Regulation of human polymorphonuclear neutrophil (PMN) activity against Candida albicans by large granular lymphocytes via release of a PMN-activating factor.
Using a 24-hr radiolabel microassay developed in our laboratory that measures [3H]glucose uptake in residual Candida, we have identified the effector cells responsible for in vitro inhibition of Candida albicans growth as mainly polymorphonuclear neutrophils (PMN) and monocytes within the human peripheral blood cells. Highly purified T cells and large granular lymphocytes (LGL) that mediate natural killer activity which were obtained by Percoll density gradient centrifugation were found to have no innate activity against C. albicans. The LGL could not be activated by interferon-alpha, interferon-gamma or interleukin 2 to inhibit Candida growth although their K562 tumor cytotoxic activity was readily enhanced by these cytokines. Stimulation with heat-killed C. albicans also did not activate fungal growth inhibitory function in LGL and the supernatant of these activated LGL had no direct fungicidal activity. However, the activated LGL supernatant had the capability to enhance PMN function against C. albicans growth. Addition of recombinant human tumor necrosis factor, affinity-purified interferon-alpha, or interferon-gamma to PMN caused increased antifungal activity in PMN. However, antibodies to these cytokines had only a partial adverse effect on the ability of the activated LGL supernatant to stimulate PMN anti-Candida function. Therefore, the activated LGL supernatant appeared to contain a potent stimulator of PMN function which is as yet unidentified. These data indicate that LGL did not directly mediate anti-Candida activity but could indirectly influence C. albicans growth by activating PMN against the fungi through the release of a specific PMN-activating factor. Our findings therefore add another role to LGL which is the regulation of PMN function, the consequence of which is regulation of fungal immunity.


A Method for Grouping Emergency Department Visits by Severity and Complexity.
Triage functions to quickly prioritize care and sort patients by anticipated resource needs. Despite widespread use of the Emergency Severity Index (ESI), there is still no universal standard for emergency department (ED) triage. Thus, it can be difficult to objectively assess national trends in ED acuity and resource requirements. We sought to derive an ESI from National Hospital Ambulatory Medical Care Survey (NHAMCS) survey items (NHAMCS-ESI) and to assess the performance of this index with respect to stratifying outcomes, including hospital admission, waiting times, and ED length of stay (LOS). We used data from the 2010-2015 NHAMCS, to create a measure of ED visit complexity based on variables within NHAMCS. We used NHAMCS data on chief complaint, vitals, resources used, interventions, and pain level to group ED visits into five levels of acuity using a stepwise algorithm that mirrored ESI. In addition, we examined associations of NHAMCS-ESI with typical indicators of acuity such as waiting time, LOS, and disposition. The NHAMCS-ESI categorization was also compared against the "immediacy" variable across all of these outcomes. Visit counts used weighted scores to estimate national levels of ED visits. The NHAMCS ED visits represent an estimated 805,726,000 ED visits over this time period. NHAMCS-ESI categorized visits somewhat evenly, with most visits (42.5%) categorized as a level 3. The categorization pattern is distinct from that of the "immediacy" variable within NHAMCS. Of admitted patients, 89% were categorized as NHAMCS-ESI level 2-3. Median ED waiting times increased as NHAMCS-ESI levels decreased in acuity (from approximately 14 minutes to 25 minutes). Median LOS decreased as NHAMCS-ESI decreased from almost 200 minutes for level 1 patients to nearly 80 minutes for level 5 patients. We derived an objective tool to measure an ED visit's complexity and resource use. This tool can be validated and used to compare complexity of ED visits across hospitals and regions, and over time.


Mannan-binding lectin in human serum, cerebrospinal fluid and brain tissue and its role in Alzheimer's disease.
Mannan-Binding lectin (MBL) is a serum lectin which can activate the classical complement pathway. Complement proteins of the classical pathway have been found in the brains of patients with Alzheimer's disease (AD) in association with AD brain pathology. To investigate the role for MBL in AD we have looked for its presence in the brain by immunohistochemistry and determined the levels of MBL in paired samples of cerebrospinal fluid and serum from AD patients and controls. MBL was detected in association with blood vessels in the brain tissue of both AD patients and control subjects. There was no apparent difference in the distribution of MBL in the brain tissue between the two groups. The mean concentration of MBL in the CSF was 44% lower in AD patients than in controls (AD 154 +/- 35 pg/ml, n = 19; non-AD 276 +/- 50 pg/ml, n = 15, p < 0.05). The levels of MBL in serum were not significantly different in the two groups. Thus, this study shows that MBL is associated with blood vessels in the brains of both AD and control subjects. Moreover, CSF levels of MBL appear to be lower in AD patients than in control subjects which may indicate a higher degree of MBL consumption connected with complement activation in the AD patients.


