Mtx toxins synergize Bacillus sphaericus and Cry11Aa against susceptible and insecticide-resistant Culex quinquefasciatus larvae.
Two mosquitocidal toxins (Mtx) of Bacillus sphaericus, which are produced during vegetative growth, were investigated for their potential to increase toxicity and reduce the expression of insecticide resistance through their interactions with other mosquitocidal proteins. Mtx-1 and Mtx-2 were fused with glutathione S-transferase and produced in Escherichia coli, after which lyophilized powders of these fusions were assayed against Culex quinquefasciatus larvae. Both Mtx proteins showed a high level of activity against susceptible C. quinquefasciatus mosquitoes, with 50% lethal concentrations (LC(50)) of Mtx-1 and Mtx-2 of 0.246 and 4.13 microg/ml, respectively. The LC(50)s were 0.406 to 0.430 microg/ml when Mtx-1 or Mtx-2 was mixed with B. sphaericus, and synergy improved activity and reduced resistance levels. When the proteins were combined with a recombinant Bacillus thuringiensis strain that produces Cry11Aa, the mixtures were highly active against Cry11A-resistant larvae and resistance was also reduced. The mixture of two Mtx toxins and B. sphaericus was 10 times more active against susceptible mosquitoes than B. sphaericus alone, demonstrating the influence of relatively low concentrations of these toxins. These results show that, similar to Cyt toxins from B. thuringiensis subsp. israelensis, Mtx toxins can increase the toxicity of other mosquitocidal proteins and may be useful for both increasing the activity of commercial bacterial larvicides and managing potential resistance to these substances among mosquito populations.


Cardiotoxic Potential of Hydroxychloroquine, Chloroquine and Azithromycin in Adult Human Primary Cardiomyocytes.
Substantial efforts have been recently committed to develop coronavirus disease-2019 (COVID-19) medications, and Hydroxychloroquine alone or in combination with Azithromycin has been promoted as a repurposed treatment. Although these drugs may increase cardiac toxicity risk, cardiomyocyte mechanisms underlying this risk remain poorly understood in humans. Therefore, we evaluated the proarrhythmia risk and inotropic effects of these drugs in the cardiomyocyte contractility-based model of the human heart. We found Hydroxychloroquine to have a low proarrhythmia risk, whereas Chloroquine and Azithromycin were associated with high risk. Hydroxychloroquine proarrhythmia risk changed to high with low level of K+, whereas high level of Mg2+ protected against proarrhythmic effect of high Hydroxychloroquine concentrations. Moreover, therapeutic concentration of Hydroxychloroquine caused no enhancement of elevated temperature-induced proarrhythmia. Polytherapy of Hydroxychloroquine plus Azithromycin and sequential application of these drugs were also found to influence proarrhythmia risk categorization. Hydroxychloroquine proarrhythmia risk changed to high when combined with Azithromycin at therapeutic concentration. However, Hydroxychloroquine at therapeutic concentration impacted the cardiac safety profile of Azithromycin and its proarrhythmia risk only at concentrations above therapeutic level. We also report that Hydroxychloroquine and Chloroquine, but not Azithromycin, decreased contractility while exhibiting multi-ion channel block features, and Hydroxychloroquine's contractility effect was abolished by Azithromycin. Thus, this study has the potential to inform clinical studies evaluating repurposed therapies, including those in the COVID-19 context. Additionally, it demonstrates the translational value of the human cardiomyocyte contractility-based model as a key early discovery path to inform decisions on novel therapies for COVID-19, malaria, and inflammatory diseases.


Inhibition of normal growth by chronic administration of delta-9-tetrahydrocannabinol.
Body weight, food and water intake, and feces weight of 20 albino rats were recorded daily for 70 days. On days 11 to 40, 12 rats received behaviorally effective doses of Delta-9-tetrahydrocannabinol, either orally or intraperitoneally. These rats ate significantly less than placebo-dosed controls during the treatment period, and gained significantly less weight. Food intake recovered in the 30-day posttreatment period, but the former drug group still weighed less than the controls on day 70. In addition, all rats who had received intraperitoneal injections of Delta-9-tetrahydrocannabinol showed evidence of chronic diffuse nonsuppurative peritonitis.


Barriers and enablers for the development and implementation of allied health clinical practice guidelines in South African primary healthcare settings: a qualitative study.
The South African allied health (AH) primary healthcare (PHC) workforce is challenged with the complex rehabilitation needs of escalating patient numbers. The application of evidence-based care using clinical practice guidelines (CPGs) is one way to make efficient and effective use of resources. Although CPGs are common for AH in high-income countries, there is limited understanding of how to do this in low- to middle-income countries. This paper describes barriers and enablers for AH CPG uptake in South African PHC. Semi-structured individual interviews were undertaken with 25 South African AH managers, policymakers, clinicians and academics to explore perspectives on CPGs. Interviews were conducted by researcher dyads, one being familiar with South African AH PHC practice and the other with CPG expertise. Rigour and transparency of data collection was ensured. Interview transcripts were analysed by structuring content into codes, categories and themes. Exemplar quotations were extracted to support themes. CPGs were generally perceived to be relevant to assist AH providers to address the challenges of consistently providing evidence-based care in South African PHC settings. CPGs were considered to be tools for managing clinical, social and economic complexities of AH PHC practice, particularly if CPG recommendations were contextusalised. CPG uptake was one way to deal with increasing pressures to make efficient use of scarce financial resources, and to demonstrate professional legitimacy. Themes comprised organisational infrastructures and capacities for CPG uptake, interactions between AH actors and interaction with broader political structures, the nature of AH evidence in CPGs, and effectively implementing CPGs into practice. CPGs contextualised to local circumstances offer South African PHC AH services with an efficient vehicle for putting evidence into practice. There are challenges to doing this, related to local barriers such as geography, AH training, workforce availability, scarce resources, an escalating number of patients requiring complex rehabilitation, and local knowledge. Concerted attempts to implement locally relevant CPGs for AH primary care in South Africa are required to improve widespread commitment to evidence-based care, as well as to plan efficient and effective service delivery models.


Matches, clips, needles, or pins.
Several methods for positioning dowel pins used in the fabrication of removable dies have been presented. Needles, pins, paper clips, hairpins, and matches are all useful in positioning dowel pins in the impression. The use of paper matches is a simple and effective method for holding the dowel pin in the proper relation while the die stone portion of the cast is being poured.


Effect of acupuncture on heart rate variability in primary dysmenorrheic women.
Primary dysmenorrhea is a common gynecological complaint among young women that is related to an autonomic nervous system (ANS) disturbance. Acupuncture is one of several therapeutic approaches for primary dysmenorrhea, since it can modulate ANS function. The heart rate variability (HRV) parameters such as high frequency (HF), low frequency (LF) and LF/HF ratio are generally accepted tools to assess ANS activity. The purpose of this study was to investigate the effects of acupuncture applied at Hegu (LI4) and Sanyinjiao (SP6) points on HRV of women with primary dysmenorrhea during the late luteal phase. The experimental design was a crossover and patient-blinded procedure. All subjects participated in Sham (SA) and Real Acupuncture (RA) procedure, separated by one month, in a crossover sequence. The participants included 38 women (mean age 22.3 years; weight 53.8 kg; height 162.6 cm). HRV measurement was 15 min before and 15 min after an acupuncture procedure. The RA procedure was performed at two bilateral acupoints, but needles were inserted subcutaneously to the acupuncture points for the SA procedure. The RA induced a significant decrease in LF/HF ratio and a significant increase in the HF power, while SA treatment caused a significant increase only in the HF power. Manual acupuncture at bilateral acupoints of LI4 and SP6 may play a role in dysmenorrhea treatment with autonomic nervous system involvement.


Temporal Dynamics of Cortical Microinfarcts in Cerebral Small Vessel Disease.
Neuropathology studies show a high prevalence of cortical microinfarcts (CMIs) in aging individuals, especially in patients with cerebrovascular disease and dementia. However, most, are invisible on T1- and T2-weighted magnetic resonance imaging (MRI), raising the question of how to explain this mismatch. Studies on small acute infarcts, detected on diffusion-weighted imaging (DWI), suggest that infarcts are largest in their acute phase and reduce in size thereafter. Therefore, we hypothesized that a subset of the CMI that are invisible on MRI can be detected on MRI in their acute phase. However, to our knowledge, a serial imaging study investigating the temporal dynamics of acute CMI (A-CMI) is lacking. To determine the prevalence of chronic CMI (C-CMI) and the cumulative incidence and temporal dynamics of A-CMI in individuals with cerebral small vessel disease (SVD). The RUN DMC-Intense study is a single-center hospital-based prospective cohort study on SVD performed between March 2016 and November 2017 and comprising 10 monthly 3-T MRI scans, including high-resolution DWI, 3-dimensional T1, 3-dimensional fluid-attenuated inversion recovery, and T2. One hundred six individuals from the previous longitudinal RUN DMC study were recruited based on the presence of progression of white matter hyperintensities on MRI between 2006 and 2015 and exclusion of causes of cerebral ischemia other than SVD. Fifty-four individuals (50.9%) participated. The median total follow-up duration was 39.5 weeks (interquartile range, 37.8-40.3). Statistical data analysis was performed between May and October 2019. We determined the prevalence of C-CMI using the baseline T1, fluid-attenuated inversion recovery, and T2 scans. Monthly high-resolution DWI scans (n = 472) were screened to determine the cumulative incidence of A-CMI. The temporal dynamics of A-CMI were determined based on the MRI scans collected during the first follow-up visit after A-CMI onset and the last available follow-up visit. The median age of the cohort at baseline MRI was 69 years (interquartile range, 66-74 years) and 34 participants (63%) were men. The prevalence of C-CMI was 35% (95% CI, 0.24-0.49). Monthly DWI detected 21 A-CMI in 7 of 54 participants, resulting in a cumulative incidence of 13% (95% CI, 0.06-0.24). All A-CMI disappeared on follow-up MRI. Acute CMI never evolved into chronically MRI-detectable lesions. We suggest that these A-CMI underlie part of the submillimeter C-CMI encountered on neuropathological examination and thereby provide a source for the high CMI burden on neuropathology.


