Early intervention for adolescents with patellofemoral pain syndrome--a pragmatic cluster randomised controlled trial.
Self-reported knee pain is highly prevalent among adolescents. As much as 50% of the non-specific knee pain may be attributed to Patellofemoral Pain Syndrome (PFPS). In the short term, exercise therapy appears to have a better effect than patient education consisting of written information and general advice on exercise or compared with placebo treatment. But the long-term effect of exercise therapy compared with patient education is conflicting. The purpose of this study is to examine the short- and long-term effectiveness of patient education compared with patient education and multimodal physiotherapy applied at a very early stage of the condition among adolescents. This study is a single blind pragmatic cluster randomised controlled trial. Four upper secondary schools have been invited to participate in the study (approximately 2500 students, aged 15-19 years). Students are asked to answer an online questionnaire regarding musculoskeletal pain. The students who report knee pain are contacted by telephone and offered a clinical examination by a rheumatologist. Subjects who fit the inclusion criteria and are diagnosed with PFPS are invited to participate in the study. A minimum of 102 students with PFPS are then cluster-randomised into two intervention groups based on which school they attend. Both intervention groups receive written information and education. In addition to patient education, one group receives multimodal physiotherapy consisting primarily of neuromuscular training of the muscles around the foot, knee and hip and home exercises. The students with PFPS fill out self-reported questionnaires at baseline, 3, 6, 12 and 24 months after inclusion in the study. The primary outcome measure is perception of recovery measured on a 7-point Likert scale ranging from "completely recovered" to "worse than ever" at 12 months. This study is designed to investigate the effectiveness of patient education compared with patient education combined with multimodal physiotherapy. If patient education and multimodal physiotherapy applied at an early stage of Patellofemoral Pain Syndrome proves effective, it may serve as a basis for optimising the clinical pathway for those suffering from the condition, where specific emphasis can be placed on early diagnosis and early treatment. clinicaltrials.gov reference: NCT01438762.


Changes in beliefs about medications during long-term care for ischemic heart disease.
Beliefs such as self-reported low necessity for medications and high concerns about medications are associated with nonadherence. Changes in these beliefs during long-term care have not previously been studied and were evaluated for purposes of this analysis. From January 2006 through September 2007, patients at 41 hospitals who met entry criteria for the CRUSADE Quality Improvement Initiative were consented for participation in a longitudinal follow-up survey study called MAINTAIN. The patients completed The Beliefs about Medicines Questionnaire at 3 and 12 months postdischarge to assess necessity and concerns related to heart medications. Internal reliability and construct validity of the survey were evaluated at both time points. Changes in necessity and concern scores from 3 to 12 months were determined, and factors associated with negative changes were explored. A total of 812 patients completed both surveys. Internal reliability and construct validity were good. From 3 to 12 months, only 9.2% of patients shifted from a high to low necessity score; however, 20.7% of patients shifted from a low to high concern score. Factors found to be statistically significantly and independently associated with increased concern were the perception that the provider did not listen carefully to the patient (odds ratio [OR] 2.63, 95% CI 1.49-4.76), depression at 12 months (OR 2.95, 95% CI 1.57-5.55), hospital discharge with > or =7 medications (OR 1.71, 95% CI 1.07-2.74), and not receiving a medication list/instructions at hospital discharge (OR 1.69, 95% CI 1.05, 2.78). Factors associated with decreased necessity included not having a cardiologist (OR 2.26, 95% CI 1.34-3.83) and nonpersistence at 12 months with lipid-lowering medication (OR 1.85, 95% CI 1.01-3.45). One third of the patients reported a negative change in their beliefs about heart medications. Although some changes were observed in perceived necessity, negative changes in concerns occurred in 1 of every 5 patients. Interventions that address these concerns may be useful in improving adherence and clinical outcomes.


Using Labour Force Survey and census data to generate denominators for occupational injury rates: an application and expansion of Haggar-Guénette's method.
The calculation of rates of occupational injury claims is essential to identify groups at high risk, yet limitations of denominator data have often restricted our capacity to do this. Haggar-Guénette's method of using Statistics Canada's data on paid workers from the Labour Force Survey as denominators has been expanded by incorporating information from the Census. The method is illustrated by calculating denominators for male construction industry workers within the province of Ontario. Information for paid workers employed in construction occupations was derived from the Labour Force Survey to produce denominators for those aged 15-64, overall and by 10-year age groups. Census data on the distribution of construction occupational roles were applied to produce denominators both overall, then simultaneously by age and by occupational role. Advantages and disadvantages, including the limitations or biases due to the differing sources for denominators and numerators are identified.


Diring Yuriakh: A lower paleolithic site in central Siberia.
Lower Paleolithic artifacts have been recovered from a single occupation surface within stratified deposits at Diring Yuriakh, an archaeological site in central Siberia. Thermoluminescence age estimates from eolian sediments indicate that the cultural horizon is greater than 260,000 years old. Diring Yuriakh is an order of magnitude older than documented Paleolithic sites in Siberia and is important for understanding the timing of human expansion into the far north, early adaptations to cold climates, and the peopling of the Americas.


Patients' perceptions of their experiences with nurse-patient communication in oncology settings: A focused ethnographic study.
The nursing shortage and its impact on patient care are well-documented global issues. Patients living with cancer as a chronic illness have many psychosocial problems and often lack adequate support as a result of ineffective nurse-patient communication. A review of the literature on factors influencing the delivery of psychosocial care to cancer patients indicates that the delivery of psychosocial care in routine cancer nursing within a biomedical healthcare system has not been widely explored. To explore patients' perceptions of their experiences with nurse-patient communication in an oncological clinical environment. A focused ethnographic study was undertaken in two oncology wards of a hospital in Hong Kong. Data were collected through observations of the ward environment, the activities and instances of nurse-patient communication, semi-structured interviews with patients, and a review of nursing documents. Two main themes were identified: 1. Nurses' workload and the environment and 2. Nurse-patient partnership and role expectations. Within these two themes were related subthemes on: Sympathy for the busy nurses; Prioritizing calls to the nurses; Partnership through relationship; Nurses' role in psychosocial care; and Reduction of psychosocial concerns through physical care. Many cancer patients do not expect to receive psychosocial care in the form of emotional talks or counseling from busy nurses, but appreciate the attention paid by nurses to their physiological and physical needs. Nurse-patient partnerships in cancer care may reduce the potential workload of nurses. The psychosocial needs of cancer patients could be optimized by providing good physical care through effective communication within a time-constrained oncology setting.


Retention and viral suppression in a cohort of HIV patients on antiretroviral therapy in Zambia: Regionally representative estimates using a multistage-sampling-based approach.
Although the success of HIV treatment programs depends on retention and viral suppression, routine program monitoring of these outcomes may be incomplete. We used data from the national electronic medical record (EMR) system in Zambia to enumerate a large and regionally representative cohort of patients on treatment. We traced a random sample with unknown outcomes (lost to follow-up) to document true care status and HIV RNA levels. On 31 July 2015, we selected facilities from 4 provinces in 12 joint strata defined by facility type and province with probability proportional to size. In each facility, we enumerated adults with at least 1 clinical encounter after treatment initiation in the previous 24 months. From this cohort, we identified lost-to-follow-up patients (defined as 90 or more days late for their last appointment), selected a random sample, and intensively reviewed their records and traced them via phone calls and in-person visits in the community. In 1 of 4 provinces, we also collected dried blood spots (DBSs) for plasma HIV RNA testing. We used inverse probability weights to incorporate sampling outcomes into Aalen-Johansen and Cox proportional hazards regression to estimate retention and viremia. We used a bias analysis approach to correct for the known inaccuracy of plasma HIV RNA levels obtained from DBSs. From a total of 64 facilities with 165,464 adults on ART, we selected 32 facilities with 104,966 patients, of whom 17,602 (17%) were lost to follow-up: Those lost to follow-up had median age 36 years, 60% were female (N = 11,241), they had median enrollment CD4 count of 220 cells/μl, and 38% had WHO stage 1 clinical disease (N = 10,690). We traced 2,892 (16%) and found updated outcomes for 2,163 (75%): 412 (19%) had died, 836 (39%) were alive and in care at their original clinic, 457 (21%) had transferred to a new clinic, 255 (12%) were alive and out of care, and 203 (9%) were alive but we were unable to determine care status. Estimates using data from the EMR only suggested that 42.7% (95% CI 38.0%-47.1%) of new ART starters and 72.3% (95% CI 71.8%-73.0%) of all ART users were retained at 2 years. After incorporating updated data through tracing, we found that 77.3% (95% CI 70.5%-84.0%) of new initiates and 91.2% (95% CI 90.5%-91.8%) of all ART users were retained (at original clinic or transferred), indicating that routine program data underestimated retention in care markedly. In Lusaka Province, HIV RNA levels greater than or equal to 1,000 copies/ml were present in 18.1% (95% CI 14.0%-22.3%) of patients in care, 71.3% (95% CI 58.2%-84.4%) of lost patients, and 24.7% (95% CI 21.0%-29.3%). The main study limitations were imperfect response rates and the use of self-reported care status. In this region of Zambia, routine program data underestimated retention, and the point prevalence of unsuppressed HIV RNA was high when lost patients were accounted for. Viremia was prevalent among patients who unofficially transferred: Sustained engagement remains a challenge among HIV patients in Zambia, and targeted sampling is an effective strategy to identify such gaps in the care cascade and monitor programmatic progress.


Amontonian frictional behaviour of nanostructured surfaces.
With nanotextured surfaces and interfaces increasingly being encountered in technological and biomedical applications, there is a need for a better understanding of frictional properties involving such surfaces. Here we report friction measurements of several nanostructured surfaces using an Atomic Force Microscope (AFM). These nanostructured surfaces provide well defined model systems on which we have tested the applicability of Amontons' laws of friction. Our results show that Amontonian behaviour is observed with each of the surfaces studied. However, no correlation has been found between measured friction and various surface roughness parameters such as average surface roughness (R(a)) and root mean squared (rms) roughness. Instead, we propose that the friction coefficient may be decomposed into two contributions, i.e., μ = μ(0) + μ(g), with the intrinsic friction coefficient μ(0) accounting for the chemical nature of the surfaces and the geometric friction coefficient μ(g) for the presence of nanotextures. We have found a possible correlation between μ(g) and the average local slope of the surface nanotextures.


