Proteoglycan interactions with Sonic Hedgehog specify mitogenic responses.
Sonic Hedgehog (Shh) has dual roles in vertebrate development, promoting progenitor cell proliferation and inducing tissue patterning. We found that the mitogenic and patterning functions of Shh can be uncoupled from one another. Using a genetic approach to selectively inhibit Shh-proteoglycan interactions in a mouse model, we found that binding of Shh to proteoglycans was required for proliferation of neural stem/precursor cells, but not for tissue patterning. Shh-proteoglycan interactions regulated both spatial and temporal features of Shh signaling. Proteoglycans localized Shh to specialized mitogenic niches and also acted at the single-cell level to regulate the duration of Shh signaling, thereby promoting a gene expression program that is important for cell division. Because activation of the Shh pathway is a feature of diverse human cancers, selective stimulation of proliferation by Shh-proteoglycan interactions may also figure prominently in neoplastic growth.


Recommendations for follow-up of colorectal cancer survivors.
Colorectal cancer (CRC) is one of the tumours with the highest incidence and mortality in the Spanish population. Nevertheless, the advances in prevention and treatment have contributed to an increased number of patients who survive for prolonged periods of time. In addition, despite recurrences, improved survival following metastasis resection is likewise on the rise. This underscores the importance of carrying out follow-up programmes even in low-risk patients for the early detection of recurrence. The main objective of this article is to provide a set of recommendations for optimising the follow-up of CRC survivors as well as for managing the sequelae that result from either pharmacological or surgical treatment.


Fetal programming of autonomic and HPA function: do people who were small babies have enhanced stress responses?
Studies in several species have demonstrated that an adverse early environment can influence the development of the autonomic nervous system and hypothalamic-pituitary-adrenal (HPA) axis. The autonomic nervous system and HPA axis are key components of the neuroendocrine response to stress and many of these animal models show altered biological responses to stress. Recent research now suggests that these processes operate in humans. An adverse early environment, as evidenced by reduced birth or infant weight, is associated with enhanced autonomic and HPA responses to experimental psychological stress. However, there appear to be marked sex differences in the mechanisms involved. Epidemiological studies demonstrate that physiological changes in these neuroendocrine systems may predispose to cardiovascular disease through their influence on risk factors such as plasma glucose and lipid concentrations and blood pressure. Thus the combination of enhanced stress susceptibility and the psychosocial stressors to which people are exposed may be an important component of the disease risk in human populations.


Improvement after cerebrospinal fluid drainage is related to levels of N-acetyl-aspartate in idiopathic normal pressure hydrocephalus.
This study uses proton magnetic resonance spectroscopy to investigate whether or not idiopathic normal pressure hydrocephalus is associated with neuronal dysfunction or ischemia in the brain. We evaluate whether or not proton magnetic resonance spectroscopy is useful for predicting improvement after long-term external lumbar drainage (ELD) of cerebrospinal fluid. Eighteen patients (mean age, 73 yr; six women) and 10 matching controls participated. Participants were characterized by clinical features, cognitive and motor function tests, and cerebrospinal fluid hydrodynamics (patients only). Signals from N-acetyl-aspartate (NAA), choline, lactate, and creatine (Cr) (reference) were sampled once in controls and twice in patients (before and after a 3-day ELD of approximately 135 mL/24 h) by proton magnetic resonance spectroscopy (1.5 T) from a 7.2-mL volume in the frontal white matter. Improvement was defined by video recordings of the patients' gait. Sixteen patients finished the ELD (one patient had meningitis, and one patient had catheter insertion failure) with a mean drain volume of 395 mL. NAA/Cr ratios were lower in patients than in controls (1.60 versus 1.84, P = 0.02), but no difference was found for choline/Cr ratios. No lactate signals were detected. Fifty percent of patients improved after ELD. They had higher NAA/Cr ratios than nonimproved patients (1.70 versus 1.51, P = 0.01), but no differences were found in choline/Cr ratios or drain volume. NAA/Cr ratios were decreased in patients with idiopathic normal pressure hydrocephalus, which is consistent with neuronal dysfunction in the frontal white matter. Improved patients had NAA/Cr ratios close to normal, indicating that enough functional neurons are a prerequisite for the cerebrospinal fluid drainage to have an effect.


Bral1, a brain-specific link protein, colocalizing with the versican V2 isoform at the nodes of Ranvier in developing and adult mouse central nervous systems.
Bral1, a brain-specific hyaluronan-binding protein, has been cloned recently. To gain insight into the role of Bral1, we generated a specific antibody against this protein. We have examined the detailed localization pattern of Bral1 protein and compared it with that of other members of the lectican proteoglycan family, such as brevican and versican, with which Bral1 is predicted to interact. The immunoreactivity of Bral1 antibody was predominantly observed in myelinated fiber tracts in the adult brain and could be detected at P20 in the white matter of the developing cerebellum, suggesting that expression starts when axonal myelination takes place. Furthermore, immunostaining demonstrated that Bral1 colocalized with the versican V2 isoform at the nodes of Ranvier. The present data suggest that Bral1 may play a pivotal role in the formation of the hyaluronan-associated matrix in the CNS that facilitates neuronal conduction by forming an ion diffusion barrier at the nodes.


A systems biology approach uncovers the core gene regulatory network governing iridophore fate choice from the neural crest.
Multipotent neural crest (NC) progenitors generate an astonishing array of derivatives, including neuronal, skeletal components and pigment cells (chromatophores), but the molecular mechanisms allowing balanced selection of each fate remain unknown. In zebrafish, melanocytes, iridophores and xanthophores, the three chromatophore lineages, are thought to share progenitors and so lend themselves to investigating the complex gene regulatory networks (GRNs) underlying fate segregation of NC progenitors. Although the core GRN governing melanocyte specification has been previously established, those guiding iridophore and xanthophore development remain elusive. Here we focus on the iridophore GRN, where mutant phenotypes identify the transcription factors Sox10, Tfec and Mitfa and the receptor tyrosine kinase, Ltk, as key players. Here we present expression data, as well as loss and gain of function results, guiding the derivation of an initial iridophore specification GRN. Moreover, we use an iterative process of mathematical modelling, supplemented with a Monte Carlo screening algorithm suited to the qualitative nature of the experimental data, to allow for rigorous predictive exploration of the GRN dynamics. Predictions were experimentally evaluated and testable hypotheses were derived to construct an improved version of the GRN, which we showed produced outputs consistent with experimentally observed gene expression dynamics. Our study reveals multiple important regulatory features, notably a sox10-dependent positive feedback loop between tfec and ltk driving iridophore specification; the molecular basis of sox10 maintenance throughout iridophore development; and the cooperation between sox10 and tfec in driving expression of pnp4a, a key differentiation gene. We also assess a candidate repressor of mitfa, a melanocyte-specific target of sox10. Surprisingly, our data challenge the reported role of Foxd3, an established mitfa repressor, in iridophore regulation. Our study builds upon our previous systems biology approach, by incorporating physiologically-relevant parameter values and rigorous evaluation of parameter values within a qualitative data framework, to establish for the first time the core GRN guiding specification of the iridophore lineage.


Community-based approaches to prevention and management of hypertension and cardiovascular disease.
Community hypertension (HTN) outreach seeks to improve public health by identifying HTN and cardiovascular disease (CVD) risks. In the 1980s, the National Heart, Lung, and Blood Institute (NHLBI) funded multiple positive community studies. Additionally, the Centers for Disease Control and Prevention's (CDC's) Racial and Ethnic Approaches to Community Health (REACH) program addresses CVD risks. In 1978, in Baltimore, MD, the Association of Black Cardiologists (ABC), organized barbershops and churches as HTN control centers, as in New Orleans, LA, since 1993, the Healthy Heart Community Prevention Project (HHCPP). Also, the NHLBI Community Health Workers and Promotores de Salud are beneficial. The American Society of Hypertension (ASH) Hypertension Community Outreach program provides free HTN and CVD screenings, digital BP monitors, multilingual and literacy-appropriate information, and videos. Contemporary major federal programs, such as the Million Hearts Initiative, are ongoing. Overall, the evidence-based Logic Model should enhance planning, implementation, and dissemination.


Association of miR-146a, miR-149, miR-196a2, and miR-499 Polymorphisms with Ossification of the Posterior Longitudinal Ligament of the Cervical Spine.
Ossification of the posterior longitudinal ligament (OPLL) of the spine is considered a multifactorial and polygenic disease. We aimed to investigate the association between four single nucleotide polymorphisms (SNPs) of pre-miRNAs [miR-146aC>G (rs2910164), miR-149T>C (rs2292832), miR-196a2T>C (rs11614913), and miR-499A>G (rs3746444)] and the risk of cervical OPLL in the Korean population. The genotypic frequencies of these four SNPs were analyzed in 207 OPLL patients and 200 controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. For four SNPs in pre-miRNAs, no significant differences were found between OPLL patients and controls. However, subgroup analysis based on OPLL subgroup (continuous: continuous type plus mixed type, segmental: segmental and localized type) showed that miR-499GG genotype was associated with an increased risk of segmental type OPLL (adjusted odds ratio = 4.314 with 95% confidence interval: 1.109-16.78). In addition, some allele combinations (C-T-T-G, G-T-T-A, and G-T-C-G of miR-146a/-149/-196a2/-499) and combined genotypes (miR-149TC/miR-196a2TT) were associated with increased OPLL risk, whereas the G-T-T-G and G-C-C-G allele combinations were associated with decreased OPLL risk. The results indicate that GG genotype of miR-499 is associated with significantly higher risks of OPLL in the segmental OPLL group. The miR-146a/-149/-196a2/-499 allele combinations may be a genetic risk factor for cervical OPLL in the Korean population.