Distinct effects of ligand-induced PDGFRα and PDGFRβ signaling in the human rhabdomyosarcoma tumor cell and stroma cell compartments.
Platelet-derived growth factor receptors (PDGFR) α and β have been suggested as potential targets for treatment of rhabdomyosarcoma, the most common soft tissue sarcoma in children. This study identifies biologic activities linked to PDGF signaling in rhabdomyosarcoma models and human sample collections. Analysis of gene expression profiles of 101 primary human rhabdomyosarcomas revealed elevated PDGF-C and -D expression in all subtypes, with PDGF-D as the solely overexpressed PDGFRβ ligand. By immunohistochemistry, PDGF-CC, PDGF-DD, and PDGFRα were found in tumor cells, whereas PDGFRβ was primarily detected in vascular stroma. These results are concordant with the biologic processes and pathways identified by data mining. While PDGF-CC/PDGFRα signaling associated with genes involved in the reactivation of developmental programs, PDGF-DD/PDGFRβ signaling related to wound healing and leukocyte differentiation. Clinicopathologic correlations further identified associations between PDGFRβ in vascular stroma and the alveolar subtype and with presence of metastases. Functional validation of our findings was carried out in molecularly distinct model systems, where therapeutic targeting reduced tumor burden in a PDGFR-dependent manner with effects on cell proliferation, vessel density, and macrophage infiltration. The PDGFR-selective inhibitor CP-673,451 regulated cell proliferation through mechanisms involving reduced phosphorylation of GSK-3α and GSK-3β. Additional tissue culture studies showed a PDGFR-dependent regulation of rhabdosphere formation/cancer cell stemness, differentiation, senescence, and apoptosis. In summary, the study shows a clinically relevant distinction in PDGF signaling in human rhabdomyosarcoma and also suggests continued exploration of the influence of stromal PDGFRs on sarcoma progression.


In vitro and in vivo activity of LS 4477 and LS 4559, novel analogues of the tubulin binder estramustine.
LS 4477 and LS 4559, two of a series of N-acyl-aminoalkyl phenyl ethers, are rationally designed compounds based on the tubulin binder estramustine. This study investigated their mechanism of action and compared their effectiveness in relation to estramustine in vitro against a panel of human and murine cell lines and in vivo against two murine colon tumour models (MAC). At biologically relevant concentrations, LS 4477 and LS 4559 caused a 59.9 and 56% reduction in tubulin assembly, respectively, compared with a 28.4% reduction in tubulin assembly by estramustine. The analogues were approximately 100 times more potent in chemosensitivity tests in vitro than the parent compound. Both analogues were orally active against the MAC 15A murine tumour model, to a greater extent than estramustine, producing significant growth delays (P<0.01). Significant activity was also shown against the slower growing MAC 26 tumour for LS 4577 (the soluble pro-drug of LS 4559). The results presented in this study suggest these compounds warrant further development with a view to assessing their clinical activity.


Segregation of vowels and consonants in human auditory cortex: evidence for distributed hierarchical organization.
The speech signal consists of a continuous stream of consonants and vowels, which must be de- and encoded in human auditory cortex to ensure the robust recognition and categorization of speech sounds. We used small-voxel functional magnetic resonance imaging to study information encoded in local brain activation patterns elicited by consonant-vowel syllables, and by a control set of noise bursts. First, activation of anterior-lateral superior temporal cortex was seen when controlling for unspecific acoustic processing (syllables versus band-passed noises, in a "classic" subtraction-based design). Second, a classifier algorithm, which was trained and tested iteratively on data from all subjects to discriminate local brain activation patterns, yielded separations of cortical patches discriminative of vowel category versus patches discriminative of stop-consonant category across the entire superior temporal cortex, yet with regional differences in average classification accuracy. Overlap (voxels correctly classifying both speech sound categories) was surprisingly sparse. Third, lending further plausibility to the results, classification of speech-noise differences was generally superior to speech-speech classifications, with the no\ exception of a left anterior region, where speech-speech classification accuracies were significantly better. These data demonstrate that acoustic-phonetic features are encoded in complex yet sparsely overlapping local patterns of neural activity distributed hierarchically across different regions of the auditory cortex. The redundancy apparent in these multiple patterns may partly explain the robustness of phonemic representations.


Shortening spin-lattice relaxation using a copper-chelated lipid at low-temperatures - A magic angle spinning solid-state NMR study on a membrane-bound protein.
Inherent low sensitivity of NMR spectroscopy has been a major disadvantage, especially to study biomolecules like membrane proteins. Recent studies have successfully demonstrated the advantages of performing solid-state NMR experiments at very low and ultralow temperatures to enhance the sensitivity. However, the long spin-lattice relaxation time, T1, at very low temperatures is a major limitation. To overcome this difficulty, we demonstrate the use of a copper-chelated lipid for magic angle spinning solid-state NMR measurements on cytochrome-b5 reconstituted in multilamellar vesicles. Our results on multilamellar vesicles containing as small as 0.5mol% of a copper-chelated lipid can significantly shorten T1 of protons, which can be used to considerably reduce the data collection time or to enhance the signal-to-noise ratio. We also monitored the effect of slow cooling on the resolution and sensitivity of (13)C and (15)N signals from the protein and (13)C signals from lipids.


Homeopathic therapy in pediatric atopic diseases: short- and long-term results.
To study the outcomes of atopic diseases in children treated with homeopathy at the Homeopathic Clinic of Lucca (Italy) and related long-term results after approximately an 8-year period. Our data derive from an observational longitudinal study carried out on 857 pediatric patients who consecutive visited from 1998 to 2014. Children with atopic diseases were 325 (37.9%), 126 (39%) suffered from atopic dermatitis, 72 (22%) from allergic rhinitis, and 127 (39%) from asthma. Moreover, a long-term study was conducted on a subset of 107/165 patients, consecutively visited from 1998 to 2006, and with ≥5 years follow-up. The study also investigated the evolution of overall symptoms in those patients with a complex atopic symptomatology. 75.8% of atopic children had moderate or major improvement (67.1% with asthma as the primary disease; 84.2% rhinitis; 84.2% dermatitis). At re-evaluation after 5-10 years, complete remission of atopic symptoms was obtained in 70.1% of the children: 84.2% in dermatitis; 48.1% in allergic rhinitis; 71.4% in asthma. Children with two or three atopic diseases at the first visit were completely cured in 40% of cases. The results seem to confirm that homeopathic medicine produces positive therapeutic response in atopic children.


Changes in Acceleration and Deceleration Capacity Throughout Professional Soccer Match-Play.
Russell, M, Sparkes, W, Northeast, J, Cook, CJ, Love, TD, Bracken, RM, and Kilduff, LP. Changes in acceleration and deceleration capacity throughout professional soccer match-play. J Strength Cond Res 30(10): 2839-2844, 2016-As the acceleration and deceleration demands of soccer are currently not well understood, this study aimed to profile markers of acceleration and deceleration capacity during professional soccer match-play. This within-player observational study required reserve team players from a Premier League club to wear 10-Hz Global Positioning System units throughout competitive matches played in the 2013-14 competitive season. Data are presented for players who completed 4 or more games during the season (n = 11), and variables are presented according to six 15-minute intervals (I1-6: 00:00-14:59 minutes, 15:00-29:59 minutes, 30:00-44:59 minutes, 45:00-59:59 minutes, 60:00-74:59 minutes, and 75:00-89:59 minutes, respectively). During I6, the distance covered (total, per minute, and at high intensity), number of sprints, accelerations (total and high intensity), decelerations (total and high intensity), and impacts were reduced compared with I1 (all p ≤ 0.05). The number of high-intensity impacts remained unchanged throughout match-play (p > 0.05). These findings indicate that high-intensity actions and markers of acceleration and deceleration capacity are reduced in the last 15 minutes of the normal duration of match-play. Such information can be used to increase the specificity of training programs designed for soccer players while also giving further insight in to the effects of 90 minutes of soccer-specific exercise. Interventions that seek to maintain the acceleration and deceleration capacity of players throughout the full duration of a soccer match warrant investigation.


Expression levels of a kinesin-13 microtubule depolymerase modulates the effectiveness of anti-microtubule agents.
Chemotherapeutic drugs often target the microtubule cytoskeleton as a means to disrupt cancer cell mitosis and proliferation. Anti-microtubule drugs inhibit microtubule dynamics, thereby triggering apoptosis when dividing cells activate the mitotic checkpoint. Microtubule dynamics are regulated by microtubule-associated proteins (MAPs); however, we lack a comprehensive understanding about how anti-microtubule agents functionally interact with MAPs. In this report, we test the hypothesis that the cellular levels of microtubule depolymerases, in this case kinesin-13 s, modulate the effectiveness of the microtubule disrupting drug colchicine. We used a combination of RNA interference (RNAi), high-throughput microscopy, and time-lapse video microscopy in Drosophila S2 cells to identify a specific MAP, kinesin-like protein 10A (KLP10A), that contributes to the efficacy of the anti-microtubule drug colchicine. KLP10A is an essential microtubule depolymerase throughout the cell cycle. We find that depletion of KLP10A in S2 cells confers resistance to colchicine-induced microtubule depolymerization to a much greater extent than depletion of several other destabilizing MAPs. Using image-based assays, we determined that control cells retained 58% (+/-2%SEM) of microtubule polymer when after treatment with 2 microM colchicine for 1 hour, while cells depleted of KLP10A by RNAi retained 74% (+/-1%SEM). Likewise, overexpression of KLP10A-GFP results in increased susceptibility to microtubule depolymerization by colchicine. Our results demonstrate that the efficacy of microtubule destabilization by a pharmacological agent is dependent upon the cellular expression of a microtubule depolymerase. These findings suggest that expression levels of Kif2A, the human kinesin-13 family member, may be an attractive biomarker to assess the effectiveness of anti-microtubule chemotherapies. Knowledge of how MAP expression levels affect the action of anti-microtubule drugs may prove useful for evaluating possible modes of cancer treatment.


Fluorescence-aided composite removal during lingual bracket debonding.
This report describes the fluorescence-aided composite removal during lingual bracket debonding with an ultraviolet light emitting diode flashlight. The purpose of this technique is to help clinicians in composite removal without enamel surface damage. The bracket debonding requires clinical attention in order to remove all composites and resins without enamel surface damage. Different protocols can be used in order to minimize the enamel damages and the excess bonding remnants. The fluorescence-aided composite removal permits to have an immediate visualization of the composites and adhesives, especially for the uneven lingual surfaces, of which the interindividual morphological variability is greater than the buccal surfaces. The fluorescence-aided composite removal during lingual brackets debonding minimizes the risks described in literature and it is an easier, more accurate, reliable, noninvasive, inexpensive, and time-saving method. The application of this technique allows, with inexpensive flashlights, to remove all the composite on the lingual surfaces during debonding, without damaging the tooth and saving time.


