regenerable calli. Tulipa species and cultivars were very recalcitrant to callus induction and plant regeneration. Agrobacterium-mediated transformation was examined in Agapanthus, Lilium, Muscari and Tricyrtis, and transgenic plants were obtained in all genera by using regenerable calli as a target material for Agrobacterium inoculation, inoculation and co-cultivation with Agrobacterium in the presence of acetosyringone, and selection of transgenic tissues and plantlets on hygromycin-containing media. Among 4 genera, Tricyrtis has several advantages for transformation studies: higher transformation efficiency, relatively small plant size, ease of cultivation, and taking only 1 year from in vitro regeneration to flowering. We are now investigating the molecular mechanisms for determining plant form, flower color and flower form by using Tricyrtis spp. as liliaceous model plants.


Use of the single strand conformation polymorphism technique and PCR to detect p53 gene mutations in small cell lung cancer.
Recent studies have suggested that the p53 oncoprotein might function normally as a tumor suppressor. Mutations in highly conserved regions of the p53 gene have been observed in numerous types of tumors and tumor cell lines. To detect in a more sensitive manner p53 gene mutations in small cell lung cancer (SCLC) we utilized the single strand conformation polymorphism (SSCP) technique of Orita et al., (1989). Using PCR primers for the most highly conserved regions of the p53 gene, including exons 4-9, we have identified p53 mutations in 5 of 9 small cell lung cancer (SCLC) tumor DNA samples and in 1 SCLC cell line. None of the mutations seen in tumor DNA samples were present in normal DNA from the same patients, indicating that mutation of the p53 gene in these tumors was a somatic event. Of the six mutations observed, two were found in exon 7, three were found in the region encompassing exons 8 and 9, and one was found in the region encompassing exons 5 and 6. Nucleotide sequencing of one of the exon 7 mutations and one of the exon 8-9 mutations indicated that each was a C to T transition. In SCLC-6 the mutation resulted in substitution of serine for proline at amino acid 278 and in SCLC-4 substitution of tryptophan for arginine at amino acid 248, both nonconservative amino acid substitutions. Both of these changes are in regions of the p53 gene where mutations have been observed in other tumors. Two additional mutations were observed in SCLC cell lines using conventional PCR techniques. One of these is a mutation which results in altered splicing of the p53 pre-mRNA.


Student abuse in medical school: a comparison of students' and faculty's perceptions.
A perceived abuse or mistreatment has been identified in the United States as a major source of stress for undergraduate medical students. The objective of this paper is to explore whether medical students in Israel have similar complaints, and if so, whether medical faculty members are aware of students' perceptions of mistreatment. Third and fifth year medical students of the 6 year undergraduate program of the Hebrew University-Hadassah Medical School were asked to respond to a self-administered questionnaire similar to those used in previous studies in the USA. An additional questionnaire was developed for faculty members. Response rates were 75% for students and 39% for faculty members. Analysis of the responses indicated that similar to the results of American studies, the number and severity of all forms of perceived incidents of abuse increased from the third (pre-clinical) to the fifth (clinical) year. Verbal abuse was the most frequently reported form of mistreatment. The most frequently identified abusers were nurses, followed by clinical faculty members, and general surgery was reported as the most abusive clinical department. Students' complaints of denial of basic privileges were more common than those reported in the USA, while verbal abuse and physical threats were more frequently reported by American medical students. Analysis of the responses of faculty members indicated that clinical faculty, residents and interns were unaware of the extent of these students' perceptions, while pre-clinical faculty overestimated the extent of verbal and psychological abuse. We conclude that the perception of abuse or mistreatment among Israeli medical students is more prevalent than either students or faculty believe.


Lumen-positive chloride transport potential in the early distal tubule of Triturus kidney: its absolute dependence on the presence of Na+ and K+ in the luminal fluid.
Microperfusion experiments with Na+-free and/or K+-free solutions revealed that the generation of Cl- -dependent lumen-positive transtubular potentials of the early distal tubule of Triturus kidney was absolutely dependent on the presence of both Na+ and K+ in the lumen. Increases in luminal Na+ or K+ concentration increased the luminal positivity in a saturable manner, and the half saturation concentration was found to be about 1 mM for both cations. The results strongly suggest that Cl- is cotransported with Na+ and K+ across the luminal membrane.