Effects of Montmorency Tart Cherry and Blueberry Juice on Cardiometabolic and Other Health-Related Outcomes: A Three-Arm Placebo Randomized Controlled Trial.
The current study aimed to investigate the influence of tart cherry and blueberry juices on cardiometabolic and other health indices following a 20-day supplementation period. Forty-five adults were randomly assigned to receive tart cherry, blueberry, or a placebo, of which they drank 60 mL per day for 20 days. The primary outcome, which was systolic blood pressure, and secondary measures, including anthropometric, energy expenditure, substrate oxidation, hematological, diastolic blood pressure/resting heart rate, psychological wellbeing, and sleep efficacy, were measured before and after the intervention. There were no statistically significant differences (p > 0.05) for systolic blood pressure; however, total and LDL cholesterol were significantly improved with blueberry intake (pre: total cholesterol = 4.36 mmol/L and LDL cholesterol = 2.71 mmol/L; post: total cholesterol = 3.79 mmol/L and LDL cholesterol = 2.23 mmol/L) compared to placebo (pre: total cholesterol = 4.01 mmol/L and LDL cholesterol = 2.45 mmol/L; post: total cholesterol = 4.34 mmol/L and LDL cholesterol = 2.67 mmol/L). Furthermore, psychological wellbeing indices measured using the Beck Depression Inventory, State Trait Anxiety Inventory, and COOP WONCA improved statistically in the blueberry arm compared to placebo. Given the clear association between lipid concentrations and the risk of cardiovascular disease as well as the importance of psychological wellbeing to health-related quality of life, this investigation indicates that it could be an effective approach to assist in managing cardiometabolic disease.


Factors that influence bidirectional long-tract homozygosis due to double-strand break repair in Candida albicans.
Genomic rearrangements have been associated with the acquisition of adaptive phenotypes, allowing organisms to efficiently generate new favorable genetic combinations. The diploid genome of Candida albicans is highly plastic, displaying numerous genomic rearrangements that are often the by-product of the repair of DNA breaks. For example, DNA double-strand breaks (DSB) repair using homologous-recombination pathways are a major source of loss-of-heterozygosity (LOH), observed ubiquitously in both clinical and laboratory strains of C. albicans. Mechanisms such as break-induced replication (BIR) or mitotic crossover (MCO) can result in long tracts of LOH, spanning hundreds of kilobases until the telomere. Analysis of I-SceI-induced BIR/MCO tracts in C. albicans revealed that the homozygosis tracts can ascend several kilobases toward the centromere, displaying homozygosis from the break site toward the centromere. We sought to investigate the molecular mechanisms that could contribute to this phenotype by characterizing a series of C. albicans DNA repair mutants, including pol32-/-, msh2-/-, mph1-/-, and mus81-/-. The impact of deleting these genes on genome stability revealed functional differences between Saccharomyces cerevisiae (a model DNA repair organism) and C. albicans. In addition, we demonstrated that ascending LOH tracts toward the centromere are associated with intrinsic features of BIR and potentially involve the mismatch repair pathway which acts upon natural heterozygous positions. Overall, this mechanistic approach to study LOH deepens our limited characterization of DNA repair pathways in C. albicans and brings forth the notion that centromere proximal alleles from DNA break sites are not guarded from undergoing LOH.


Extrathymic development of V alpha 14-positive T cells.
It is known that rearrangement of the T cell antigen receptor (TCR) gene occurs in the thymus during T cell development and consequently results both in the deletion of DNA between the variable (V) and diversity/joining segments and in the formation of a circular DNA with recombination signal sequences. Here, we provide evidence that V alpha 14+ TCR gene rearrangements take place in extrathymic sites, such as bone marrow, liver, and intestine, but not in spleen, because we were able to detect frequent productive and nonproductive V alpha 14+ coding and signal sequences as a result of TCR rearrangements in extrathymic sites. Similar findings were also detected in athymic mice. Quantitative analysis shows that the relative amounts of V alpha 14 gene-mediated signal sequences in extrathymic tissues are higher than those in thymus. On the contrary, TCR rearrangements of V alpha 1.1 T cells, which are known to develop in the thymus, were mainly detected in the thymus, Peyer's patch, and spleen, but not in other extrathymic tissues, showing patterns distinct from V alpha 14 TCR rearrangements. These findings are evidence of extrathymic development of V alpha 14+ T cells. Differential characteristic TCR rearrangement patterns also indicate that distinct TCR repertoires are generated in different lymphoid tissues.


Male students' perceptions about gender imbalances in a speech-language pathology and audiology training programme of a South African institution of higher education.
The professions of speech-language pathology and audiology (SLPA) are characterised by occupational gender segregation. Reasons given by men are a lack of awareness of SLPA; a perception of poor salaries; a perception of poor working conditions; a stereotype that the profession is a female occupation; and a perception that working with children is feminine. Objectives: The aim of this study was to describe the perceptions of male students in a specific SLPA training programme regarding gender imbalances. The objectives were to describe the influences of gender on the career orientation of students, the factors shaping students' career choices and the experiences of students during their undergraduate training. Method: This study followed a quantitative descriptive survey design. Thirty-one conveniently sampled male SLPA students, across different years of study, completed a self-administered questionnaire. Results: Participants indicated that they did not believe in the gendered nature of occupations and that SLPA are not 'female only' professions. They (65%) recorded that they considered salary and career prospects for choosing a career, and 74% specified that they chose SLPA because of a desire to help people. The perception of male students regarding their training appears to be positive as participants are comfortable with the clinical (71%) and learning (77%) activities. However, barriers were reported and some lecture information may be more obvious or applicable to female students. A need was identified to address the issue of working alone with paediatric clients and pertains to the societal awareness regarding sexual harassment of children, which may impact spontaneity in engagements. Conclusion: The recruitment of male students should be prioritised to better reflect the client population served by the professions. Recommendations include career counselling in rural schools, recruitment campaigns to identify interested students, public awareness regarding the professions and guidance regarding working with paediatric clients.


TRANSPORTATION BARRIERS TO ACCESS HEALTH CARE FOR SURGICAL CONDITIONS IN MALAWI a cross sectional nationwide household survey.
It is estimated that nearly five billion people worldwide do not have access to safe surgery. This access gap disproportionately affects low-and middle-income countries (LMICs). One of the barriers to healthcare in LMICs is access to transport to a healthcare facility. Both availability and affordability of transport can be issues delaying access to health care. This study aimed to describe the main transportation factors affecting access and delay in reaching a facility for health care in Malawi. This was a multi-stage, clustered, probability sampling with systematic sampling of households for transportation access to general health and surgical care. Malawi has an estimated population of nearly 18 million people, with a total of 48,233 registered settlements spread over 28 administrative districts. 55 settlements per district were randomly selected for data collection, and 2-4 households were selected, depending on the size. Two persons per household were interviewed. The Surgeons Overseas Assessment of Surgical need (SOSAS) tool was used by trained personnel to collect data during the months of July and August 2016. Analysis of data from 1479 households and 2958 interviewees was by univariate and multivariate methods. Analysis showed that 90.1% were rural inhabitants, and 40% were farmers. No formal employment was reported for 24.9% persons. Animal drawn carts prevailed as the most common mode of transport from home to the primary health facility - normally a health centre. Travel to secondary and tertiary level health facilities was mostly by public transport, 31.5 and 43.4% respectively. Median travel time from home to a health centre was 1 h, and 2.5 h to a central hospital. Thirty nine percent of male and 59% of female head of households reported lack financial resources to go to a hospital. In Malawi, lack of suitable transport, finances and prolonged travel time to a health care centre, all pose barriers to timely access of health care. Improving the availability of transport between rural health centres and district hospitals, and between the district and central hospitals, could help overcome the transportation barriers to health care.


Using informant reports to detect cognitive decline in mild cognitive impairment.
The use of informant rating scales in older adults at risk of dementia may assist with early detection and intervention strategies. This study aims to evaluate whether informants rate greater cognitive change in patients with mild cognitive impairment (MCI) compared to cognitively intact individuals, and to determine the relationship between informant ratings of cognitive change and neuropsychological performance. One hundred and nine health-seeking older adults underwent clinical and neuropsychological assessments, and informants completed the Cambridge Behavioral Inventory-Revised (CBI-R). Patients were rated according to MCI criteria, including amnestic and non-amnestic subtypes, or as being cognitively intact. CBI-R ratings were evaluated with respect to MCI diagnosis and neuropsychological performance. Compared to cognitively intact individuals, informants rated patients with MCI as having significantly more change in overall functioning (p < 0.05) as well as in specific domains of memory and orientation (p < 0.01), everyday skills (p < 0.05), and motivation (p < 0.05), even after controlling for depressive symptom severity. In further analyses, the non-amnestic MCI subgroup only had more informant-rated mood changes compared to the amnestic subgroup. In relation to neuropsychological performance, informant ratings were related to poorer visual memory, verbal learning and memory, language, and psychomotor speed, with correlations ranging from -0.19 to -0.43 (p < 0.05). These findings indicate that informants are sensitive to subtle early cognitive change in individuals with MCI, and that their ratings are related to objectively measured neuropsychological performance. Thus, the CBI-R may be valuable in assisting early screening and intervention processes.


Altered surfactant homeostasis and alveolar type II cell morphology in mice lacking surfactant protein D.
Surfactant protein D (SP-D) is one of two collectins found in the pulmonary alveolus. On the basis of homology with other collectins, potential functions for SP-D include roles in innate immunity and surfactant metabolism. The SP-D gene was disrupted in embryonic stem cells by homologous recombination to generate mice deficient in SP-D. Mice heterozygous for the mutant SP-D allele had SP-D concentrations that were approximately 50% wild type but no other obvious phenotypic abnormality. Mice totally deficient in SP-D were healthy to 7 months but had a progressive accumulation of surfactant lipids, SP-A, and SP-B in the alveolar space. By 8 weeks the alveolar phospholipid pool was 8-fold higher than wild-type littermates. There was also a 10-fold accumulation of alveolar macrophages in the null mice, and many macrophages were both multinucleated and foamy in appearance. Type II cells in the null mice were hyperplastic and contained giant lamellar bodies. These alterations in surfactant homeostasis were not associated with detectable changes in surfactant surface activity, postnatal respiratory function, or survival. The findings in the SP-D-deficient mice suggest a role for SP-D in surfactant homeostasis.


Inferring Multidimensional Rates of Aging from Cross-Sectional Data.
Modeling how individuals evolve over time is a fundamental problem in the natural and social sciences. However, existing datasets are often cross-sectional with each individual observed only once, making it impossible to apply traditional time-series methods. Motivated by the study of human aging, we present an interpretable latent-variable model that learns temporal dynamics from cross-sectional data. Our model represents each individual's features over time as a nonlinear function of a low-dimensional, linearly-evolving latent state. We prove that when this nonlinear function is constrained to be order-isomorphic, the model family is identifiable solely from cross-sectional data provided the distribution of time-independent variation is known. On the UK Biobank human health dataset, our model reconstructs the observed data while learning interpretable rates of aging associated with diseases, mortality, and aging risk factors.


