A musculoskeletal model of the equine forelimb for determining surface stresses and strains in the humerus-part II. Experimental testing and model validation.
The first objective of this study was to experimentally determine surface bone strain magnitudes and directions at the donor site for bone grafts, the site predisposed to stress fracture, the medial and cranial aspects of the transverse cross section corresponding to the stress fracture site, and the middle of the diaphysis of the humerus of a simplified in vitro laboratory preparation. The second objective was to determine whether computing strains solely in the direction of the longitudinal axis of the humerus in the mathematical model was inherently limited by comparing the strains measured along the longitudinal axis of the bone to the principal strain magnitudes and directions. The final objective was to determine whether the mathematical model formulated in Part I [Pollock et al., 2008, ASME J. Biomech. Eng., 130, p. 041006] is valid for determining the bone surface strains at the various locations on the humerus where experimentally measured longitudinal strains are comparable to principal strains. Triple rosette strain gauges were applied at four locations circumferentially on each of two cross sections of interest using a simplified in vitro laboratory preparation. The muscles included the biceps brachii muscle in addition to loaded shoulder muscles that were predicted active by the mathematical model. Strains from the middle grid of each rosette, aligned along the longitudinal axis of the humerus, were compared with calculated principal strain magnitudes and directions. The results indicated that calculating strains solely in the direction of the longitudinal axis is appropriate at six of eight locations. At the cranial and medial aspects of the middle of the diaphysis, the average minimum principal strain was not comparable to the average experimental longitudinal strain. Further analysis at the remaining six locations indicated that the mathematical model formulated in Part I predicts strains within +/-2 standard deviations of experimental strains at four of these locations and predicts negligible strains at the remaining two locations, which is consistent with experimental strains. Experimentally determined longitudinal strains at the middle of the diaphysis of the humerus indicate that tensile strains occur at the cranial aspect and compressive strains occur at the caudal aspect while the horse is standing, which is useful for fracture fixation.


How can patient experience scores be used to predict quality inspection ratings? A retrospective cross-sectional study of national primary care datasets in the UK.
The relationship between patient feedback in the General Practice Patient Survey (GPPS) and Care Quality Commission (CQC) inspections of practices was investigated to understand whether there is an association between patient views and regulator ratings of quality. The specific aims were to understand whether patients' self-reported experiences of primary care can predict CQC inspection ratings of GP practices by: (i) Measuring the association between GPPS results and CQC inspection ratings of GP practices; (ii) Building a predictive model of GP practice quality ratings that use GPPS results; and (iii) Evaluating the predictive model for risk stratification. Retrospective analysis of routinely collected data using decision tree modelling. Primary care: GP practices in England. GPPS scores and GP practice CQC inspection ratings during 2018. Most GP practices (72%, 974/1350) were rated as 'Good' overall by CQC. Simply assuming that all practices will be rated as 'Good' results in a correct prediction 72% of the time, and it was not possible to improve on this overall level of predictive accuracy using decision tree modelling (correct in 73% of cases). However, a set of GPPS questions were found to have value in identifying practices at elevated risk of a poor inspection rating. Although there were some associations between GPPS data and CQC inspection ratings, there were limitations to the use of GPPS data for predictive analysis. This is a likely result of the majority of CQC inspections of GPs resulting in a 'Good' or 'Outstanding' rating. However, some GPPS questions were found to have value in identifying practices at higher risk of an 'Inadequate' or 'Requires Improvement' rating, and this may be valuable for surveillance purposes. For example, the CQC could use key questions from the survey to target inspection planning.


Random drug excess.
If a patient is treated in a hospital, drugs are administered at 'deterministic', known times. However, if the patient is not supervised the times of drug intake may be less 'deterministic' and less well known. It seems therefore worthwhile to make a theoretical experiment in which one assumes that drugs are applied according to a random process. In the present article it is proposed that shot noise models could be used to represent concentration curves in such situations. By means of Monte Carlo methods different measures of random drug excess are described. An included program may encourage readers to consider these methods and to perform computer experiments.


Evaluation of the CRUSADE Risk Score for Predicting Major Bleeding in Patients with Concomitant Kidney Dysfunction and Acute Coronary Syndromes.
Kidney dysfunction (KD) has been associated with increased risk for major bleeding (MB) in patients with acute coronary syndromes (ACS) and may be in part related to an underuse of evidence-based therapies. Our aim was to assess the predictive ability of the Can Rapid risk stratification of Unstable angina patients Suppress ADverse outcomes with Early implementation of the ACC/AHA guidelines (CRUSADE) risk score in patients with concomitant ACS and chronic kidney disease. We conducted a retrospective analysis of a prospective registry including 1,587 ACS patients. In-hospital MB was prospectively recorded according to the CRUSADE and Bleeding Academic Research Consortium (BARC) criteria. KD was defined as an estimated glomerular filtration rate <60 mL/min/1.73 m2. The predictive ability of the CRUSADE risk score was assessed by discrimination and calibration analyses. A total of 465 (29%) subjects had KD. In multivariate logistic regression analyses, we found high CRUSADE risk score values to be associated with a higher rate of in-hospital MB; however, among patients with KD, it was not associated with BARC MB. Regardless of the MB definition, the predictive ability of the CRUSADE score in patients with KD was lower: area under the curve (AUC) 0.71 versus 0.79, p = 0.03 for CRUSADE MB and AUC 0.65 versus 0.75, p = 0.02 for BARC MB. Hosmer-Lemeshow analyses showed a good calibration in all renal function subgroups for both MB definitions (all p values >0.3). The CRUSADE risk score shows a lower accuracy for predicting in-hospital MB in KD patients compared to those without KD.


Modifications induced by gestational diabetes mellitus on cellular membrane properties.
Alterations in erythrocyte plasma membrane properties (enzymatic activities and membrane fluidity) have been observed in patients affected by insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM). In order to verify whether these alterations are present also in gestational diabetes mellitus (GDM) we studied the plasma membranes obtained from two different cellular types (erythrocyte from both mother and cord blood and placenta syncytiothrophoblast cell) of 16 healthy pregnant women and 15 women affected by GDM. The following determinations were performed on the membrane preparations: Na+/K(+)-ATPase activity, acetyl-cholinesterase (AchE) activity, membrane fluidity and cholesterol:phospholipid ratio. We observed a reduction of both enzymatic activities and a decrease of membrane fluidity in maternal and cord blood erythrocytes and in syncytiotrophoblast plasma membranes in GDM pregnant women in comparison with controls. The cholesterol to phospholipid ratio was significantly lower in the erythrocyte membranes of women affected by GDM than in normal pregnant women, while it was increased in the cord blood erythrocyte membranes and in placental membranes in GDM in comparison with controls. The present study found, in GDM patients, a membrane alteration similar to the abnormality reported in IDDM and NIDDM (i.e. decreased Na+/K(+)-ATPase activity), while opposite modifications were observed with regard to other membrane activities and properties. The different membrane alterations observed in GDM with respect to IDDM and NIDDM might be linked to the different degree of metabolic control, on the contrary the reduced Na+/K(+)-ATPase activity might be a primary event in the pathogenesis of diabetes mellitus per se and might constitute a signal of high risk of developing the disease later in the women affected by GDM during pregnancy.


The Pandemic's Toll on Young Adolescents: Prevention and Intervention Targets to Preserve Their Mental Health.
Adolescence is characterized by dramatic physical, social, and emotional changes, making teens particularly vulnerable to the mental health effects of the COVID-19 pandemic. This longitudinal study identifies young adolescents who are most vulnerable to the psychological toll of the pandemic and provides insights to inform strategies to help adolescents cope better in times of crisis. A data-driven approach was applied to a longitudinal, demographically diverse cohort of more than 3,000 young adolescents (11-14 years) participating in the ongoing Adolescent Brain Cognitive Development Study in the United States, including multiple prepandemic visits and three assessments during the COVID-19 pandemic (May-August 2020). We fitted machine learning models and provided a comprehensive list of predictors of psychological distress in individuals. Positive affect, stress, anxiety, and depressive symptoms were accurately detected with our classifiers. Female sex and prepandemic internalizing symptoms and sleep problems were strong predictors of psychological distress. Parent- and youth-reported pandemic-related psychosocial factors, including poorer quality and functioning of family relationships, more screen time, and witnessing discrimination in relation to the pandemic further predicted youth distress. However, better social support, regular physical activities, coping strategies, and healthy behaviors predicted better emotional well-being. Findings highlight the importance of social connectedness and healthy behaviors, such as sleep and physical activity, as buffering factors against the deleterious effects of the pandemic on adolescents' mental health. They also point to the need for greater attention toward coping strategies that help the most vulnerable adolescents, particularly girls and those with prepandemic psychological problems.


Family food work: lessons learned from urban Aboriginal women about nutrition promotion.
This article reports on ethnographic study of urban Aboriginal family food and implications for nutrition promotion. Data were collected over 2 years through in-depth interviews and participant observation in groups conducted through Indigenous organisations in a suburb of Brisbane. Issues when organising family food include affordability, keeping family members satisfied and being able to share food, a lack of cooking ideas, the accessibility of nutrition information, additional work involved in ensuring healthy eating, and a desire for convenience. Many different health professionals provide nutrition advice, often directing it towards individuals and not providing adequate guidance to facilitate implementation. The easiest advice to implement worked from existing household food practices, skills and budget. Cooking workshops helped to provide opportunities to experiment with recommended foods so that women could confidently introduce them at home. Aboriginal women are concerned about healthy eating for their families. Disadvantage can limit dietary change and the complexity of family food work is often underestimated in nutrition promotion. Household, rather than individual, framing of nutrition promotion can lead to more sustainable healthy eating changes.