Ectopic Premolar Tooth in the Sigmoid Notch.
Impaction of a mandibular premolar is relatively uncommon. Ectopic placement is more unusual and there has been no discussion in the literature of an ectopic mandibular premolar in the coronoid process. In this case report, we present an impacted ectopic mandibular permanent premolar in the sigmoid notch (incisura mandibulae) region. Etiology of the tooth and treatment options are discussed and illustrated by Cone Beam Computed Tomography (CBCT) images.


Colonization and diversification of Galápagos terrestrial fauna: a phylogenetic and biogeographical synthesis.
Remote oceanic islands have long been recognized as natural models for the study of evolutionary processes involved in diversification. Their remoteness provides opportunities for isolation and divergence of populations, which make islands remarkable settings for the study of diversification. Groups of islands may share a relatively similar geological history and comparable climate, but their inhabitants experience subtly different environments and have distinct evolutionary histories, offering the potential for comparative studies. A range of organisms have colonized the Galápagos Islands, and various lineages have radiated throughout the archipelago to form unique assemblages. This review pays particular attention to molecular phylogenetic studies of Galápagos terrestrial fauna. We find that most of the Galápagos terrestrial fauna have diversified in parallel to the geological formation of the islands. Lineages have occasionally diversified within islands, and the clearest cases occur in taxa with very low vagility and on large islands with diverse habitats. Ecology and habitat specialization appear to be critical in speciation both within and between islands. Although the number of phylogenetic studies is continuously increasing, studies of natural history, ecology, evolution and behaviour are essential to completely reveal how diversification proceeded on these islands.


Intracellular transport and storage of secretory proteins in relation to cytodifferentiation in neoplastic pancreatic acinar cells.
The pancreatic acinar carcinoma established in rat by Reddy and Rao (1977, Science 198:78-80) demonstrates heterogeneity of cytodifferentiation ranging from cells containing abundant well-developed secretory granules to those with virtually none. We examined the synthesis intracellular transport and storage of secretory proteins in secretory granule-enriched (GEF) and secretory granule-deficient (GDF) subpopulations of neoplastic acinar cells separable by Percoll gradient centrifugation, to determine the secretory process in cells with distinctly different cytodifferentiation. The cells pulse-labeled with [3H]leucine for 3 min and chase incubated for up to 4 h were analyzed by quantitative electron microscope autoradiography. In GEF neoplastic cells, the results of grain counts and relative grain density estimates establish that the label moves successively from rough endoplasmic reticulum (RER) leads to the Golgi apparatus leads to post-Golgi vesicles (vacuoles or immature granules) leads to mature secretory granules, in a manner reminiscent of the secretory process in normal pancreatic acinar cells. The presence of approximately 40% of the label in association with secretory granules at 4 h postpulse indicates that GEF neoplastic cells retain (acquire) the essential regulatory controls of the secretory process. In GDF neoplastic acinar cells the drainage of label from RER is slower, but the peak label of approximately 20% in the Golgi apparatus is reached relatively rapidly (10 min postpulse). The movement of label from the Golgi to the post-Golgi vesicles is evident; further delineation of the secretory process in GDF neoplastic cells, however, was not possible due to lack of secretory granule differentiation. The movement of label from RER leads to the Golgi apparatus leads to the post-Golgi vesicles suggests that GDF neoplastic cells also synthesize secretory proteins, but to a lesser extent than the GEF cells. The reason(s) for the inability of GDF cells to concentrate and store exportable proteins remain to be elucidated.