Space-Selective Fabrication of Light-Emitting Carbon Dots in Polymer Films Using Electron-Beam-Induced Chemical Reactions.
Nanocarbon-based materials have excellent properties, including high electrical conductivity as well as charity-dependent optical absorption and luminescence; therefore, the materials are promising in applications for nanoelectric devices, nanophotonics, and so on. Carbon dots (CDs) are one of the carbon materials recently fabricated. Optical properties of CDs have been reported to be similar to those of polycyclic aromatic hydrocarbons (PAHs). For this reason, the CDs are considered to be composed of PAH. Synthesis of CDs has previously been accomplished through hydrothermal synthesis and microwave irradiation. These methods require a long synthesis time, and the processes involve multiple steps. In this study, we developed a fabrication method of CDs in simple and spatially selective ways, by using radical reactions in an organic polymer film with focused electron-beam irradiation. We investigated various organic polymers as reaction materials and found that polystyrene has a higher efficiency for CD formation than other organic polymers investigated. Absorption, photoluminescence, and Raman scattering properties of the electron-beam-irradiated sample were in good agreement with those reported for the CDs. The technique developed in this study is promising for fabricating light-emitting CDs and photonic crystals in a simple and flexible manner.


Saccharification of biomass using whole solid-state fermentation medium to avoid additional separation steps.
The enzymatic hydrolysis of steam-exploded sugarcane bagasse (SESB) was investigated using enzymatic extracts (EE) and whole fermentation media (WM), produced in-house, from Aspergillus niger 3T5B8 and Trichoderma reesei Rut-C30 cultivated on wheat bran under solid-state fermentation (SSF). A detailed and quantitative comparison of the different hydrolysis conditions tested was carried out using the Chrastil approach for modeling enzymatic reactions by fitting the experimental data of total reducing sugar (TRS) released according to hydrolysis time. Conversion of SESB using A. niger enzymatic complex were up to 3.2-fold higher (in terms of TRS) than T. reesei at similar enzyme loadings, which could be correlated to the higher β-glucosidase levels (up to 35-fold higher) of A. niger enzymatic complex. Conversion yields after 72 h exceeded 40% in terms of TRS when the WM was supplemented with a low dosage of a commercial enzyme preparation. When the combination of WM (from either T. reesei or A. niger) and commercial cellulase was used, the dosage of the commercial enzyme could be reduced by half, while still providing a hydrolysis that was up to 36% more efficient. Furthermore, SESB hydrolysis using either EE or WM resulted in similar yields, indicating that the enzyme extraction/filtration steps could be eliminated from the overall process. This procedure is highly advantageous in terms of reduced enzyme and process costs, and also avoids the generation of unnecessary effluent streams. Thus, the enzymatic conversion of SESB using the WM from SSF is cost-effective and compatible with the biorefinery concept.


Umbilical cord blood banking: an update.
Umbilical cord blood is a potential vast source of primitive hematopoietic stem and progenitor cells available for clinical application to reconstitute the hematopoietic system and/or restore immunological function in affected individuals requiring treatment. Cord blood can be used as an alternative source for bone marrow transplantation and its use is developing into a new field of treatment for pediatric and adult patients presenting with hematological disorders, immunological defects and specific genetic diseases. More than 25,000 allogeneic cord blood transplantations have been performed worldwide since the first cord blood transplantation in 1988. There are two banking options for storing umbilical cord blood [private (family) and public]. Cord blood stored in private banks are used for either autologous or allogeneic transplants for the infant donor or related family members but private cord blood banks are not searchable or available to the public. More than 780,000 cord blood units are stored in over 130 private cord blood banks, worldwide, and over 400,000 units in more than 100 quality controlled public cord blood banks. Researchers continue to evaluate the usefulness of cord blood cells in treating human diseases or disorders for purposes other than hematological disorders including heart disease, strokes, brain or spinal cord injuries and cancer. This review summarizes the status of umbilical cord blood banking, its history and current and potential use in the treatment of human disease.


The anthropocene: from global change to planetary stewardship.
Over the past century, the total material wealth of humanity has been enhanced. However, in the twenty-first century, we face scarcity in critical resources, the degradation of ecosystem services, and the erosion of the planet's capability to absorb our wastes. Equity issues remain stubbornly difficult to solve. This situation is novel in its speed, its global scale and its threat to the resilience of the Earth System. The advent of the Anthropence, the time interval in which human activities now rival global geophysical processes, suggests that we need to fundamentally alter our relationship with the planet we inhabit. Many approaches could be adopted, ranging from geoengineering solutions that purposefully manipulate parts of the Earth System to becoming active stewards of our own life support system. The Anthropocene is a reminder that the Holocene, during which complex human societies have developed, has been a stable, accommodating environment and is the only state of the Earth System that we know for sure can support contemporary society. The need to achieve effective planetary stewardship is urgent. As we go further into the Anthropocene, we risk driving the Earth System onto a trajectory toward more hostile states from which we cannot easily return.


NSCLC molecular testing in Central and Eastern European countries.
The introduction of targeted treatments for subsets of non-small cell lung cancer (NSCLC) has highlighted the importance of accurate molecular diagnosis to determine if an actionable genetic alteration is present. Few data are available for Central and Eastern Europe (CEE) on mutation rates, testing rates, and compliance with testing guidelines. A questionnaire about molecular testing and NSCLC management was distributed to relevant specialists in nine CEE countries, and pathologists were asked to provide the results of EGFR and ALK testing over a 1-year period. A very high proportion of lung cancer cases are confirmed histologically/cytologically (75-100%), and molecular testing of NSCLC samples has been established in all evaluated CEE countries in 2014. Most countries follow national or international guidelines on which patients to test for EGFR mutations and ALK rearrangements. In most centers at that time, testing was undertaken on request of the clinician rather than on the preferred reflex basis. Immunohistochemistry, followed by fluorescent in situ hybridization confirmation of positive cases, has been widely adopted for ALK testing in the region. Limited reimbursement is a significant barrier to molecular testing in the region and a disincentive to reflex testing. Multidisciplinary tumor boards are established in most of the countries and centers, with 75-100% of cases being discussed at a multidisciplinary tumor board at specialized centers. Molecular testing is established throughout the CEE region, but improved and unbiased reimbursement remains a major challenge for the future. Increasing the number of patients reviewed by multidisciplinary boards outside of major centers and access to targeted therapy based on the result of molecular testing are other major challenges.


Differential regulation of embryonic and adult β cell replication.
Diabetes results from an inadequate functional β cell mass, either due to autoimmune destruction (Type 1 diabetes) or insulin resistance combined with β cell failure (Type 2 diabetes). Strategies to enhance β cell regeneration or increase cell proliferation could improve outcomes for patients with diabetes. Research conducted over the past several years has revealed that factors regulating embryonic β cell mass expansion differ from those regulating replication of β cells post-weaning. This article aims to compare and contrast factors known to control embryonic and postnatal β cell replication. In addition, we explore the possibility that connective tissue growth factor (CTGF) could increase adult β cell replication. We have already shown that CTGF is required for embryonic β cell proliferation and is sufficient to induce replication of embryonic β cells. Here we examine whether adult β cell replication and expansion of β cell mass can be enhanced by increased CTGF expression in mature β cells.


Inflammatory cytokine levels and breast cancer risk factors: racial differences of healthy Caucasian and African American women.
To examine racial differences in inflammatory cytokine levels (interleukin [IL]-6 and interferon-gamma [IFN-γ]) and breast cancer (BC) risk factors between healthy Caucasian and African American women; to examine differences in relationships of inflammatory cytokine levels with BC risk factors between these groups of women; and to determine the independent contribution of race to IL-6 and IFN-γ after controlling for relevant covariates. Cross-sectional and correlational descriptive design. Community surrounding a state university health system in the southeastern United States. 113 healthy women (65 Caucasians and 48 African Americans) aged 20 years or older and not pregnant. Secondary analysis of data collected from self-report questionnaires and blood samples. Inflammatory cytokine levels, BC risk factors (age, age at menarche, age at first live birth, family history of BC, breast biopsy, breastfeeding history and duration, body mass index, and physical activity), and race. Significant racial differences were noted in IL-6 and IFN-γ levels, reproductive or hormonal and lifestyle BC risk factors, and relationships between African American and Caucasian women. Controlling for all other effects, race appeared to be a significant predictor for IL-6 and IFN-γ. Racial differences in inflammatory cytokines and BC risk factors may provide partial evidence for existing racial disparities in BC for African American and Caucasian women. Additional studies are needed to confirm that potential. Additional biobehavioral research in racial disparities in BC may help to inform nurses to target race-specific modifications of lifestyle and behavioral factors to reduce BC health disparity between African American and Caucasian women. Being an African American woman predicted a higher level of inflammatory cytokine production after controlling for selected BC risk factors. Great potential exists for inflammatory responses as one of the underlying biologic mechanisms for existing BC disparity and for culturally tailored lifestyle or behavioral modification interventions for reducing BC risk and racial disparity.


Effect of Pseudomonas aeruginosa elastase B on level and activity of immune proteins/peptides of Galleria mellonella hemolymph.
Abstract Susceptibility of proteins and peptides present in immune hemolymph of Galleria mellonella Fabricius (Lepidoptera: Pyralidae) larvae to proteolytic degradation by purified elastase B of Pseudomonas aeruginosa was studied. Results showed that apoLp-III protein was gradually digested by elastase B in vitro. Additionally, polipeptides with molecular mass 6.5 and 4 kDa were degraded after treatment with the studied enzyme. The lack of these peptides and the decrease in anti-Escherichia coli activity could indicate that inducible antimicrobial peptides were digested by elastase B. On the contrary, no change in the lysosome activity level was observed in immune hemolymph incubated with elastase B. Thus, elastase B might contribute to the pathogenesis of P. aeruginosa.