Neonatal severe hyperparathyroidism: genotype/phenotype correlation and the use of pamidronate as rescue therapy.
Familial hypocalciuric hypercalcaemia (FHH) is an autosomal dominant condition due to heterozygous loss of function calcium sensing receptor (CaSR) mutations. However, individuals who are homozygous for CaSR mutations have neonatal severe hyperparathyroidism (NSHPT), which unlike the relatively benign and asymptomatic FHH can be fatal without parathyroidectomy. We report three patients with NSHPT associated with marked hypercalcaemia and severe hyperparathyroidism with related skeletal demineralisation. We describe the novel use of intravenous pamidronate in NSHPT, to control severe hypercalcaemia in these patients prior to parathyroidectomy and in one individual as a rescue therapy to stabilise life-threatening demineralisation. Furthermore, a marked phenotypic heterogeneity was observed amongst four members from a large kindred with the same homozygous CaSR mutations: one patient would have died without parathyroidectomy in infancy; a second patient survived infancy but underwent parathyroidectomy in early childhood following severe symptomatic hypercalcaemic episodes; whilst the other two patients have survived to adolescence without parathyroidectomy. Additionally, in contrast to the literature these two individuals suffered minimal morbidity. We commend the short-term use of pamidronate in neonatal severe hyperparathyroidism to treat extreme hypercalcaemia and halt hyperparathyroid-driven skeletal demineralisation in preparation for parathyroidectomy. The remarkable degree of phenotypic variation demonstrated remains unexplained without functional studies; this variability highlights the challenge of treating this rare condition.


Cross-Sectional Associations of Reallocating Time Between Sedentary and Active Behaviours on Cardiometabolic Risk Factors in Young People: An International Children's Accelerometry Database (ICAD) Analysis.
Sedentary time and time spent in various intensity-specific physical activity are co-dependent, and increasing time spent in one behaviour requires decreased time in another. The aim of the present study was to examine the theoretical associations with reallocating time between categories of intensities and cardiometabolic risk factors in a large and heterogeneous sample of children and adolescents. We analysed pooled data from 13 studies comprising 18,200 children and adolescents aged 4-18 years from the International Children's Accelerometry Database (ICAD). Waist-mounted accelerometers measured sedentary time, light physical activity (LPA) and moderate-to-vigorous physical activity (MVPA). Cardiometabolic risk factors included waist circumference (WC), systolic blood pressure (SBP), fasting high- and low-density lipoprotein cholesterol (HDL-C and LDL-C), triglycerides, insulin, and glucose. Associations of reallocating time between the various intensity categories with cardiometabolic risk factors were explored using isotemporal substitution modelling. Replacing 10 min of sedentary time with 10 min of MVPA showed favourable associations with WC, SBP, LDL-C, insulin, triglycerides, and glucose; the greatest magnitude was observed for insulin (reduction of 2-4%), WC (reduction of 0.5-1%), and triglycerides (1-2%). In addition, replacing 10 min of sedentary time with an equal amount of LPA showed beneficial associations with WC, although only in adolescents. Replacing sedentary time and/or LPA with MVPA in children and adolescents is favourably associated with most markers of cardiometabolic risk. Efforts aimed at replacing sedentary time with active behaviours, particularly those of at least moderate intensity, appear to be an effective strategy to reduce cardiometabolic risk in young people.


Behavioural interventions for urinary incontinence in community-dwelling seniors: an evidence-based analysis.
In early August 2007, the Medical Advisory Secretariat began work on the Aging in the Community project, an evidence-based review of the literature surrounding healthy aging in the community. The Health System Strategy Division at the Ministry of Health and Long-Term Care subsequently asked the secretariat to provide an evidentiary platform for the ministry's newly released Aging at Home Strategy.After a broad literature review and consultation with experts, the secretariat identified 4 key areas that strongly predict an elderly person's transition from independent community living to a long-term care home. Evidence-based analyses have been prepared for each of these 4 areas: falls and fall-related injuries, urinary incontinence, dementia, and social isolation. For the first area, falls and fall-related injuries, an economic model is described in a separate report.Please visit the Medical Advisory Secretariat Web site, http://www.health.gov.on.ca/english/providers/program/mas/mas_about.html, to review these titles within the Aging in the Community series.AGING IN THE COMMUNITY: Summary of Evidence-Based AnalysesPrevention of Falls and Fall-Related Injuries in Community-Dwelling Seniors: An Evidence-Based AnalysisBehavioural Interventions for Urinary Incontinence in Community-Dwelling Seniors: An Evidence-Based AnalysisCaregiver- and Patient-Directed Interventions for Dementia: An Evidence-Based AnalysisSocial Isolation in Community-Dwelling Seniors: An Evidence-Based AnalysisThe Falls/Fractures Economic Model in Ontario Residents Aged 65 Years and Over (FEMOR) OBJECTIVE: To assess the effectiveness of behavioural interventions for the treatment and management of urinary incontinence (UI) in community-dwelling seniors. TARGET POPULATION AND CONDITION Urinary incontinence defined as "the complaint of any involuntary leakage of urine" was identified as 1 of the key predictors in a senior's transition from independent community living to admission to a long-term care (LTC) home. Urinary incontinence is a health problem that affects a substantial proportion of Ontario's community-dwelling seniors (and indirectly affects caregivers), impacting their health, functioning, well-being and quality of life. Based on Canadian studies, prevalence estimates range from 9% to 30% for senior men and nearly double from 19% to 55% for senior women. The direct and indirect costs associated with UI are substantial. It is estimated that the total annual costs in Canada are $1.5 billion (Cdn), and that each year a senior living at home will spend $1,000 to $1,500 on incontinence supplies. Interventions to treat and manage UI can be classified into broad categories which include lifestyle modification, behavioural techniques, medications, devices (e.g., continence pessaries), surgical interventions and adjunctive measures (e.g., absorbent products). The focus of this review is behavioural interventions, since they are commonly the first line of treatment considered in seniors given that they are the least invasive options with no reported side effects, do not limit future treatment options, and can be applied in combination with other therapies. In addition, many seniors would not be ideal candidates for other types of interventions involving more risk, such as surgical measures. It is recognized that the terms "senior" and "elderly" carry a range of meanings for different audiences; this report generally uses the former, but the terms are treated here as essentially interchangeable. Behavioural interventions can be divided into 2 categories according to the target population: caregiver-dependent techniques and patient-directed techniques. Caregiver-dependent techniques (also known as toileting assistance) are targeted at medically complex, frail individuals living at home with the assistance of a caregiver, who tends to be a family member. These seniors may also have cognitive deficits and/or motor deficits. A health care professional trains the senior's caregiver to deliver an intervention such as prompted voiding, habit retraining, or timed voiding. The health care professional who trains the caregiver is commonly a nurse or a nurse with advanced training in the management of UI, such as a nurse continence advisor (NCA) or a clinical nurse specialist (CNS). The second category of behavioural interventions consists of patient-directed techniques targeted towards mobile, motivated seniors. Seniors in this population are cognitively able, free from any major physical deficits, and motivated to regain and/or improve their continence. A nurse or a nurse with advanced training in UI management, such as an NCA or CNS, delivers the patient-directed techniques. These are often provided as multicomponent interventions including a combination of bladder training techniques, pelvic floor muscle training (PFMT), education on bladder control strategies, and self-monitoring. Pelvic floor muscle training, defined as a program of repeated pelvic floor muscle contractions taught and supervised by a health care professional, may be employed as part of a multicomponent intervention or in isolation. Education is a large component of both caregiver-dependent and patient-directed behavioural interventions, and patient and/or caregiver involvement as well as continued practice strongly affect the success of treatment. Incontinence products, which include a large variety of pads and devices for effective containment of urine, may be used in conjunction with behavioural techniques at any point in the patient's management. A comprehensive search strategy was used to identify systematic reviews and randomized controlled trials that examined the effectiveness, safety, and cost-effectiveness of caregiver-dependent and patient-directed behavioural interventions for the treatment of UI in community-dwelling seniors (see Appendix 1). Are caregiver-dependent behavioural interventions effective in improving UI in medically complex, frail community-dwelling seniors with/without cognitive deficits and/or motor deficits?Are patient-directed behavioural interventions effective in improving UI in mobile, motivated community-dwelling seniors?Are behavioural interventions delivered by NCAs or CNSs in a clinic setting effective in improving incontinence outcomes in community-dwelling seniors? The quality of the evidence was assessed as high, moderate, low, or very low according to the GRADE methodology and GRADE Working Group. As per GRADE the following definitions apply: HighFurther research is very unlikely to change confidence in the estimate of effect.ModerateFurther research is likely to have an important impact on confidence in the estimate of effect and may change the estimate.LowFurther research is very likely to have an important impact on confidence in the estimate of effect and is likely to change the estimate.Very LowAny estimate of effect is very uncertain Executive Summary Table 1 summarizes the results of the analysis. The available evidence was limited by considerable variation in study populations and in the type and severity of UI for studies examining both caregiver-directed and patient-directed interventions. The UI literature frequently is limited to reporting subjective outcome measures such as patient observations and symptoms. The primary outcome of interest, admission to a LTC home, was not reported in the UI literature. The number of eligible studies was low, and there were limited data on long-term follow-up. Executive Summary Table 1:Summary of Evidence on Behavioural Interventions for the Treatment of Urinary Incontinence in Community-Dwelling SeniorsInterventionTarget PopulationInterventionsConclusionsGRADE quality of the evidence1. Caregiver-dependent techniques (toileting assistance)Medically complex, frail individuals at home with/without cognitive deficits and/or motor deficitsDelivered by informal caregivers who are trained by a nurse or a nurse with specialized UI training (NCA/CNS)Prompted voidingHabit retrainingTimed voidingThere is no evidence of effectiveness for habit retraining (n=1 study) and timed voiding (n=1 study).Prompted voiding may be effective, but effectiveness is difficult to substantiate because of an inadequately powered study (n=1 study).Resource implications and caregiver burden (usually on an informal caregiver) should be considered.Low2. Patient-directed techniquesMobile, motivated seniorsDelivered by a nurse or a nurse with specialized UI training (NCA/CNS)Multicomponent behavioural interventionsInclude a combination ofBladder trainingPFMT (with or without biofeedback)Bladder control strategiesEducationSelf-monitoringSignificant reduction in the mean number of incontinent episodes per week (n=5 studies, WMD 3.63, 95% CI, 2.07-5.19)Significant improvement in patient's perception of UI (n=3 studies, OR 4.15, 95% CI, 2.70-6.37)Suggestive beneficial impact on patient's health-related quality of lifeModerate PFMT aloneSignificant reduction in the mean number of incontinent episodes per week (n=1 study, WMD 10.50, 95% CI, 4.30-16.70)Moderate3. Behavioural interventions led by an NCA/CNS in a clinic settingCommunity-dwelling seniorsBehavioural interventions led by NCA/CNSOverall, effective in improving incontinence outcomes (n=3 RCTs + 1 Ontario-based before/after study)Moderate*CI refers to confidence interval; CNS, clinical nurse specialist; NCA, nurse continence advisor; PFMT, pelvic floor muscle training; RCT, randomized controlled trial; WMD, weighted mean difference; UI, urinary incontinence. A budget impact analysis was conducted to forecast costs for caregiver-dependent and patient-directed multicomponent behavioural techniques delivered by NCAs, and PFMT alone delivered by physiotherapists. (ABSTRACT TRUNCATED)