Entropy in an expanding universe.
The question of how the observed evolution of organized structures from initial chaos in the expanding universe can be reconciled with the laws of statistical mechanics is studied, with emphasis on effects of the expansion and gravity. Some major sources of entropy increase are listed. An expanding "causal" region is defined in which the entropy, though increasing, tends to fall further and further behind its maximum possible value, thus allowing for the development of order. The related questions of whether entropy will continue increasing without limit in the future, and whether such increase in the form of Hawking radiation or radiation from positronium might enable life to maintain itself permanently, are considered. Attempts to find a scheme for preserving life based on solid structures fail because events such as quantum tunneling recurrently disorganize matter on a very long but fixed time scale, whereas all energy sources slow down progressively in an expanding universe. However, there remains hope that other modes of life capable of maintaining themselves permanently can be found.


Severe neurotoxic envenoming by the Malayan krait Bungarus candidus (Linnaeus): response to antivenom and anticholinesterase.
Five patients were bitten by the Malayan krait Bungarus candidus (Linnaeus) in eastern Thailand or north western Malaya. Two patients were not envenomed but the other three developed generalised paralysis which progressed to respiratory paralysis in two cases, one of which ended fatally. One patient showed parasympathetic abnormalities. Anticholinesterase produced a dramatic improvement in one patient. Another patient probably benefited from paraspecific antivenom. The efficacy of antivenoms and adjuvants such as anticholinesterases in patients with neurotoxic envenoming requires further study.


A functional "knockout" of human keratin 14.
The importance of keratins and other intermediate filaments in the maintenance of tissue structure is emphasized by the discovery that many hereditary skin-blistering diseases are caused by mutations in keratin genes. Here, we describe a situation in which keratin 14 (K14) is missing altogether in the epidermis: A homozygous 2-nucleotide deletion in exon I of the K14 gene causes premature termination of the mRNA transcripts upstream from the start of the rod domain and results in a K14 null phenotype. In this individual no keratin intermediate filaments are visible in basal epidermal cells, although filaments are present in the upper layers of the epidermis. No compensating keratin expression is detected in vivo, and K14 mRNA is down-regulated. The individual, diagnosed as Köbner (generalized) EBS, suffers from severe widespread keratinocyte fragility and blistering at many body sites, but although the phenotype is severe, it is not lethal. This K14-/- phenotype confirms that only one K14 gene is expressed in human epidermis and provides an important model system for examining the interdependence of different keratin filament systems and their associated structures in the skin.


Evaluation of the effect of uremia on peritoneal permeability in an experimental model of continuous ambulatory peritoneal dialysis in anephric rats.
Anephric rats were maintained on continuous ambulatory peritoneal dialysis (CAPD). Peritoneal permeability was assessed during a standard 4-hour peritoneal equilibration test (PET) performed with Dianeal 3.86% (Baxter Healthcare, Deerfield, Illinois, U.S.A.). The effect of uremia on peritoneal permeability was evaluated in an experimental protocol in which each animal served as its own control. In each rat, PET1 (control) was performed before removal of kidneys and PET2 (uremia) was performed four days after removal of kidneys. Net ultrafiltration during a 4-hour exchange with Dianeal 3.86% was higher during PET1 (3.8 +/- 2.3 mL) than during PET2 (-1.3 +/- 3.3 mL), p < 0.05. Peritoneal permeability to urea and glucose was similar in both series. Transperitoneal equilibration of creatinine concentration was faster in uremic animals: D/P at 4 hours was 0.94 +/- 0.06 during PET2 versus 0.77 +/- 0.08 during PET1, p < 0.001. The opposite difference was seen for total protein: D/Px 1000 after a 4-hour dwell was 51.4 +/- 19.8 during PET2 versus 70.3 +/- 12.9 during PET1, p < 0.05. Our results show that uremia modifies the permeability of the peritoneum to both water and solutes.


Directed glial differentiation and transdifferentiation for neural tissue regeneration.
Glial cells which are indispensable for the central nervous system development and functioning, are proven to be vulnerable to a harmful influence of pathological cues and tissue misbalance. However, they are also highly sensitive to both in vitro and in vivo modulation of their commitment, differentiation, activity and even the fate-switch by different types of bioactive molecules. Since glial cells (comprising macroglia and microglia) are an abundant and heterogeneous population of neural cells, which are almost uniformly distributed in the brain and the spinal cord parenchyma, they all create a natural endogenous reservoir of cells for potential neurogenerative processes required to be initiated in response to pathophysiological cues present in the local tissue microenvironment. The past decade of intensive investigation on a spontaneous and enforced conversion of glial fate into either alternative glial (for instance from oligodendrocytes to astrocytes) or neuronal phenotypes, has considerably extended our appreciation of glial involvement in restoring the nervous tissue cytoarchitecture and its proper functions. The most effective modulators of reprogramming processes have been identified and tested in a series of pre-clinical experiments. A list of bioactive compounds which are potent in guiding in vivo cell fate conversion and driving cell differentiation includes a selection of transcription factors, microRNAs, small molecules, exosomes, morphogens and trophic factors, which are helpful in boosting the enforced neuro-or gliogenesis and promoting the subsequent cell maturation into desired phenotypes. Herein, an issue of their utility for a directed glial differentiation and transdifferentiation is discussed in the context of elaborating future therapeutic options aimed at restoring the diseased nervous tissue.


Discovery of a novel species, Theileria haneyi n. sp., infective to equids, highlights exceptional genomic diversity within the genus Theileria: implications for apicomplexan parasite surveillance.
A novel apicomplexan parasite was serendipitously discovered in horses at the United States - Mexico border. Phylogenetic analysis based on 18S rDNA showed the erythrocyte-infective parasite to be related to, but distinct from, Theileria spp. in Africa, the most similar taxa being Theileria spp. from waterbuck and mountain zebra. The degree of sequence variability observed at the 18S rDNA locus also suggests the likely existence of additional cryptic species. Among described species, the genome of this novel equid Theileria parasite is most similar to that of Theileria equi, also a pathogen of horses. The estimated divergence time between the new Theileria sp. and T. equi, based on genomic sequence data, is greater than 33 million years. Average protein sequence divergence between them, at 23%, is greater than that of Theileria parva and Theileria annulata proteins, which is 18%. The latter two represent highly virulent Theileria spp. of domestic cattle, as well as of African and Asian wild buffalo, respectively, which differ markedly in pathology, host cell tropism, tick vector and geographical distribution. The extent of genome-wide sequence divergence, as well as significant morphological differences, relative to T. equi justify the classification of Theileria sp. as a new taxon. Despite the overall genomic divergence, the nine member equi merozoite antigen (EMA) superfamily, previously found as a multigene family only in T. equi, is also present in the novel parasite. Practically, significant sequence divergence in antigenic loci resulted in this undescribed Theileria sp. not being detectable using currently available diagnostic tests. Discovery of this novel species infective to equids highlights exceptional diversity within the genus Theileria, a finding with serious implications for apicomplexan parasite surveillance.


Multimodal registration of retinal images using self organizing maps.
In this paper, an automatic method for registering multimodal retinal images is presented. The method consists of three steps: the vessel centerline detection and extraction of bifurcation points only in the reference image, the automatic correspondence of bifurcation points in the two images using a novel implementation of the self organizing maps and the extraction of the parameters of the affine transform using the previously obtained correspondences. The proposed registration algorithm was tested on 24 multimodal retinal pairs and the obtained results show an advantageous performance in terms of accuracy with respect to the manual registration.


Characteristics and outcome of 16 periprosthetic shoulder joint infections.
Shoulder arthroplasties are increasingly performed, but data on periprosthetic joint infections (PJI) in this anatomical position are limited. We retrospectively investigated the characteristics and outcome of shoulder PJI after primary arthroplasty from 1998 to 2010 in a single centre. Periprosthetic joint infection was defined as periprosthetic purulence, presence of sinus tract or microbial growth. A Kaplan-Meier survival method was used to estimate relapse-free survival of prosthesis. From 1,571 primary shoulder prostheses, we evaluated 16 patients with a PJI at different stages, i.e, early (n = 4), delayed (n = 6) and late (n = 6) infections. The median patient age was 67 (range 53-86) years, and 69 % were females. The most commonly isolated microorganism was Propionibacterium acnes in 38 % of patients (monobacterial in four and polymicrobial in two patients). In 14 of the 16 patients, surgical interventions consisting of debridement and implant retention (6 patients), exchange (7) and explantation (1) were performed. Four patients had a relapse of infection with P. acnes (n = 3) or Bacteroides fragilis (n = 1). The relapse-free survival of the prosthesis was 75 % (95 % confidence interval 46-90 %) after 1 and 2 years, 100 % in six patients following the treatment algorithm for hip and knee PJI and 60 % in 10 patients not followed up. All but one of the relapses were previously treated without exchange of the prosthesis. As recommended for hip and knee PJI, we suggest treating shoulder PJI with a low-grade infection by microorganisms such as P. acnes with an exchange of the prosthesis. Cohort studies are needed to verify our results.


Dimension selection for feature selection and dimension reduction with principal and independent component analysis.
This letter is concerned with the problem of selecting the best or most informative dimension for dimension reduction and feature extraction in high-dimensional data. The dimension of the data is reduced by principal component analysis; subsequent application of independent component analysis to the principal component scores determines the most nongaussian directions in the lower-dimensional space. A criterion for choosing the optimal dimension based on bias-adjusted skewness and kurtosis is proposed. This new dimension selector is applied to real data sets and compared to existing methods. Simulation studies for a range of densities show that the proposed method performs well and is more appropriate for nongaussian data than existing methods.