Changing climate change: The carbon budget and the modifying-work of the IPCC.
Over the last 10 years, the concept of a global 'carbon budget' of allowable CO2 emissions has become ubiquitous in climate science and policy. Since it was brought to prominence by the Fifth Assessment Report of the IPCC, the carbon budget has changed how climate change is enacted as an issue of public concern, from determining the optimal rate of future emissions to establishing a fixed limit for how much emissions should be allowed before they must be stopped altogether. Exploring the emergence of the carbon budget concept, this article shows how the assessment process of the IPCC has offered scientific experts the means to modify how the climate issue is problematized, and discusses the implications of this 'modifying-work' for the politics of climate change. It finds that the 'modified climate issue' must be seen as an outcome of the ordinary work within established scientific and political institutions, and the agency these institutions afford scientists to enact the issue differently. On this basis, it argues that the case of the carbon budget holds important insights not only for the relationship between climate science and policy, but also for the pragmatist literature on 'issue formation' in STS.


Retention of total carotenoid and β-carotene in yellow sweet cassava (Manihot esculenta Crantz) after domestic cooking.
Over the last decade, considerable efforts have been made to identify cassava cultivars to improve the vitamin A nutritional status of undernourished populations, especially in northeast Brazil, where cassava is one of the principal and essentially only nutritional source. The aim of this study was to evaluate the total carotenoid, β-carotene, and its all-E-, 9-, and 13-Z-β-carotene isomers content in seven yellow sweet cassava roots and their retention after three boiling cooking methods. The total carotenoid, β-carotene, and its all-E-, 9-, and 13-Z-β-carotene isomers in yellow sweet cassava samples were determined by ultraviolet/visible spectrometry and high-performance liquid chromatography, respectively, before and after applying the cooking methods. All analyses were performed in triplicate. The total carotenoid in raw roots varied from 2.64 to 14.15 µg/g and total β-carotene from 1.99 to 10.32 µg/g. The β-carotene predominated in all the roots. The Híbrido 2003 14 08 cultivar presented the highest β-carotene content after cooking methods 1 and 3. The 1153 - Klainasik cultivar presented the highest 9-Z-β-carotene content after cooking by method 3. The highest total carotenoid retention was observed in cultivar 1456 - Vermelhinha and that of β-carotene for the Híbrido 2003 14 11 cultivar, both after cooking method 1. Evaluating the real retention percentage (RR%) in sweet yellow cassava after home cooking methods showed differences that can be attributed to the total initial carotenoid contents. However, no cooking method uniformly provided a higher total carotenoid or β-carotene retention in all the cultivars. Differences were found in the cooking methods among the samples regarding total carotenoid or β-carotene retention, suggesting that the different behaviors of the cultivars need to be further analyzed. However, high percentages of total carotenoid or β-carotene retention were observed and can minimize vitamin A deficiency in low-income populations.


The influence of espresso coffee on neurocognitive function in HIV-infected patients.
The aim of our study was to evaluate the impact of coffee intake on cognitive function in persons living with HIV (PLWH). 130 PLWH with CD4 > 200 cells/mm(3), undetectable viral load, treated with HAART were included. A structured interview was applied and relevant clinical and laboratory data were assessed, including coffee intake. For neuropsychological assessment, the HIV Neurobehavioral Research Center Battery was chosen. Univariate nonparametric statistics and multivariate regression model were used. A significant association between espresso coffee use and a better cognitive function was verified in five of the eight psychometric measurements. In the multivariate analysis, after variable adjustment, linear regression analysis showed that coffee intake was a positive predictor for attention/working memory, executive functions and Global Deficit Score. Although the mechanisms behind the influence of caffeine on cognitive functioning are controversial, regular espresso coffee intake may have favourable effects on cognitive deterioration caused by HIV.


Zwitterionic 3D-Printed Non-Immunogenic Stealth Microrobots.
Microrobots offer transformative solutions for non-invasive medical interventions due to their small size and untethered operation inside the human body. However, they must face the immune system as a natural protection mechanism against foreign threats. Here, non-immunogenic stealth zwitterionic microrobots that avoid recognition from immune cells are introduced. Fully zwitterionic photoresists are developed for two-photon polymerization 3D microprinting of hydrogel microrobots with ample functionalization: tunable mechanical properties, anti-biofouling and non-immunogenic properties, functionalization for magnetic actuation, encapsulation of biomolecules, and surface functionalization for drug delivery. Stealth microrobots avoid detection by macrophage cells of the innate immune system after exhaustive inspection (>90 hours), which has not been achieved in any microrobotic platform to date. These versatile zwitterionic materials eliminate a major roadblock in the development of biocompatible microrobots, and will serve as a toolbox of non-immunogenic materials for medical microrobot and other device technologies for bioengineering and biomedical applications.


Blood flow restricted exercise for athletes: A review of available evidence.
This study aimed to collate current evidence regarding the efficacy of various blood flow restriction (BFR) strategies for well-trained athletes, and to provide insight regarding how such strategies can be used by these populations. Review article. Studies that had investigated the acute or adaptive responses to BFR interventions in athletic participants were identified from searches in MEDLINE (PubMed), SPORTDiscus (EBSCO) and Google Scholar databases up to April 2015. The reference lists of identified papers were also examined for relevant studies. Twelve papers were identified from 11 separate investigations that had assessed acute and adaptive responses to BFR in athletic cohorts. Of these, 7 papers observed enhanced hypertrophic and/or strength responses and 2 reported alterations in the acute responses to low-load resistance exercise when combined with BFR. One paper had examined the adaptive responses to moderate-load resistance training with BFR, 1 noted improved training responses to low-work rate BFR cardiovascular exercise, and 1 reported on a case of injury following BFR exercise in an athlete. Current evidence suggests that low-load resistance training with BFR can enhance muscle hypertrophy and strength in well-trained athletes, who would not normally benefit from using light loads. For healthy athletes, low-load BFR resistance training performed in conjunction with normal high-load training may provide an additional stimulus for muscular development. As low-load BFR resistance exercise does not appear to cause measureable muscle damage, supplementing normal high-load training using this novel strategy may elicit beneficial muscular responses in healthy athletes.


Preservation of rainbow trout (Oncorhynchus mykiss) eyed eggs using a perfluorochemical as an oxygen carrier.
The objective of this study was to maintain the viability of chilled rainbow trout (Oncorhynchus mykiss) eyed eggs during storage using oxygenated perfluorochemical (PFC). Three trials were conducted using eggs at 161, 180 or 217 degree days (days from fertilization x incubation temperature in degrees C). A separate trial was conducted for 147 degree day eggs that were not at the eyed stage. For each trial, eggs were stored in a moisture-saturated atmosphere at 1 degrees C in PFC, water, and 1:1 combinations of PFC and PBS, PFC and 0.3 M glucose, PFC and mineral oil, or PFC and water. The PFC was oxygenated before each trial and all media were oxygenated at weekly intervals during the storage period. Eggs from each trial were also incubated without storage to provide Day 0 results. After 3 and 5 weeks of storage, eggs from each medium were incubated at 10 degrees C until hatch. Hatching percentage was expressed as a percentage of Day 0 results. The percentage of normal alevins that hatched was also determined. There were interactions (P < 0.01) between stage of development and treatment for hatching percentage after 3 and 5 weeks of storage. After 3 weeks of storage, eggs stored at 161, 180, or 217 degree days without PFC had hatching rates of 0-14.3% but eggs stored in any medium with PFC had hatching percentages from 75.1 to 106.4% of Day 0 values. After 5 weeks of storage, eggs stored at 161 degree days in PFC plus PBS or PFC plus water, and eggs stored at 217 degree days in PFC or PFC plus water, had higher (P < 0.05) hatching percentages than eggs stored in any of the other media. Eggs stored at 161 degree days for 5 weeks in PFC and water had a higher (P < 0.05) percentage of normal alevins hatching than eggs stored in PFC and PBS. Because of their early developmental stage, eggs stored at 147 degree days had low hatching percentages, except eggs stored for 3 weeks in PFC or PFC plus PBS. Chilling eyed eggs of rainbow trout to 1 degrees C and storing them in water with PFC as an oxygen carrier can preserve their viability for 5 weeks.


Role of carnitine esters in brain neuropathology.
L-Carnitine (L-C) is a naturally occurring quaternary ammonium compound endogenous in all mammalian species and is a vital cofactor for the mitochondrial oxidation of fatty acids. Fatty acids are utilized as an energy substrate in all tissues, and although glucose is the main energetic substrate in adult brain, fatty acids have also been shown to be utilized by brain as an energy substrate. L-C also participates in the control of the mitochondrial acyl-CoA/CoA ratio, peroxisomal oxidation of fatty acids, and the production of ketone bodies. Due to their intrinsic interaction with the bioenergetic processes, they play an important role in diseases associated with metabolic compromise, especially mitochondrial-related disorders. A deficiency of carnitine is known to have major deleterious effects on the CNS. Several syndromes of secondary carnitine deficiency have been described that may result from defects in intermediary metabolism and alterations principally involving mitochondrial oxidative pathways. Mitochondrial superoxide formation resulting from disturbed electron transfer within the respiratory chain may affect the activities of respiratory chain complexes I, II, III, IV, and V and underlie some CNS pathologies. This mitochondrial dysfunction may be ameliorated by L-C and its esters. In addition to its metabolic role, L-C and its esters such as acetyl-L-carnitine (ALC) poses unique neuroprotective, neuromodulatory, and neurotrophic properties which may play an important role in counteracting various disease processes. Neural dysfunction and metabolic imbalances underlie many diseases, and the inclusion of metabolic modifiers may provide an alternative and early intervention approach, which may limit further developmental damage, cognitive loss, and improve long-term therapeutic outcomes. The neurophysiological and neuroprotective actions of L-C and ALC on cellular processes in the central and peripheral nervous system show such effects. Indeed, many studies have shown improvement in processes, such as memory and learning, and are discussed in this review.