Murine junctional adhesion molecules JAM-B and JAM-C mediate endothelial and stellate cell interactions during hepatic fibrosis.
Classical junctional adhesion molecules JAM-A, JAM-B and JAM-C influence vascular permeability, cell polarity as well as leukocyte recruitment and immigration into inflamed tissue. As the vasculature becomes remodelled in chronically injured, fibrotic livers we aimed to determine distribution and role of junctional adhesion molecules during this pathological process. Therefore, livers of naïve or carbon tetrachloride-treated mice were analyzed by immunohistochemistry to localize all 3 classical junctional adhesion molecules. Hepatic stellate cells and endothelial cells were isolated and subjected to immunocytochemistry and flow cytometry to determine localization and functionality of JAM-B and JAM-C. Cells were further used to perform contractility and migration assays and to study endothelial tubulogenesis and pericytic coverage by hepatic stellate cells. We found that in healthy tissue, JAM-A was ubiquitously expressed whereas JAM-B and JAM-C were restricted to the vasculature. During fibrosis, JAM-B and JAM-C levels increased in endothelial cells and JAM-C was de novo generated in myofibroblastic hepatic stellate cells. Soluble JAM-C blocked contractility but increased motility in hepatic stellate cells. Furthermore, soluble JAM-C reduced endothelial tubulogenesis and endothelial cell/stellate cell interaction. Thus, during liver fibrogenesis, JAM-B and JAM-C expression increase on the vascular endothelium. More importantly, JAM-C appears on myofibroblastic hepatic stellate cells linking them as pericytes to JAM-B positive endothelial cells. This JAM-B/JAM-C mediated interaction between endothelial cells and stellate cells stabilizes vessel walls and may control the sinusoidal diameter. Increased hepatic stellate cell contraction mediated by JAM-C/JAM-C interaction may cause intrahepatic vasoconstriction, which is a major complication in liver cirrhosis.


Health behaviors of mandated and voluntary students in a motivational intervention program.
College students engage in many unhealthy behaviors, one of these, heavy alcohol use, is a major global public health problem. This longitudinal study examined whether students' mandated/voluntary status in a program to reduce college drinking was associated with overall health, ethnicity, gender, and personality traits. Both mandated and voluntary groups participated in the Motivational Intervention (MI) program to prevent high risk drinking. Freshmen (710 voluntary, 190 mandated, n = 900) between the ages of 18 and 21, received the MI at baseline and again at 2 weeks, with boosters at 3, 6 and 12 months. Participants completed three measures: the Daily Drinking Questionnaire (DDQ); the Substance Use Risk Profile Scale (SURPS), and the Health Promoting Lifestyle Profile II (HPLPII). Mandated and voluntary participants were compared at baseline and following the intervention using two sample t-tests for continuous variables (overall health, personality traits, drinking measures), and chi-square for categorical variables (gender, ethnicity). Linear mixed models were used to identify associations between HPLP II scores and mandated/voluntary status, time, ethnicity gender and SURPS scale scores. In both groups, alcohol consumption dropped significantly by 12 months. Overall health-promoting behaviors, physical activity, stress management, and interpersonal relations improved in both groups between baseline and 12 months. Associations were found between alcohol consumption, personality traits, gender, and lifestyle health-promoting behaviors. In particular, impulsivity and hopelessness were associated with poor health behaviors. Intervention programs to reduce drinking by college students need to address developmental dynamics of freshmen students, including gender, psychosocial factors, personality, and lifestyle health-promoting behaviors.


Antiinflammatory effects of reconstituted high-density lipoprotein during human endotoxemia.
High-density lipoprotein (HDL) has been found to neutralize LPS activity in vitro and in animals in vivo. We sought to determine the effects of reconstituted HDL (rHDL) on LPS responsiveness in humans in a double-blind, randomized, placebo-controlled, cross-over study. rHDL, given as a 4-h infusion at 40 mg/kg starting 3.5 h before endotoxin challenge (4 ng/kg), reduced flu-like symptoms during endotoxemia, but did not influence the febrile response. rHDL potently reduced the endotoxin-induced release of TNF, IL-6, and IL-8, while only modestly attenuating the secretion of proinflammatory cytokine inhibitors IL-1ra, soluble TNF receptors and IL-10. In addition, rHDL attenuated LPS-induced changes in leukocyte counts and the enhanced expression of CD11b/CD18 on granulocytes. Importantly, rHDL infusion per se, before LPS administration, was associated with a downregulation of CD14, the main LPS receptor, on monocytes. This effect was biologically relevant, since monocytes isolated from rHDL-treated whole blood showed reduced expression of CD14 and diminished TNF production upon stimulation with LPS. These results suggest that rHDL may inhibit LPS effects in humans in vivo not only by binding and neutralizing LPS but also by reducing CD14 expression on monocytes.


An evolutionary transcriptomics approach links CD36 to membrane remodeling in replicative senescence.
Cellular senescence, the irreversible ceasing of cell division, has been associated with organismal aging, prevention of cancerogenesis, and developmental processes. As such, the evolutionary basis and biological features of cellular senescence remain a fascinating area of research. In this study, we conducted comparative RNAseq experiments to detect genes associated with replicative senescence in two different human fibroblast cell lines and at different time points. We identified 841 and 900 genes (core senescence-associated genes) that are significantly up- and downregulated in senescent cells, respectively, in both cell lines. Our functional enrichment analysis showed that downregulated core genes are primarily involved in cell cycle processes while upregulated core gene enrichment indicated various lipid-related processes. We further demonstrated that downregulated genes are significantly more conserved than upregulated genes. Using both transcriptomics and genetic variation data, we identified one of the upregulated, lipid metabolism genes, CD36, as an outlier. We found that overexpression of CD36 induces a senescence-like phenotype and, further, the media of CD36-overexpressing cells alone can induce a senescence-like phenotype in proliferating young cells. Moreover, we used a targeted lipidomics approach and showed that phosphatidylcholines accumulate during replicative senescence in these cells, suggesting that upregulation of CD36 could contribute to membrane remodeling during senescence. Overall, these results contribute to the understanding of evolution and biology of cellular senescence and identify several targets and questions for future studies.


Genomic relationships between selected phage types of Salmonella enterica subsp. enterica serotype typhimurium defined by ribotyping, IS200 typing and PFGE.
The genomic relationship between isolates representing 17 definitive phage types (DTs) of Salmonella enterica subsp. enterica serotype typhimurium (S. typhimurium) were analysed using three different typing methods: IS200 typing using the restriction enzymes EcoRI and PvuII, ribotyping using SmaI and EcoRI, and PFGE using XbaI. These methods were used to study four DTs in greater detail; in all 18 (DT 49), 10 (DT 110), five (DT 120) and seven (DT 135) isolates were studied. The combined data generated two large clusters, which could be divided into five groups. Within the first cluster, a close similarity was indicated between isolates of the following phage types: group A-DTs 44, 49, 135 and 204c, with DT 9 distantly related; group B-DTs 95 and 99; and group C-DTs 104a, 110 and 120. The other large cluster contained group D-DTs 10, 20 and 146, with DT 12 distantly related, and group E-DTs 69, 103 and 153. The same grouping was observed by principal component analysis, but a minimum spanning tree linked DT 12 to group E and not group D in this analysis. Among the typing methods used, IS200 gave the best representation of the overall similarity between the S. typhimurium isolates. Five different IS200 profiles were obtained among isolates belonging to DT 49. Only one profile was observed within each of the phage types DT 110, 120 and 135. All isolates within each of these four phage types were of one ribotype. Isolates of DT 49 showed four PFGE patterns, while one pattern was present within isolates of the three other phage types. Members of these four phage types were found to be clonally related as they formed tight subclusters separated from isolates of other phage types.


Course of self-reported symptoms of attention deficit and hyperactivity in substance abusers during early treatment.
Attention deficit and hyperactivity disorder has been associated with poor outcome in studies of substance use disorders. This study aimed to assess the course of self-reported symptoms of both attention deficit and hyperactivity among adults presenting for treatment for substance use disorders. A sample of 75 substance abusers were assessed after they were admitted to a centralized intake unit, and followed at 3 and 6 months after intake by independent interviewers (follow-up rate 81%). Symptoms of attention deficit and hyperactivity were assessed with the Adult Self-report Scale for ADHD (ASRS). Both types of symptoms declined significantly during follow-up, but attention symptoms had a high intraclass correlation (0.79), and hyperactivity had a moderate intraclass correlation (0.64). Both baseline attention deficit and hyperactivity symptoms were associated with worse work and social adjustment after controlling for baseline functioning. Hyperactivity was associated with poorer substance use outcomes at the trend level. In conclusion, both dimensions of ADHD contribute to worse functioning during early treatment for substance abusers, and the ASRS is a reasonably stable measure of ADHD symptoms during early recovery.