Neurotrophic factor for central neurons.
A serum-free culture of dissociated neurons from embryonic rat hippocampus has been established as a rapid and quantitative in vitro test system for neurotrophic signals in the mammalian brain. By means of this cell culture bioassay, a novel low molecular weight neurotrophic factor (NTF) could be identified. NTF is essential for in vitro brain neuron development, promoting survival and neurite outgrowth. The diffusible factor is synthesized and secreted into serum-free defined medium by cultured astrocytes from rat cerebral hemispheres. The number of viable neurons responding to NTF by neurite outgrowth is dependent on the concentration of the factor. Fractionation of astroglial conditioned medium by gel filtration on columns of Sephadex G-10 recovered biological activity of NTF in a single sharp peak corresponding to an apparent molecular weight of approximately equal to 500. NTF is stable to heat and cold and resistant to trypsin and pronase. Unlike nerve growth factor, NTF has no apparent effect on the neurite outgrowth of peripheral neurons. NTF-like activity is present in situ in the mammalian brain, in certain other nonneural tissues, and in C6 and B12 glioma cell conditioned media.


Magnesium level correlation with clinical status and quality of life in women with hormone related conditions and pregnancy based on real world data.
This study was aimed to assess the effectiveness of magnesium (Mg)-vitamin B 6 replenishment and its correlation with clinical status in pregnant women (PW), and quality of life in women with hormone-related conditions (HRCW) and hypomagnesemia (HME). Data collected in four observational studies were pooled and analysed. All women received Mg supplementation for 4 weeks. The proportion of women with normalized Mg level, and the correlation between serum Mg dynamics and number of symptoms/complaints (PW) or changes in World Health Organization quality of life questionnaire scores (WHOQOL; HRCW) were evaluated. 869 PW and 957 HRCW were included in the study. Normalization of serum Mg level to ≥ 0.66 mmol/L occurred in 92.1% of PW and 78.4% of HRCW, and to ≥ 0.8 mmol/L in 73.8% and 58.9%, respectively. Mg normalization was accompanied by a median decrease of 1 symptom and 1 complaint in PW. Serum Mg level increase by 0.1 mmol/L was associated to significant changes in the WHOQOL scores in HRCW. Treatment of HME with the Mg for approximately 4 weeks provided a high response rate of Mg serum level, was associated with an improvement in symptom severity and complaints in PW, and WHOQOL score in HRCW. A 0.8 mmol/L cut-off appeared to be more relevant in terms of patient-reported outcomes.


The natural course of COVID-19 patients without clinical intervention.
The natural course of coronavirus disease 2019 (COVID-19) patients without clinical intervention has not yet been documented. One hundred and fifty-eight patients from two hospitals were enrolled to identify the indicators of severe COVID-19 and observe the natural course of COVID-19 patients without clinical intervention. The total computed tomography (CT) score, a quantitative score based on assessment of the number, quadrant, and area of the lesions in CT, tended to perform better than assessment based only on the number or area of the lesions (p = 0.0004 and p = 0.0887, respectively). Multivariate logistic regression showed that the total CT score, chest tightness, lymphocyte, and lactate dehydrogenase (LDH) were independent factors for severe COVID-19. For patients admitted in 2 weeks from onset to hospitalization, the frequency of severe COVID-19 was gradually increased with the delayed hospitalization. The symptoms of fatigue, dry cough, sputum production, chest tightness, and polypnea were gradually more frequent. The levels of C-reactive protein, alanine aminotransferase, aspartate aminotransferase, total bilirubin, direct bilirubin, γ-glutamyl transpeptidase, LDH, and d-dimer were also gradually increased, as well as the scores based on CT. Conversely, the lymphocyte count and the albumin level were gradually decreased with the delayed hospitalization. Detail turning points of the above alterations were observed after 10-14 days from onset to hospitalization. Total CT score was a simple and feasible score for identifying severe COVID-19. COVID-19 patients without clinical intervention deteriorated gradually during the initial 10-14 days but gradually improved thereafter.


Interactions among Ecological Factors That Explain the Psychosocial Quality of Life of Children with Complex Needs.
Purpose. To explore the associations and interactions among ecological factors and explain the psychosocial quality of life of children with complex needs. Methods. In this cross-sectional survey consenting parents were identified by the Children's Treatment Network. Families were eligible if the child from 0 to 19 years, resided in Simcoe/York, and there were multiple family needs. Regression analysis was used to explore associations and interactions. n = 429. Results. Younger children, without conduct disorder, without hostile and punitive parenting and with low adverse family impact demonstrated the highest levels of psychosocial quality of life. Statistically significant interactions between processes of care and parent variables highlight the complexity of real life situations. Conclusions. It is not possible to fully understand the child's psychosocial quality of life in complex needs families by considering only simple associations between ecological factors. A multitude of factors and interactions between these factors are simultaneously present and the care of these families requires a holistic approach.


NIH State-of-the-Science Conference Statement on Tobacco Use: Prevention, Cessation, and Control.
To provide health care providers, patients, and the general public with a responsible assessment of currently available data on tobacco use: prevention, cessation and control. A non-DHHS, non-advocate 14-member panel included experts in the fields of medicine, general and pediatric psychiatry, addiction medicine, nursing, social work, population science, cancer prevention, minority health and health disparities, clinical study methodology, clinical epidemiology, and a public representative. A listing of the panel members and their institutional affiliations is included in the draft conference statement. In addition, 15 experts from pertinent fields presented data to the panel and conference audience. Presentations by experts and a systematic review of the literature prepared by the RTI International-University of North Carolina Evidence-based Practice Center, through the Agency for Healthcare Research and Quality. Scientific evidence was given precedence over anecdotal experience. The panel drafted its statement based on scientific evidence presented in open forum and on published scientific literature. The draft statement was presented on the final day of the conference and circulated to the audience for comment. The panel released a revised statement later that day at http://consensus.nih.gov. This statement is an independent report of the panel and is not a policy statement of the NIH or the Federal Government. Tobacco use remains a very serious public health problem. Coordinated national strategies for tobacco prevention, cessation, and control are essential if the United States is to achieve the Healthy People 2010 goals. Most adult smokers want to quit, and effective interventions exist. However, only a small proportion of tobacco users try treatment. This gap represents a major national quality-of-care problem. Many cities and states have implemented effective policies to reduce tobacco use; public health and government leaders should learn from these experiences. Because smokeless tobacco use may increase in the United States, it will be increasingly important to understand net population harms related to use of smokeless tobacco. Prevention, especially among youth, and cessation are the cornerstones of strategies to reduce tobacco use. Tobacco use is a critical and chronic problem that requires close attention from health care providers, health care organizations, and research support organizations.


"Hidden" dUTPase sequence in human immunodeficiency virus type 1 gp120.
A coding region homologous to the sequence for essential eukaryotic enzyme dUTPase has been identified in different genomic regions of several viral lineages. Unlike the nonprimate lentiviruses (caprine arthritis- encephalitis virus, equine infectious anemia virus, feline immunodeficiency virus, and visna virus), where dUTPase is integrated into the pol coding region, this enzyme has never been demonstrated to be present in the primate lentivirus genomes (human immunodeficiency virus type 1 [HIV-1], HIV-2, or the related simian immunodeficiency virus). A novel approach allowed us to identify a weak but significant sequence similarity between HIV-1 gp120 and the human dUTPase. This finding was then extended to all of the primate lentivirus lineages. Together with the recently reported fragmentary structural similarity between the V3 loop region and the Escherichia coli dUTPase (P. D. Kwong, R. Wyatt, J. Robinson, R. W. Sweet, J. Sodroski, and W. A. Hendrickson, Nature 393:648-659, 1998), our results strongly suggest that an ancestral dUTPase gene has evolved into the present primate lentivirus CD4 and cytokine receptor interacting region of gp120.


Radiation dose estimates in humans for (11)C-acetate whole-body PET.
11C-Acetate is currently being investigated as a new tracer for imaging neoplasms, most notably prostate cancer and its metastases. Previously reported dose estimates for (11)C-acetate prepared by the Oak Ridge Institute for Science and Education (ORISE) were based on a simple 3-compartment model in which all activity not measured in blood or excretion via breath was assumed to reside in the heart. Because all organs are involved in acetate metabolism to some extent, these estimates might overestimate heart and underestimate other organ dosimetry. Dynamic whole-body (11)C-acetate PET was therefore performed on 6 healthy human volunteers. Measured dose estimates for all target organs were compared with the existing ORISE values. After transmission scanning had been performed for measured attenuation, 525 MBq of (11)C-acetate were injected intravenously, and 5 sequential whole-body emission scans were obtained from the head to mid thighs. Regions of interest were drawn to encompass the entire activity in all visible organs at each time point. Time-activity data were fit in a least-squares sense to obtain residence times. Absorbed dose estimates were determined using MIRDOSE3.1 software. The effective dose was 0.0049 mSv/MBq. The organs receiving the highest absorbed doses were the pancreas (0.017 mGy/MBq), bowel (0.011 mGy/MBq), kidneys (0.0092 mGy/MBq), and spleen (0.0092 mGy/MBq). No urinary excretion of tracer was measurable. Using these new estimates for (11)C-acetate dosimetry, the maximum injected activity under Radioactive Drug Research Committee limits can be raised up to 5-fold over the limit imposed by the previous ORISE estimates. A higher injected activity would improve counting statistics and, it is hoped, overall image quality and tumor detection with whole-body (11)C-acetate PET.


Metabolic Profiles of Cancer Stem Cells and Normal Stem Cells and Their Therapeutic Significance.
Cancer stem cells (CSCs) are a rare cancer cell population, responsible for the facilitation, progression, and resistance of tumors to therapeutic interventions. This subset of cancer cells with stemness and tumorigenic properties is organized in niches within the tumor microenvironment (TME) and presents altered regulation in a variety of metabolic pathways, including glycolysis, oxidative phosphorylation (OXPHOS), as well as lipid, amino acid, and iron metabolism. CSCs exhibit similarities as well as differences when comparedto normal stem cells, but also possess the ability of metabolic plasticity. In this review, we summarize the metabolic characteristics of normal, non-cancerous stem cells and CSCs. We also highlight the significance and implications of interventions targeting CSC metabolism to potentially achieve more robust clinical responses in the future.