Orthogonal Gelations to Synthesize Core-Shell Hydrogels Loaded with Nanoemulsion-Templated Drug Nanoparticles for Versatile Oral Drug Delivery.
Hydrophobic active pharmaceutical ingredients (APIs) are ubiquitous in the drug development pipeline, but their poor bioavailability often prevents their translation into drug products. Industrial processes to formulate hydrophobic APIs are expensive, difficult to optimize, and not flexible enough to incorporate customizable drug release profiles into drug products. Here, a novel, dual-responsive gelation process that exploits orthogonal thermo-responsive and ion-responsive gelations is introduced. This one-step "dual gelation" synthesizes core-shell (methylcellulose-alginate) hydrogel particles and encapsulates drug-laden nanoemulsions in the hydrogel matrices. In situ crystallization templates drug nanocrystals inside the polymeric core, while a kinetically stable amorphous solid dispersion is templated in the shell. Drug release is explored as a function of particle geometry, and programmable release is demonstrated for various therapeutic applications including delayed pulsatile release and sequential release of a model fixed-dose combination drug product of ibuprofen and fenofibrate. Independent control over drug loading between the shell and the core is demonstrated. This formulation approach is shown to be a flexible process to develop drug products with biocompatible materials, facile synthesis, and precise drug release performance. This work suggests and applies a novel method to leverage orthogonal gel chemistries to generate functional core-shell hydrogel particles.


Surface salt bridges modulate the DNA site size of bacterial histone-like HU proteins.
Bacterial histone-like DNA-binding proteins are best known for their role in compacting the genomic DNA. Of these proteins, HU is ubiquitous and highly conserved across the eubacterial kingdom. Using the HBsu (Bacillus subtilis-encoded HU homologue) as a model, we explore here the molecular basis for the ability of some HU homologues to engage a longer approx. 35 bp DNA site as opposed to the much shorter sites reported for other homologues. Using electrophoretic mobility-shift assays, we show that the DNA site size for HBsu is approx. 10-13 bp and that a specific surface salt bridge limits the DNA site size for HBsu. Surface exposure of the highly conserved Lys3, achieved by substitution of its salt-bridging partner Asp26 with Ala, leads to enhanced DNA compaction by HBsu-D26A (where D26A stands for the mutant Asp26-->Ala), consistent with the interaction of Lys3 with the ends of a 25 bp duplex. Both HBsu and HBsu-D26A bend DNA, as demonstrated by their equivalent ability to promote ligase-mediated DNA cyclization, indicating that residues involved in mediating DNA kinks are unaltered in the mutant protein. We suggest that Lys3 is important for DNA wrapping due to its position at a distance from the DNA kinks where it can exert optimal leverage on flanking DNA and that participation of Lys3 in a surface salt bridge competes for its interaction with DNA phosphates, thereby reducing the occluded site size.


A combination of plant-derived odors reduces corticosterone and oxidative indicators of stress.
In this study, we measured typical stress markers in addition to oxidative status and reduced glutathione in erythrocytes, and plasma lipid peroxidation of restraint-stressed animals exposed to a combination of plant-derived odors (0.03% Z-3-hexen-1-ol, 0.03% E-2-hexenal, and 0.015% α-pinene in triethyl citrate). Male Wistar rats aged 6-7 weeks postnatal were exposed to vehicle (triethyl citrate, n = 12), plant-derived odors (n = 12), or 1% propionic acid odor (n = 12) under control or stress conditions, and blood samples were collected. Restraint stress increased plasma glucose and plasma corticosterone concentrations by approximately 10% (P < 0.01) and 125% (P < 0.001), respectively, in vehicle-exposed animals. Similar increases were observed in animals exposed to a 1% propionic acid odor, indicating the novelty of odor exposure does not alter stress responsiveness. There was also an increase of approximately 15% in both erythrocytic oxidative status (P < 0.001) and plasma lipid peroxidation (P < 0.05), and a decrease of approximately the same magnitude in reduced glutathione (P < 0.05) in restrained animals with vehicle exposure. There were no differences observed between control and stress treatment with plant-derived odor exposure in any of the measured parameters. It was concluded that exposure to plant-derived odors reduce corticosterone, glucose, and redox responses elicited by psychological stress.