Radioiodine-induced oxidative stress in patients with differentiated thyroid carcinoma and effect of supplementation with vitamins C and E and selenium (antioxidants).
The objective of this study, in addition to confirming that therapy with 131I causes oxidative stress, was to evaluate the effect of supplementation with vitamins C and E and selenium on this phenomenon by measuring plasma 8-epi-PGF2a, a marker of lipid peroxidation. Forty patients with thyroid cancer submitted to thyroidectomy, who received 3.7 GBq 131I after levothyroxine withdrawal, were selected; 20 patients did not receive (control group) and 20 patients received (intervention group) daily supplementation consisting of 2000 mg vitamin C, 1000 mg vitamin E and 400 µg selenium for 21 days before 131I. Plasma 8-epi-PGF2a was measured immediately before and 2 and 7 days after 131I. A significant increase in plasma 8-epi-PGF2a after 131I was observed in the two groups. The concentrations of 8-epi-PGF2α were significantly higher in the control group before and 2 and 7 days after 131I. The percentage of patients with elevated 8-epi-PGF2α was also significantly higher in the control group before and after 131I. Furthermore, the increase (percent) in 8-epi-PGF2α was significantly greater in the control group (average of 112.3% versus 56.3%). Only two patients (10%) reported side effects during supplementation. Ablation with 131I causes oxidative stress which can be minimized by the use of antioxidants.


A neuro-mimetic dynamic scheduling algorithm for control: analysis and applications.
A simple neuronal network model of the baroreceptor reflex is analyzed. From a control perspective, the analysis suggests a dynamic scheduled control mechanisms by which the baroreflex may perform regulation of the blood pressure. The main objectives of this work are to investigate the static and dynamic response characteristics of the single neurons and the network, to analyze the neuromimetic dynamic scheduled control function of the model, and to apply the algorithm to nonlinear process control problems. The dynamic scheduling activity of the network is exploited in two control architectures. Control structure I is drawn directly from the present model of the baroreceptor reflex. An application of this structure for level control in a conical tank is described. Control structure II employs an explicit set point to determine the feedback error. The performance of this control structure is illustrated on a nonlinear continuous stirred tank reactor with van de Vusse kinetics. The two case studies validate the dynamic scheduled control approach for nonlinear process control applications.


The discrimination of facial sex in developmental prosopagnosia.
Developmental prosopagnosia (DP) is a neurodevelopmental condition characterised by difficulties recognising and discriminating faces. It is currently unclear whether the perceptual impairments seen in DP are restricted to identity information, or also affect the perception of other facial characteristics. To address this question, we compared the performance of 17 DPs and matched controls on two sensitive sex categorisation tasks. First, in a morph categorisation task, participants made binary decisions about faces drawn from a morph continuum that blended incrementally an average male face and an average female face. We found that judgement precision was significantly lower in the DPs than in the typical controls. Second, we used a sex discrimination task, where female or male facial identities were blended with an androgynous average face. We manipulated the relative weighting of each facial identity and the androgynous average to create four levels of signal strength. We found that DPs were significantly less sensitive than controls at each level of difficulty. Together, these results suggest that the visual processing difficulties in DP extend beyond the extraction of facial identity and affects the extraction of other facial characteristics. Deficits of facial sex categorisation accord with an apperceptive characterisation of DP.


Systematic delays in antibiotic administration in the emergency department for adult patients admitted with pneumonia.
The authors sought to determine the contribution of delays in care on time to antibiotics for patients admitted from the emergency department (ED) with pneumonia and to identify patients at risk for delayed antibiotics. This was a retrospective cohort study of patients admitted to the Hospital of the University of Pennsylvania (HUP) and to Pennsylvania Presbyterian Hospital (Presbyterian) with an admission diagnosis of pneumonia in 2004. A total of 393 patients were included. Ninety percent had antibiotics documented as given in the ED. Eighty-three (43%) of 209 at HUP and 104 (64%) of 161 patients at Presbyterian received antibiotics within four hours. Patients who received antibiotics more than four hours after ED arrival experienced longer waits for radiograph orders (HUP, 54 min [95% confidence interval {CI} = 33 to 76 min]; Presbyterian, 43 min [95% CI = 29 to 58 min]), for radiograph performance (HUP, 21 min [95% CI = 4 to 39 min], Presbyterian, 24 min [95% CI = 8 to 47 min]), for antibiotic orders (HUP, 56 min [95% CI = 38 to 95 min]; Presbyterian, 67 min [95% CI = 33 to 103 min]), and for antibiotic administration (HUP, 28 min [95% CI = 17 to 39 min]; Presbyterian, 30 min [95% CI = 21 to 38 min]). Patients with lower severity scores (p = 0.005) and patients with nonclassic clinical presentations for pneumonia were at increased risk for delayed antibiotics (odds ratio, 2.2; 95% CI = 1.1 to 4.4). Antibiotic delays for patients admitted with pneumonia occur across multiple care processes. Less severely ill patients and patients with nonclassic presentations are at higher risk for delayed antibiotic administration. Hospitals should consider performing a similar analysis to evaluate hospital-specific and patient-specific care delays.


Host-pathogen interactions and virulence-associated genes during Candida albicans oral infections.
Oral infections with Candida albicans are very common diseases in even only mildly immunocompromised patients. By using genome-wide microarrays, in vitro infection models and samples from patients with pseudomembranous candidiasis, several genes have been identified which encode known and unknown fungal factors associated with oral infection. The expression of selected genes has been investigated via qRT-PCR in both in vitro models and in vivo samples from patients. Several lines of evidence suggest that fungal morphology plays a key role in adhesion to and invasion into oral epithelial cells and mutants lacking regulators of hyphal formation are attenuated in their ability to invade and damage epithelial cells. Adhesion is mediated by hyphal-associated factors such as Hwp1 and the Als adhesin family. Hyphal formation facilitates epithelial invasion via two routes: active penetration and induced endocytosis. While induced endocytosis is predominantly mediated by the adhesin and invasin Als3, active penetration seems to be supported by hydrolase activity and mechanical pressure. Expression profiles reflect the morphological switch and an adaptive response to neutral pH, non-glucose carbon sources, and nitrosative stress.


High-resolution mapping of the X-linked hypohidrotic ectodermal dysplasia (EDA) locus.
The X-linked hypohidrotic ectodermal dysplasia (EDA) locus has been previously localized to the subchromosomal region Xq11-q21.1. We have extended our previous linkage studies and analyzed linkage between the EDA locus and 10 marker loci, including five new loci, in 41 families. Four of the marker loci showed no recombination with the EDA locus, and six other loci were also linked to the EDA locus with recombination fractions of .009-.075. Multipoint analyses gave support to the placement of the PGK1P1 locus proximal to the EDA locus and the DXS453 and PGK1 loci distal to EDA. Further ordering of the loci could be inferred from a human/rodent somatic cell hybrid derived from an affected female with EDA and an X;9 translocation and from studies of an affected male with EDA and a submicroscopic deletion. Three of the proximal marker loci, which showed no recombination with the EDA locus, when used in combination, were informative in 92% of females. The closely linked flanking polymorphic loci DXS339 and DXS453 had heterozygosities of 72% and 76%, respectively, and when used jointly, they were doubly informative in 52% of females. The human DXS732 locus was defined by a conserved mouse probe pcos169E/4 (DXCrc169 locus) that cosegregates with the mouse tabby (Ta) locus, a potential homologue to the EDA locus. The absence of recombination between EDA and the DXS732 locus lends support to the hypothesis that the DXCrc169 locus in the mouse and the DXS732 locus in humans may contain candidate sequences for the Ta and EDA genes, respectively.


Structural and functional characterization of mycobactericidal ubiquitin-derived peptides in model and bacterial membranes.
The mycobactericidal properties of macrophages include the delivery of bacteria to a hydrolytic lysosome enriched in bactericidal ubiquitin-derived peptides (Ub-peptides). To improve our understanding of interactions of ubiquitin-derived peptides with mycobacteria, we further characterized the structure and function of bactericidal Ub-peptide Ub2. We found that Ub2 adopts a β-sheet conformation in the context of sodium dodecyl sulfate micelles and phospholipid (1:1 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine) vesicles that was dependent upon the primary sequence of the peptide. Point mutations in Ub2 that reduced the net charge of the peptide decreased Ub2 bactericidal activity. We investigated Ub-peptide function in the context of model membranes and intact bacteria. Differential scanning calorimetry analysis demonstrated that Ub2 inserts into and perturbs model phospholipid vesicles. In addition, we demonstrate that Ub2 disrupts the integrity of the mycobacterial membrane, equilibrates the transmembrane potential, and is localized within both the mycobacterial membrane and cytoplasm of treated bacteria. Finally, we identified additional bactericidal Ub-peptides and characterized their activity and structure. This study provides new insight into the mycobactericidal mechanisms of Ub-peptides.


Ouabain induces endocytosis of plasmalemmal Na/K-ATPase in LLC-PK1 cells by a clathrin-dependent mechanism.
We have demonstrated that ouabain causes dose- and time-dependent decreases in (86)Rb uptake in porcine proximal tubular (LLC-PK1) cells. The present study addresses the molecular mechanisms involved in this process. Studies were performed with cultured LLC-PK1 and Src family kinase deficient (SYF) cells. We found that 50 nmol/L ouabain applied to the basal, but not apical, aspect for 12 hours caused decreases in the plasmalemmal Na/K-ATPase. This loss of plasmalemmal Na/K-ATPase reverses completely within 12 to 24 hours after removal of ouabain. Ouabain also increased the Na/K-ATPase content in both early and late endosomes, activated phosphatidylinositol 3-kinase (PI(3)K), and also caused a translocation of some Na/K-ATPase to the nucleus. Immunofluorescence demonstrated that the Na/K-ATPase colocalized with clathrin both before and after exposure to ouabain, and immunoprecipitation experiments confirmed that ouabain stimulated interactions among the Na/K-ATPase, adaptor protein-2 (AP-2), and clathrin. Potassium (K) depletion, chlorpromazine, or PI(3)K inhibition all significantly attenuated this ouabain-induced endocytosis. Inhibition of the ouabain-activated signaling process through Src by 4-Amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2) significantly attenuated ouabain-induced endocytosis. Moreover, experiments performed in SYF cells demonstrated that ouabain induced increases in the endocytosis of the Na/K-ATPase when Src was reconstituted (SYF+), but not in the Src-deficient (SYF-) cells. These data demonstrate that ouabain stimulates a clathrin-dependent endocytosis pathway that translocates the Na/K-ATPase to intracellular compartments, thus suggesting a potential role of endocytosis in ouabain-induced signal transduction as well as proximal tubule sodium handling.