DETERMINATION OF FACTORS ASSOCIATED WITH LONG-TERM ENDOTHELIAL LOSS AND REFRACTIVE RESULT IN PATIENTS WITH ARTISAN PHAKIC LENS.
To determine the changes in endothelial cell density, refractive results, and risk factors associated with endothelial loss in patients with irisclaw phakic intraocular lenses for myopia/myopic astigmatism (Artisan). Data collection was obtained from an existing database with information on patients with a phakic Artisan lens implant between 1998 and 2011 at the Virgilio Galvis Ophthalmology Centre, with at least 5 years of follow-up. As a second stage, an analysis was carried out to identify the change in endothelial cell density and its potential associated factors. A total of 80 eyes with myopic errors were included with a follow-up of 11.9 + 3.48 years. The percentage of total loss of endothelial cells was greater than 25% of the preoperative density in 43.8% of the eyes. A postoperative annual loss > 1.6% was found in 47.0% of the eyes with that information available. 41 eyes (51%) had final endothelial density < 2000 cells/mm2, and 7 (8.8%) eyes had endothelial cell density < 1000 cells/mm2. Among the variables studied, no associated factors for long-term endothelial loss were found. During the course of the study, 8 (10%) phakic intraocular lenses were explanted, including 3 with accelerated endothelial loss, and another 2 with cataract associated and a significantly low endothelial density. The last mean spherical equivalent was -0.81 (±1.01 D), and the final uncorrected distance visual acuity was 0.45 logMar (Snellen 20/56). Artisan-type phakic lenses are a good alternative for the correction of high myopic defects, with predictable refractive results in the long term. However, there is an increased loss of endothelial cells in the long term in a high percentage of patients. Strict postoperative follow-up, including endothelial evaluation, is required, and further studies are warranted.


Increased corrosion resistance of stent materials by converting current surface film of polycrystalline oxide into amorphous oxide.
Current efforts of new stent technology have been aimed largely at the improvement of intravascular stent biocompatibility. Among the chemical characteristics of metallic stents, surface oxide corrosion properties are paramount. Using our unique technique, the currently marketed 316 L stainless steel and nitinol stent wires covered with polycrystalline oxide were chemically etched and then passivated to form amorphous oxide. Excellent metallic-stent corrosion resistance with an amorphous oxide surface was demonstrated in our previous in vitro study. For in vivo validation, we compared the corrosion behavior of different oxide surfaces on various forms of test wires in the abdominal aorta of mongrel dogs using open-circuit potential and cyclic anodic polarization measurements. After conduction, the retrieved test wires were observed under scanning electron microscope. No passivity breakdown was found for wires covered with amorphous oxide, while wires with polycrystalline oxide showed breakdown at potentials between +0.2 to + 0.6 V. It has been proven that severe pitting or crevice corrosion occurred on the surface of polycrystalline oxide, while the surface of amorphous oxide was free of degradations in our experiment. We have demonstrated that this amorphous oxide coating on metallic material provides better corrosion resistance, not only in vitro but also in vivo, and it is superior not only in strength safety but also in medical device biocompatibility.


Functional evaluation before and after interventional bronchoscopy in patients with malignant central airway obstruction.
Rigid or flexible interventional bronchoscopy is increasingly accepted for treating central tracheal or bronchial obstruction caused by incurable pulmonary malignancy. Immediate and sustained symptomatic relief and improvement of quality of life can be offered to patients suffering from dyspnoea and facing death by suffocation. Selection of patients based on functional aspects is of central importance for success in endoscopic treatment. A thorough workup is conducted with close cooperation between the various specialists involved in thoracic oncology (pulmonologist, oncologist, radio-oncologist, thoracic and upper airway surgeon). Careful history taking, clinical examination, pulmonary function testing (PFT) and radiologic imaging are needed to assess the general condition of the patient and give information of type and stage of the underlying malignancy. In eligible patients, timing and method of the intervention depend on the acuteness of the presentation and the endoscopic assessment of the lesion responsible for the obstruction. Regular postinterventional follow-up should be conducted by the patient's doctor and consist of clinical examination, PFT and chest radiography. Close cooperation with the specialist centre is necessary to refer patients for repeat bronchoscopy and management of suspected recurrences or complications.


Babesia equi-induced anemia in a Quarter Horse and subsequent regulatory response.
A 7-year-old Quarter Horse gelding used for unsanctioned racing was examined because of fever and anorexia. Physical examination revealed fever, tachycardia, and tachypnea. Results of a CBC indicated anemia and mild thrombocytopenia. Results of microscopic examination of a blood smear indicated piroplasms in erythrocytes, consistent with Babesia spp. Regulatory authorities were contacted, and results of serologic testing at the National Veterinary Services Laboratories confirmed acute Babesia equi infection. Equids on the home premises of the index horse were placed under quarantine. Those equids were tested for piroplasmosis, and 6 of 63 horses had positive results for B equi. Another horse that had previously been housed on the index premises also had positive results for B equi. Competent tick vectors for piroplasmosis organisms were not identified. All 8 horses with piroplasmosis were Quarter Horses that participated in unsanctioned racing and were trained by the same person. Two of the horses were illegally removed from the index premises; these 2 horses and the other horse with piroplasmosis that was previously housed on the index premises could not be found. The other 5 horses with piroplasmosis were euthanized. Investigators concluded that transmission of B equi among horses was most likely iatrogenic. The United States has been considered piroplasmosis free. However, veterinarians should consider piroplasmosis in horses with signalments and clinical signs similar to those of the index horse of this report. Regulatory authorities should be contacted regarding horses in which piroplasmosis is suspected.


Glucocorticoids increase skeletal muscle NF-κB inducing kinase (NIK): links to muscle atrophy.
Glucocorticoids (GC) are a frontline therapy for numerous acute and chronic diseases because of their demonstrated efficacy at reducing systemic inflammation. An unintended side effect of GC therapy is the stimulation of skeletal muscle atrophy. Pathophysiological mechanisms responsible for GC-induced skeletal muscle atrophy have been extensively investigated, and the ability to treat patients with GC without unintended muscle atrophy has yet to be realized. We have reported that a single, standard-of-care dose of Methylprednisolone increases in vivo expression of NF-κB-inducing kinase (NIK), an important upstream regulatory kinase controlling NF-κB activation, along with other key muscle catabolic regulators such as Atrogin-1 and MuRF1 that induce skeletal muscle proteolysis. Here, we provide experimental evidence that overexpressing NIK by intramuscular injection of recombinant human NIK via adenoviral vector in mouse tibialis anterior muscle induces a 30% decrease in the average fiber cross-sectional area that is associated with increases in mRNA expression of skeletal muscle atrophy biomarkers MuRF1, Atrogin-1, myostatin and Gadd45. A single injection of GC induced NIK mRNA and protein within 2 h, with the increased NIK localized to nuclear and sarcolemmal locations within muscle fibers. Daily GC injections induced skeletal muscle fore limb weakness as early as 3 days with similar atrophy of muscle fibers as observed with NIK overexpression. NIK overexpression in primary human skeletal muscle myotubes increased skeletal muscle atrophy biomarkers, while NIK knockdown significantly attenuated GC-induced increases in NIK and Atrogin-1. These results suggest that NIK may be a novel, previously unrecognized mediator of GC-induced skeletal muscle atrophy.


Evaluating glioma therapies: modeling treatments and predicting outcomes.
Intra-arterial chemotherapy with carmustine (BCNU) and interstitial radiation therapy with the use of stereotactically placed 125I sources are aggressive local therapies for malignant glioma. These therapies emerged in the 1980s and both appeared promising in phase II studies but yielded disappointing results in subsequent randomized controlled trials by the Brain Tumor Cooperative Group (BTCG). Florell and colleagues had prepared us for the possibility that brachytherapy would have less impact on survival than anticipated from the phase II experience by demonstrating that patients who were judged eligible for interstitial radiation, but treated conventionally, lived significantly longer than those who were ineligible and had better than average outcomes. To further examine the impact of patient selection on outcome, we used the database of Florell et al. to assess the survival of patients with malignant glioma who were eligible or ineligible for chemotherapy by three intra-arterial methods, one of which was similar to that employed by the BTCG in its randomized, controlled trial evaluating intra-arterial BCNU. The medical records and computed tomography (CT) scans of 102 consecutive patients with malignant glioma receiving standard treatment (i.e., maximum feasible surgical resection, external-beam radiotherapy, and often adjuvant systemic chemotherapy) at a single cancer center in Canada during the calendar years 1988 and 1989 were used for this analysis. Based on CT imaging and blind to outcome, an interventional neuroradiologist decided which patients were eligible or ineligible for intra-arterial chemotherapy via injection of two major arteries, via injection of one major artery, or via selective middle-cerebral artery injection. A Karnofsky performance score of greater than or equal to 60 was required. The percent of eligible patients, the median survival time, and the distribution of prognostic factors were analyzed for each group of eligible and ineligible patients. Median survival times were compared with the use of the generalized Wilcoxon (Breslow) test. All P values were based on two-tailed tests. For two-vessel treatment, 72.5% of the patients (74 of 102) were eligible; the eligible patients on average lived longer than the ineligible patients (14.8 versus 3.5 months; P < .00001). For one-vessel treatment, 48% of the patients (49 of 102) were eligible; again, the eligible patients lived longer than the ineligible patients (18.4 versus 5.1 months; P < .00001). For middle-cerebral artery treatment, 30% of the patients (31 of 102) were eligible, and these eligible patients did live somewhat longer than the ineligible patients, but this result did not reach statistical significance (13.6 versus 9.9 months; P = .1304). Trends were similar for patients with glioblastoma multiforme and anaplastic glioma. The median duration of survival was 11.4 months for all patients. Patients who were eligible for intra-arterial chemotherapy lived significantly longer or somewhat longer (depending on the selection criteria used) than patients who were ineligible and had better than expected outcomes. Patients who were judged eligible for intra-arterial chemotherapy by the two-vessel method and the control group in the BTCG phase III trial of intra-arterial chemotherapy had similar median survival times (14.8 versus 14.0 months). Modeling treatments with the use of a comprehensive clinical and imaging database of unselected, conventionally treated patients may help investigators decide if new therapies warrant definitive evaluation in randomized trials by measuring the degree to which patient selection may have enhanced phase II study outcomes.


Nineteen-channel receive array and four-channel transmit array coil for cervical spinal cord imaging at 7T.
To design and validate a radiofrequency (RF) array coil for cervical spinal cord imaging at 7T. A 19-channel receive array with a four-channel transmit array was developed on a close-fitting coil former at 7T. Transmit efficiency and specific absorption rate were evaluated in a B1 (+) mapping study and an electromagnetic model. Receive signal-to-noise ratio (SNR) and noise amplification for parallel imaging were evaluated and compared with a commercial 3T 19-channel head-neck array and a 7T four-channel spine array. The performance of the array was qualitatively demonstrated in human volunteers using high-resolution imaging (down to 300 μm in-plane). The transmit and receive arrays showed good bench performance. The SNR was approximately 4.2-fold higher in the 7T receive array at the location of the cord with respect to the 3T coil. The g-factor results showed an additional acceleration was possible with the 7T array. In vivo imaging was feasible and showed high SNR and tissue contrast. The highly parallel transmit and receive arrays were demonstrated to be fit for spinal cord imaging at 7T. The high sensitivity of the receive coil combined with ultra-high field will likely improve investigations of microstructure and tissue segmentation in the healthy and pathological spinal cord.