Targeted disruption of the mouse homologue of the Drosophila polyhomeotic gene leads to altered anteroposterior patterning and neural crest defects.
The rae28 gene is a mouse homologue of the Drosophila polyhomeotic gene (Nomura, M., Takihara, Y. and Shimada, K. (1994) Differentiation 57, 39-50), which is a member of the Polycomb group (Pc-G) of genes (DeCamillis, M., Cheng, N., Pierre, D. and Brock, H.W. (1992) Genes Dev. 6, 223-232). The Pc-G genes are required for the correct expression of the Homeotic complex genes and segment specification during Drosophila embryogenesis and larval development. To study the role of the rae28 gene in mouse development, we generated rae28-deficient mice by gene targeting in embryonic stem cells. The rae28-/- homozygous mice exhibited perinatal lethality, posterior skeletal transformations and defects in neural crest-related tissues, including ocular abnormalities, cleft palate, parathyroid and thymic hypoplasia and cardiac anomalies. The anterior boundaries of Hoxa-3, a-4, a-5, b-3, b-4 and d-4 expression were shifted rostrally in the paraxial mesoderm of the rae28-/- homozygous embryos, and those of Hoxb-3 and b-4 expression were also similarly altered in the rhombomeres and/or pharyngeal arches. These altered Hox codes were presumed to be correlated with the posterior skeletal transformations and neural crest defects observed in the rae28-/- homozygous mice. These results indicate that the rae28 gene is involved in the regulation of Hox gene expression and segment specification during paraxial mesoderm and neural crest development.


Free energy simulations of a GTPase: GTP and GDP binding to archaeal initiation factor 2.
Archaeal initiation factor 2 (aIF2) is a protein involved in the initiation of protein biosynthesis. In its GTP-bound, "ON" conformation, aIF2 binds an initiator tRNA and carries it to the ribosome. In its GDP-bound, "OFF" conformation, it dissociates from tRNA. To understand the specific binding of GTP and GDP and its dependence on the ON or OFF conformational state of aIF2, molecular dynamics free energy simulations (MDFE) are a tool of choice. However, the validity of the computed free energies depends on the simulation model, including the force field and the boundary conditions, and on the extent of conformational sampling in the simulations. aIF2 and other GTPases present specific difficulties; in particular, the nucleotide ligand coordinates a divalent Mg(2+) ion, which can polarize the electronic distribution of its environment. Thus, a force field with an explicit treatment of electronic polarizability could be necessary, rather than a simpler, fixed charge force field. Here, we begin by comparing a fixed charge force field to quantum chemical calculations and experiment for Mg(2+):phosphate binding in solution, with the force field giving large errors. Next, we consider GTP and GDP bound to aIF2 and we compare two fixed charge force fields to the recent, polarizable, AMOEBA force field, extended here in a simple, approximate manner to include GTP. We focus on a quantity that approximates the free energy to change GTP into GDP. Despite the errors seen for Mg(2+):phosphate binding in solution, we observe a substantial cancellation of errors when we compare the free energy change in the protein to that in solution, or when we compare the protein ON and OFF states. Finally, we have used the fixed charge force field to perform MDFE simulations and alchemically transform GTP into GDP in the protein and in solution. With a total of about 200 ns of molecular dynamics, we obtain good convergence and a reasonable statistical uncertainty, comparable to the force field uncertainty, and somewhat lower than the predicted GTP/GDP binding free energy differences. The sign and magnitudes of the differences can thus be interpreted at a semiquantitative level, and are found to be consistent with the experimental binding preferences of ON- and OFF-aIF2.