CYP2C8 Is a Novel Target of Peroxisome Proliferator-Activated Receptor α in Human Liver.
Human cytochrome P450 (CYP) 2C enzymes metabolize ∼30% of clinically prescribed drugs and various environmental chemicals. CYP2C8, an important member of this subfamily, metabolizes the anticancer drug paclitaxel, certain antidiabetic drugs, and endogenous substrates, including arachidonic acid, to physiologically active epoxyeicosatrienoic acids. Previous studies from our laboratory showed that microRNA 107 (miR107) and microRNA 103 downregulate CYP2C8 post-transcriptionally. miR107 is located in intron 5 of the pantothenate kinase 1 (PANK1) gene. p53 has been reported to coregulate the induction of PANK1 and miR107. Here, we examine the possible downregulation of CYP2C8 by drugs capable of inducing miR107. Hypolipidemic drugs, such as bezafibrate, known activators of the peroxisome proliferator-activated receptor α (PPARα), induce both the PANK1 gene and miR107 (∼2.5-fold) in primary human hepatocytes. Surprisingly, CYP2C8 mRNA and protein levels were induced by bezafibrate. CYP2C8 promoter activity was increased by ectopic expression of PPARα in HepG2 cells, with a further increase after bezafibrate (∼18-fold), 4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio acetic acid (∼10-fold) treatment, or the antidiabetic drug rosiglitazone, all known PPAR activators. Promoter sequence analyses, deletion studies, mutagenesis studies, and electrophoretic mobility shift assays identified a PPARα response element located at position -2109 base pair relative to the translation start site of CYP2C8. Chromatin immunopreciptation assay analysis confirmed recruitment of PPARα to this PPARα response element after bezafibrate treatment of human hepatocytes. Thus, we show for the first time that CYP2C8 is transcriptionally regulated by PPARα, suggesting the potential for drug-drug interactions due to upregulation of CYP2C8 by PPAR activators.


A new chimeric triple reporter fusion protein as a tool for in vitro and in vivo multimodal imaging to monitor the development of African trypanosomes and Leishmania parasites.
Trypanosomiases and leishmaniases, caused by a group of related protist parasites, are Neglected Tropical Diseases currently threatening >500 million people worldwide. Reporter proteins have revolutionised the research on infectious diseases and have opened up new advances in the understanding of trypanosomatid-borne diseases in terms of both biology, pathogenesis and drug development. Here, we describe the generation and some applications of a new chimeric triple reporter fusion protein combining the red-shifted firefly luciferase PpyREH9 and the tdTomato red fluorescent protein, fused by the TY1 tag. Expressed in both Trypanosoma brucei brucei and Leishmania major transgenic parasites, this construct was successfully assessed on different state-of-the-art imaging technologies, at different scales ranging from whole organism to cellular level, both in vitro and in vivo in murine models. For T. b. brucei, the usefulness of this triple marker to monitor the entire parasite cycle in both tsetse flies and mice was further demonstrated. This stable reporter allows to qualitatively and quantitatively scrutinize in real-time several crucial aspects of the parasite's development, including the development of African trypanosomes in the dermis of the mammalian host. We briefly discuss developments in bio-imaging technologies and highlight how we could improve our understanding of parasitism by combining the genetic engineering of parasites to the one of the hosting organisms in which they complete their developmental program.


Prostate cancer: correlation of MR imaging and MR spectroscopy with pathologic findings after radiation therapy-initial experience.
To prospectively evaluate magnetic resonance (MR) imaging and MR spectroscopy for depiction of local prostate cancer recurrence after external-beam radiation therapy, with step-section pathologic findings as the standard of reference. Study received institutional approval, and written informed consent was obtained. Study was compliant with Health Insurance Portability and Accountability Act. Sextant biopsy, digital rectal examination, MR imaging, MR spectroscopy, and salvage radical prostatectomy with step-section pathologic examination were performed in nine patients with increasing prostate-specific antigen levels after external-beam radiation therapy. MR imaging criterion for tumor was a focal nodular region of reduced signal intensity at T2-weighted imaging. MR spectroscopic criteria for tumor were voxels with choline (Cho) plus creatine (Cr) to citrate (Cit) ratio ([Cho + Cr]/Cit) of at least 0.5 or voxels with detectable Cho and no Cit in the peripheral zone. Sensitivity and specificity of sextant biopsy, digital rectal examination, MR imaging, and MR spectroscopy were determined by using a prostate sextant as the unit of analysis. For feature analysis, MR imaging and MR spectroscopic findings were correlated with step-section pathologic findings. MR imaging and MR spectroscopy showed estimated sensitivities of 68% and 77%, respectively, while sensitivities of biopsy and digital rectal examination were 48% and 16%, respectively. MR spectroscopy appears to be less specific (78%) than the other three tests, each of which had a specificity higher than 90%. MR spectroscopic feature analysis showed that a metabolically altered benign gland could be falsely identified as tumor by using MR spectroscopic criteria; further analysis of MR spectroscopic features did not lead to improved MR spectroscopic criteria for recurrent tumor. In summary, MR imaging and MR spectroscopy may be more sensitive than sextant biopsy and digital rectal examination for sextant localization of cancer recurrence after external-beam radiation therapy.


Replication of breast cancer susceptibility loci in whites and African Americans using a Bayesian approach.
Genome-wide association studies (GWAS) and candidate gene analyses have led to the discovery of several dozen genetic polymorphisms associated with breast cancer susceptibility, many of which are considered well-established risk factors for the disease. Despite attempts to replicate these same variant-disease associations in African Americans, the evaluable populations are often too small to produce precise or consistent results. We estimated the associations between 83 previously identified single nucleotide polymorphisms (SNPs) and breast cancer among Carolina Breast Cancer Study (1993-2001) participants using maximum likelihood, Bayesian, and hierarchical methods. The selected SNPs were previous GWAS hits (n = 22), near-hits (n = 19), otherwise well-established risk loci (n = 5), or located in the same genes as selected variants (n = 37). We successfully replicated 18 GWAS-identified SNPs in whites (n = 2,352) and 10 in African Americans (n = 1,447). SNPs in the fibroblast growth factor receptor 2 gene (FGFR2) and the TOC high mobility group box family member 3 gene (TOX3) were strongly associated with breast cancer in both races. SNPs in the mitochondrial ribosomal protein S30 gene (MRPS30), mitogen-activated protein kinase kinase kinase 1 gene (MAP3K1), zinc finger, MIZ-type containing 1 gene (ZMIZ1), and H19, imprinted maternally expressed transcript gene (H19) were associated with breast cancer in whites, and SNPs in the estrogen receptor 1 gene (ESR1) and H19 gene were associated with breast cancer in African Americans. We provide precise and well-informed race-stratified odds ratios for key breast cancer-related SNPs. Our results demonstrate the utility of Bayesian methods in genetic epidemiology and provide support for their application in small, etiologically driven investigations.


Prognostic significance of cell cycle- and invasion-related molecular markers and genomic instability in primary carcinoma of the vagina.
This study aimed to analyze the prognostic value of DNA content and biological markers for cell cycle regulation and invasion in primary carcinoma of the vagina (PCV). Seventy-two consecutive patients with PCV, categorized as short-term (≤ 2 years) and long-term (≥ 8 years) survivors, were evaluated for DNA content by image cytometry, and for expression of p53, p21, cyclin A, Ki67, E-cadherin, and laminin-5γ2 chain by immunohistochemistry. The relationship between these biological markers and histopathological and clinical parameters was assessed. All PCV showed aneuploid DNA content. Most of the PCV patients showed no overexpression of p53 and high expression of p21, cyclin A, and Ki67. Loss or underexpression of E-cadherin was found in 94% (68/72) of PCV patients, and all patients showed immunopositivity for the laminin-5γ2 chain. Tumors with a vaginal longitudinal location in the lower third or in the entire vagina more often had overexpression of p53, high expression of Ki67 (P = 0.044), and underexpression of E-cadherin (P = 0.038), than tumors confined only to the upper third. Overexpression of p53 was significantly associated with short-term survival in the univariate analysis, but not in the multivariate analysis adjusted for age at diagnosis and tumor size. The expression level of some markers was related to tumor location, which might be indicative of different genesis. Overexpression of p53 was associated with short-term survival, but the only independent predictors of survival were age at diagnosis and tumor size.


First in vitro fertilization baby-this is how it happened.
On July 25, 1978, the first human was born following extracorporeal fertilization, an event that opened up a new medical science: expanding our knowledge of and developing novel treatments for infertility, radically changing the opportunities for families with inherited monogenic disorders, generating the new discipline of clinical embryology, and paving the way for studies into stem cell biology. In vitro fertilization (IVF), as it became known in its simplest form, went even further: it engaged the myriad of minds in human society. Not only were books written on the moral status of the human embryo, the ethics of IVF practice, and exercising governments on appropriate-which turned out to be disparate-regulation, it redefined family life! The prediction I made in 1985 that one day we may see five "parents" for one child became a reality quicker than we could have imagined at the time. The new medical science marched inexorably on in almost all countries of the world-a universal human plight at last had an opportunity for remedy: more than 10 million couples seeking a resolution to their infertility became parents, men who had no option to have their own genetic child became genetic fathers, and ever-increasing monogenic conditions were not being passed on to the next generation. The future may well bring to bear the opportunity for in vitro-developed viable gametes to generate successful pregnancies, and other "futuristic" opportunities for IVF science. But its story began over a century ago with seeking an understanding on how an egg matures and how to achieve successful fertilization-a fundamental scientific inquiry. It took one man to go beyond that scientific endeavor, to take head on a society unprepared and unwilling to accept human fertilization in vitro and unempathetic to the plight of the infertile; one man to see what prospects lay ahead for humanity should IVF become a reality, and for that man to battle every step of the way for nearly 2 decades to achieve that dream.


Role of the core DNA polymerase III subunits at the replication fork. Alpha is the only subunit required for processive replication.
The DNA polymerase III holoenzyme is composed of 10 subunits. The core of the polymerase contains the catalytic polymerase subunit, alpha, the proofreading 3'-->5' exonuclease, epsilon, and a subunit of unknown function, theta. The availability of the holoenzyme subunits in purified form has allowed us to investigate their roles at the replication fork. We show here that of the three subunits in the core polymerase, only alpha is required to form processive replication forks that move at high rates and that exhibit coupled leading- and lagging-strand synthesis in vitro. Taken together with previous data this suggests that the primary determinant of replication fork processivity is the interaction between another holoenzyme subunit, tau, and the replication fork helicase, DnaB.