An exploration of the experience of trainee integrative psychotherapists on therapeutic alliance in the light of their attachment style.
Therapeutic Relationship has been regarded as a profound element of the psychotherapeutic procedure and crucial for the outcome of therapy. Research evidence so far claims that both therapist's and client's personality can impact the Therapeutic Relationship. Based on the well-researched Attachment Theory there has been some research mostly focused on how clients' attachment patterns can affect the Alliance. Limited research, though, on how therapists experience the Relationship in relation with their Attachment Styles do exist. The current study investigates trainee therapists' experience of the Therapeutic Relationship in the light of their Attachment Style. A qualitative approach was used for this research to investigate in depth the experience of five trainee Integrative psychotherapists who were recruited to take part in semi-structured interviews. Interviews were transcribed and verbatim was divided into superordinate and subordinate themes and analyzed by Interpretative Phenomenological Analysis method. Four master themes were identified. Findings were then compared and discussed according to existing literature. All participants referred to the relevance of Attachment Theory in psychotherapy. They also agreed on the activation of their Attachment Styles during the therapeutic process. The securely attached therapist confirms previous studies on her capability to offer a secure base for her clients. On the other hand therapists with insecure Attachment Styles appeared sensitive in their collaboration mostly with insecure clients. Anxiously attached therapists proved to experience difficulty in the initial stages of the Therapeutic Relationship while the combination of avoidant therapist-avoidant client appeared to be the most demanding one, regarding the concept of trust and relationship ruptures. We suggest that the exploration of trainee therapists' Attachment Styles as well as the study of the Attachment Theory should be added to all Counseling and Psychotherapy curricula regardless the therapeutic orientation because of its contribution to forming positive Therapeutic Relationships.


Pre-illness diet as risk factor in pediatric inflammatory bowel disease in Saudi Arabia.
Inflammatory bowel disease (IBD) is increasing in Saudi Arabia but there's little knowledge about its risk factors. Our aim is to investigate the dietary risk factors of this disease in Saudi children. The children investigated for IBD were prospectively enrolled. Those with confirmed IBD were designated cases and the others were controls. The average food frequency intake at least 3 months before onset of illness was obtained by direct interview and recorded on a modified food frequency questionnaire. Fast food, sweet gaseous soft drinks, fruits, and vegetables were selected. Statistical analysis included descriptive statistics, multivariate logistic regression was used, and the odds ratio was calculated to report the statistical significance of the results. A total of 91 children, 52 IBD (38 CD, 14 UC) and 39 controls were enrolled. Consumption of fast food and sweet gaseous drinks was more frequent in children with IBD than controls, whereas consumption of fruits and vegetables was less frequent in children with IBD. Only less fruits consumption was significantly associated with IBD as the odds of children getting IBD was 2.89 (1.06, 7.87). We report statistically significant positive association between less fruits consumption and IBD. Awaiting the results of larger sample size and more quantitative studies, the benefits of frequent fruit consumptions should be part of public health education.


The UGA3-GLT1 intergenic region constitutes a promoter whose bidirectional nature is determined by chromatin organization in Saccharomyces cerevisiae.
Transcription of an important number of divergent genes of Saccharomyces cerevisiae is controlled by intergenic regions, which constitute factual bidirectional promoters. However, few of such promoters have been characterized in detail. The analysis of the UGA3-GLT1 intergenic region has provided an interesting model to study the joint action of two global transcriptional activators that had been considered to act independently. Our results show that Gln3p and Gcn4p exert their effect upon cis-acting elements, which are shared in a bidirectional promoter. Accordingly, when yeast is grown on a low-quality nitrogen source, or under amino acid deprivation, the expression of both UGA3 and GLT1 is induced through the action of both these global transcriptional modulators that bind to a region of the bidirectional promoter. In addition, we demonstrate that chromatin organization plays a major role in the bidirectional properties of the UGA3-GLT1 promoter, through the action of an upstream Abf1p-binding consensus sequence and a polydAdT(tract). Mutations in these cis-elements differentially affect transcription of UGA3 and GLT1, and thus alter the overall relative expression. This is the first example of an intergenic region constituting a promoter whose bidirectional character is determined by chromatin organization.


The Polycomb protein RING1B enables estrogen-mediated gene expression by promoting enhancer-promoter interaction and R-loop formation.
Polycomb complexes have traditionally been prescribed roles as transcriptional repressors, though increasing evidence demonstrate they can also activate gene expression. However, the mechanisms underlying positive gene regulation mediated by Polycomb proteins are poorly understood. Here, we show that RING1B, a core component of Polycomb Repressive Complex 1, regulates enhancer-promoter interaction of the bona fide estrogen-activated GREB1 gene. Systematic characterization of RNA:DNA hybrid formation (R-loops), nascent transcription and RNA Pol II activity upon estrogen administration revealed a key role of RING1B in gene activation by regulating R-loop formation and RNA Pol II elongation. We also found that the estrogen receptor alpha (ERα) and RNA are both necessary for full RING1B recruitment to estrogen-activated genes. Notably, RING1B recruitment was mostly unaffected upon RNA Pol II depletion. Our findings delineate the functional interplay between RING1B, RNA and ERα to safeguard chromatin architecture perturbations required for estrogen-mediated gene regulation and highlight the crosstalk between steroid hormones and Polycomb proteins to regulate oncogenic programs.


Implant surgery complications: etiology and treatment.
Implant surgery complications are frequent occurrences in dental practice and knowledge in the management of these cases is essential. The aim of this review was to highlight the challenges of treatment plan-related, anatomy-related, and procedure-related surgical complications as well as to discuss the etiology, management and treatment options to achieve a satisfactory treatment outcome.


Determinants of catheter loss following continuous ambulatory peritoneal dialysis peritonitis.
Few patients are able to resume peritoneal dialysis (PD) therapy after an episode of peritonitis that requires catheter removal. PD catheter loss is therefore regarded as an important index of patient morbidity. The aim of the present study was to evaluate factors influencing catheter loss in patients suffering from continuous ambulatory PD (CAPD) peritonitis. We retrospectively reviewed 579 episodes of CAPD peritonitis from 1999 to 2006 in a tertiary-care referral hospital. Demographic, biochemical, and microbiological characteristics were recorded. Episodes resulting in PD catheter removal (n = 68; 12%) were compared by both univariate and multivariate analyses with those in which PD catheters were preserved. The incidence of PD catheter loss increased as the number of organisms cultured increased (p = 0.001). Also, PD catheter removal was more likely to occur after peritonitis episodes with low serum albumin level (p = 0.004), those with long duration of PD effluent leukocyte count remaining above 100/microL (p < 0.001), those with concomitant tunnel infection (p < 0.001), those with concomitant exit-site infection (p = 0.005), and those with presence of catastrophic intra-abdominal visceral events (p < 0.001). Duration on PD preceding the peritonitis episode was of borderline significance (p = 0.080). On the contrary, initial PD effluent leukocyte count and serum level of C-reactive protein were not predictive of PD catheter loss. Micro-organisms of the Enterobacteriaceae family were the major pathogens responsible for PD catheter loss following polymicrobial peritonitis. Furthermore, we found that there was no association between polymicrobial peritonitis and the catastrophic intra-abdominal visceral event, although both resulted in a greater incidence of PD catheter loss. Among the single-organism group in our population, the microbiological determinants of PD catheter loss included fungi (p < 0.001), anaerobes (p = 0.018), and Pseudomonas sp (borderline significance: p = 0.095). PD catheter loss as a consequence of peritonitis is related primarily to hypoalbuminemia, longer duration of PD effluent leukocyte count remaining above 100/muL, the etiologic source of the infection, and the organism causing the infection. Peritonitis associated with concomitant tunnel or exit-site infections and abdominal catastrophes were more likely to proceed to PD catheter loss. The microbiological determinants of PD catheter loss in the present study included polymicrobial infections caused by Enterobacteriaceae as well as monomicrobial pseudomonal, anaerobic, and fungal infections.


Choline PET or PET/CT and biochemical relapse of prostate cancer: a systematic review and meta-analysis.
The increase of prostate-specific antigen (PSA) after radical retropubic prostatectomy (RP) or external beam radiotherapy (EBRT) is the most sensitive tool for detecting prostate cancer (PCa) recurrence, although this measure cannot distinguish between local, regional, or distant recurrence. The aim of this meta-analysis was to evaluate the diagnostic performance of 18F-choline and 11C-choline PET or PET/CT in detection of locoregional or distant metastases in PCa. Medline, Web of Knowledge, and Google Scholar search was carried out in order to select English-language articles dealing with diagnostic performance of both 18F-choline and 11C-choline PET for the detection of PCa recurrence after RP or EBRT. Articles were included only if absolute numbers of true-positive, true-negative, false-positive, and false-negative test results were available or derivable from the text and regarded local, lymph node, and distant metastases. Reviews, clinical reports, and editorial articles were excluded. All complete studies were re-analyzed thus performing a quantitative analysis. From the years 2000 to 2012, we found 53 complete articles that critically evaluated the role of choline PET in restaging patients with PCa recurrence. The meta-analysis was carried out and dealt with 19 selected studies (12 studies for all sites of disease, 3 for lymph node metastases, and 4 for local recurrence), with a total of 1555 patients. The meta-analysis provided a pooled sensitivity of 85.6% (95% CI: 82.9%-88.1%) and pooled specificity of 92.6% (95% CI: 90.1%-94.6%) for all sites of disease (prostatic fossa, lymph nodes, and bone), a pooled sensitivity of 75.4% (95% CI: 66.9%-82.6%) and pooled specificity of 82% (95% CI: 68.6%-91.4%) for prostatic fossa recurrence, and a pooled sensitivity of 100% (95% CI: 90.5%-100%) and pooled specificity of 81.8% (95% CI: 48.2%-97.7%) for lymph node metastases. The heterogeneity ranged between 0.00% and 88.6%. The diagnostic odds ratios were 62.123 (95% CI: 24.783-155.72), 5.869 (95% CI: 1.818-18.946), and 138.57 (95% CI: 11.27-1703.8), respectively, for all sites of disease, local recurrence, and lymph node disease. Choline PET and PET/CT represent high sensitivity and specificity techniques for the detection of locoregional and distant metastases in PCa patients with recurrence of disease. Moreover, a high diagnostic odds ratio was found for the identification of lymph node disease in patients with biochemical recurrence of PCa.