A Stability-indicating High Performance Liquid Chromatographic Assay for the Simultaneous Determination of Atenolol and Lercanidipine Hydrochloride in Tablets.
A simple, rapid, precise and accurate isocratic reversed phase stability indicating HPLC method was developed and validated for the simultaneous determination of atenolol and lercanidipine hydrochloride in commercial tablets. The chromatographic separation was achieved on phenomenex Gemini C18 (250×4.6 mm, 5 μm) column using a mobile phase consisting of acetonitrile and buffer (20 mM potassium dihydrogen phosphate pH 3.5) in the ratio of (55:45, v/v) at a flow rate of 1.0 ml/min and UV detection at 235 nm. The linearity of the proposed method was investigated in the range of 40-160 μg/ml (r(2)=0.9995) for atenolol and 8-32 μg/ml (r(2)=0.9993) for lercanidipine. Degradation products produced as a result of stress studies did not interfere with the detection of atenolol and lercanidipine and the assay can thus be considered stability-indicating.


Regulation of heat shock protein synthesis in human skin fibroblasts in response to oxidative stress: role of vitamin E.
Skin plays an important role in protection against oxidative stressors such as ultraviolet radiation, ozone and chemicals. Chronic sun exposure causes degenerative changes in the skin that are recognized as photoaging. Oxidative stress has been shown to alter the expression of mammalian antioxidant enzymes as well as to enhance numerous transcription factors, including nuclear factor kappaB, stress-activated protein kinase and heat shock factor This latter is the transcription factor for the synthesis of heat shock proteins, which have been known to protect against a wide variety of toxic conditions, including extreme temperatures, oxidative stress and cytotoxic drugs. In this study we investigated the role of oxidative stress in the induction of heat shock protein (HSP) 70 in human skin fibroblasts and the effect of vitamin E. We found that significant HSP70 induction occurred after exposure to HOOH and that this was associated with a significant perturbation in protein and nonprotein sulfhydryl groups, and with a significant increase in protein carbonyl levels. Treatment with vitamin E conferred significant protection against stress-induced modifications of cellular sulfhydryl and carbonyl content, while maintaining functional levels of cytoprotective HSP70. Our results point to the possible involvement of redox mechanisms in the heat shock signal transduction pathway, which may play an important regulatory role in the genetic mechanisms of tolerance to oxidative stress. Exogenous antioxidant supplementation with vitamin E could have cosmetic benefits and may be an efficient tool to mitigate the consequences of free radical-induced skin damage.


Association Between Circulating Troponin Concentrations, Left Ventricular Systolic and Diastolic Functions, and Incident Heart Failure in Older Adults.
Cardiac troponin is associated with incident heart failure and greater left ventricular (LV) mass. Its association with LV systolic and diastolic functions is unclear. To define the association of high-sensitivity cardiac troponin T (hs-cTnT) with LV systolic and diastolic functions in the general population, and to evaluate the extent to which that association accounts for the correlation between hs-cTnT concentration and incident heart failure overall, heart failure with preserved LV ejection fraction (LVEF; HFpEF), and heart failure with LVEF less than 50%. This analysis of the Atherosclerosis Risk in Communities (ARIC) Study, an ongoing epidemiologic cohort study in US communities, included participants without cardiovascular disease (n = 4111). Available hs-cTnT measurements for participants who attended ARIC Study visits 2 (1990 to 1992), 4 (1996 to 1998), and 5 (2011 to 2013) were assessed cross-sectionally against echocardiographic measurements taken at visit 5 and against incident health failure after visit 5. Changes in hs-cTnT concentrations from visits 2 and 4 were also examined. Data analyses were performed from August 2017 to July 2018. Cardiac structure and function by echocardiography at visit 5, and incident heart failure during a median 4½ years follow-up after visit 5. Of the 6538 eligible participants, 4111 (62.9%) without cardiovascular disease were included. Among these participants, 2586 (62.9%) were female, and the mean (SD) age was 75 (5) years. Median (interquartile range) hs-cTnT concentration at visit 5 was 9 (7-14) ng/L and was detectable in 3946 participants (96.0%). After adjustment for demographic and clinical covariates, higher hs-cTnT levels were associated with greater LV mass index (adjusted mean [SE] for group 1: 33.8 [0.5] vs group 5: 40.1 [0.4]; P for trend < .001) and with worse diastolic function, including lower tissue Doppler imaging e' (6.00 [0.07] vs 5.54 [0.06]; P for trend < .001), higher E/e' ratio (11.4 [0.2] vs 12.9 [0.1]; P for trend < .001), and greater left atrial volume index (23.4 [0.4] vs 26.4 [0.3]; P for trend < .001), independent of LV mass index; hs-cTnT level was not associated with measures of LV systolic function. Accounting for diastolic function attenuated the association of hs-cTnT concentration with incident HFpEF by 41% and the association with combined heart failure with midrange and reduced ejection fraction combined (LVEF <50) by 17%. Elevated hs-cTnT concentration and diastolic dysfunction were additive risk factors for incident heart failure. For any value of late-life hs-cTnT levels, longer duration of detectable hs-cTnT from midlife to late life was associated with greater LV mass in late life but not with worse LV systolic or diastolic function. This study shows that higher hs-cTnT concentrations were associated with worse diastolic function, irrespective of LV mass, but not with systolic function; these findings suggest that high levels of hs-cTnT may serve as an early marker of subclinical alterations in diastolic function that may lead to a predisposition to heart failure.


Theories explaining racial differences in the utilization of diagnostic and therapeutic procedures for cerebrovascular disease.
Despite a higher risk of stroke, blacks are less likely than whites to receive the invasive procedures that are used to diagnose and treat cerebrovascular disease, particularly carotid endarterectomy. Explanations for the lower rate of procedural use include racial bias, racial differences in pathophysiology of cerebrovascular disease, affordability, and racial variation in patient decisions regarding care. Studies consistently indicate that blacks are less likely to have severe atherosclerotic lesions of the carotid arteries, reducing their likelihood of being appropriate candidates for carotid endarterectomy. Although ability to pay does not explain the observed variation in use of carotid endarterectomy, it may influence the evaluation process prior to hospitalization. A racial difference in patients' decisions about health care for cerebrovascular disease has never been investigated; it should be a topic of future studies.


Effect of dose rate on the radiation-induced bystander response.
Radiation-induced biological bystander effects have become a well-established phenomenon associated with the interaction of radiation with cells. These so-called bystander effects have been seen across a variety of end points for both high and low linear energy transfer (LET) radiations, utilizing a variety of dose rates and radiation sources. In this study, the effect of dose rate and different low LET sources on the bystander cell survival fraction (SF) was examined. The cell line investigated was the human keratinocyte HPV-G. The bystander response was measured via clonogenic assay after medium transfer protocol. Cells were irradiated using (60)Co gamma-rays and 20 MeV electrons at doses of 0.5, 5 and 10 Gy with varying dose rates. Both gamma and electron irradiation decreased recipient SF at 0.5 Gy and 5 Gy, respectively. Subsequent recovery of the SF to control levels for 10 Gy was observed. There was no dose rate dependence for (60)Co irradiation. A significant difference in the survival fraction was observed for electron irradiation at 10 Gy and a high dose rate. Furthermore, survival fractions were compared between (60)Co and 20 MeV electron irradiations. This showed a significant increase in the survival fraction 'recovery' at 10 Gy for a (60)Co dose rate of 1.1 Gy min(-1) compared to 20 MeV electrons at 1.0 Gy min(-1). No such difference was observed when comparing at higher dose rates. Lastly, increases in survival fraction at 10 Gy were abolished and the SF decreased by the plating of increased numbers of recipient cells. Such evidence may help gain insight into the nature and mechanism(s) surrounding bystander signal production, how these phenomena are tested and their eventual application in a clinical setting.


Tolerability of Capsaicinoids from Capsicum Extract in a Beadlet Form: A Pilot Study.
A single center, open-label, dose-finding adaptive study was conducted in twelve healthy overweight female subjects. The study was to evaluate the safety and tolerability of the capsaicinoids (CAPs) from Capsicum extract in a beadlet form compared to placebo in a healthy overweight population. The investigational product capsaicinoids (CAPs) from Capsicum extract in a beadlet form (Capsimax®) a proprietary encapsulated form of Capsicum extract in beadlet form supplemented at 2 mg, 4 mg, 6 mg, 8 mg and 10 mg of CAPs. An ascending dose protocol evaluated a total dose of 10 mg daily given in five divided doses (2 mg, 4 mg, 6 mg, 8 mg and 10 mg of CAPs). Each dose was given for a week. Safety and tolerability were assessed. Primary outcomes were tolerability assessments and reports of adverse events. Tolerability assessments were observed on skin color and any changes in skin, bowel movement, digestion, mouth or throat, hair color or changes in hair color, urination includes frequency and burning sensations, breathing, any changes in their health. Secondary outcomes were body weight, body mass index (BMI), blood pressure (SBP/DBP), vital signs, electrocardiograms, clinical chemistry parameters including liver function tests, lung function tests and kidney function tests and complete blood count (CBC). No dose effective changes were observed. The escalating dose levels of CAPs in a beadlet form product found was tolerable and safe for weight management studies. Tolerability assessments and safety blood markers showed no significant changes from baseline. No significant serious adverse events were reported throughout the duration of the study. Further longer term studies are required to explore the tolerability of the product. This trial is registered with # ISRCTN10975080.