Qigong for the treatment of tinnitus: a prospective randomized controlled study.
Tinnitus is a frequent disorder which is very difficult to treat. Qigong is a mindful exercise and an important constituent of traditional Chinese medical practice. Here we performed a randomized controlled trial to evaluate the effect of a Qigong intervention on patients with tinnitus. We hypothesized that especially tinnitus patients with somatosensoric components may benefit from the mind-body technique of Qigong. Eighty patients with tinnitus of at least 3 months duration were randomly assigned to an intervention group (n=40) consisting of 10 Qigong training sessions in 5 weeks or a waiting-list control group (n=40). Tinnitus severity was assessed with a visual analogue scale (VAS) and with a tinnitus questionnaire (TBF-12) before treatment, immediately after treatment, and 1 and 3 months after treatment. Qigong did not cause any side effects and was completed by 80% of the assigned patients. Compared with the control group, Qigong participants experienced improvement in tinnitus severity, as reflected by a significant reduction in both the VAS and the TBF-12. In the subgroup of patients with somatosensoric tinnitus, Qigong effects were more pronounced, resulting in a highly significant improvement in both scales compared to the waiting-list group. These findings suggest that Qigong interventions could be a useful complement to the therapeutic management of patients with tinnitus and especially for those with somatosensoric components. Satisfaction with the intervention, a high degree of completion, and stability of the effects for at least 3 months after the intervention further underscore the potential of Qigong in the treatment of tinnitus.


Predominant contribution of cis-regulatory divergence in the evolution of mouse alternative splicing.
Divergence of alternative splicing represents one of the major driving forces to shape phenotypic diversity during evolution. However, the extent to which these divergences could be explained by the evolving cis-regulatory versus trans-acting factors remains unresolved. To globally investigate the relative contributions of the two factors for the first time in mammals, we measured splicing difference between C57BL/6J and SPRET/EiJ mouse strains and allele-specific splicing pattern in their F1 hybrid. Out of 11,818 alternative splicing events expressed in the cultured fibroblast cells, we identified 796 with significant difference between the parental strains. After integrating allele-specific data from F1 hybrid, we demonstrated that these events could be predominately attributed to cis-regulatory variants, including those residing at and beyond canonical splicing sites. Contrary to previous observations in Drosophila, such predominant contribution was consistently observed across different types of alternative splicing. Further analysis of liver tissues from the same mouse strains and reanalysis of published datasets on other strains showed similar trends, implying in general the predominant contribution of cis-regulatory changes in the evolution of mouse alternative splicing.


Clinico-embryological submission of an unilateral anomalous presentation of axillary artery.
Descriptions of the variant arterial pattern of upper limb are not exceptional and are therefore frequently reported in anatomy archives. A noteworthy deviation from the usual branching pattern was observed unilaterally in a single cadaver. This unique division of axillary artery (AA) was present on the right side in an adult human cadaver of Indian origin. The first part of axillary artery gave off the superior thoracic and thoraco-acromial arteries. Just proximal to the upper border of pectoralis minor the AA was observed to divide into two trunks a medial and a lateral. The lateral trunk continued into the brachium as the usual axillary artery where as the medial trunk displayed the other branches deep and distal to the pectoralis minor muscle. The remarkable feature was the wide caliber of the axillary artery where it bifurcated into two branches. An attempt has been made to dwell upon the embryological basis of the present anomaly. The relevance of anomalous arterial pattern of upper limb (U.L.) is realized while performing percutaneous arterial venous catheter insertion into subclavian vein via the infraclavicular route. We advocate a meticulous familiarization of the anatomy of axillary artery and its topographical relationship to other neurovascular structures for the operating plastic surgeon, anesthetist and radiologist (Fig. 1, Ref. 12).


Highly Efficient Luminescent Metal-Organic Framework for the Simultaneous Detection and Removal of Heavy Metals from Water.
We have designed and synthesized an isoreticular series of luminescent metal-organic frameworks (LMOFs) by incorporating a strongly emissive molecular fluorophore and functionally diverse colinkers into Zn-based structures. The three-dimensional porous networks of LMOF-261, -262, and -263 represent a unique/new type of nets, classified as a 2-nodal, (4,4)-c net (mot-e type) with 4-fold, class IIIa interpenetration. All compounds crystallize in a body-centered tetragonal crystal system (space group I41/a). A systematic study has been implemented to analyze their interactions with heavy metals. LMOF-263 exhibits impressive water stability, high porosity, and strong luminescence, making it an excellent candidate as a fluorescent chemical sensor and adsorbent for aqueous contaminants. It is extremely responsive to toxic heavy metals at a parts per billion level (3.3 ppb Hg2+, 19.7 ppb Pb2+) and demonstrates high selectivity for heavy metals over light metals, with detection ratios of 167.4 and 209.5 for Hg2+/Ca2+ and Hg2+/Mg2+, respectively. Mixed-metal adsorption experiments also show that LMOF-263 selectively adsorbs Hg2+ over other heavy metal ions in addition to light metals. The Pb2+ KSV value for LMOF-263 (55,017 M-1) is the highest among LMOFs reported to date, and the Hg2+ KSV value is the second highest (459,446 M-1). LMOF-263 exhibits a maximum adsorption capacity of 380 mg Hg2+/g. The Hg2+ adsorption process follows pseudo-second-order kinetics, removing 99.1% of the metal within 30 min. An in situ XPS study provides insight to help understand the interaction mechanism between Hg2+ and LMOF-263. No other MOFs have demonstrated such a high performance in both the detection and the capture of Hg2+ from aqueous solution.


A Novel Report on the Compassionate Use of Baricitinib in Treating a Pediatric Patient With Severe Symptoms of COVID-19 Infection.
Since the outbreak of the pandemic coronavirus disease 2019 (COVID-19), there has been an increasing need for treatment to decrease morbidity and mortality of patients presenting with severe disease symptoms. There has been increasing evidence to suggest that the pathophysiological basis is a severe inflammatory response that resembles the cytokine release syndrome. Current strategies to counteract this involve modifiers of the immune response such as interleukin (IL)-6 receptor blockers and Janus kinase (JAK) inhibitors. An example of a JAK inhibitor is baricitinib. In this case, we present a 17-year-old female admitted with severe COVID-19 symptoms, who was placed on high-flow nasal cannula and started on azithromycin and hydroxychloroquine, which were standard of care at the time. Due to the worsening of symptoms, she was given baricitinib for compassionate use. There was a rapid improvement in clinical and imaging findings, and the patient was discharged from the hospital within 8 days of admission. This study is fascinating because there are very limited studies published on the benefits of baricitinib in managing patients with severe symptoms of COVID-19 especially in the pediatric population, and the rapidity in recovery time was remarkable.


Injuries of the trochanteric region: can analysis of radiographic indices help in prediction of recurrent osteoporotic hip fractures?
In a context of bone fragility, primitive and subsequent fractures are a growing problem in the industrialized countries where the mean age of the population is constantly increasing. Among the various factors that favor a fragility fracture, the most important is osteoporosis, a pathology that can be prevented through diagnostic screenings and treated by pharmacological and rehabilitative therapies. The aim of this study is to identify the subjects who are likely to have a higher risk of subsequent fractures of the trochanteric region through a retrospective radiographic evaluation of patients affected by low-energy trochanteric fractures and operated by intramedullary fixation between June 2013 and June 2015, so they can be targeted for prevention interventions. Three hundred and sixty-one patients yet alive were analyzed 2 years after surgery. Fifty-one (group 1), characterized by another contralateral trochanteric femoral fracture, were included. All subjects were retrospectively examined with the analysis of contralateral femur X-ray performed at the time of initial trauma in order to detect a condition of bone fragility and a predisposition to fractures by evaluating three radiographic indices (Singh index, Dorr's classification and Cortical Thickness Index). Patients of group 1 were compared to the other 310 patients (group 2) affected by isolated trochanteric fracture. Group 1 had all radiographic indices worse than group 2. The results observed suggest that orthopedists can use radiographic indices, in particular Cortical Thickness Index, as a valuable, simple and inexpensive screening tool for prevention of recurrent osteoporotic fractures.


Use of multivariate analysis to suggest a new molecular classification of colorectal cancer.
Molecular classification of colorectal cancer (CRC) is currently based on microsatellite instability (MSI), KRAS or BRAF mutation and, occasionally, chromosomal instability (CIN). Whilst useful, these categories may not fully represent the underlying molecular subgroups. We screened 906 stage II/III CRCs from the VICTOR clinical trial for somatic mutations. Multivariate analyses (logistic regression, clustering, Bayesian networks) identified the primary molecular associations. Positive associations occurred between: CIN and TP53 mutation; MSI and BRAF mutation; and KRAS and PIK3CA mutations. Negative associations occurred between: MSI and CIN; MSI and NRAS mutation; and KRAS mutation, and each of NRAS, TP53 and BRAF mutations. Some complex relationships were elucidated: KRAS and TP53 mutations had both a direct negative association and a weaker, confounding, positive association via TP53-CIN-MSI-BRAF-KRAS. Our results suggested a new molecular classification of CRCs: (1) MSI(+) and/or BRAF-mutant; (2) CIN(+) and/or TP53(-) mutant, with wild-type KRAS and PIK3CA; (3) KRAS- and/or PIK3CA-mutant, CIN(+) , TP53-wild-type; (4) KRAS(-) and/or PIK3CA-mutant, CIN(-) , TP53-wild-type; (5) NRAS-mutant; (6) no mutations; (7) others. As expected, group 1 cancers were mostly proximal and poorly differentiated, usually occurring in women. Unexpectedly, two different types of CIN(+) CRC were found: group 2 cancers were usually distal and occurred in men, whereas group 3 showed neither of these associations but were of higher stage. CIN(+) cancers have conventionally been associated with all three of these variables, because they have been tested en masse. Our classification also showed potentially improved prognostic capabilities, with group 3, and possibly group 1, independently predicting disease-free survival.