Geographic disparity, area poverty, and human papillomavirus vaccination.
A human papillomavirus (HPV) vaccine was approved by the Food and Drug Administration for use among women/girls in 2006. Since that time, limited research has examined HPV vaccine uptake among adolescent girls and no studies have examined the role of geographic disparities in HPV vaccination. The purpose of this study is to examine geographic disparity in the prevalence of human papillomavirus (HPV) vaccination and to examine individual-, county-, and state-level correlates of vaccination. Three-level random intercept multilevel logistic regression models were fitted to data from girls aged 13-17 years living in six U.S. states using data from the 2008 Behavioral Risk Factor Surveillance System (BRFSS) and the 2000 U.S. census. Data from 1709 girls nested within 274 counties and six states were included. Girls were predominantly white (70.6%) and insured (74.5%). Overall, 34.4% of girls were vaccinated. Significant geographic disparity across states (variance=0.134, SE=0.065) and counties (variance=0.146, SE=0.063) was present, which was partially explained by state and county poverty levels. Independent of individual-level factors, poverty had differing effects at the state and county level: girls in states with higher levels of poverty were less likely whereas girls in counties with higher poverty levels were more likely to be vaccinated. Household income demonstrated a similar pattern to that of county-level poverty: Compared to girls in the highest-income families, girls in the lowest-income families were more likely to be vaccinated. The results of this study suggest geographic disparity in HPV vaccination. Although higher state-level poverty is associated with a lower likelihood of vaccination, higher county-level poverty and lower income at the family level is associated with a higher likelihood of vaccination. Research is needed to better understand these disparities and to inform interventions to increase vaccination among all eligible girls.


Tropomyosin Ser-283 pseudo-phosphorylation slows myofibril relaxation.
Tropomyosin (Tm) is a central protein in the Ca(2+) regulation of striated muscle. The αTm isoform undergoes phosphorylation at serine residue 283. While the biochemical and steady-state muscle function of muscle purified Tm phosphorylation have been explored, the effects of Tm phosphorylation on the dynamic properties of muscle contraction and relaxation are unknown. To investigate the kinetic regulatory role of αTm phosphorylation we expressed and purified native N-terminal acetylated Ser-283 wild-type, S283A phosphorylation null and S283D pseudo-phosphorylation Tm mutants in insect cells. Purified Tm's regulate thin filaments similar to that reported for muscle purified Tm. Steady-state Ca(2+) binding to troponin C (TnC) in reconstituted thin filaments did not differ between the 3 Tm's, however disassociation of Ca(2+) from filaments containing pseudo-phosphorylated Tm was slowed compared to wild-type Tm. Replacement of pseudo-phosphorylated Tm into myofibrils similarly prolonged the slow phase of relaxation and decreased the rate of the fast phase without altering activation kinetics. These data demonstrate that Tm pseudo-phosphorylation slows deactivation of the thin filament and muscle force relaxation dynamics in the absence of dynamic and steady-state effects on muscle activation. This supports a role for Tm as a key protein in the regulation of muscle relaxation dynamics.


Anthropometric and physical performance profiles of elite karate kumite and kata competitors.
KARATE TOURNAMENTS CONSIST OF TWO EQUALLY IMPORTANT KARATE DISCIPLINES: the kumite and kata competitions. Due to being based both on the distinctive selection of movement techniques and their kinematic and kinetic patterns, we hypothesized that the elite kumite and kata competitors could differ regarding their anthropometric and physical performance profiles. Thirty-one senior male karate competitors of the national karate team (kumite n = 19; kata n = 12) participated in this study. The tests applied included both the assessment of anthropometric (body height, mass and body mass index) and the following physical performance measurements: the adductor and hamstring flexibility (sideward leg splits test), speed and acceleration (20-m sprint test with 10-m acceleration time), explosive power (countermovement and standing triple jump), agility ("T"- test) and aerobic endurance (20-m multistage shuttle run test). The kumite competitors revealed a larger body size through body height (p = 0.01) and mass (p = 0.03), while the differences in body composition were non-significant. The kumite competitors also demonstrated higher acceleration (p = 0.03) and explosive power (standing triple jump; p = 0.03). A 6-7° higher flexibility of the kata competitors remained somewhat below the level of significance (p = 0.09). The findings could be interpreted by the distinctive differences in the movement techniques. Specifically, a higher explosive power could be beneficial for kumite, while both a smaller stature and higher flexibility (particularly of the lower extremity) could be important for the exceptionally low postures of the kata competitors. Although further elucidation is apparently needed, the obtained finding could be of importance for both the early selection and training of karate competitors.