A systems approach identifies time-dependent associations of multimorbidities with pancreatic cancer risk.
Pancreatic ductal adenocarcinoma (PDAC) is usually diagnosed in late adulthood; therefore, many patients suffer or have suffered from other diseases. Identifying disease patterns associated with PDAC risk may enable a better characterization of high-risk patients. Multimorbidity patterns (MPs) were assessed from 17 self-reported conditions using hierarchical clustering, principal component, and factor analyses in 1705 PDAC cases and 1084 controls from a European population. Their association with PDAC was evaluated using adjusted logistic regression models. Time since diagnosis of morbidities to PDAC diagnosis/recruitment was stratified into recent (<3 years) and long term (≥3 years). The MPs and PDAC genetic networks were explored with DisGeNET bioinformatics-tool which focuses on gene-diseases associations available in curated databases. Three MPs were observed: gastric (heartburn, acid regurgitation, Helicobacter pylori infection, and ulcer), metabolic syndrome (obesity, type-2 diabetes, hypercholesterolemia, and hypertension), and atopic (nasal allergies, skin allergies, and asthma). Strong associations with PDAC were observed for ≥2 recently diagnosed gastric conditions [odds ratio (OR), 6.13; 95% confidence interval CI 3.01-12.5)] and for ≥3 recently diagnosed metabolic syndrome conditions (OR, 1.61; 95% CI 1.11-2.35). Atopic conditions were negatively associated with PDAC (high adherence score OR for tertile III, 0.45; 95% CI, 0.36-0.55). Combining type-2 diabetes with gastric MP resulted in higher PDAC risk for recent (OR, 7.89; 95% CI 3.9-16.1) and long-term diagnosed conditions (OR, 1.86; 95% CI 1.29-2.67). A common genetic basis between MPs and PDAC was observed in the bioinformatics analysis. Specific multimorbidities aggregate and associate with PDAC in a time-dependent manner. A better characterization of a high-risk population for PDAC may help in the early diagnosis of this cancer. The common genetic basis between MP and PDAC points to a mechanistic link between these conditions.


Buprenorphine and norbuprenorphine quantification in human plasma by simple protein precipitation and ultra-high performance liquid chromatography tandem mass spectrometry.
A highly sensitive ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantification of buprenorphine and its major metabolite norbuprenorphine in human plasma. In order to speed up the process and decrease costs, sample preparation was performed by simple protein precipitation with acetonitrile. To the best of our knowledge, this is the first application of this extraction technique for the quantification of buprenorphine in plasma. Matrix effects were strongly reduced and selectivity increased by using an efficient chromatographic separation on a sub-2 μm column (Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm) in 5 min with a gradient of ammonium formate 20 mM pH 3.05 and acetonitrile as mobile phase at a flow rate of 0.4 ml/min. Detection was made using a tandem quadrupole mass spectrometer operating in positive electrospray ionization mode, using multiple reaction monitoring. The procedure was fully validated according to the latest Food and Drug Administration guidelines and the Société Française des Sciences et Techniques Pharmaceutiques. Very good results were obtained by using a stable isotope-labeled internal standard for each analyte, to compensate for the variability due to the extraction and ionization steps. The method was very sensitive with lower limits of quantification of 0.1 ng/ml for buprenorphine and 0.25 ng/ml for norbuprenorphine. The upper limit of quantification was 250 ng/ml for both drugs. Trueness (98.4-113.7%), repeatability (1.9-7.7%), intermediate precision (2.6-7.9%) and internal standard-normalized matrix effects (94-101%) were in accordance with international recommendations. The procedure was successfully used to quantify plasma samples from patients included in a clinical pharmacogenetic study and can be transferred for routine therapeutic drug monitoring in clinical laboratories without further development.


The contribution of ice cover to sediment resuspension in a shallow temperate lake: possible effects of climate change on internal nutrient loading.
The effect of ice cover on sediment resuspension and internal total P (Tot-P) loading was studied in the northern temperate Kirkkojärvi basin in Finland. The gross sedimentation and resuspension rates were estimated with sediment traps during ice-cover and ice-free periods. After ice break, the average gross sedimentation rate increased from 1.4 to 30.0 g dw m(-2) d(-1). Resuspension calculations showed clearly higher values after ice break as well. Under ice cover, resuspension ranged from 50 to 78% of the gross sedimentation while during the ice-free period it constituted from 87 to 97% of the gross sedimentation. Consequently, the average resuspension rate increased from 1.0 g dw m(-2) d(-1) under ice-cover to 27.0 g dw m(-2) d(-1) after thaw, indicating the strong effect of ice cover on sediment resuspension. To estimate the potential effect of climate change on internal P loading caused by resuspension we compared the Tot-P loading calculations between the present climate and the climate with doubled atmospheric CO2 concentration relative to the present day values (ice cover reduced from current 165 to 105 d). The annual load increased from 7.4 to 9.4 g m(-2). In conclusion, the annual internal Tot-P loading caused by resuspension will increase by 28% in the Kirkkojärvi basin if the 2xCO2 climate scenario comes true.


Modest spontaneous recovery of ventilation following chronic high cervical hemisection in rats.
Following C2 spinal hemisection (C2HS) in adult rats, ipsilateral phrenic motoneuron (PhMN) recovery occurs through a time-dependent activation of latent, crossed-spinal collaterals (i.e., spontaneous crossed phrenic phenomenon; sCPP) from contralateral bulbospinal axons. Ventilation is maintained during quiet breathing after C2HS, but the ability to increase ventilation during a respiratory stimulation (e.g. hypercapnia) is impaired. We hypothesized that long-term expression of the sCPP would correspond to a progressive normalization in ventilatory patterns during respiratory challenge. Breathing was assessed via plethsymography in unanesthetized animals and phrenic motor output was measured in urethane-anesthetized, paralyzed and vagotomized rats. At 2-week post-C2HS, minute ventilation (VE) was maintained during baseline (room air) conditions as expected but was substantially blunted during hypercapnic challenge (68+/-3% of VE in uninjured, weight-matched rats). However, by 12 weeks the spinal-lesioned rats achieved a hypercapnic VE response that was 85+/-7% of control (p=0.017 vs. 2 wks). These rats also exhibited augmented breaths (AB's) or "sighs" more frequently (p<0.05) than controls; however, total AB volume was significantly less than control at 2- and 12-week post-injury (69+/-4% and 80+/-5%, p<0.05, respectively). We also noted that phrenic neurograms demonstrated a consistent delay in onset of the ipsilateral vs. contralateral inspiratory phrenic burst at 2-12-week post-injury. Finally, the ipsilateral phrenic response to respiratory challenge (hypoxia) was greater, though not normalized, at 4-12- vs. 2-week post-injury. We conclude that recovery of ventilation deficits occurs over 2-12-week post-C2HS; however, intrinsic neuroplasticity remains insufficient to concurrently restore a normal level of ipsilateral phrenic output.


Turkish Health Policies: Past, Present, and Future.
In this article, health policies in Turkey from the 1900s to the present are reviewed in light of the available data on the number of health institutions, life expectancy, infant mortality, and state budget allocation. While a significant quantitative development is observed in the provision of health services, the state by far maintained a leading role in the provision of health services until the 1980s, when an initially weak but steady deviation began, significantly increasing its pace from 2002 and creating a new path where the private sector raised its share with the extensive implementation of neoliberal economic policies. The emergence of public-private partnership projects as a financing tool in the health sector, with various models from the 1990s around the world, found its place in the Turkish health system in the form of city hospitals from 2013, creating a new deviation in the provision of health services by the state.


Mammalian Synthetic Biology: Time for Big MACs.
The enabling technologies of synthetic biology are opening up new opportunities for engineering and enhancement of mammalian cells. This will stimulate diverse applications in many life science sectors such as regenerative medicine, development of biosensing cell lines, therapeutic protein production, and generation of new synthetic genetic regulatory circuits. Harnessing the full potential of these new engineering-based approaches requires the design and assembly of large DNA constructs-potentially up to chromosome scale-and the effective delivery of these large DNA payloads to the host cell. Random integration of large transgenes, encoding therapeutic proteins or genetic circuits into host chromosomes, has several drawbacks such as risks of insertional mutagenesis, lack of control over transgene copy-number and position-specific effects; these can compromise the intended functioning of genetic circuits. The development of a system orthogonal to the endogenous genome is therefore beneficial. Mammalian artificial chromosomes (MACs) are functional, add-on chromosomal elements, which behave as normal chromosomes-being replicating and portioned to daughter cells at each cell division. They are deployed as useful gene expression vectors as they remain independent from the host genome. MACs are maintained as a single-copy and can accommodate multiple gene expression cassettes of, in theory, unlimited DNA size (MACs up to 10 megabases have been constructed). MACs therefore enabled control over ectopic gene expression and represent an excellent platform to rapidly prototype and characterize novel synthetic gene circuits without recourse to engineering the host genome. This review describes the obstacles synthetic biologists face when working with mammalian systems and how the development of improved MACs can overcome these-particularly given the spectacular advances in DNA synthesis and assembly that are fuelling this research area.


A numerical investigation of the effect of external resistance and applied potential on the distribution of periodicity and chaos in the anodic dissolution of nickel.
The oscillatory electrodissolution of nickel is one among several reactions utilized as a model-system to study the emergence of oscillations and pattern formation in electrochemical interfaces, in addition to frequently providing experimental proofs for theoretical predictions in synchronization engineering. The reaction was modeled in 1992 by Haim and co-workers [J. Phys. Chem. 1992, 96, 2676] and since then the model has been used with great success. Although some numerical studies have been done in this regard, there is apparently no detailed investigation of the effect of control parameters on the complex dynamics of nickel dissolution. Here, we provide a well-detailed and rigorous analysis of the effect of the external resistance and applied potential by simulating high-resolution phase diagrams based on the calculation of Lyapunov exponents and isospike diagrams. Our findings clearly indicate a strong dependence of the self-similar periodic islands, the so-called shrimps (i.e., periodic islands within chaotic domains in the parameter space), with the control parameters. Overall, we have observed a low density of periodic structures in the phase diagrams, being completely suppressed for large values of resistance and potential. The shrimp-like structures become gradually elongated with an increase of the control parameters to the point where only diagonally aligned periodic bands intertwined with chaotic domains are present. Interestingly, period-doubling cascades were observed not only on the shrimps but also on the periodic bands. The detailed distribution of chaos and periodicity of oscillatory electrodissolution reactions in resistance-potential phase diagrams can bring, for instance, important information to experimentalists to set a desired dynamic behavior and, therefore, to create novel nanostructured self-organized materials.