Inhibition of TNF receptor 1 internalization by adenovirus 14.7K as a novel immune escape mechanism.
The adenoviral protein E3-14.7K (14.7K) is an inhibitor of TNF-induced apoptosis, but the molecular mechanism underlying this protective effect has not yet been explained exhaustively. TNF-mediated apoptosis is initiated by ligand-induced recruitment of TNF receptor-associated death domain (TRADD), Fas-associated death domain (FADD), and caspase-8 to the death domain of TNF receptor 1 (TNFR1), thereby establishing the death-inducing signaling complex (DISC). Here we report that adenovirus 14.7K protein inhibits ligand-induced TNFR1 internalization. Analysis of purified magnetically labeled TNFR1 complexes from murine and human cells stably transduced with 14.7K revealed that prevention of TNFR1 internalization resulted in inhibition of DISC formation. In contrast, 14.7K did not affect TNF-induced NF-kappaB activation via recruitment of receptor-interacting protein 1 (RIP-1) and TNF receptor-associated factor 2 (TRAF-2). Inhibition of endocytosis by 14.7K was effected by failure of coordinated temporal and spatial assembly of essential components of the endocytic machinery such as Rab5 and dynamin 2 at the site of the activated TNFR1. Furthermore, we found that the same TNF defense mechanisms were instrumental in protecting wild-type adenovirus-infected human cells expressing 14.7K. This study describes a new molecular mechanism implemented by a virus to escape immunosurveillance by selectively targeting TNFR1 endocytosis to prevent TNF-induced DISC formation.


Neurotoxicity and behavioral effects of thiram in rats.
Eight of 24 female rats fed 66.9 mg/kg-day of thiram developed neurotoxicity. The neurotoxic effects were characterized by ataxia and paralysis of the hind legs. There were demyelination, degeneration of the axis cylinders, and presence of macrophages in the nerve bundle of the sciatic nerve. Degeneration in the ventral horn of the lower lumbar region of the spinal cord was evidenced by chromatolysis of motorneurons, pyknosis, and satellitosis. During a second experiment, 4 of 24 females fed 65.8 mg/kg--day also developed ataxia and paralysis. An additional 9 females showed clasping of the hind feet when picked up by the tail. Nerve conduction could not be measured for one severely ataxic rat and the electromyogram indicated a loss of motor unit function. Histopathology of this rat, along with the others, suggests the peripheral nerve as the primary site of the lesion. Thiram also caused behavioral changes in apparently normal rats. The walking pattern of the hind legs was altered with decreases in stride width and the angle between contralateral steps. These rats required significantly more shock-motivations and cleared a lower height in a jump/climb ability test. An open-field study indicated that thiram caused hyperactivity in the nonataxic rats of both sexes. Three of 24 rats fed 95.8 mg/kg-day of ferbam also developed ataxia or paralysis.


Galectin-3 blockade inhibits cardiac inflammation and fibrosis in experimental hyperaldosteronism and hypertension.
Hypertensive cardiac remodeling is accompanied by molecular inflammation and fibrosis, 2 mechanisms that finally affect cardiac function. At cardiac level, aldosterone promotes inflammation and fibrosis, although the precise mechanisms are still unclear. Galectin-3 (Gal-3), a β-galactoside-binding lectin, is associated with inflammation and fibrosis in the cardiovascular system. We herein investigated whether Gal-3 inhibition could block aldosterone-induced cardiac inflammation and fibrosis and its potential role in cardiac damage associated with hypertension. Aldosterone-salt-treated rats presented hypertension, cardiac inflammation, and fibrosis that were prevented by the pharmacological inhibition of Gal-3 with modified citrus pectin. Cardiac inflammation and fibrosis presented in spontaneously hypertensive rats were prevented by modified citrus pectin treatment, whereas Gal-3 blockade did not modify blood pressure levels. In the absence of blood pressure modifications, Gal-3 knockout mice were resistant to aldosterone-induced cardiac inflammation. In human cardiac fibroblasts, aldosterone increased Gal-3 expression via its mineralocorticoid receptor. Gal-3 and aldosterone enhanced proinflammatory and profibrotic markers, as well as metalloproteinase activities in human cardiac fibroblasts, effects that were not observed in Gal-3-silenced cells treated with aldosterone. In experimental hyperaldosteronism, the increase in Gal-3 expression was associated with cardiac inflammation and fibrosis, alterations that were prevented by Gal-3 blockade independently of blood pressure levels. These data suggest that Gal-3 could be a new molecular mechanism linking cardiac inflammation and fibrosis in situations with high-aldosterone levels, such as hypertension.


Biochemical and immunologic characterization of a major surface antigen of Dirofilaria immitis infective larvae.
A 35 kD major surface antigen of Dirofilaria immitis third-stage larvae was characterized biochemically and immunologically. Living larvae were iodinated by using Iodo-gen, iodosulfanilic acid, lactoperoxidase-glucose oxidase, and Bolton-Hunter reagents. Detergent extracts of larvae labeled by the first three methods showed one major 35 kD component and a number of smaller components of about 6 kD, as analyzed by one-dimensional SDS-PAGE. In contrast, extracts from larvae labeled with the Bolton-Hunter reagent showed multiple bands on gels. The 35kD molecule was shown to be exposed on the larval surface, insofar as it was accessible to trypsin-proteolysis on living radiolabeled larvae. Two-dimensional gel electrophoresis resolved the 35 kD band into two components: a major one with a pI of 3.8, and a minor one of pI 7.3. The lower m.w. bands were resolved into about 12 constituents with pI values from 3.5 to 8.0. Of all these surface molecules, the only one that was antigenic was the 35 kD component. It could be immunoprecipitated with sera from dogs carrying an occult experimental D. immitis infection or with sera from dogs immunized with irradiated third-stage larvae of this parasite. Similarly, sera from rabbits immunized repeatedly with normal unirradiated larvae also precipitated the 35 kD antigen. None of these sera, however, contained detectable antibodies to the surface-labeled low m.w. molecules. Sera from rabbits immunized with D. immitis adult worms and microfilariae precipitated the 35 kD antigen, which is therefore not stage specific. In contrast, sera from dogs experimentally infected with Toxocara canis and Ancylostoma caninum or with Uncinaria stenocephala (a canine hookworm) did not contain antibodies to the 35 kD antigen, but did cross-react with many other D. immitis adult and microfilarial antigens. This molecule may therefore be species specific. Evidence for glycosylation of the 35 kD molecule was not found: it did not bind to peanut, wheat germ, lentil, or Ulex europeus lectins, and its electrophoretic mobility was not altered after treatment with endoglycosidase-F or mild alkali solutions.


Iterative optimization calibration method for stereo deflectometry.
An accurate system calibration method is presented in this paper to calibrate stereo deflectometry. A corresponding iterative optimization algorithm is also proposed to improve the system calibration accuracy. This merges CCD parameters and geometrical relation between CCDs and the LCD into one cost function. In this calibration technique, an optical flat acts as a reference mirror and simultaneously reflect sinusoidal fringe patterns into the two CCDs. The normal vector of the reference mirror is used as an intermediate variable to implement this iterative optimization algorithm until the root mean square of the reprojection errors converge to a minimum. The experiment demonstrates that this method can optimize all the calibration parameters and can effectively reduce reprojection error, which correspondingly improves the final reconstruction accuracy.


Extracorporeal Membrane Oxygenation as a Bridge to Cardiac Transplantation in Children.
The feasibility and efficacy of extracorporeal membrane oxygenation (ECMO) as a bridge to cardiac transplantation was examined in 6 pediatric patients who suffered irreversible myocardial failure after undergoing surgery for congenital heart defects. The mean time of ECMO support was 260.5 h, range, 101-402 h. Three patients underwent transplantation, 2 of whom are long-term survivors. Progressive hypotension as a result of capillary leak syndrome precluded further ECMO support in the other 3 patients. Overall, 2 of the 6 patients survived. Major complications were encountered in 4 patients including bleeding in 2, a seizure in 1, and renal failure in 3, 2 of whom recovered renal function after transplantation. Infection did not occur in any of the 6 patients. Exchanging ECMO components was performed with no difficulties; these exchanges included a centrifugal pump once for 2 patients and a membrane oxygenator once for 3 patients. Our results indicate that ECMO can safely keep critically ill pediatric transplant candidates alive for more than 1 week with a low incidence of multiple organ failure.


Caffeine, Energy Beverage Consumption, Fitness, and Sleep in U.S. Army Aviation Personnel.
BACKGROUND: Caffeine-containing products and dietary supplements are widely used by military populations, but little is known about their use by aviation personnel. This study assessed self-reported sleep, fitness, work-schedules, and caffeine/energy drink use.METHODS: A standardized survey was conducted in person by study personnel using tablet computers. A total of 188 aircrew members from the Combat Aviation Brigade at Fort Campbell, KY, participated in the survey. Focus groups were conducted with a subset of 47 subjects.RESULTS: The majority of subjects reported their physical fitness, health, and diets were good. They reported sleeping about 6 h per day and stated they needed additional sleep to feel fully rested. Their caffeine consumption averaged 346 ± 23 mg · d-1 with most derived from coffee (139 ± 12 mg · d-1) and energy drinks (110 ± 13 mg · d-1). About half (55%) of participants used energy drinks at least once per week and they consumed greater amounts of caffeine than nonusers. Focus group data indicated crewmembers primarily consumed energy drinks to enhance performance degraded by variations in work schedules and lack of sufficient sleep. Participants expressed a desire for additional education on diets and energy drinks as well as on aeromedical policies governing energy drink and supplement use.CONCLUSIONS: Caffeinated products, including coffee and energy drinks, are routinely used by Army aircrews to increase alertness. Aircrew personnel consider them generally safe, but would like to receive education about these beverages, other dietary issues, and Army policies governing their use in aircrew.Bukhari AS, Caldwell JA, DiChiara AJ, Merrill EP, Wright AO, Cole RE, Hatch-McChesney A, McGraw SM, Lieberman HR. Caffeine, energy beverage consumption, fitness, and sleep in U.S. Army aviation personnel. Aerosp Med Hum Perform. 2020; 91(8):641-650.