Sex-specific differences in HPA axis activity in VLBW preterm newborns.
Sex-specific differences in hypothalamic-pituitary-adrenal axis activity might explain why male preterm infants are at higher risk of neonatal mortality and morbidity than their female counterparts. We examined whether male and female preterm infants differed in cortisol production and metabolism at 10 days post-partum. This prospective study included 36 preterm born infants (18 boys) with a very low birth weight (VLBW) (<1.500 g). At 10 days postnatal age, urine was collected over a 4- to 6-h period. Glucocorticoid metabolites were measured using gas chromatography-mass spectrometry. Main outcome measures were: (1) cortisol excretion rate, (2) sum of all glucocorticoid metabolites, as an index of corticosteroid excretion rate, and (3) ratio of 11-OH/11-OXO metabolites, as an estimate of 11B-hydroxysteroid dehydrogenase (11B-HSD) activity. Differences between sexes, including interaction with Score of Neonatal Acute Physiology Perinatal Extension-II (SNAPPE II), sepsis and bronchopulmonary dysplasia (BPD), were assessed. No differences between sexes were found for cortisol excretion rate, corticosteroid excretion rate or 11B-HSD activity. Interaction was observed between: sex and SNAPPE II score on 11B-HSD activity (P = 0.04) and sex and BPD on cortisol excretion rate (P = 0.04). This study did not provide evidence for sex-specific differences in adrenocortical function in preterm VLBW infants on a group level. However, in an interaction model, sex differences became manifest under stressful circumstances. These patterns might provide clues for the male disadvantage in neonatal mortality and morbidity following preterm birth. However, due to the small sample size, the data should be seen as hypothesis generating.


Error rates in buccal-dental microwear quantification using scanning electron microscopy.
Dental microwear, usually analyzed using scanning electron microscopy (SEM) techniques, is a good indicator of the abrasive potential of past human population diets. Scanning electron microscopy secondary electrons provide excellent images of dental enamel relief for characterizing striation density, average length, and orientation. However, methodological standardization is required for interobserver comparisons since semiautomatic counting procedures are still used for micrograph characterization. The analysis of normally distributed variables allows the characterization of small interpopulation differences. However, the interobserver error rates associated with SEM experience and the degree of expertise in measuring striations are critical to population dietary interpretation. The interobserver comparisons made here clearly indicate that the precision of SEM buccal microwear measurements depends heavily on variable definition and the researcher's expertise. Moreover, error rates are not the only concern for dental microwear research. Low error rates do not guarantee that all researchers are measuring the same magnitudes of the variables considered. The results obtained show that researchers tend to maintain high intrapopulation homogeneity and low measurement error rates, whereas significant interobserver differences appear. Such differences are due to a differential interpretation of SEM microwear features and variable definitions that require detailed and precise agreement among researchers. The substitution of semiautomatic with fully automated procedures will completely avoid interobserver error rate differences.


Genes and causation.
Relating genotypes to phenotypes is problematic not only owing to the extreme complexity of the interactions between genes, proteins and high-level physiological functions but also because the paradigms for genetic causality in biological systems are seriously confused. This paper examines some of the misconceptions, starting with the changing definitions of a gene, from the cause of phenotype characters to the stretches of DNA. I then assess whether the 'digital' nature of DNA sequences guarantees primacy in causation compared to non-DNA inheritance, whether it is meaningful or useful to refer to genetic programs, and the role of high-level (downward) causation. The metaphors that served us well during the molecular biological phase of recent decades have limited or even misleading impacts in the multilevel world of systems biology. New paradigms are needed if we are to succeed in unravelling multifactorial genetic causation at higher levels of physiological function and so to explain the phenomena that genetics was originally about. Because it can solve the 'genetic differential effect problem', modelling of biological function has an essential role to play in unravelling genetic causation.


Enterotoxigenic Escherichia coli in developing countries: epidemiology, microbiology, clinical features, treatment, and prevention.
ETEC is an underrecognized but extremely important cause of diarrhea in the developing world where there is inadequate clean water and poor sanitation. It is the most frequent bacterial cause of diarrhea in children and adults living in these areas and also the most common cause of traveler's diarrhea. ETEC diarrhea is most frequently seen in children, suggesting that a protective immune response occurs with age. The pathogenesis of ETEC-induced diarrhea is similar to that of cholera and includes the production of enterotoxins and colonization factors. The clinical symptoms of ETEC infection can range from mild diarrhea to a severe cholera-like syndrome. The effective treatment of ETEC diarrhea by rehydration is similar to treatment for cholera, but antibiotics are not used routinely for treatment except in traveler's diarrhea. The frequency and characterization of ETEC on a worldwide scale are inadequate because of the difficulty in recognizing the organisms; no simple diagnostic tests are presently available. Protection strategies, as for other enteric infections, include improvements in hygiene and development of effective vaccines. Increases in antimicrobial resistance will dictate the drugs used for the treatment of traveler's diarrhea. Efforts need to be made to improve our understanding of the worldwide importance of ETEC.


Anti-inflammatory property of the cannabinoid agonist WIN-55212-2 in a rodent model of chronic brain inflammation.
Cannabinoid receptors (CBr) stimulation induces numerous central and peripheral effects. A growing interest in the beneficial properties of manipulating the endocannabinoid system has led to the possible involvement of CBr in the control of brain inflammation. In the present study we examined the effect of the CBr agonist, (R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)-pyrrolo[1,2,3-de]-1,4benzoxazin-6-yl]-1-naphthalenyl-methanone mesylate (WIN-55212-2), on microglial activation and spatial memory performance, using a well-characterized animal model of chronic brain inflammation produced by the infusion of lipopolysaccharide (LPS, 250 ng/h for 3 weeks) into the fourth ventricle of young rats. WIN-55212-2 (0.5 or 1.0 mg/kg/day, i.p.) was administered for 3 weeks. During the third week of treatment, spatial memory ability was examined using the Morris water-maze task. We found that 0.5 and 1 mg/kg WIN-55212-2 reduced the number of LPS-activated microglia, while 1 mg/kg WIN-55212-2 potentiated the LPS-induced impairment of performance in the water maze task. Cannabinoid receptors 1 were not expressed by microglia and astrocytes, suggesting an indirect effect of WIN-55212-2 on microglia activation and memory impairment. Our results emphasize the potential use of CBr agonists in the regulation of inflammatory processes within the brain; this knowledge may lead to the use of CBr agonists in the treatment of neurodegenerative diseases associated with chronic neuroinflammation, such as Alzheimer disease.


N-terminal pro-brain natriuretic peptide (NT-proBNP) serum concentrations in apparently healthy Bosnian women.
Brain natriuretic peptide (BNP) is a cardiac hormone secreted predominantly from the ventricles. This hormone is produced as pre-prohormone BNP (pro BNP), than cleaved by corine to biologically active 32-aminoacid BNP and non-biologically active N-terminal-pro brain natriuretic peptide (NTproBNP). NTproBNP has been found to be a useful marker for the diagnosis of heart failure and left ventricular systolic dysfunction. Recent studies showed that concentration of BNP and NTproBNP predict cardiovascular disease in apparently healthy individuals but their full screening characteristics are not firmly established. As NTproBNP serum concentration is altered by numerous factors there are also interindividual variations in NTproBNP values. There are no previous results for Bosnian population so the aim of this study was to asses normal range of NTproBNP serum concentrations in apparently healthy women using electrochemiluminescence immunoassay (Elecsys, Roche Diagnostic). A group of 45, healthy females, aged 39.19 (+/-6.62), were enrolled in this study. Mean serum concentration of NTproBNP was 60.32 (+/-36.25) pg/ml, with the range of 112,60 pg/ml (minimum-maximum: 13.6-126.00 pg/ml). We conclude that NTproBNP serum concentration in apparently healthy Bosnian women was not different from the average values of NTproBNP obtained on Europen's population. Thus, we suggest that the NTproBNP serum upper cut off values measured by using electrochemiluminescence immunoassay "ECLIA" (Elecsys 2010, Roche Diagnostic) for Bosnian females, aged < or =50 years, should be 155 pg/ml as reported by Roche Diagnostic.


Longitudinal Dynamic Analyses of Depression and Academic Achievement in the Hawaiian High Schools Health Survey using Contemporary Latent Variable Change Models.
The scientific literature consistently supports a negative relationship between adolescent depression and educational achievement, but we are certainly less sure on the causal determinants for this robust association. In this paper we present multivariate data from a longitudinal cohort-sequential study of high school students in Hawai'i (following McArdle, 2009; McArdle, Johnson, Hishinuma, Miyamoto, & Andrade, 2001). We first describe the full set of data on academic achievements and self-reported depression. We then carry out and present a progression of analyses in an effort to determine the accuracy, size, and direction of the dynamic relationships among depression and academic achievement, including gender and ethnic group differences. We apply three recently available forms of longitudinal data analysis: (1) Dealing with Incomplete Data -- We apply these methods to cohort-sequential data with relatively large blocks of data which are incomplete for a variety of reasons (Little & Rubin, 1987; McArdle & Hamagami, 1992). (2) Ordinal Measurement Models (Muthén & Muthén, 2006) -- We use a variety of statistical and psychometric measurement models, including ordinal measurement models to help clarify the strongest patterns of influence. (3) Dynamic Structural Equation Models (DSEMs; McArdle, 2009). We found the DSEM approach taken here was viable for a large amount of data, the assumption of an invariant metric over time was reasonable for ordinal estimates, and there were very few group differences in dynamic systems. We conclude that our dynamic evidence suggests that depression affects academic achievement, and not the other way around. We further discuss the methodological implications of the study.


Functional analysis of light-regulated promoter region of AtPolλ gene.
Genetic and molecular analyses mainly in Arabidopsis and in some other plants have demonstrated involvement of light signaling in cell cycle regulation. In this report, we show light-mediated activation of the promoter of AtPolλ gene, a homolog of mammalian DNA polymerase λ in Arabidopsis thaliana and an important component of DNA damage repair/recombination machinery in plants. Analyses of the light-mediated promoter activity using various deletion versions of AtPolλ promoter in transformed Arabidopsis and tobacco (Nicotiana tabaccum) plants indicate that a 130-bp promoter region between -536 and -408 of AtPolλ promoter is essential for light-induced regulation of AtPolλ expression. DNA-protein interaction studies reveal that an ATCT-motif and AE-box light-responsive elements in the light-regulated promoter region confer light responsiveness of AtPolλ promoter. DNA-binding analysis has identified a 63-kDa trans-acting protein factor which showed specific binding to ATCT-motif, while another trans-acting factor of ~52 kDa was found to bind specifically to both ATCT and AE-box sequences. The 52-kDa protein has been identified as B3-domain transcription factor by MALDI-TOF/MS analysis. Overall, our results provide novel information on the role of light signaling in regulation of expression of an important component of DNA repair machinery in plants.