The anatomy of the fourth and fifth lumbar arteries in humans: an arteriographic study in one hundred live subjects.
The anatomy of the arteries on the anterolateral surfaces of the lower two lumbar vertebral bodies has been described in 100 living subjects, using radiographs obtained at aortography. The classic description of this anatomy is commonly correct in the young subject. In addition, a precostal anastomosis was seen in 5 out of 7 subjects aged 10 years or younger. Distortion of this pattern was seen in older subjects; most of this distortion was due to degenerative disease. The average number of observed arteries around the fifth lumbar vertebral body declined steadily in decennial age groups over 30 years, but a statistically significant difference could not be found between the categories of indications for aortography, nor between those judged grossly atherosclerotic and those not so judged as a result of aortography. The precostal anastomosis is an important collateral supply in some older subjects with arterial disease. In addition, a common artery of origin for the fourth lumbar arteries was observed in 15 subjects, and an origin from the median sacral artery for the fourth lumbar arteries was observed in two subjects. These features have not previously been described in healthy subjects.


Rituximab-dependent cytotoxicity by natural killer cells: influence of FCGR3A polymorphism on the concentration-effect relationship.
The FCGR3A gene dimorphism generates two allotypes: FcgammaRIIIa-158V and FcgammaRIIIa-158F. The genotype homozygous for FcgammaRIIIa-158V (VV) is associated with higher clinical response to rituximab, a chimeric anti-CD20 IgG1 used in the treatment of B lymphoproliferative malignancies. Our objective was to determine whether this genetic association relates to rituximab-dependent cytotoxicity mediated by FcgammaRIIIa/CD16a+ cells. The number of CD16+ circulating monocytes, T cells, and natural killer (NK) cells in 54 donors was first shown to be unrelated to FCGR3A polymorphism. We then demonstrated that FcgammaRIIIa-158V displays higher affinity for rituximab than FcgammaRIIIa-158F by comparing rituximab concentrations inhibiting the binding of 3G8 mAb (anti-CD16) with VV NK cells and NK cells homozygous for FcgammaRIIIa-158F (FF). VV and FF NK cells killed Daudi cells similarly after FcgammaRIIIa engagement by saturating concentrations of rituximab or 3G8. However, the rituximab concentration resulting in 50% lysis (EC(50)) observed with NK cells from VV donors was 4.2 times lower than that observed with NK cells from FF donors (on average 0.00096 and 0.00402 microg/ml, respectively, P = 0.0043). Finally, the functional difference between VV and FF NK cells was restricted to rituximab concentrations weakly sensitizing CD20. This study supports the conclusion that FCGR3A genotype is associated with response to rituximab because it affects the relationship between rituximab concentration and NK cell-mediated lysis of CD20+ cells. Rituximab administration could therefore be adjusted according to FCGR3A genotype.


Pregnancy-related outcomes among women with physical disabilities: A systematic review.
Disability among women of reproductive age is common; many of these women desire children and do not have impaired fertility. To examine the epidemiological literature on perinatal health outcomes among women with physical disabilities. We searched Medline and CINAHL for articles published January 2009-April 2020 following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Eligible studies were observational, quantitative, and reported on physical disabilities in association with prenatal, perinatal, postpartum, and/or infant health outcomes. We included studies that grouped physical and non-physical disabilities, such as surveys that queried only about general daily life limitations. We excluded case reports, descriptive studies without comparison groups, and studies conducted in low- or middle-income countries. Data extraction was done using predefined data fields. All authors were involved in screening activities, data extraction, and/or quality assessment (rating and areas for bias). A total of 2650 articles were evaluated, of which sixteen met inclusion criteria (8 cross-sectional studies and 8 retrospective cohort studies). Assessments of disability status and perinatal outcomes widely varied across studies. Studies were rated as poor (n = 8) or fair quality (n = 8). Findings suggested that women with physical disabilities were at risk of several adverse outcomes, including caesarean delivery, infections, preterm complications, and maternal post-delivery hospitalisations, while their infants may be at risk of low birthweight and small-for-gestational age. Women classified as having complex/severe disabilities were often observed to be at higher risk of adverse outcomes compared to women with less severe disabilities. Research assessing how physical, functional, and medical restrictions influence health outcomes among women with physical disabilities, from preconception through postpartum, is limited. Longitudinal studies with comprehensive data collection that accurately identify women with physical disabilities are critical to understanding their reproductive health risks and outcomes.


Unpacking disparities in substance-related outcomes among racial, ethnic, sexual, and gender minoritized groups during adolescence and emerging adulthood.
Alcohol and other drug (AOD) use increases substantially from adolescence to emerging adulthood, and recent longitudinal studies show disparities in AOD-related outcomes by racial and ethnic, as well as sexual and gender minority (SGM), identities. Greater insight is needed into how individual, social, and environmental contexts interact and affect such disparities, as well as why disparate outcomes are found across different domains (e.g., social, educational, economic), even after accounting for intensity of use. This commentary addresses these important and timely issues. We provide a brief overview of the literature, including our own team's work over the last 14 years, to identify and understand disparities in AOD-related outcomes during adolescence and emerging adulthood across individuals with different racial and ethnic, and sexual and gender, identities. We then discuss paths forward to advance research and build a stronger evidence base, leading to the development and identification of effective interventions that can help mitigate disparities among historically marginalized adolescents and emerging adults. Existing research highlights the need for further longitudinal work in several areas, including addressing contextual factors at various levels (e.g., individual, social, environmental) that may contribute to outcomes for different groups of individuals, developing and testing culturally appropriate AOD-related services, giving greater consideration to intersectionality of multiple minority identities, and using novel statistical approaches to help improve the estimation of differences across smaller subgroups of individuals in existing cohorts. To inform prevention programming and policy for improving health and well-being of historically marginalized populations, it is important to continue our efforts to understand disparities in AOD-related outcomes using multidisciplinary, equity, and intersectionality lenses. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


The role of fractalkine in the recruitment of monocytes to the endothelium.
Recombinant fractalkine possesses both chemoattractive and adhesive properties in vitro. Previous studies have demonstrated an upregulation of this molecule on the membranes of activated human endothelial cells and hypothesised that fractalkine plays a role in the recruitment and adherence of monocytes to the activated endothelium. Here we present data analysing both the adhesive and chemoattractive properties of this chemokine expressed by activated human umbilical vein endothelial cells. We demonstrate that both recombinant fractalkine and endogenously produced fractalkine function as adhesion molecules, tethering monocytes to the endothelium. However, our data demonstrate that although recombinant fractalkine has the potential to function as a potent monocyte chemoattractant, the endogenous fractalkine cleaved from activated human umbilical vein endothelial cells is not responsible for the observed chemotaxis in this model. Instead, we show that monocyte chemoattractant protein-1 (MCP-1), secreted from the activated human umbilical vein endothelial cells, is responsible for the chemotaxis of these monocytes.


Impact of Quercetin Encapsulation with Added Phytosterols on Bilayer Membrane and Photothermal-Alteration of Novel Mixed Soy Lecithin-Based Liposome.
This study used highly lipophilic agents with an aim to increase the oxidant inhibitory activity and enhance photothermal stability of a novel mixed soy lecithin (ML)-based liposome by changing the composition of formulation within the membrane. Specifically, the development and optimization of the liposome intended for improving Trolox equivalent antioxidant capacity (TEAC) value and %TEAC loss was carried out by incorporating a natural antioxidant, quercetin (QU). In this context, a focus was set on QU encapsulation in ML-based liposomes and the concentration-dependent solubility of QU was investigated and calculated as encapsulation efficiency (EE). To explore the combined effects of the incorporation of plant sterols on the integrity and entrapment capacity of mixed phospholipid vesicles, conjugation of two types of phytosterols (PSs), namely β-sitosterol (βS) and stigmasterol (ST), to mixed membranes at different ratios was also performed. The EE measurement revealed that QU could be efficiently encapsulated in the stable ML-based liposome using 0.15 and 0.1 g/100 mL of βS and ST, respectively. The aforementioned liposome complex exhibited a considerable TEAC (197.23%) and enhanced TEAC loss (30.81%) when exposed to ultraviolet (UV) light (280-320 nm) over a 6 h duration. It appeared that the presence and type of PSs affect the membrane-integration characteristics as well as photodamage transformation of the ML-based liposome. The association of QU with either βS or ST in the formulation was justified by their synergistic effects on the enhancement of the EE of liposomes. Parallel to this, it was demonstrated that synergistic PS effects could be in effect in the maintenance of membrane order of the ML-based liposome. The findings presented in this study provided useful information for the development and production of stable QU-loaded ML-based liposomes for food and nutraceutical applications and could serve as a potential mixed lipids-based delivery system in the disease management using antioxidant therapy.


Photodynamic therapy: oncologic horizons.
Photodynamic therapy (PDT) is a light-based intervention with a long and successful clinical track record for both oncology and non-malignancies. In cancer patients, a photosensitizing agent is intravenously, orally or topically applied and allowed time to preferentially accumulate in the tumor region. Light of the appropriate wavelength and intensity to activate the particular photosensitizer employed is then introduced to the tumor bed. The light energy will activate the photosensitizer, which in the presence of oxygen should allow for creation of the toxic photodynamic reaction generating reactive oxygen species. The photodynamic reaction creates a cascading series of events including initiation of apoptotic and necrotic pathways both in tumor and neovasculature, leading to permanent lesion destruction often with upregulation of the immune system. Cutaneous phototoxicity from unintentional sunlight exposure remains the most common morbidity from PDT. This paper will highlight current research and outcomes from the basic science and clinical applications of oncologic PDT and interpret how these findings may lead to enhanced and refined future PDT.