Effects of the type 2 diabetes-associated PPARG P12A polymorphism on progression to diabetes and response to troglitazone.
The common P12A polymorphism in PPARG (a target for thiazolidinedione medications) has been consistently associated with type 2 diabetes. We examined whether PPARG P12A affects progression from impaired glucose tolerance to diabetes, or responses to preventive interventions (lifestyle, metformin, or troglitazone vs. placebo). This study included 3548 Diabetes Prevention Program participants. We performed Cox regression analysis using genotype at PPARG P12A, intervention, and their interactions as predictors of diabetes incidence. We also genotyped five other PPARG variants implicated in the response to troglitazone and assessed their effect on insulin sensitivity at 1 yr. Consistent with prior cross-sectional studies, P/P homozygotes at PPARG P12A appeared more likely to develop diabetes than alanine carriers (hazard ratio, 1.24; 95% confidence interval, 0.99-1.57; P=0.07) with no interaction of genotype with intervention. There was a significant interaction of genotype with body mass index and waist circumference (P=0.03 and 0.002, respectively) with the alanine allele conferring less protection in more obese individuals. Neither PPARG P12A nor five other variants significantly affected the impact of troglitazone on insulin sensitivity in 340 participants at 1 yr. The proline allele at PPARG P12A increases risk for diabetes in persons with impaired glucose tolerance, an effect modified by body mass index. In addition, PPARG P12A has little or no effect on the beneficial response to troglitazone.


Hurling alone? How social capital failed to save the Irish from cardiovascular disease in the United States.
We performed a historical review of cardiovascular risk profiles of Irish immigrants to the United States, 1850-1970, in regard to lifestyle, socio-economic circumstances, and social capital. We analyzed US Census data from 1850-1970, area-based social and epidemiological data from Boston, data from Ireland's National Nutrition Surveillance Centre, and literature on Irish migration. The Irish were consistently at increased risk of cardiovascular diseases, a risk that related initially to material deprivation, across the life course of at least 2 generations. The principal difference between the Irish and other disadvantaged immigrant groups, such as the Italians, was dietary habits influenced by experiences during the Irish famine. Although there was a psychosocial component to the disadvantage and discrimination they experienced as an ethnic group, the Irish also exhibited strong community networks and support structures that might have been expected to counteract discrimination's negative effects. However, the Irish's high levels of social capital were not protective for cardiovascular disease.


The structure and substrate specificity of human Cdk12/Cyclin K.
Phosphorylation of the RNA polymerase II C-terminal domain (CTD) by cyclin-dependent kinases is important for productive transcription. Here we determine the crystal structure of Cdk12/CycK and analyse its requirements for substrate recognition. Active Cdk12/CycK is arranged in an open conformation similar to that of Cdk9/CycT but different from those of cell cycle kinases. Cdk12 contains a C-terminal extension that folds onto the N- and C-terminal lobes thereby contacting the ATP ribose. The interaction is mediated by an HE motif followed by a polybasic cluster that is conserved in transcriptional CDKs. Cdk12/CycK showed the highest activity on a CTD substrate prephosphorylated at position Ser7, whereas the common Lys7 substitution was not recognized. Flavopiridol is most potent towards Cdk12 but was still 10-fold more potent towards Cdk9. T-loop phosphorylation of Cdk12 required coexpression with a Cdk-activating kinase. These results suggest the regulation of Pol II elongation by a relay of transcriptionally active CTD kinases.


Polysome trafficking of transcripts and microRNAs in regenerating liver after partial hepatectomy.
Liver regeneration after 70% partial hepatectomy (PH) in rats induces >95% of hepatocytes to undergo two rounds of semisynchronous cell replication. Gene expression is controlled primarily by posttranscriptional processing, including changes in mRNA stability. However, the translational activity of a specific mRNA can also be modulated after PH, resulting in significant uncoupling of protein and transcript levels relative to quiescent liver for many genes including c-myc and p53. Although the precise mechanism by which this uncoupling occurs is unknown, the polysomal association of mRNA and microRNA (miRNA) can significantly modulate rate of decay as well as translational activity. Thus we characterized the association of c-myc and p53 mRNAs and miRNAs in free and cytoskeleton- and membrane-bound polysome populations 3, 6, and 24 h after PH. The transcripts for c-myc and p53 were differentially distributed in the three discrete polysome populations, and this was dramatically modulated during liver regeneration. Nascent polysome-associated p53 and c-myc proteins were also differentially expressed in the free and cytoskeleton- and membrane-bound polysomes and significantly uncoupled from transcript levels relative to nonresected liver. At least 85 miRNAs were associated with the three polysome populations, and their abundance and distribution changed significantly during liver regeneration. These data suggest that posttranscriptional control of c-myc and p53 protein expression is associated with the translocation of transcripts between the different polyribosomes. The alteration of expression for the same transcript in different polysome populations may, in part, be due to the action of miRNAs.


Human and Drosophila homeodomain proteins that enhance the DNA-binding activity of serum response factor.
Cells with distinct developmental histories can respond differentially to identical signals, suggesting that signals are interpreted in a fashion that reflects a cell's identity. How this might occur is suggested by the observation that proteins of the homeodomain family, including a newly identified human protein, enhance the DNA-binding activity of serum response factor, a protein required for the induction of genes by growth and differentiation factors. Interaction with proteins of the serum response factor family may allow homeodomain proteins to specify the transcriptional response to inductive signals. Moreover, because the ability to enhance the binding of serum response factor to DNA residues within the homeodomain but is independent of homeodomain DNA-binding activity, this additional activity of the homeodomain may account for some of specificity of action of homeodomain proteins in development.


Using an agent-based model to analyze the dynamic communication network of the immune response.
The immune system behaves like a complex, dynamic network with interacting elements including leukocytes, cytokines, and chemokines. While the immune system is broadly distributed, leukocytes must communicate effectively to respond to a pathological challenge. The Basic Immune Simulator 2010 contains agents representing leukocytes and tissue cells, signals representing cytokines, chemokines, and pathogens, and virtual spaces representing organ tissue, lymphoid tissue, and blood. Agents interact dynamically in the compartments in response to infection of the virtual tissue. Agent behavior is imposed by logical rules derived from the scientific literature. The model captured the agent-to-agent contact history, and from this the network topology and the interactions resulting in successful versus failed viral clearance were identified. This model served to integrate existing knowledge and allowed us to examine the immune response from a novel perspective directed at exploiting complex dynamics, ultimately for the design of therapeutic interventions. Analyzing the evolution of agent-agent interactions at incremental time points from identical initial conditions revealed novel features of immune communication associated with successful and failed outcomes. There were fewer contacts between agents for simulations ending in viral elimination (win) versus persistent infection (loss), due to the removal of infected agents. However, early cellular interactions preceded successful clearance of infection. Specifically, more Dendritic Agent interactions with TCell and BCell Agents, and more BCell Agent interactions with TCell Agents early in the simulation were associated with the immune win outcome. The Dendritic Agents greatly influenced the outcome, confirming them as hub agents of the immune network. In addition, unexpectedly high frequencies of Dendritic Agent-self interactions occurred in the lymphoid compartment late in the loss outcomes. An agent-based model capturing several key aspects of complex system dynamics was used to study the emergent properties of the immune response to viral infection. Specific patterns of interactions between leukocyte agents occurring early in the response significantly improved outcome. More interactions at later stages correlated with persistent inflammation and infection. These simulation experiments highlight the importance of commonly overlooked aspects of the immune response and provide insight into these processes at a resolution level exceeding the capabilities of current laboratory technologies.


Synthesis of reverse osmosis membranes by plasma polymerization of allylamine.
The polymerization of allylamine in a radio-frequency electrodeless plasma to form thin polymer films on microporous filter media provides very effective dry composite membranes for reverse osmosis. Salt and urea rejections as high as 98 percent and 46 percent, respectively, have been achieved from a solution containing 10 grams of sodium chloride per liter and 10 grams of urea per liter. The plasma conditions and polymerization technique are discussed.


Protection by 20-5,14-HEDGE against surgically induced ischemia reperfusion lung injury in rats.
We previously reported that the cytochrome P450 product 20-hydroxyeicosatetraenoic acid has prosurvival effects in pulmonary artery endothelial cells and ex vivo pulmonary arteries. We tested the potential of a 20-hydroxyeicosatetraenoic acid analog N-[20-hydroxyeicosa-5(Z),14(Z)-dienoyl]glycine (20-5,14-HEDGE) to protect against lung ischemic reperfusion injury in rats. Furthermore, we examined activation of innate immune system components, high mobility group box 1 (HMGB1) and toll-like receptor 4 (TLR4), in this model as well as the effect of 20-5,14-HEDGE on this signaling pathway. Sprague-Dawley rats treated with 20-5,14-HEDGE or vehicle were subjected to surgically induced, unilateral lung ischemia for 60 minutes followed by reperfusion for 2 hours in vivo. Injury was assessed histologically by hematoxylin and eosin, and with identification of myeloperoxidase immunohistochemically. The HMGB1 and TLR4 proteins were identified by Western blot. Caspase 3 activity or 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a yellow tetrazole, incorporation were used to measure apoptosis and cell survival. The ischemia reperfusion injury evoked atelectasis and hemorrhage, an influx of polymorphonuclear cells, and increased TLR4 and HMGB1 expression. Caspase 3 activity was increased, and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide incorporation was decreased. The 20-5,14-HEDGE protected against each of these endpoints, including infiltration of polymorphonuclear cells, with no changes in caspase 3 activity in other organs. Lung ischemia reperfusion produces apoptosis and activation of the innate immune system including HMGB1 and TLR4 within 2 hours of reperfusion. Treatment with 20-5,14-HEDGE decreases activation of this response system, and salvages lung tissue.


Assessment of Risk Communication about Undercooked Hamburgers by Restaurant Servers.
According to the U.S. Food and Drug Administration 2013 Model Food Code, it is the duty of a food establishment to disclose and remind consumers of risk when ordering undercooked food such as ground beef. The purpose of this study was to explore actual risk communication behaviors of food establishment servers. Secret shoppers visited 265 restaurants in seven geographic locations across the United States, ordered medium rare burgers, and collected and coded risk information from chain and independent restaurant menus and from server responses. The majority of servers reported an unreliable method of doneness (77%) or other incorrect information (66%) related to burger doneness and safety. These results indicate major gaps in server knowledge and risk communication, and the current risk communication language in the Model Food Code does not sufficiently fill these gaps. The question is "should servers even be acting as risk communicators?" There are numerous challenges associated with this practice, including high turnover rates, limited education, and the high stress environment based on pleasing a customer. If servers are designated as risk communicators, food establishment staff should be adequately trained and provided with consumer advisory messages that are accurate, audience appropriate, and delivered in a professional manner so that customers can make informed food safety decisions.