Sepsis induces long-term metabolic and mitochondrial muscle stem cell dysfunction amenable by mesenchymal stem cell therapy.
Sepsis, or systemic inflammatory response syndrome, is the major cause of critical illness resulting in admission to intensive care units. Sepsis is caused by severe infection and is associated with mortality in 60% of cases. Morbidity due to sepsis is complicated by neuromyopathy, and patients face long-term disability due to muscle weakness, energetic dysfunction, proteolysis and muscle wasting. These processes are triggered by pro-inflammatory cytokines and metabolic imbalances and are aggravated by malnutrition and drugs. Skeletal muscle regeneration depends on stem (satellite) cells. Herein we show that mitochondrial and metabolic alterations underlie the sepsis-induced long-term impairment of satellite cells and lead to inefficient muscle regeneration. Engrafting mesenchymal stem cells improves the septic status by decreasing cytokine levels, restoring mitochondrial and metabolic function in satellite cells, and improving muscle strength. These findings indicate that sepsis affects quiescent muscle stem cells and that mesenchymal stem cells might act as a preventive therapeutic approach for sepsis-related morbidity.


Effects of lipid nanocarriers on the performance of topical vehicles in vivo.
Nanocarrier systems have been extensively studied for their suitability in personal care formulations. Theoretically, they could enhance skin delivery of active compounds, thereby improving in vivo efficacy of the products. As such the aim of this study was to evaluate the effect of a lipid nanocarrier (LNC) system loaded with tocopheryl acetate (TA) on the hydration, biomechanical properties, and antioxidant capacity of human skin, when used in two different vehicles, and compare it with a non-LNC formulation. TA-loaded lipid nanocarriers (TA-LNCs) were produced by the phase inversion method, using physiological lipids and purified by ultra-centrifugation. They were incorporated into a hydrophilic gel and foam, and their performance compared with a saturated TA solution in silicon oil. Skin hydration and biomechanical properties were measured by means of a corneometer and a cutometer, respectively, while a high-resolution spectrophotometer was used to assess skin redness after stimulation by methyl nicotinate in a micro-inflammatory test. Both short-term (3 h) and long-term trials (4 weeks) were performed. The results confirmed that the LNCs enhanced skin hydration in both studies, while skin viscoelastic parameters remained practically unchanged during the 4-week study. The antioxidant assessment failed to show significant difference between the test sites. TA-loaded LNCs exhibited the ability to enhance skin hydration, while their effect on skin biomechanical properties and on antioxidant efficacy could not be statistically proved.


Adverse effects of medical cannabinoids: a systematic review.
The therapeutic use of cannabis and cannabis-based medicines raises safety concerns for patients, clinicians, policy-makers, insurers, researchers and regulators. Although the efficacy of cannabinoids is being increasingly demonstrated in randomized controlled trials, most safety information comes from studies of recreational use. We performed a systematic review of safety studies of medical cannabinoids published over the past 40 years to create an evidence base for cannabis-related adverse events and to facilitate future cannabis research initiatives. We critically evaluated the quality of published studies with a view to identifying ways to improve future studies. A total of 321 articles were eligible for evaluation. After excluding those that focused on recreational cannabis use, we included 31 studies (23 randomized controlled trials and 8 observational studies) of medical cannabis use in our analysis. In the 23 randomized controlled trials, the median duration of cannabinoid exposure was 2 weeks (range 8 hours to 12 months). A total of 4779 adverse events were reported among participants assigned to the intervention. Most (4615 [96.6%]) were not serious. Of the 164 serious adverse events, the most common was relapse of multiple sclerosis (21 events [12.8%]), vomiting (16 events [9.8%]) and urinary tract infection (15 events [9.1%]). The rate of nonserious adverse events was higher among participants assigned to medical cannabinoids than among controls (rate ratio [RR] 1.86, 95% confidence interval [CI] 1.57-2.21); the rates of serious adverse events did not differ significantly between these 2 groups (RR 1.04, 95% CI 0.78-1.39). Dizziness was the most commonly reported nonserious adverse event (714 events [15.5%]) among people exposed to cannabinoids. Short-term use of existing medical cannabinoids appeared to increase the risk of nonserious adverse events. The risks associated with long-term use were poorly characterized in published clinical trials and observational studies. High-quality trials of long-term exposure are required to further characterize safety issues related to the use of medical cannabinoids.


The Role of Metabotropic Glutamate Receptor Genes in Schizophrenia.
Genomic studies revealed two main components in the genetic architecture of schizophrenia, one constituted by common variants determining a distributed polygenic effect and one represented by a large number of heterogeneous rare and highly disruptive mutations. These gene modifications often affect neural transmission and different studies proved an involvement of metabotropic glutamate receptors in schizophrenia phenotype. Through the combination of literature information with genomic data from public repositories, we analyzed the current knowledge on the involvement of genetic variations of the human metabotropic glutamate receptors in schizophrenia and related endophenotypes. Despite the analysis did not reveal a definitive connection, different suggestive associations have been identified and in particular a relevant role has emerged for GRM3 in affecting specific schizophrenia endophenotypes. This supports the hypothesis that these receptors are directly involved in schizophrenia disorder.


Prediction of deleterious non-synonymous SNPs based on protein interaction network and hybrid properties.
Non-synonymous SNPs (nsSNPs), also known as Single Amino acid Polymorphisms (SAPs) account for the majority of human inherited diseases. It is important to distinguish the deleterious SAPs from neutral ones. Most traditional computational methods to classify SAPs are based on sequential or structural features. However, these features cannot fully explain the association between a SAP and the observed pathophysiological phenotype. We believe the better rationale for deleterious SAP prediction should be: If a SAP lies in the protein with important functions and it can change the protein sequence and structure severely, it is more likely related to disease. So we established a method to predict deleterious SAPs based on both protein interaction network and traditional hybrid properties. Each SAP is represented by 472 features that include sequential features, structural features and network features. Maximum Relevance Minimum Redundancy (mRMR) method and Incremental Feature Selection (IFS) were applied to obtain the optimal feature set and the prediction model was Nearest Neighbor Algorithm (NNA). In jackknife cross-validation, 83.27% of SAPs were correctly predicted when the optimized 263 features were used. The optimized predictor with 263 features was also tested in an independent dataset and the accuracy was still 80.00%. In contrast, SIFT, a widely used predictor of deleterious SAPs based on sequential features, has a prediction accuracy of 71.05% on the same dataset. In our study, network features were found to be most important for accurate prediction and can significantly improve the prediction performance. Our results suggest that the protein interaction context could provide important clues to help better illustrate SAP's functional association. This research will facilitate the post genome-wide association studies.


Comparison Among Aerobic Exercise and Other Types of Interventions to Treat Depression: A Systematic Review.
Depression is a common and disabling disease that affects over 100 million people worldwide and can have a significant impact on physical and mental health, reducing their quality of life. Thus, the aim of this article was to provide information on research results and key chains related to the therapeutic effects of chronic aerobic exercise compared with other types of interventions to treat depression, which may become a useful clinical application in a near future. Researches have shown the effectiveness of alternative treatments, such as physical exercise, minimizing high financial costs and minimizing side effects. In this review, the data analyzed allows us to claim that alternative therapies, such as exercise, are effective on controlling and reducing symptoms. 69.3% of the studies that investigated the antidepressant effects of exercise on depressive were significant, and the other 30.7% of the studies improved only in general physiological aspects, such as increased oxygen uptake, increased use of blood glucose and decreased body fat percentage, with no improvement on symptoms of depression. From the sample analyzed, 71.4% was composed of women, and regarding the severity of symptoms, 85% had mild to moderate depression and only 15% had moderate to severe depression. However, there is still disagreement regarding the effect of exercise compared to the use of antidepressants in symptomatology and cognitive function in depression, this suggests that there is no consensus on the correct intensity of aerobic exercise as to achieve the best dose-response, with intensities high to moderate or moderate to mild.


In vitro osteoblast response to ferritic stainless steel fiber networks for magneto-active layers on implants.
The use of a porous coating on prosthetic components to encourage bone ingrowth is an important way of improving uncemented implant fixation. Enhanced fixation may be achieved by the use of porous magneto-active layers on the surface of prosthetic implants, which would deform elastically on application of a magnetic field, generating internal stresses within the in-growing bone. This approach requires a ferromagnetic material able to support osteoblast attachment, proliferation, differentiation, and mineralization. In this study, the human osteoblast responses to ferromagnetic 444 stainless steel networks were considered alongside those to nonmagnetic 316L (medical grade) stainless steel networks. While both networks had similar porosities, 444 networks were made from coarser fibers, resulting in larger inter-fiber spaces. The networks were analyzed for cell morphology, distribution, proliferation, and differentiation, extracellular matrix production and the formation of mineralized nodules. Cell culture was performed in both the presence of osteogenic supplements, to encourage cell differentiation, and in their absence. It was found that fiber size affected osteoblast morphology, cytoskeleton organization and proliferation at the early stages of culture. The larger inter-fiber spaces in the 444 networks resulted in better spatial distribution of the extracellular matrix. The addition of osteogenic supplements enhanced cell differentiation and reduced cell proliferation thereby preventing the differences in proliferation observed in the absence of osteogenic supplements. The results demonstrated that 444 networks elicited favorable responses from human osteoblasts, and thus show potential for use as magnetically active porous coatings for advanced bone implant applications.


The diastolic viscous properties of cat papillary muscle.
The measurement of ventricular diastolic compliance is complicated by the presence of viscous properties. To approach this problem, cat papillary muscles contracting isometrically at 15/min at 26 degrees C were stretched at different velocities during diastole. At a given length there was no excess of force above that defined by the static force-length curve until velocity of stretch exceeded 1 muscle length/sec. At greater velocities of stretch there was an increase of force with increasing velocity of stretch denoting viscous resistance to stretch. The viscous characteristic was alinear and increased with muscle length. At a given muscle length potentiation of contractile state by paired pulse stimulation caused a change in the diastolic force-length curve but there was no apparent relationship between viscous resistance to stretch and contractile state or the length of the "series viscous element." These results lead one to expect viscous resistance to stretch during rapid filling of the intact left ventricle.