DCE and DW MRI in monitoring response to androgen deprivation therapy in patients with prostate cancer: a feasibility study.
Androgen deprivation therapy (ADT) is a key primary treatment for advanced and metastatic prostate cancer and is an important neoadjuvant before radiotherapy. We evaluated 3.0 T dynamic contrast-enhanced MRI and diffusion-weighted (DW) MRI in monitoring ADT response. Twenty-three consecutive patients with prostate cancer treated by primary ADT were included. Imaging was performed at baseline and 3 months posttreatment with ADT. After 3 months therapy there was a significant reduction in all dynamic contrast-enhanced MRI parameters measured in tumor regions of interest (K(trans), k(ep), v(p), IAUGC-90); P < 0.001. Areas of normal-appearing peripheral zone showed no significant change; P = 0.285-0.879. Post-ADT, there was no significant change in apparent diffusion coefficient values in tumors, whilst apparent diffusion coefficient values significantly decreased in areas of normal-appearing peripheral zone, from 1.786 × 10(-3) mm(2) /s to 1.561 × 10(-3) mm(2) /s; P = 0.007. As expected the median Prostate-Specific Antigen (PSA) significantly reduced from 30 ng/mL to 1.5 ng/mL posttreatment, and median prostate volume dropped from 47.6 cm(3) to 24.9 cm(3) ; P < 0.001. These results suggest that dynamic contrast-enhanced MRI and diffusion-weighted MRI offer different information but that both could prove useful adjuncts to the anatomical information provided by T2-weighted imaging. dynamic contrast-enhanced as a marker of angiogenesis may help demonstrate ADT resistance and diffusion-weighted imaging may be more accurate in determining presence of tumor cell death versus residual tumor.


Naltrexone in the treatment of opioid-dependent pregnant women: the case for a considered and measured approach to research.
The present paper considers naltrexone to treat opioid dependence during pregnancy. The public health problem of opioid dependence and its treatment during pregnancy is reviewed first. Next, the naltrexone and opioid dependence treatment literature is summarized, with overviews of the pre-clinical and clinical research on prenatal naltrexone exposure. Finally, considerations and recommendations for future medication research for the treatment of opioid dependence in pregnant women are provided. The efficacy of long-acting injectable naltrexone relative to placebo, its blockade of opioid agonist euphoric effects, its lack of abuse and tolerance development and its modest adverse effect profile make it a potential medication for opioid-dependent pregnant women. However, it is not without seriously concerning potential drawbacks, including the difficulty surrounding medication induction that may lead to vulnerability with regard to relapse, physical dependence re-establishment, increased risk behaviors, treatment dropout and resulting opioid overdose. Before embarking on future research with this medication, the benefits and risks for the mother-embryo/fetus/child dyad should be weighed carefully. Should future research be conducted, a multi-level commitment to proactive ethical research is needed to reach the ultimate goal of improving the lives of women and children affected by opioid dependence.


Mutarotation of aldoses: Getting a deeper knowledge of a classic equilibrium enabled by computational analyses.
The mutarotation equilibrium, by which reducing carbohydrates exist in solution as the α and β anomers of cyclic (furanoid and pyranoid) structures, along with open-chain (aldehyde and hydrate) forms, and whose ratios are depending on factors such as temperature, pH and solvent, portraits a phenomenon involved in numerous processes of chemical and biological importance. Herein, we have developed a DFT-based rationale that provides a broader landscape for anomerizations and ring-open chain interconversions, together with the pivotal role exerted not only by the aldehyde intermediate (essentially the only acyclic structure taken into account so far), but also the hydrate form (often more abundant at the equilibrium). These calculations reveal a more complex and richer scenario than was thought, and identify different mutarotation mechanisms that hinge on every monosaccharide. It is noteworthy that pyranose-furanose interconversion may actually occur without the intermediacy of open-chain forms. For the aldoses evaluated, namely d-glucose, d-ribose, and d-xylose, all structures involved in mutarotation undergo interconversion pathways, whose energy barriers calculated at the M06-2X/6-311++G(d,p) level, are in good agreement with previous experimental measurements.


Bidi and hookah use among Canadian youth: findings from the 2010 Canadian Youth Smoking Survey.
Although cigarette use among Canadian youth has decreased significantly in recent years, alternative forms of tobacco use are becoming increasingly popular. Surveillance of youth tobacco use can help inform prevention programs by monitoring trends in risk behaviors. We examined the prevalence of bidi and hookah use and factors associated with their use among Canadian youth by using data from the 2010-2011 Youth Smoking Survey (YSS). We analyzed YSS data from 28,416 students (2006-2007) and 31,396 students (2010-2011) in grades 9 through 12 to examine prevalence of bidi and hookah use. We conducted multivariate logistic regression analyses of 2010-2011 YSS data to examine factors associated with bidi and hookah use. From 2006 through 2010, prevalence of hookah use among Canadian youth increased by 6% (P = .02). Marijuana use emerged as a consistent predictor of bidi and hookah use. Males, youth of black, Latin, or other descent, and youth of Asian descent were more likely to use bidis (odds ratio [OR], 1.5; OR, 15.6; OR, 14.9) or hookah (OR, 1.3; OR, 2.4; OR, 1.5). Current cigarette smokers were more likely than nonsmokers to be current users of bidis (OR, 6.7) and hookahs (OR, 3.0), and occasional and frequent alcohol drinkers were also more likely than nondrinkers to be current hookah users (OR, 2.8; OR, 3.6). Although bidi use has not changed significantly among Canadian youth, the increase in hookah use warrants attention. Understanding the factors associated with use of bidis and hookahs can inform the development of tobacco use prevention programs to address emerging at-risk youth populations.


Response to conjugate Haemophilus influenzae B vaccine among infants in Niamey, Niger.
Despite near elimination of Haemophilus influenzae b (Hib) meningitis from several industrialized countries following introduction of conjugate Hib vaccines into infant immunization schedules, Hib remains a major cause of meningitis and pneumonia in resource-poor countries. In Niger, Hib causes nearly 200 cases of meningitis per 100,000 children < one year of age, and > 40% of cases are fatal. We evaluated the immunogenicity of Hib polysaccharide-tetanus toxoid conjugate vaccine (PRP-T) administered in the same syringe as diphtheria-tetanus-pertussis (DTP) vaccine among infants in Niger. Infants were randomized into group 1 (PRP-T at six, 10, and 14 weeks), group 2 (PRP-T at 10 and 14 weeks), or a control group (meningococcal A/C polysaccharide vaccine). By 14 weeks of age, all subjects in groups land 2 had > or = 0.15 microg/ml of anti-PRP antibody, and 82% versus 76% had > or = 1.0 microg/ml of antibody (P=not significant). By nine months of age the proportion of infants with > or = 0.15 and > or = 1.0 microg/ml was group I=97% and 76%; group 2=93% and 67%; controls=10% and 2.6%. Four weeks after the first, second, and third doses of PRP-T, infants in group 1 showed geometric mean titers (GMTs) of 0.19, 3.97, and 6.09 microg/ml while infants in group 2 had GMTs of 2.40 and 4.41 microg/ml four weeks after the delayed first and second doses. Both PRP-T groups had significantly higher GMTs at 18 weeks and nine months of age than infants in the control group. The Hib PRP-T vaccine was immunogenic in infants in Niger. The strong response after PRP-T was initiated one month after the first DTP vaccination may reflect carrier priming. Two dose schedules of PRP-T should be given serious consideration, particularly if their reduced cost permits vaccine introduction that would be otherwise unaffordable.


Ring Attractor Dynamics Emerge from a Spiking Model of the Entire Protocerebral Bridge.
Animal navigation is accomplished by a combination of landmark-following and dead reckoning based on estimates of self motion. Both of these approaches require the encoding of heading information, which can be represented as an allocentric or egocentric azimuthal angle. Recently, Ca2+ correlates of landmark position and heading direction, in egocentric coordinates, were observed in the ellipsoid body (EB), a ring-shaped processing unit in the fly central complex (CX; Seelig and Jayaraman, 2015). These correlates displayed key dynamics of so-called ring attractors, namely: (1) responsiveness to the position of external stimuli; (2) persistence in the absence of external stimuli; (3) locking onto a single external stimulus when presented with two competitors; (4) stochastically switching between competitors with low probability; and (5) sliding or jumping between positions when an external stimulus moves. We hypothesized that ring attractor-like activity in the EB arises from reciprocal neuronal connections to a related structure, the protocerebral bridge (PB). Using recent light-microscopy resolution catalogs of neuronal cell types in the PB (Lin et al., 2013; Wolff et al., 2015), we determined a connectivity matrix for the PB-EB circuit. When activity in this network was simulated using a leaky-integrate-and-fire model, we observed patterns of activity that closely resemble the reported Ca2+ phenomena. All qualitative ring attractor behaviors were recapitulated in our model, allowing us to predict failure modes of the putative PB-EB ring attractor and the circuit dynamics phenotypes of thermogenetic or optogenetic manipulations. Ring attractor dynamics emerged under a wide variety of parameter configurations, even including non-spiking leaky-integrator implementations. This suggests that the ring-attractor computation is a robust output of this circuit, apparently arising from its high-level network properties (topological configuration, local excitation and long-range inhibition) rather than fine-scale biological detail.


Clinical aspects of diabetic neuropathies.
Diabetic neuropathy is a common complication of diabetes that may be associated both with considerable morbidity (painful polyneuropathy, neuropathic ulceration) and mortality (autonomic neuropathy). The epidemiology and natural history of diabetic neuropathy is clouded with uncertainty, largely due to confusion in the definition and measurement of this disorder. We have reviewed a variety of the clinical manifestations associated with somatic and autonomic neuropathy and discussed current views related to the management of the different abnormalities. Although unproven, the best evidence suggests that near normal control of blood glucose in the early years following onset of diabetes may help delay the development of clinically significant nerve impairment. Intensive therapy to achieve normalization of blood glucose may also lead to reversibility of early diabetic neuropathy, but again this is unproven. Our ability to manage successfully the many different manifestations of diabetic neuropathy depends ultimately on our success in uncovering the pathogenic processes underlying this disorder. The recent resurgence of interest in the vascular hypothesis, for example, has opened up new avenues of investigation for therapeutic intervention. Paralleling our increased understanding of the pathogenesis of diabetic neuropathy, there must be refinements in our ability to measure quantitatively the different types of defects that occur in this disorder. These tests must be validated and standardized to allow comparability between studies and more meaningful interpretation of study results.