Evidence for monophyly and arthropod affinity of cambrian giant predators.
The Chinese Early Cambrian Chengjiang fauna includes three different anomalocaridids, a globally spread, extinct marine group including the largest known Cambrian animals. Anomalocaridids were active predators, and their presence implies that a complex ecosystem appeared abruptly in the earliest Phanerozoic. Complete specimens display several sets of characters shared only with some other exclusively Cambrian forms. This evidence indicates that anomalocaridids, Opabinia, and Kerygmachela form a monophyletic clade. Certain features indicate arthropod affinities of the lade, and for this group an unnamed (sub)phylum-level taxon within an arthropod (super)phylum is proposed.


Acid aspiration-induced lung injury in rabbits is mediated by interleukin-8-dependent mechanisms.
Acid aspiration lung injury may be mediated primarily by neutrophils recruited to the lung by acid-induced cytokines. We hypothesized that a major acid-induced cytokine was IL-8 and that a neutralizing anti-rabbit-IL-8 monoclonal antibody (ARIL8.2) would attenuate acid-induced lung injury in rabbits. Hydrochloric acid (pH = 1.5 in 1/3 normal saline) or 1/3 normal saline (4 ml/kg) was instilled into the lungs of ventilated, anesthetized rabbits. The rabbits were studied for 6 or 24 h. In acid-instilled rabbits without the anti-IL-8 monoclonal antibody, severe lung injury developed in the first 6 h; in the long-term experiments, all rabbits died with lung injury between 12 and 14 h. In acid-instilled rabbits given the anti-IL-8 monoclonal antibody (2 mg/kg, intravenously) either as pretreatment (5 min before the acid) or as treatment (1 h after the acid), acid-induced abnormalities in oxygenation and extravascular lung water were prevented and extravascular protein accumulation was reduced by 70%; in the long-term experiments, anti-IL-8 treatment similarly protected lung function throughout the 24-h period. The anti-IL-8 monoclonal antibody also significantly reduced air space neutrophil counts and IL-8 concentrations. This study establishes IL-8 as a critical cytokine for the development of acid-induced lung injury. Neutralization of IL-8 may provide the first useful therapy for this clinically important form of acute lung injury.


[Results of the Cilostazol Stroke Prevention Study II (CSPS II): a randomized controlled trial for the comparison of cilostazol and aspirin in stroke patients].
We compared the efficacy and safety of cilostazol and aspirin in 2,672 Japanese patients with non-cardioembolic ischemic stroke. The patients were randomized to be allocated either on cilostazol (200 mg/day) group or aspirin (81 mg/day) group, and were followed up for one to five years (average 29 months). The primary endpoint was any stroke, and safety endpoint was hemorrhagic stroke or hemorrhage requiring hospitalization. Annual incidence of stroke was significantly lower in the cilostazol group (2.76%) than in the aspirin group (3.71%) (relative risk reduction [RRR] 25.7%, p=0.0357) and annual incidence of hemorrhagic stroke or hemorrhage requiring hospitalization was 0.77% in the cilostazol group and 1.77% in the aspirin group (RRR 54.2%, p=0.0004). The sub-analyses between subtypes of ischemic stroke showed that annual incidence of hemorrhagic stroke was much lower in the cilostazol group (0.36%) than in the aspirin group (1.20%) among patients with lacunar stroke (p=0.003). The results suggest that cilostazol has a favorable risk-benefit profile alternative to aspirin for secondary stroke prevention in patients with non-cardioembolic ischemic stroke, particularly in patients with lacunar stroke, who are at high risk of hemorrhagic stroke.


Basal Ganglia activity in pathological gambling: a fluorodeoxyglucose-positron emission tomography study.
Pathological gambling (PG) is a disorder classified as an impulse control disorder (DSM-IV) bridging impulsive, compulsive and addictive behaviors. The striatum and thalamus are supposed to be involved in the pathophysiological substrate of these behaviors. An increased relative glucose metabolic rate (rGMR) in patients with a diagnosis of PG had previously been reported in the medial and orbitofrontal cortex. We extended our studies to include functional alterations of the striatum and thalamus in a cohort of patients with PG before and after treatment with lithium. Twenty-one patients with PG who met lifetime comorbid bipolar spectrum diagnoses and a comparison group of 21 age- and sex-matched controls underwent a baseline positron emission tomography (PET) scan. Sixteen of these patients entered a randomized double-blind placebo-controlled parallel-group-design trial of lithium and underwent a follow-up PET scan at week 10. Anatomical MRI were obtained and the structures outlined on consecutive axial slices. These individual hand-drawn templates were used to identify structures on the PET scan of each patient, and the rGMR was measured. The PG patients had a decrement of the rGMR in the ventral parts of the striatum and thalamus, and an increment of the rGMR in the dorsal parts as compared with the controls. Lithium treatment increased the ventral caudate rGMR to a trend level in the patients, but had no effect on the metabolism of either the putamen or the thalamus. Because of their extensive connectivity to the frontal cortex, striatal and thalamic functional alteration may contribute to faulty decision making processes in PG patients. By increasing the ventral rGMR of the caudate nucleus, lithium treatment may reduce cognitive dysfunction and symptoms in PG patients.


[Adult smoking trends in Mexico between 1988 and 2008].
To describe the changes in several smoking indicators occurred in Mexico over the past two decades and to explore if the tobacco control policies implemented in Mexico, since 2004, show a favorable impact on tobacco consumption by 2008. We analyze trends in comparable data on the prevalence of never and daily smokers, using the five National Addiction Surveys conducted between 1988 and 2008. The analysis is restricted to persons aged 18 through 65 years. Data are adjusted for age, sex and marginality index. Between 2002 and 2008 the percentage of never smokers has increased by 19.6% and the percentage of daily smokers has decreased by 24.8%. These changes were more important in men, but in women the average number of cigarettes consumed decreased by 21.1% over the compared period and there was also a 13.9% rise in the percentage of women attempting to quit. Daily smokers prevalence has declined more rapidly since 2005, coincidently with an increase in cigarette taxation. Over the past two decades there has been in Mexico an increase in the percentage of never smokers and a decline in the percentage of daily smokers. A positive impact is observed, few years after the implementation of more effective tobacco control policies in Mexico, notably higher taxation of tobacco products.


Ultraendurance exercise increases the production of reactive oxygen species in isolated mitochondria from human skeletal muscle.
Exercise-induced oxidative stress is important for the muscular adaptation to training but may also cause muscle damage. We hypothesized that prolonged exercise would increase mitochondrial production of reactive oxygen species (ROS) measured in vitro and that this correlates with oxidative damage. Eight male athletes (24-32 yr) performed ultraendurance exercise (kayaking/running/cycling) with an average work intensity of 55% V(O(2peak)) for 24 h. Muscle biopsies were taken from vastus lateralis before exercise, immediately after exercise, and after 28 h of recovery. The production of H(2)O(2) was measured fluorometrically in isolated mitochondria with the Amplex red and peroxidase system. Succinate-supported mitochondrial H(2)O(2) production was significantly increased after exercise (73% higher, P = 0.025) but restored to the initial level at recovery. Plasma level of free fatty acids (FFA) increased fourfold and exceeded 1.2 mmol/l during the last 6 h of exercise. Plasma FFA at the end of exercise was significantly correlated to mitochondrial ROS production (r = 0.74, P < 0.05). Mitochondrial content of 4-hydroxy-nonenal-adducts (a marker of oxidative damage) was increased only after recovery and was not correlated with mitochondrial ROS production. Total thiol group level and glutathione peroxidase activity were elevated after recovery. In conclusion, ultraendurance exercise increases ROS production in isolated mitochondria, but this is reversed after 28 h recovery. Mitochondrial ROS production was not correlated with oxidative damage of mitochondrial proteins, which was increased at recovery but not immediately after exercise.


Assessing the human footprint on the sea-floor of coastal systems: the case of the Venice Lagoon, Italy.
Coastal systems are among the most studied, most vulnerable, and economically most important ecosystems on Earth; nevertheless, little attention has been paid, so far, to the consequences of human activities on the shallow sea-floor of these environments. Here, we present a quantitative assessment of the effects of human actions on the floor of the tidal channels from the Venice Lagoon using 2500 kilometres of full coverage multibeam bathymetric mapping. Such extended dataset provides unprecedented evidence of pervasive human impacts, which extend far beyond the well known shrinking of salt marshes and artificial modifications of inlet geometries. Direct and indirect human imprints include dredging marks and fast-growing scours around anthropogenic structures built to protect the historical city of Venice from flooding. In addition, we document multiple effects of ship traffic (propeller-wash erosion, keel ploughing) and diffuse littering on the sea-floor. Particularly relevant, in view of the ongoing interventions on the lagoon morphology, is the evidence of the rapid morphological changes affecting the sea-floor and threatening the stability of anthropogenic structures.


Distribution of CWG and CCWGG in the human genome.
Expression of the bacterial CG methyltransferase M*HhaI in mammalian cells appears to generate significant biological effects, while biological effects of the expression of the non-CG methyltransferase M*EcoRII in human cells have not been detected. The association of cytosine methylation with the CG site in mammals is also associated with clustering of CG sites near 5' control regions (CG-islands) of human genes.Moreover spontaneous deamination of 5-methylcytosine at these sites is thought to lead to the well known deficiency of CG sites in genomes where endogenous CG methyltransferases are expressed. Since these associations are generally taken to imply a biological function for the CG dinucleotide that is associated with its selective methylation by endogenous DNA methylation systems, we have asked whether or not CWG or CCWGG sites are clustered in regions flanking human genes and whether or not an overall deficiency of CWG or CCWGG occurs in the human genome. Using build 36.1, of the human genome, we inspected the regions flanking the 28,501 well known gene loci in the human genome. Our analysis confirmed the expected clustering of CG sites near the 5' region of known genes and open reading frames. In contrast to the CG site, neither the CWG site nor the CCWGG site recognized by the bacterial methyltransferase M*EcoRII were clustered in any particular region near known genes and open reading frames. Moreover, neither the CCWGG nor the CWG site was depleted in the human genome, again in sharp contrast to the known genomic deficiency of CpG sites. Our findings suggest that in contrast to CG site recognition, human cytosine methyltransferases recognize CWG and CCWGG only at very low frequency if at all.