Administration of zoledronic acid enhances the effects of docetaxel on growth of prostate cancer in the bone environment.
After development of hormone-refractory metastatic disease, prostate cancer is incurable. The recent history of chemotherapy has shown that with difficult disease targets, combinatorial therapy frequently offers the best chance of a cure. In this study we have examined the effects of a combination of zoledronic acid (ZOL), a new-generation bisphosphonate, and docetaxel on LuCaP 23.1, a prostate cancer xenograft that stimulates the osteoblastic reaction when grown in the bone environment. Intra-tibial injections of LuCaP 23.1 cells were used to generate tumors in the bone environment, and animals were treated with ZOL, docetaxel, or a combination of these. Effects on bone and tumor were evaluated by measurements of bone mineral density and histomorphometrical analysis. ZOL decreased proliferation of LuCaP 23.1 in the bone environment, while docetaxel at a dose that effectively inhibited growth of subcutaneous tumors did not show any effects in the bone environment. The combination of the drugs significantly inhibited the growth of LuCaP 23.1 tumors in the bone. In conclusion, the use of the osteolysis-inhibitory agent ZOL in combination with docetaxel inhibits growth of prostate tumors in bone and represents a potential treatment option.


Food supply and seawater pCO2 impact calcification and internal shell dissolution in the blue mussel Mytilus edulis.
Progressive ocean acidification due to anthropogenic CO(2) emissions will alter marine ecosystem processes. Calcifying organisms might be particularly vulnerable to these alterations in the speciation of the marine carbonate system. While previous research efforts have mainly focused on external dissolution of shells in seawater under saturated with respect to calcium carbonate, the internal shell interface might be more vulnerable to acidification. In the case of the blue mussel Mytilus edulis, high body fluid pCO(2) causes low pH and low carbonate concentrations in the extrapallial fluid, which is in direct contact with the inner shell surface. In order to test whether elevated seawater pCO(2) impacts calcification and inner shell surface integrity we exposed Baltic M. edulis to four different seawater pCO(2) (39, 142, 240, 405 Pa) and two food algae (310-350 cells mL(-1) vs. 1600-2000 cells mL(-1)) concentrations for a period of seven weeks during winter (5°C). We found that low food algae concentrations and high pCO(2) values each significantly decreased shell length growth. Internal shell surface corrosion of nacreous ( = aragonite) layers was documented via stereomicroscopy and SEM at the two highest pCO(2) treatments in the high food group, while it was found in all treatments in the low food group. Both factors, food and pCO(2), significantly influenced the magnitude of inner shell surface dissolution. Our findings illustrate for the first time that integrity of inner shell surfaces is tightly coupled to the animals' energy budget under conditions of CO(2) stress. It is likely that under food limited conditions, energy is allocated to more vital processes (e.g. somatic mass maintenance) instead of shell conservation. It is evident from our results that mussels exert significant biological control over the structural integrity of their inner shell surfaces.


Predicting age at onset of menopause: testing the "adaptive onset" hypothesis.
This study tests the "adaptive onset" hypothesis (AOH) proposed by Kuhle and explores the potential adaptive origins of menopause (Kuhle (2007) [3]). The AOH posits that both menopause and the timing of its onset are adaptive, and that age at onset may be mediated according to the likelihood of successful continued reproduction. Twelve factors were hypothesized to predict whether continued reproduction would be successful. Eight factors were tested using data extracted from the NLS Mature Women cohort. Statistical analyses reveal no support for predictions of the AOH; in fact, a majority of analyses suggest trends opposite from those predicted.


Transcriptional analysis of South African cassava mosaic virus-infected susceptible and tolerant landraces of cassava highlights differences in resistance, basal defense and cell wall associated genes during infection.
Cassava mosaic disease is caused by several distinct geminivirus species, including South African cassava mosaic virus-[South Africa:99] (SACMV). To date, there is limited gene regulation information on viral stress responses in cassava, and global transcriptome profiling in SACMV-infected cassava represents an important step towards understanding natural host responses to plant geminiviruses. A RNA-seq time course (12, 32 and 67 dpi) study, monitoring gene expression in SACMV-challenged susceptible (T200) and tolerant (TME3) cassava landraces, was performed using the Applied Biosystems (ABI) SOLiD next-generation sequencing platform. The multiplexed paired end sequencing run produced a total of 523 MB and 693 MB of paired-end reads for SACMV-infected susceptible and tolerant cDNA libraries, respectively. Of these, approximately 50.7% of the T200 reads and 55.06% of TME3 reads mapped to the cassava reference genome available in phytozome. Using a log2 fold cut-off (p<0.05), comparative analysis between the six normalized cDNA libraries showed that 4181 and 1008 transcripts in total were differentially expressed in T200 and TME3, respectively, across 12, 32 and 67 days post infection, compared to mock-inoculated. The number of responsive transcripts increased dramatically from 12 to 32 dpi in both cultivars, but in contrast, in T200 the levels did not change significantly at 67 dpi, while in TME3 they declined. GOslim functional groups illustrated that differentially expressed genes in T200 and TME3 were overrepresented in the cellular component category for stress-related genes, plasma membrane and nucleus. Alterations in the expression of other interesting genes such as transcription factors, resistance (R) genes, and histone/DNA methylation-associated genes, were observed. KEGG pathway analysis uncovered important altered metabolic pathways, including phenylpropanoid biosynthesis, sucrose and starch metabolism, and plant hormone signalling. Molecular mechanisms for TME3 tolerance are proposed, and differences in patterns and levels of transcriptome profiling between T200 and TME3 with susceptible and tolerant phenotypes, respectively, support the hypothesis that viruses rearrange their molecular interactions in adapting to hosts with different genetic backgrounds.


Good participatory practice for coronavirus disease 2019 (COVID-19) research: the case of a COVID-19 prevention study.
Background: The COPCOV study (chloroquine/ hydroxychloroquine prevention of coronavirus disease), which started recruitment in April 2020, is a multi-country double-blind, randomised, placebo-controlled trial which is being conducted in healthcare facilities involved in COVID-19 case management. Participants are staff employed in facilities managing people with proven or suspected COVID-19. As part of the study, we conducted a series of engagement sessions. The aims were to assess the feasibility of the study, to identify context-specific ethical issues, to understand possible concerns, to fine tune research procedures and to refine the COPCOV information materials. Methods: The COPCOV study was approved by relevant institutional review boards. The sessions described in this paper were part of the study. We conducted a series of engagement sessions, each involving a short presentation of the study, a section where attendees were asked to express their willingness to participate in such a study, which information they would need to change their view and an open Q&A section. Answers were transcribed and coded into themes by two independent investigators. Themes were derived from the data. They complemented other site-specific engagement, communication, and public relation activities such as press releases and websites. Results and conclusions: From 16 th March 2020 to 20 th January 2021, 12 engagement sessions were conducted in Thailand, Laos, Vietnam, Nepal and the UK involving 213 attendees in total. Issues raised revolved around the social value and study rationale; safety of trial medications and risk-benefit balance; study design and commitments. These sessions helped us identify concerns people had, which helped us refine information materials as well as complement site feasibility assessments. Our experience strongly supports the use of participatory practices prior to conducting clinical trials.


Burnout, career satisfaction, and well-being among US neurologists in 2016.
To study prevalence of and factors that contribute to burnout, career satisfaction, and well-being in US neurologists. A total of 4,127 US American Academy of Neurology member neurologists who had finished training were surveyed using validated measures of burnout, career satisfaction, and well-being from January 19 to March 21, 2016. Response rate was 40.5% (1,671 of 4,127). Average age of participants was 51 years, with 65.3% male and nearly equal representation across US geographic regions. Approximately 60% of respondents had at least one symptom of burnout. Hours worked/week, nights on call/week, number of outpatients seen/week, and amount of clerical work were associated with greater burnout risk. Effective support staff, job autonomy, meaningful work, age, and subspecializing in epilepsy were associated with lower risk. Academic practice (AP) neurologists had a lower burnout rate and higher rates of career satisfaction and quality of life than clinical practice (CP) neurologists. Some factors contributing to burnout were shared between AP and CP, but some risks were unique to practice setting. Factors independently associated with profession satisfaction included meaningfulness of work, job autonomy, effectiveness of support staff, age, practicing sleep medicine (inverse relationship), and percent time in clinical practice (inverse relationship). Burnout was strongly associated with decreased career satisfaction. Burnout is common in all neurology practice settings and subspecialties. The largest driver of career satisfaction is the meaning neurologists find in their work. The results from this survey will inform approaches needed to reduce burnout and promote career satisfaction and well-being in US neurologists.


Polarization Sagnac interferometer with postmodulation for gravitational-wave detection.
We describe a polarization Sagnac interferometer with an in-loop half-wave plate that allows signal detection at the reciprocal port of the beam splitter while maintaining the ability to detect the signal at a dark fringe. Postmodulation and balanced heterodyne detection are used to recover the signal. This topology is simple to control because of its common-path characteristics and its collinear signal and local oscillator. The robustness of this scheme to amplitude and frequency fluctuations of the laser is demonstrated. Intraloop birefringence in this interferometer acts as a loss, reducing the power on the detector. The magnitude of this loss is discussed and experimentally verified.


Neuroprotective efficacy of methylene blue in ischemic stroke: an MRI study.
Methylene blue (MB) has unique energy-enhancing and antioxidant properties and is FDA-approved drug to treat methemoglobinemia and cyanide poisoning. This study evaluated the efficacy of MB to treat ischemic stroke in rats using longitudinal MRI and behavioral measures. Rats were subjected to 60-minute middle-cerebral-artery occlusion. In a randomized double-blinded design, vehicle or MB was administered after reperfusion. The initial lesion volumes at 30 minutes post-ischemia were not significantly different between the two groups (P = 0.92). The final infarct volumes two days after stroke increased in the vehicle group but decreased in the MB group, yielding a 30% difference in infarct volume (P = 0.03). Tracking tissue fate on a pixel-by-pixel basis showed that MB salvaged more initial core pixels compared to controls (22±3% versus 11±3%, P = 0.03), and more mismatch pixels compared to controls (83±3% versus 61±8%, P = 0.02). This study demonstrates MB treatment minimizes ischemic brain injury and improves functional outcomes.