Estrogen-related receptor γ (ERRγ) regulates oxygen-dependent expression of voltage-gated potassium (K+) channels and tissue kallikrein during human trophoblast differentiation.
Estrogen-related receptor γ (ERRγ) serves a critical O2-dependent regulatory role in the differentiation of human cytotrophoblasts to syncytiotrophoblast. In this study, we investigated expression of genes encoding tissue kallikrein (KLK1) and voltage-gated K(+) channels (KV7) during differentiation of human trophoblasts in culture and the roles of ERRγ and O2 tension in their regulation. Expression of KLK1 and the KV7 channel subunits, KCNQ1, KCNE1, KCNE3, and KCNE5, increased during differentiation of cultured human trophoblast cells in a 20% O2 environment. Notably, together with ERRγ, expression of KLK1, KCNQ1, KCNE1, KCNE3, and KCNE5 was markedly reduced when cells were cultured in a hypoxic environment (2% O2). Moreover, upon transduction of trophoblast cells with short hairpin RNAs for endogenous ERRγ, KLK1, KCNQ1, KCNE1, and KCNE3 expression was significantly decreased. Promoter and site-directed mutagenesis studies in transfected cells identified putative ERRγ response elements within the KLK1 and KCNE1 5'-flanking regions required for ERRγ-stimulated transcriptional activity. Binding of endogenous ERRγ to these ERRγ response elements increased during trophoblast differentiation in culture and was inhibited by hypoxia. The KV7 blocker linopirdine reduced human chorionic gonadotropin secretion and aggregation of cultured human trophoblasts, suggesting a possible role of KV7 channels in cell fusion and differentiation. Illumina gene expression arrays of cultured human trophoblast cells revealed several genes upregulated during syncytiotrophoblast differentiation and downregulated upon ERRγ knockdown involved in cell differentiation, adhesion, and synthesis of steroid and peptide hormones required for placental development and function. Collectively, these findings suggest that ERRγ mediates O2-dependent expression of genes involved in human trophoblast differentiation, function, and vascular homeostasis.


Comparative study of the neuronal plasticity along the neuraxis of the vibrissal sensory system of adult rat following unilateral infraorbital nerve damage and subsequent regeneration.
The aim of the present study was to examine the physiological consequences of a unilateral infraorbital nerve lesion and its regeneration at different levels of the somatosensory neuraxis. In animals whose right infraorbital nerve had been crushed, a large unresponsive area was found in the main brainstem trigeminal nucleus (Pr5). Responses evoked by ipsilateral vibrissal deflection in the middle of Pr5 reappeared only on days 22-35 after the nerve had been transected, whereas recovery from the nerve crush took only 7-9 days. However, no sign of short-term neuronal plasticity was observed in Pr5 after peripheral nerve injury. An enlargement of the receptive fields in two-thirds of the units and a lengthening in the delay of the evoked responses were observed as long-term plastic changes in Pr5 neurons after peripheral-nerve regeneration. In the ventral posteromedial nucleus of the thalamus (VPM) of partly denervated animals, however, only minutes or hours after the nerve crush, certain units were found to respond in some cases not only to the vibrissae, but also to mechanical stimulation of the face over the eye (two units), the nose (one unit), and the midline (one unit). Apart from the experiments involving incomplete denervation, the vibrissal representation areas of the VPM were unresponsive to stimulation of both the vibrissae and other parts of the face until nerve regeneration had occurred. In the somatosensory cortex, an infraorbital nerve crush immediately resulted in a large cortical area being unresponsive to vibrissal deflection. It was noteworthy, however, that shortly after the nerve crush, this large unresponsive whisker representation cortical area was invaded from the rostromedial direction by responses evoked by stimulation of the forepaw digits. In spite of the reappearance of vibrissa-evoked responses 7-10 days after the nerve crush, an expanded digital representation could still be observed 3 weeks after the nerve crush, resulting in an overlapping area of digital and vibrissal representations. The withdrawal of the expanded representation of forepaw digits was completed by 60 days after the nerve crush. The results obtained in Pr5, the VPM, and the cortex strongly suggest that the higher the station in the neuraxis, the greater the degree of plasticity after infraorbital nerve injury.


Denaturing high performance liquid chromatography analysis of the DPYD gene in patients with lethal 5-fluorouracil toxicity.
Dihydropyrimidine dehydrogenase (DPD) enzyme deficiency is a pharmacogenetic syndrome with possible fatal outcome following 5-fluorouracil (5-FU) treatment. Several studies examining the molecular basis for DPD deficiency have identified over 30 sequence variations in the DPYD gene (which codes for the DPD enzyme). Our laboratory has recently developed and validated a denaturing high performance liquid chromatography method capable of identifying both known and unknown sequence variations in the DPYD gene. In the present study, we used this denaturing high performance liquid chromatography approach to examine the DPYD genotype of three patients who experienced lethal toxicity after administration of 5-FU. DPD enzyme activity could only be measured in one patient before death and demonstrated that lethal toxicity can occur in a partially DPD-deficient individual. Multiple heterozygous sequence variations (both known and unknown) were detected in all three patients including the novel variants 545T>A, M182K and 2329G>T, A777S. We conclude that (a) lethal toxicity can occur in partially DPD-deficient individuals after administration of 5-FU and is not exclusive to profoundly DPD-deficient individuals as suggested previously, (b) the complicated heterozygote genotype seen in these patients, combined with DPD deficiency being an autosomal codominant inherited syndrome, precludes the use of simple genotyping assays that identify only one or two mutations as a method for identifying DPD-deficient individuals; and (c) these multiple heterozygote genotypes (which are more difficult to accurately characterize) may be responsible for some of the conflicting reports which suggests a lack of correlation between phenotype and genotype.


What items should be included in an early warning score for remote assessment of suspected COVID-19? qualitative and Delphi study.
To develop items for an early warning score (RECAP: REmote COVID-19 Assessment in Primary Care) for patients with suspected COVID-19 who need escalation to next level of care. The study was based in UK primary healthcare. The mixed-methods design included rapid review, Delphi panel, interviews, focus groups and software development. Participants were 112 primary care clinicians and 50 patients recovered from COVID-19, recruited through social media, patient groups and snowballing. Using rapid literature review, we identified signs and symptoms which are commoner in severe COVID-19. Building a preliminary set of items from these, we ran four rounds of an online Delphi panel with 72 clinicians, the last incorporating fictional vignettes, collating data on R software. We refined the items iteratively in response to quantitative and qualitative feedback. Items in the penultimate round were checked against narrative interviews with 50 COVID-19 patients. We required, for each item, at least 80% clinician agreement on relevance, wording and cut-off values, and that the item addressed issues and concerns raised by patients. In focus groups, 40 clinicians suggested further refinements and discussed workability of the instrument in relation to local resources and care pathways. This informed design of an electronic template for primary care systems. The prevalidation RECAP-V0 comprises a red flag alert box and 10 assessment items: pulse, shortness of breath or respiratory rate, trajectory of breathlessness, pulse oximeter reading (with brief exercise test if appropriate) or symptoms suggestive of hypoxia, temperature or fever symptoms, duration of symptoms, muscle aches, new confusion, shielded list and known risk factors for poor outcome. It is not yet known how sensitive or specific it is. Items on RECAP-V0 align strongly with published evidence, clinical judgement and patient experience. The validation phase of this study is ongoing. NCT04435041.


Different effects of hyperlipidic diets in human lactation and adulthood: growth versus the development of obesity.
After birth, the body shifts from glucose as primary energy substrate to milk-derived fats, with sugars from lactose taking a secondary place. At weaning, glucose recovers its primogeniture and dietary fat role decreases. In spite of human temporary adaptation to a high-fat (and sugars and protein) diet during lactation, the ability to thrive on this type of diet is lost irreversibly after weaning. We could not revert too the lactating period metabolic setting because of different proportions of brain/muscle metabolism in the total energy budget, lower thermogenesis needs and capabilities, and absence of significant growth in adults. A key reason for change was the limited availability of foods with high energy content at weaning and during the whole adult life of our ancestors, which physiological adaptations remain practically unchanged in our present-day bodies. Humans have evolved to survive with relatively poor diets interspersed by bouts of scarcity and abundance. Today diets in many societies are largely made up from choice foods, responding to our deeply ingrained desire for fats, protein, sugars, salt etc. Consequently our diets are not well adjusted to our physiological needs/adaptations but mainly to our tastes (another adaptation to periodic scarcity), and thus are rich in energy roughly comparable to milk. However, most adult humans cannot process the food ingested in excess because our cortical-derived craving overrides the mechanisms controlling appetite. This is produced not because we lack the biochemical mechanisms to use this energy, but because we are unprepared for excess, and wholly adapted to survive scarcity. The thrifty mechanisms compound the effects of excess nutrients and damage the control of energy metabolism, developing a pathologic state. As a consequence, an overflow of energy is generated and the disease of plenty develops.


NC1 domain of type VII collagen binds to the beta3 chain of laminin 5 via a unique subdomain within the fibronectin-like repeats.
Type VII collagen, the major component of anchoring fibrils, consists of a central collagenous triple-helical domain flanked by two noncollagenous, globular domains, NC1 and NC2. Approximately 50% of the molecular mass of the molecule is consumed by the NC1 domain. We previously demonstrated that NC1 binds to various extracellular matrix components including a complex of laminin 5 and laminin 6 (Chen et al, 1997a). In this study, we examined the interaction of NC1 with laminin 5 (a component of anchoring filaments). Both authentic and purified recombinant NC1 bound to human and rat laminin 5 as measured by enzyme-linked immunosorbant assay and by binding of 125I-radiolabeled NC1 to laminin 5-coated wells, but not to laminin 1 or albumin. NC1 bound predominantly to the beta3 chain of laminin 5, but also to the gamma2 chain when examined by a protein overlay assay. The binding of 125I-NC1 to laminin 5 was inhibited by a 50-fold excess of unlabeled NC1 or de-glycosylated NC1, as well as a polyclonal antibody to laminin 5 or a monoclonal antibody to the beta3 chain. In contrast, the NC1-laminin 5 interaction was not affected by a monoclonal antibody to the alpha3 chain. Using NC1 deletion mutant recombinant proteins, a 285 AA (residues 760-1045) subdomain of NC1 was identified as the binding site for laminin 5. IgG from an epidermolysis bullosa acquisita serum containing autoantibodies to epitopes within NC1 that colocalized with the laminin 5 binding site inhibited the binding of NC1 to laminin 5. Thus, perturbation of the NC1-laminin 5 interaction may contribute to the pathogenesis of epidermolysis bullosa acquisita.