Acceptance and patient functioning in chronic pain: the mediating role of physical activity.
Activity engagement, which refers to pursuing activities despite pain, is conceptually related to pain acceptance. A recent diary study suggested that pain acceptance precedes and directly influences physical activity, and thus, pain acceptance appears to be an important aspect of eliciting better functioning by allowing patients with chronic pain (PCPs) to participate in physical activity despite the pain. This retrospective, cross-sectional cohort study was conducted to investigate the mediating effect of physical activity on the association between pain acceptance and functioning and the association between physical activity level and functioning in PCPs. The study participants were 176 patients seeking treatment for chronic pain at a tertiary pain center in Korea. The Chronic Pain Acceptance Questionnaire, International Physical Activity Questionnaire-Short Form, and Short Form-36 were used to assess pain acceptance, physical activity, and patient functioning. The mediating effects of physical activity were tested using procedures described by Baron and Kenny. Also, three hierarchical multiple linear regression analyses were conducted to identify levels of physical activity that predict measures of patient functioning. Physical activity partially mediated the association between pain acceptance and patient functioning except for pain intensity. Furthermore, walking and moderate activity were found to be associated with improvements in all areas of patient functioning except for pain intensity. To improve the functioning of PCPs, health professionals may need to help patients accept private experiences related to pain and plan activity pacing programs that focus on value-based activities.


Qualitative and Quantitative Characteristics of the Electroencephalogram in Normal Horses during Administration of Inhaled Anesthesia.
The effects of anesthesia on the equine electroencephalogram (EEG) after administration of various drugs for sedation, induction, and maintenance are known, but not that the effect of inhaled anesthetics alone for EEG recording. To determine the effects of isoflurane and halothane, administered as single agents at multiple levels, on the EEG and quantitative EEG (qEEG) of normal horses. Six healthy horses. Prospective study. Digital EEG with video and quantitative EEG (qEEG) were recorded after the administration of one of the 2 anesthetics, isoflurane or halothane, at 3 alveolar doses (1.2, 1.4 and 1.6 MAC). Segments of EEG during controlled ventilation (CV), spontaneous ventilation (SV), and with peroneal nerve stimulation (ST) at each MAC multiple for each anesthetic were selected, analyzed, and compared. Multiple non-EEG measurements were also recorded. Specific raw EEG findings were indicative of changes in the depth of anesthesia. However, there was considerable variability in EEG between horses at identical MAC multiples/conditions and within individual horses over segments of a given epoch. Statistical significance for qEEG variables differed between anesthetics with bispectral index (BIS) CV MAC and 95% spectral edge frequency (SEF95) SV MAC differences in isoflurane only and median frequency (MED) differences in SV MAC with halothane only. Unprocessed EEG features (background and transients) appear to be beneficial for monitoring the depth of a particular anesthetic, but offer little advantage over the use of changes in mean arterial pressure for this purpose.


Sharing the burden: antigen transport and firebreaks in immune responses.
Communication between cells is crucial for immune responses. An important means of communication during viral infections is the presentation of viral antigen on the surface of an infected cell. Recently, it has been shown that antigen can be shared between infected and uninfected cells through gap junctions, connexin-based channels, that allow the transport of small molecules. The uninfected cell receiving antigen can present it on its surface. Cells presenting viral antigen are detected and killed by cytotoxic T lymphocytes. The killing of uninfected cells can lead to increased immunopathology. However, the immune response might also profit from killing those uninfected bystander cells. One benefit might be the removal of future 'virus factories'. Another benefit might be through the creation of 'firebreaks', areas void of target cells, which increase the diffusion time of free virions, making their clearance more likely. Here, we use theoretical models and simulations to explore how the mechanism of gap junction-mediated antigen transport (GMAT) affects the dynamics of the virus and immune response. We show that under the assumption of a well-mixed system, GMAT leads to increased immunopathology, which always outweighs the benefit of reduced virus production due to the removal of future virus factories. By contrast, a spatially explicit model leads to quite different results. Here we find that the firebreak mechanism reduces both viral load and immunopathology. Our study thus shows the potential benefits of GMAT and illustrates how spatial effects may be crucial for the quantitative understanding of infection dynamics and immune responses.


Cell-mediated infection of human placental trophoblast with HIV in vitro.
In order to investigate how human immunodeficiency virus (HIV) gains entry to the placenta, we have performed in vitro experiments in which highly purified trophoblast cells isolated from term human placentas were examined for their susceptibility to HIV infection. Trophoblast cells were exposed to cell-free HIV-1 for up to 24 h, after which the cultures were monitored by p24 antigen capture assay, reverse transcriptase assay, and electron microscopy for evidence of virus uptake and replication. None was found. In the second series of experiments, trophoblast cells were cocultured with HIV-infected MOLT-4 cells for 24 h, stained using an anti-HIV antibody, and examined by immunofluorescence microscopy. The MOLT cells were strongly positive, as expected, but many trophoblast colonies also showed a punctate staining pattern. Examination of similar cultures using the electron microscope revealed MOLT cells adherent to trophoblast but no evidence of cell-cell fusion. Virions were observed in coated pits at the trophoblast cell surface and in endosomes or multivesicular bodies in the cytoplasm. These observations are consistent with an endocytosis-mediated mechanism of virus entry. Virions were also observed budding from the trophoblast plasma membrane, indicating that these cells can support HIV replication. To our knowledge, these results show for the first time that HIV can infect placental trophoblast cells in vitro. The results suggest that the placenta could become infected with HIV by the interaction of virus-infected maternal lymphocytes with syncytiotrophoblast bordering the maternal blood in the intervillous space.


Political will, traditional leaders and the fight against HIV/AIDS: a South African case study.
"Political will" and leadership are increasingly considered key contextual influences on the outcomes of HIV/AIDS programmes in sub-Saharan Africa. Such debates tend to focus on the role of national leadership in shaping responses to the epidemic, with little attention to local leaders. Yet many of the settings in which HIV/AIDS flourishes are geographically distant from the reach of national leadership and policies. Furthermore, local leaders often play a key role in shaping how national policies and decisions are interpreted and implemented in local areas. Against this background, we present a case study of the impact of the leadership style of a traditional Chief on a community-based AIDS programme in a South African rural community, which sought to build community-level "AIDS competence", using the "empowerment via participation" approach. The case study involved 134 interviews and 57 focus groups conducted over three years. Thematic content analysis revealed a number of direct and indirect ways in which his leadership style impacted on project outcomes. Despite his strong support for the programme, the Chief's "traditional" attitudes towards women and youth, his celebration of polygamy, and his authoritarian governance style undermined the project's "empowerment via participation" agenda - especially the programme's attempts to reduce AIDS stigma, to build female and youth capacity to control their sexual health, and to encourage men to take responsibility for their role in tackling AIDS.


T. brucei infections abrogate diverse plasma cell-mediated effector B cell responses, independently of their specificity, affinity and host genetic background.
Antibody-mediated parasite killing is considered the most effective host immune response against extracellular trypanosome parasites. However, due to host-parasite co-evolution pressure, these parasites have "learned" how to hijack the host immune system via the development of immune evasion strategies. Hereby they prevent elimination and promote transmission. In the past, our group has shown that African trypanosome parasites are able to "shut down" the host B cell compartment, via the abolishment of the homeostatic B cell compartment. In line with this, we have reported that trypanosome infections result in detrimental outcomes on auto-reactive and cancer B cells. To unravel the immune mechanisms involved in these processes we adopted here a well-defined B cell vaccine model, i.e. the thymo-dependent hapten-carrier NP-CGG (4-Hydroxy-3-nitrophenylacetyl-Chicken Gamma Globulin) emulsified in Alum adjuvant. Results show that T. brucei infections abrogate the circulating titres of vaccine-induced CGG-specific as well as NP-specific IgG1+ antibodies, a hallmark of memory B cell responses in this model. This happens independently of their affinity and IFNɣ signalling. Next, we demonstrate that T. brucei infections also induce a decrease of anti-NP IgG3+ antibodies induced by the administration of NP coupled to Ficoll, a thymo-independent antigen. Confirming the non-specificity of the infection-associated immunopathology, this report also shows that trypanosome infections abolish vaccine-induced memory response against malaria parasite in BALB/c mice. Together, these data indicates that T. brucei infections impair every stages of B cell development, including effector plasma B cells, independently of their specificity and affinity as well as the host genetic background.


Management of common sports-related injuries about the foot and ankle.
Foot and ankle injuries are commonplace in competitive sports. Improvements in injury surveillance programs and injury reporting have enabled physicians to better recognize and manage specific foot and ankle injuries, with a primary goal of efficient and safe return to play. Athletes are becoming stronger, faster, and better conditioned, and higher-energy injuries are becoming increasingly common. Close attention is required during examination to accurately identify such injuries as turf toe, ankle injuries, tarsometatarsal (ie, Lisfranc) injuries, and stress fractures. Early diagnosis and management of these injuries are critical. Ultimately, however, pressure to return to play must not compromise appropriate care and long-term outcomes.


In vitro circadian period is associated with circadian/sleep preference.
Evaluation of circadian phenotypes is crucial for understanding the pathophysiology of diseases associated with disturbed biological rhythms such as circadian rhythm sleep disorders (CRSDs). We measured clock gene expression in fibroblasts from individual subjects and observed circadian rhythms in the cells (in vitro rhythms). Period length of the in vitro rhythm (in vitro period) was compared with the intrinsic circadian period, τ, measured under a forced desynchrony protocol (in vivo period) and circadian/sleep parameters evaluated by questionnaires, sleep log, and actigraphy. Although no significant correlation was observed between the in vitro and in vivo periods, the in vitro period was correlated with chronotype, habitual sleep time, and preferred sleep time. Our data demonstrate that the in vitro period is significantly correlated with circadian/sleep preference. The findings suggest that fibroblasts from individual patients can be utilized for in vitro screening of therapeutic agents to provide personalized therapeutic regimens for CRSD patients.