Intoxications par l'aldicarbe : revue des cas survenus dans le nord de la france entre 1998 et 2001.
Among the 2 726 cases of human pesticide poisoning collected at the Poison Centre of Lille from January 1998 to May 2001, 39 cases were related to the ingestion of aldicarb. Analysis of the circumstances found a suicide attempts in 33 cases and an accidental ingestion in 6 cases. The sex-ratio was 31 men for 8 women, mean age was of 36,6 years (15 months - 77 years). Thirty one poisoning were symptomatic with muscarinic signs (20 cases), digestive (15 cases), neurological ( 8 cases), nicotinic signs (6 cases). Treatment was based on digestive evacuation (14 cases), administration of activated charcoal (14 cases), atropine (7 cases), pralidoxime (1 case), preservative vital functions by intubation and ventilation (7 cases). Sedation was necessary in 4 cases. Hospitalization was necessary in 34 cases. The Poison Severity Score was estimated at 0 (any gravity) in 5 cases, 1 (weak) in 12 cases, 2 (moderate) in 7 cases, 3 (severe) in 8 cases and 4 (lethal) in 2 cases. In one of deaths, aldicarb was determined by HPLC-DAD in blood (6,04 μg / ml), urines (1,88 μg / ml) and gastric contents (3,98 μg / ml). These concentrations are the most important ever described in the litterature. Aldicarb is the most toxic carbamate insecticide for human.


Murine caspase-11, an ICE-interacting protease, is essential for the activation of ICE.
We report here the inactivation of a member of the Ice/Ced-3 (caspase) family of cell death genes, casp-11, by gene targeting. Like Ice-deficient mice, casp-11 mutant mice are resistant to endotoxic shock induced by lipopolysaccharide. Production of both IL-1alpha and IL-1beta after lipopolysaccharide stimulation, a crucial event during septic shock and an indication of ICE activation, is blocked in casp-11 mutant mice. casp-11 mutant embryonic fibroblast cells are resistant to apoptosis induced by overexpression of ICE. Furthermore, we found that pro-caspase-11 physically interacts with pro-ICE in cells, and the expression of casp-11 is essential for activation of ICE. Our data suggest that caspase-11 is a component of ICE complex and is required for the activation of ICE.


The role of polyphenols in modern nutrition.
Polyphenols are found in plant-based foods and beverages, notably apples, berries, citrus fruit, plums, broccoli, cocoa, tea and coffee and many others. There is substantial epidemiological evidence that a diet high in polyphenol-rich fruit, vegetables, cocoa and beverages protects against developing cardiovascular disease and type 2 diabetes. The absorption and metabolism of these compounds have been well described and, for many, the gut microbiota play a critical role in absorption; taking into consideration the parent compound and the metabolites from colon bacteria catabolism, more than 80% of a dose can be absorbed and ultimately excreted in the urine. Common polyphenols in the diet are flavanols (cocoa, tea, apples, broad beans), flavanones (hesperidin in citrus fruit), hydroxycinnamates (coffee, many fruits), flavonols (quercetin in onions, apples and tea) and anthocyanins (berries). Many intervention studies, mechanistic in vitro data and epidemiological studies support a role for polyphenols against the development of chronic diseases. For example, flavanols decrease endothelial dysfunction, lower blood pressure and cholesterol, and modulate energy metabolism. Coffee and tea both reduce the risk of developing type 2 diabetes, through action of their constituent polyphenols. Despite extensive research, the exact mechanisms of action of polyphenols in the human body have not been decisively proven, but there is strong evidence that some targets such as nitric oxide metabolism, carbohydrate digestion and oxidative enzymes are important for health benefits. Consumption of polyphenols as healthy dietary components is consistent with the advice to eat five or more portions of fruit and vegetables per day, but it is currently difficult to recommend what 'doses' of specific polyphenols should be consumed to derive maximum benefit.