Microchimerism in female bone marrow and bone decades after fetal mesenchymal stem-cell trafficking in pregnancy.
Fetal cells enter maternal blood during pregnancy and persist in women with autoimmune disease. The frequency of subsequent fetomaternal microchimerism in healthy women and its cell type is unknown. To test the hypothesis that fetal mesenchymal stem cells persist in maternal organs, we studied female bone marrow and ribs. Male cells were identified by XY fluorescence in-situ hybridisation in marrow-derived mesenchymal stem cells and in rib sections from all women with male pregnancies, but not in controls (9/9 vs 0/5, p=0.0005). We conclude that fetal stem cells transferred into maternal blood engraft in marrow, where they remain throughout life. This finding has implications for normal pregnancy, for obstetric complications that increase fetomaternal trafficking, and for graft survival after transplantation.


The Composition and Biological Activity of Honey: A Focus on Manuka Honey.
Honey has been used as a food and medical product since the earliest times. It has been used in many cultures for its medicinal properties, as a remedy for burns, cataracts, ulcers and wound healing, because it exerts a soothing effect when initially applied to open wounds. Depending on its origin, honey can be classified in different categories among which, monofloral honey seems to be the most promising and interesting as a natural remedy. Manuka honey, a monofloral honey derived from the manuka tree (Leptospermum scoparium), has greatly attracted the attention of researchers for its biological properties, especially its antimicrobial and antioxidant capacities. Our manuscript reviews the chemical composition and the variety of beneficial nutritional and health effects of manuka honey. Firstly, the chemical composition of manuka honey is described, with special attention given to its polyphenolic composition and other bioactive compounds, such as glyoxal and methylglyoxal. Then, the effect of manuka honey in wound treatment is described, as well as its antioxidant activity and other important biological effects.


Long-range GABAergic projections contribute to cortical feedback control of sensory processing.
In the olfactory system, the olfactory cortex sends glutamatergic projections back to the first stage of olfactory processing, the olfactory bulb (OB). Such corticofugal excitatory circuits - a canonical circuit motif described in all sensory systems- dynamically adjust early sensory processing. Here, we uncover a corticofugal inhibitory feedback to OB, originating from a subpopulation of GABAergic neurons in the anterior olfactory cortex and innervating both local and output OB neurons. In vivo imaging and network modeling showed that optogenetic activation of cortical GABAergic projections drives a net subtractive inhibition of both spontaneous and odor-evoked activity in local as well as output neurons. In output neurons, stimulation of cortical GABAergic feedback enhances separation of population odor responses in tufted cells, but not mitral cells. Targeted pharmacogenetic silencing of cortical GABAergic axon terminals impaired discrimination of similar odor mixtures. Thus, corticofugal GABAergic projections represent an additional circuit motif in cortical feedback control of sensory processing.


U.S.-Based National Surveillance for Fidaxomicin Susceptibility of Clostridioides difficile-Associated Diarrheal Isolates from 2013 to 2016.
In 2011, we initiated a sentinel surveillance network to assess changes in Clostridioides (formerly Clostridium) difficile antimicrobial susceptibility to fidaxomicin from 6 geographically dispersed medical centers in the United States. This report summarizes data from 2013 to 2016. C. difficile isolates or toxin-positive stools from patients were referred to a central laboratory. Antimicrobial susceptibility was determined by agar dilution. CLSI, EUCAST, or FDA breakpoints were used, where applicable. Toxin gene profiles were characterized by multiplex PCR on each isolate. A random sample of approximately 40% of isolates, stratified by institution and year, was typed by restriction endonuclease analysis (REA). Among 1,889 isolates from 2013 to 2016, the fidaxomicin MIC90 was 0.5 μg/ml; all isolates were inhibited at ≤1 μg/ml. There were decreases in metronidazole and vancomycin MICs over time. Clindamycin resistance remained unchanged (27.3%). An increase in imipenem resistance was observed. By 2015 to 2016, moxifloxacin resistance decreased in all centers. The proportion of BI isolates decreased from 25.5% in 2011 to 2012 to 12.8% in 2015 to 2016 (P < 0.001). The BI REA group correlated with moxifloxacin resistance (BI 84% resistant versus non-BI 12.5% resistant). Fidaxomicin MICs have not changed among C. difficile isolates of U.S. origin over 5 years post licensure. There has been an overall decrease in MICs for vancomycin, metronidazole, moxifloxacin, and rifampin and an increase in isolates resistant to imipenem. Moxifloxacin resistance remained high among the BI REA group, but the proportion of BI isolates has decreased. Continued geographic variations in REA groups and antimicrobial resistance persist.


The natural caesarean: a woman-centred technique.
Although much effort has gone into promoting early skin-to-skin contact and parental involvement at vaginal birth, caesarean birth remains entrenched in surgical and resuscitative rituals, which delay parental contact, impair maternal satisfaction and reduce breastfeeding. We describe a 'natural' approach that mimics the situation at vaginal birth by allowing (i) the parents to watch the birth of their child as active participants (ii) slow delivery with physiological autoresuscitation and (iii) the baby to be transferred directly onto the mother's chest for early skin-to-skin. Studies are required into methods of reforming caesarean section, the most common operation worldwide.


Ultrastructural morphometric analysis of myocardium from dogs, rats, hamsters, mice, and from human hearts.
Volume densities of mitochondria, myofibrils, and unspecified cytoplasm were measured by ultrastructural morphometry in myocardium from dogs, rats, hamsters, mice, and in biopsied tissue from human hearts. Human myocardium was composed of 23% mitochondria, 59% myofibrils, and 18% cytoplasm. Volume densities for mitochondria were 22% for dogs, 28% for rats and hamsters, and 32% for mice. Myofibrillar volume densities were highest in dogs with 63%, 57% for rats and hamsters, and 49% for mice. Differences were significant between all except man and dog, and rat and hamster. In an extensive analysis of canine myocardium, it could be shown that the quantitative composition of tissue from the left ventricular free wall (anterior, lateral, posterior) and the papillary muscles was identical. There were also no differences between subepi- and subendocardium as well as the midmyocardium. Volume densities from longitudinal sections were identical to those from transversal sections. Fixation with glutaraldehyde by perfusion or immersion provided identical results. There were no differences between volume densities in samples from the left ventricular free wall (anterior, lateral, and posterior) in rats, hamsters, and mice. It is concluded that each mammalian species is characterized by a very typical quantitative composition of the myocardium. The increase in mitochondrial volume correlated well with the increase in heart rate and oxygen consumption in smaller animals. These quantitative data are regarded as the morphological correlate of the differing functional capacity of hearts from different species.


Comparison of clinical manifestations and outcome of community-acquired bloodstream infections among the oldest old, elderly, and adult patients.
Valid studies comparing the clinical characteristics among adult, elderly, and the oldest old bacteremic patients are lacking. We conducted a prospective, observational study in the emergency department (ED) of a university medical center between June 2001 and June 2002. All patients >18 years of age who registered in the ED with a clinically significant, culture-positive, bloodstream infection (BSI) were enrolled. Patients were divided into 3 groups based on age: 1) oldest old (> or =85 yr), 2) elderly (65-84 yr), and 3) adult (18-64 yr). The clinical and laboratory manifestations and 30-day mortality were recorded. Group comparisons were performed using the chi-square test or analysis of variance (ANOVA) test, as indicated. Survival was analyzed using the Kaplan-Meier method and the Cox-regression model, adjusted for potential confounders.A total of 890 cases of community-acquired BSI were eligible for analysis. Compared to the adult group, both the elderly and the oldest old patients had more atypical clinical manifestations, a higher propensity to develop organ failure, and a worse prognosis. Elderly patients had significantly less tachycardia (p = 0.001), but more acute respiratory (p = 0.007) and renal failure (p = 0.037); the oldest old patients had more afebrile episodes (p = 0.006), leukocytosis (p = 0.012), and more patients developed respiratory failure (p = 0.009), acute renal failure (p = 0.011), septic shock (p = 0.022), and altered mental status (p = 0.013). Urinary tract infections were the main source of BSI for both the elderly and oldest old, while the oldest old patients had significantly more pneumonia than the elderly or adults. As a group, older patients had fewer signs and symptoms of BSI, but a higher risk of organ failure and a worse prognosis than younger patients.


Training in the prevention of cervical cancer: advantages of e-learning.
Cervical cancer remains the second most common cancer for women worldwide and is the cancer priority in most low- and middle-income countries (LMIC). The development of vaccines against the human papilloma virus (HPV) and the impact of technology both for the detection of HPV and cervical cancer represent milestones and new opportunities in prevention. New internet-based technologies are generating mass access to training programmes. This article presents the methodology for developing an online training programme for the prevention of cervical cancer as well as the results obtained during the four year period wherein the same programme was delivered in Latin America.


[Adequacy of medication in patients 65 years or older in teaching health centers in Cáceres, Spain].
The appropriateness of drug prescription is objective of the National Health System for the sanitary and economic repercussions linked. The main objective of this paper is to analyze the appropriateness of drug prescription on patients 65 years or older and identify the most prevalent prescribing errors. Observational, descriptive, transversal, multicenter study in Health Centers doctors in training in Cáceres. 471 patients 65 years or older were included by convenience samplin.The sample size was calculated to an accuracy 3.5%-5% and a confidence level 95%. The measured variable was the criteria STOPP/START. We calculated mean and standard deviation for quantitative variables and relative frequencies for qualitative variables. The most frequent STOPP criteria were: duplication of drugs (9.76% CI 95% 7.40 to 12.78), prolonged use of long-acting benzodiazepines (7.22%, 95% CI 5.21 to 9, 92) and the use of aspirin in patients without cardiovascular secondary prevention (7% 95% CI 5.03 to 9.68). The most frequent START criteria were: omission of antiplatelet agents and statins in diabetes coexisting one or more cardiovascular risk factors (9.34 and 7.03 to 12.31% 95% 95% 4.46% 2.93 to 6, 72), calcium and vitamin D in osteoporosis (5.31%, 95% CI 3.61 to 7.72) and metformin in type 2 diabetes metabolic syndrome ± (4.46%, 95% CI 2.93 - 6.72). The main inappropriate prescribing concerns treatments that should be suspended or modify. The duplications were the most common cause of inappropriate prescribing, being more frequently involved drugs like benzodiazepines, NSAIDs and drugs in drugs associations. The second cause of inadequacy most prevalent was the prescription of benzodiazepines with long half life. The third most common cause was acetylsalicylic acid, both through overuse or omission.