A human relevant mixture of persistent organic pollutants (POPs) and perfluorooctane sulfonic acid (PFOS) differentially affect glutamate induced excitotoxic responses in chicken cerebellum granule neurons (CGNs) in vitro.
Primary cultures of cerebellar granule neurons (CGNs) derived from chicken embryos were used to explore the effects on developmental neurotoxicity by a complex defined mixture of persistent organic pollutants (POPs). Its chemical composition and concentrations were based on blood levels in the Norwegian/Scandinavian population. Perfluorooctane sulfonic acid (PFOS) alone, its most abundant compound was also evaluated. Different stages of CGNs maturation, between day in vitro (DIV) 1, 3, and 5 were exposed to the POP mixture, or PFOS alone. Their combination with glutamate, an excitatory endogenous neurotransmitter important in neurodevelopment, also known to cause excitotoxicity was evaluated. Outcomes with the mixture at 500x blood levels were compared to PFOS at its corresponding concentration of 20 μM. The POP mixture reduced tetrazolium salt (MTT) conversion at earlier stages of maturation, compared to PFOS alone. Glutamate-induced excitotoxicity was enhanced above the level of that induced by glutamate alone, especially in mature CGNs at DIV5. Glutathione (GSH) concentrations seemed to set the level of sensitivity for the toxic insults from exposures to the pollutants. The role of N-methyl-D-aspartate receptor (NMDA-R) mediated calcium influx in pollutant exposures was investigated using the non-competitive and competitive receptor antagonists MK-801 and CGP 39551. Observations indicate a calcium-independent, but still NMDA-R dependent mechanism in the absence of glutamate, and a calcium- and NMDA-R dependent one in the presence of glutamate. The outcomes for the POP mixture cannot be explained by PFOS alone, indicating that other chemicals in the mixture contribute its overall effect.


Hip Arthroscopy for Femoroacetabular Impingement-Associated Labral Tears: Current Status and Future Prospects.
Femoroacetabular impingement (FAI) has emerged as a common cause of hip pain, especially in young patients. While the exact cause of FAI is unknown, it is thought to result from repetitive microtrauma to the proximal femoral epiphysis leading to abnormal biomechanics. Patients typically present with groin pain that is exacerbated by hip flexion and internal rotation. Diagnosis of FAI is made through careful consideration of patient presentation as well as physical exam and diagnostic imaging. Use of radiographs can help diagnose both cam and pincer lesions, while the use of MRI can diagnose labral tears and cartilage damage associated with FAI. Both non-operative and surgical options have their role in the treatment of FAI and its associated labral tears; however, hip arthroscopy has had successful outcomes when compared with physical therapy alone. Unfortunately, chondral lesions associated with FAI have had poorer outcomes with a higher conversion rate to arthroplasty. Capsular closure following hip arthroscopy has shown superior clinical outcomes and therefore should be performed if possible. More recently, primary labral reconstruction has emerged in the literature as a good option for irreparable labral tears. While non-operative management may have its role in treating patients with FAI, hip arthroscopy has developed a successful track record in being able to treat cam and pincer lesions, chondral damage, and labral injuries.


Hormesis and health: a little of what you fancy may be good for you.
The term hormesis refers to beneficial effects from low doses of potentially harmful substances. Although there are many laboratory examples of this phenomenon, it remains controversial and has never become widely accepted by the health community. This review goes beyond the laboratory and describes many clinical and common sense, real-world examples of hormesis that often go unrecognized. Many vitamins and minerals are essential for life at low doses but toxic at higher ones. Similarly, exercise, caloric restriction, and alcohol consumption are examples of processes that are harmful in the extreme but beneficial in moderation. This review also highlights possible reasons why acceptance of the hormetic paradigm has lagged. These include high-dose toxicologic testing that precludes the demonstration of low-level effects and the threat posed by hormesis to the currently accepted precautionary principle, which assumes that any dose of a chemical is potentially harmful.


Cross sectional survey of multicentre clinical databases in the United Kingdom.
To describe the multicentre clinical databases that exist in the United Kingdom, to report on their quality, to explore which organisational and managerial features are associated with high quality, and to make recommendations for improvements. Cross sectional survey, with interviews with database custodians and search of electronic bibliographic database (PubMed). 105 clinical databases across the United Kingdom. Clinical databases existed in all areas of health care, but their distribution was uneven-cancer and surgery were better covered than mental health and obstetrics. They varied greatly in age, size, growth rate, and geographical areas covered. Their scope (and thus their potential uses) and the quality of the data collected also varied. The latter was not associated with any organisational characteristics. Despite impressive achievements, many faced substantial financial uncertainty. Considerable scope existed for improvements: greater use of nationally approved codes; more support from relevant professional organisations; greater involvement by nurses, allied health professionals, managers, and laypeople in database management teams; and more attention to data security and ensuring patient confidentiality. With some notable exceptions, the audit and research potential of most databases had not been realised: half the databases had each produced only four or fewer peer reviewed research articles. At least one clinical database support unit is needed in the United Kingdom to provide assistance in organisation and management, information technology, epidemiology, and statistics. Without such an initiative, the variable picture of databases reported here is likely to persist and their potential not be realised.


Design, synthesis, and structure-activity relationship studies of novel thioether pleuromutilin derivatives as potent antibacterial agents.
A series of novel thioether pleuromutilin derivatives incorporating various heteroaromatic substituents into the C14 side chain have been reported. Structure-activity relationship (SAR) studies resulted in compounds 52 and 55 with the most potent in vitro antibacterial activity among the series (MIC = 0.031-0.063 μg/mL). Further optimization to overcome the poor water solubility of compound 55 resulted in compounds 87, 91, 109, and 110 possessing good in vitro antibacterial activity with increased hydrophilicity. Compound 114, the water-soluble phosphate prodrug of compound 52, was also prepared and evaluated. Among the derivatives, compound 110 showed moderate pharmacokinetic profiles and good in vivo efficacy in both MSSA and MRSA systemic infection models. Compound 110 was further evaluated in CYP450 inhibition assay and displayed intermediate in vitro inhibition of CYP3A4.


Interventions Within the Scope of Occupational Therapy Addressing Behavioral and Psychological Symptoms of Dementia (2018-2022).
Systematic review briefs provide a summary of the findings from systematic reviews evaluated in conjunction with the American Occupational Therapy Association's Evidence-Based Practice Program. Each systematic review brief summarizes the evidence on a theme related to a systematic review topic. This systematic review brief presents finding related to interventions within the scope of occupational therapy that address behavioral and psychological symptoms of dementia.


Tailoring and evaluating treatment with the Patient-Specific Needs Evaluation: A Patient-Centered Approach.
No patient-reported instrument assesses patient-specific information needs, treatment goals, and Personal Meaningful Gain (PMG, a novel construct evaluating individualized, clinically relevant improvement). This study reports the development of the Patient-Specific Needs Evaluation (PSN) and examines its discriminative validity (i.e., its ability to distinguish satisfied from dissatisfied patients) and test-retest reliability in patients with hand or wrist conditions. A mixed-methods approach was used to develop and validate the PSN, following COSMIN guidelines, including pilot testing, a survey (pilot: n=223, final PSN: n=275), cognitive debriefing (n=16), expert input, and validation. Discriminative validity was assessed by comparing the satisfaction level of patients who did or did not achieve their PMG (n=1,985) and test-retest reliability using absolute agreement, Cohen's kappa, and ICCs (n=102). We used a sample of 2,860 patients to describe responses to the final PSN. The PSN has only five questions (completion time ±3 minutes) and is freely accessible online. The items and response options were considered understandable by 90-92% and complete by 84-89% of the end-users. The PSN had excellent discriminative validity (Cramer's V: 0.48, p<0.001) and moderate to high test-retest reliability (Kappa: 0.46-0.68, ICCs: 0.53-0.73). The PSN is a freely available patient-centered decision-support tool that helps clinicians tailor their consultations to the patient's individual needs and goals. It contains the PMG, a novel construct evaluating individualized, clinically relevant treatment outcomes. The PSN may function as a conversation starter, facilitate expectation management, and aid shared decision-making. The PSN is implementation-ready and can be readily adapted to other patient populations. I.


Continued transmission of hepatitis B and C viruses, but no transmission of human immunodeficiency virus among intravenous drug users participating in a syringe/needle exchange program.
The virological efficacy of a syringe/needle exchange program was evaluated in a cohort incidence study. Of 698 intravenous drug users (IVDUs) initially recruited, 15 (2.1%) were HIV-positive at baseline. Adequate follow-up was possible in 515 (74%) and showed no new cases of HIV infection during a median of 31 months. Most IVDUs had been previously exposed to HBV (anti-HBc-positive 70.1%) and HCV (anti-HCV-positive 90.7%). Of those 159 IVDUs negative at baseline for anti-HBc and/or anti-HCV, 56 (35%) seroconverted to one or both viruses during follow-up, corresponding to 11.7 seroconversions/100 y at risk for HBV and 26.3 seroconversions/100 y for HCV. Multiple logistic regression analysis showed hepatitis seroconversion to correlate with imprisonment during the study (OR 2.2; 95% CI 1.04-4.74), absence of drug-free periods (OR 5.7; CI 1.44-22.3) and frequent syringe/needle exchanges (OR 1.31; CI 1.02-1.7). The absence of HIV spread was probably partly due to the low prevalence of HIV-infected IVDUs in the city. Despite free syringes and needles, both HBV and HCV continued to spread at high rates. Nevertheless, syringe/needle exchange programs, coupled with monitoring of serostatus provide good surveillance and are valuable for further assessment of remaining risks.