Determination of lead in fine particulates by slurry sampling electrothermal atomic absorption spectrometry.
A simple method for determining lead in fine particulates (PM2.5) by using electrothermal atomic absorption spectrometry (ETAAS) has been developed. Particulates collected on Nuclepore filter by using a dichotomous sampler were suspended in diluted nitric acid after ultrasonic agitation. The dislodging efficiency is nearly 100% after agitation for 5 min. In order to study the suspension behavior of PM2.5 in solvents, a Brookhaven ZetaPlus Particle Size Analyzer was used to determine the particle size distribution and suspension behavior of air particulates in the solvent. The pre-digestion and modification effect of nitric acid would be discussed. Palladium was added as a chemical modifier and the temperature program of ETAAS was changed in order to improve the recovery. The slurry was introduced directly into a graphite tube for atomization. The metal content in the sample was determined by the standard addition method. In addition, a conventional acid digestion procedure was applied to verify the efficiency of the slurry sampling method. It offers a quick and efficient alternative method for heavy metal characterization in fine particulates.


Veillonella seminalis sp. nov., a novel anaerobic Gram-stain-negative coccus from human clinical samples, and emended description of the genus Veillonella.
Ten isolates of unknown, Gram-stain-negative, anaerobic cocci were recovered from human clinical samples, mainly from semen. On the basis of their phenotypic features, including morphology, main metabolic end products, gas production, nitrate reduction and decarboxylation of succinate, the strains were identified as members of the genus Veillonella. Multi-locus sequence analysis and corresponding phylogenies were based on 16S rRNA, dnaK and rpoB genes, and on the newly proposed gltA gene. The strains shared high levels of genetic sequence similarity and were related most closely to Veillonella ratti. The strains could not be differentiated from V. ratti on the basis of 16S rRNA gene sequence analysis while gltA, rpoB and dnaK gene sequences showed 85.1, 93.5 and 90.2% similarity with those of the type strain of V. ratti, respectively. Phylogenetic analyses revealed that the isolates formed a robust clade in the V. ratti-Veillonella criceti-Veillonella magna subgroup of the genus Veillonella. As observed for V. criceti, the isolates were able to ferment fructose. In contrast to other members of the genus Veillonella, the 10 strains were not able to metabolize lactate. Cellular fatty acid composition was consistent with that of other species of the genus Veillonella. From these data, the 10 isolates are considered to belong to a novel species in the genus Veillonella, for which the name Veillonella seminalis sp. nov. is proposed. The type strain is ADV 4313.2(T) ( = CIP 107810(T) = LMG 28162(T)). Veillonella strain ACS-216-V-Col6b subjected to whole genome sequencing as part as the Human Microbiome Project is another representative of V. seminalis sp. nov. An emended description of the genus Veillonella is also proposed.


Effect of intestinal microbiota on exercise performance in mice.
The antioxidant enzyme system helps protect against intense exercise-induced oxidative damage and is related to the physical status of athletes. Evidence suggests that intestinal microbiota may be an important environmental factor associated with host metabolism, physiology, and antioxidant endogenous defense. However, evidence of the effect of gut microbiota status on exercise performance and physical fatigue is limited. We investigated the association of intestinal bacteria and exercise performance in specific pathogen-free (SPF), germ-free (GF), and Bacteroides fragilis (BF) gnotobiotic mice. Endurance swimming time was longer for SPF and BF than GF mice, and the weight of liver, muscle, brown adipose, and epididymal fat pads was higher for SPF and BF than GF mice. The serum levels of glutathione peroxidase (GPx) and catalase were greater in SPF than GF mice. Serum superoxide dismutase activity was lower in BF than SPF and GF mice. In addition, hepatic GPx level was higher in SPF than GF and BF mice. Gut microbial status could be crucial for exercise performance and its potential action linked with the antioxidant enzyme system in athletes.