Sex-dependent changes in cerebellar thyroid hormone-dependent gene expression following perinatal exposure to thimerosal in rats.
Mammalian brain development is regulated by the action of thyroid hormone (TH) on target genes. We have previously shown that the perinatal exposure to thimerosal (TM, metabolized to ethylmercury) exerts neurotoxic effects on the developing cerebellum and is associated with a decrease in cerebellar D2 activity, which could result in local brain T3 deficiency. We have also begun to examine TM effect on gene expression. The objective of this study was to expand on our initial observation of altered cerebellar gene expression following perinatal TM exposure and to examine additional genes that include both TH-dependent as well as other genes critical for cerebellar development in male and female neonates exposed perinatally (G10-G15 and P5 to P10) to TM. We report here for the first time that expression of suppressor-of-white-apricot-1 (SWAP-1), a gene negatively regulated by T3, was increased in TM-exposed males (61.1% increase), but not in females; (p<0.05). Positively regulated T3-target genes, Purkinje cell protein 2 (Pcp2; p=0.07) and Forkhead box protein P4 (FoxP4; p=0.08), showed a trend towards decreased expression in TM-exposed males. The expression of deiodinase 2 (DIO2) showed a trend towards an increase in TM-exposed females, while deiodinase 3 (DIO3), transthyretin (TTR), brain derived neurotrophic factor (BDNF) and reelin (RELN) was not significantly altered in either sex. Since regulation of gene splicing is vital to neuronal proliferation and differentiation, altered expression of SWAP-1 may exert wide ranging effects on multiple genes involved in the regulation of cerebellar development. We have previously identified activation of another TH-dependent gene, outer dense fiber of sperm tails 4, in the TM exposed male pups. Together, these results also show sex-dependent differences between the toxic impacts of TM in males and females. Interestingly, the genes that were activated by TM are negatively regulated by TH, supporting our hypothesis of local brain hypothyroidism being induced by TM and suggesting a novel mechanism of action TM in the developing brain.


[Sexual dimorphism and asymmetry of digit ratio (2D:4D) – pilot study].
Numerous studies have demonstrated sexual dimorphism in 2D:4D digit ratio. Shall be regarded that the value of the 2D:4D ratio is determined in fetal life by the action of hormones. The aim of the study was to evaluate asymmetry and sexual dimorphism in 2D:4D digit ratio. The study was conducted on a group of 115 women and 46 men, aged 19–25 years. Individuals, which were included in the study, are students of two universities in Cracow – Academy of Physical Education and the Jagiellonian University. The measurement of the length the fingers II and IV both hands was made in each person and the value of digit ratio was calculated for all obtained data. The women had the higher value of the index 2D:4D in both hands, compared with men covered by the survey. In males it was demonstrated higher average value of the length of the finger IV than the finger II.


Production and assay of antibodies to an activator of adenylate cyclase, forskolin.
Forskolin is a unique diterpene activator of adenylate cyclase which has been extensively used in the study of cAMP generating systems. This report describes the production of antibodies to forskolin and the optimization of two sensitive assay methods for such antibodies. 7-0-Hemisuccinyl 7-deacetyl forskolin, coupled to either human serum albumin or goat IgG, was injected into goats to elicit antibodies to the forskolin hapten. Two assay methods, a radioimmunoassay with [12-3H]forskolin as a tracer and a colorimetric enzyme-linked immunosorbent assay (ELISA) with horse radish peroxidase-labelled rabbit anti-goat IgG as an indicator, were optimized to test for the presence of forskolin antibodies in antisera and isolated IgG fractions. The titers for forskolin antisera were 4000-10000. Both assay methods can be adapted to quantify forskolin and its protein conjugates. The availability of antibodies to this diterpene will be useful in accelerating the understanding of the mechanism of adenylate cyclase activation by forskolin.


Exposure to household air pollution from solid cookfuels and childhood stunting: a population-based, cross-sectional study of half a million children in low- and middle-income countries.
Household air pollution from the incomplete combustion of solid cookfuels in low- and middle-income countries (LMICs) has been largely ignored as a potentially important correlate of stunting. Our objective was to examine the association between solid cookfuel use and stunting in children aged <5 y. We used data from 59 LMICs' population-based cross-sectional demographic and health surveys; 557 098 children aged <5 y were included in our analytical sample. Multilevel logistic regression was used to examine the association between exposure to solid cookfuel use and childhood stunting, adjusting for child sex, age, maternal education and number of children living in the household. We explored the association across key subgroups. Solid cookfuel use was associated with child stunting (adjusted OR 1.58, 95% CI 1.55 to 1.61). Children living in households using solid cookfuels were more likely to be stunted if they lived in rural areas, the poorest households, had a mother who smoked tobacco or were from the Americas. Focused strategies to reduce solid cookfuel exposure might contribute to reductions in childhood stunting in LMICs. Trial evidence to assess the effect of reducing solid cookfuel exposure on childhood stunting is urgently needed.


Structural aspects of leg-to-gonopod metamorphosis in male helminthomorph millipedes (Diplopoda).
In the adult males of helminthomorph millipedes, one or two pairs of legs in the anterior part of the trunk are strongly modified into sexual appendages (gonopods) used for sperm transfer during the copula. Gonopods differentiate in an advanced phase of post-embryonic development, in most cases as replacement for the walking legs of the seventh trunk ring, as these first regress to tiny primordia, to eventually develop into gonopods at a subsequent stadium. These extremely localized but dramatic changes have been described as a non-systemic metamorphosis. In the present study we describe morphological and anatomical changes of trunk ring VII associated with non-systemic metamorphosis in four helminthomorph species. As documented here for the first time by means of traditional histology methods and new techniques based on confocal laser scanning microscopy, the external modifications caused by non-systemic metamorphosis are associated to a huge rearrangement of internal anatomy, mostly due to the development of gonopod apodemes and extrinsic muscles. Internal changes in the seventh trunk ring, locally leading to the dorsal displacement of the ventral nerve cord and the digestive tract, are modulated in a taxon-specific manner, and are very conspicuous in the blaniulids Nopoiulus kochii and Blaniulus guttulatus, with likely major functional consequences.


Benefits and Toxicity of Disulfiram in Preclinical Models of Nephropathic Cystinosis.
Nephropathic cystinosis is a rare disease caused by mutations of the CTNS gene that encodes for cystinosin, a lysosomal cystine/H+ symporter. The disease is characterized by early-onset chronic kidney failure and progressive development of extra-renal complications related to cystine accumulation in all tissues. At the cellular level, several alterations have been demonstrated, including enhanced apoptosis, altered autophagy, defective intracellular trafficking, and cell oxidation, among others. Current therapy with cysteamine only partially reverts some of these changes, highlighting the need to develop additional treatments. Among compounds that were identified in a previous drug-repositioning study, disulfiram (DSF) was selected for in vivo studies. The cystine depleting and anti-apoptotic properties of DSF were confirmed by secondary in vitro assays and after treating Ctns-/- mice with 200 mg/kg/day of DSF for 3 months. However, at this dosage, growth impairment was observed. Long-term treatment with a lower dose (100 mg/kg/day) did not inhibit growth, but failed to reduce cystine accumulation, caused premature death, and did not prevent the development of renal lesions. In addition, DSF also caused adverse effects in cystinotic zebrafish larvae. DSF toxicity was significantly more pronounced in Ctns-/- mice and zebrafish compared to wild-type animals, suggesting higher cell toxicity of DSF in cystinotic cells.


Normal Skin Cells Increase Aggressiveness of Cutaneous Melanoma by Promoting Epithelial-to-Mesenchymal Transition via Nodal and Wnt Activity.
Melanoma is a lethal form of skin cancer triggered by genetic and environmental factors. Excision of early-stage, poorly aggressive melanoma often leads to a successful outcome; however, left undiagnosed these lesions can progress to metastatic disease. This research investigates whether the exposure of poorly aggressive melanoma to certain normal skin cells can explain how non-metastatic melanoma becomes more aggressive while still confined to the skin. To this end, we used a serial co-culture approach to sequentially expose cells from two different, poorly aggressive human melanoma cell lines against normal cells of the skin beginning with normal melanocytes, then epidermal keratinocytes, and finally dermal fibroblasts. Protein extraction of melanoma cells occurred at each step of the co-culture sequence for western blot (WB) analysis. In addition, morphological and functional changes were assessed to detect differences between the serially co-cultured melanoma cells and non-co-cultured cells. Results show that the co-cultured melanoma cells assumed a more mesenchymal morphology and displayed a significant increase in proliferation and invasiveness compared to control or reference cells. WB analysis of protein from the co-cultured melanoma cells showed increased expression of Snail and decreased levels of E-cadherin suggesting that epithelial-to-mesenchymal transition (EMT) is occurring in these co-cultured cells. Additional WB analysis showed increased levels of Nodal protein and signaling and signs of increased Wnt activity in the co-cultured melanoma cells compared to reference cells. These data suggest that interaction between poorly aggressive melanoma cells with normal cells of the skin may regulate the transition from localized, poorly aggressive melanoma to invasive, metastatic disease via Nodal and/or Wnt induced EMT.


Extraction of Active Compounds from Mixtures of Hemp (Cannabis sativa) with Plants of the Zingiberaceae Family.
Hemp is probably one of the most studied plants for its health-promoting properties, with countless documented and patented extraction methods, but literature is scarce on the simultaneous extraction of mixture of raw materials. Hemp, along with other plant materials, could represent a potentially highly valuable source material with resulting reciprocal effects. In this study, hemp (Cannabis sativa) and three members of the Zingiberaceae family, ginger (Zingiber officinale), turmeric (Curcuma longa), and cardamom (Elettaria cardamomum), were extracted simultaneously, and their bioactive component values were investigated. Two extraction methods were used, namely ultrasound-assisted extraction with ethanol and supercritical fluid extraction with carbon dioxide. First, extracts were obtained from separate plant materials. Then, hemp was extracted in combination with ginger, turmeric, and cardamom in a 1:1 ratio. The extracts obtained were evaluated for their antioxidant activity and total phenolic content using UV/VIS spectrophotometry; cannabinoid content, 6-gingerol, and 6-shogaol were measured using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS); volatile components such as 1,8-cineole, alpha-terpinyl acetate, linalool, and aR-turmerone were measured using gas chromatography with mass spectrometry (GC/MS).