17-AAG and Apoptosis, Autophagy, and Mitophagy in Canine Osteosarcoma Cell Lines.
Canine osteosarcoma is highly resistant to current chemotherapy; thus, clarifying the mechanisms of tumor cell resistance to treatments is an urgent need. We tested the geldanamycin derivative 17-AAG (17-allylamino-17-demethoxygeldanamycin) prototype of Hsp90 (heat shock protein 90) inhibitors in 2 canine osteosarcoma cell lines, D22 and D17, derived from primary and metastatic tumors, respectively. With the aim to understand the interplay between cell death, autophagy, and mitophagy, in light of the dual effect of autophagy in regulating cancer cell viability and death, D22 and D17 cells were treated with different concentrations of 17-AAG (0.5 μM, 1 μM) for 24 and 48 hours. 17-AAG-induced apoptosis, necrosis, autophagy, and mitophagy were assessed by transmission electron microscopy, flow cytometry, and immunofluorescence. A simultaneous increase in apoptosis, autophagy, and mitophagy was observed only in the D22 cell line, while D17 cells showed low levels of apoptotic cell death. These results reveal differential cell response to drug-induced stress depending on tumor cell type. Therefore, pharmacological treatments based on proapoptotic chemotherapy in association with autophagy regulators would benefit from a predictive in vitro screening of the target cell type.


Mirror-symmetrical populations of wide-field amacrine cells of the macaque monkey retina.
Retinas of macaque monkeys were immunostained for glycogen phosphorylase (glypho). Glypho was localized to regular and displaced amacrine cells. Their processes occupied two narrow strata within the inner plexiform layer (IPL). The labeling pattern is reminiscent of cholinergic amacrine cells; however, double immunostaining of the retinas for choline acetyltransferase and glypho revealed two different cell populations. Intracellular injection of DiI showed that glypho-immunoreactive amacrine cells are wide-field amacrine cells with straight, radially oriented, and sparsely branched dendrites. The density of the cells increased from approximately 70/mm(2) in the peripheral retina to approximately 700/mm(2) in the central retina. The regular glypho-immunoreactive amacrine cells branch in sublamina 2 of the IPL, where they receive input from OFF-cone bipolar cells. The displaced cells branch in sublamina 3/4 and receive input from ON-cone bipolar cells. This suggests that the regular cells are OFF-cells and the displaced cells are ON-cells. The cells express gamma-aminobutyric acid (GABA)-like immunoreactivity and receive glycinergic input through synapses expressing preferentially the glycine receptor alpha2 subunit. The close proximity of the dendritic strata of glypho-immunoreactive amacrine cells, cholinergic amacrine cells, and direction-selective ganglion cells suggests a possible role of the cells in the generation of direction-selective light responses of the monkey retina.


[Early diagnosis of autism: Phenotype-endophenotype].
Autism Spectrum Disorders have for some time been the focus of intense interest for clinicians and researchers because of the high prevalence of the disorders among children in the community (approximately 1%), their severity and pervasiveness. Particular attention has been paid to the early diagnosis of the disorder and to the intensive therapeutic intervention. Currently the best prognosis for autism lays in the early diagnosis and intervention. Postponing the diagnosis and the intervention beyond infancy is considered loss of precious time. The diagnosis of autism, which begins early in life, was until recently considered that could be reliability made at the age of 3 years. Recent follow up studies however on children at risk for autism (children who had an older sibling with autism) have shown that the clinical signs of autism emerge at the end of the first year and become distinct by the end of the second year when the diagnosis can reliably be made. From a clinical perspective it is noted that the early clinical signs of risk for autism are related to social communication (e.g. limited or absent response when calling his/her name and to joint attention), stereotype behaviours and body movements or unusual handling of objects (e.g. intensive observation of objects and stereotype movements of hands and tapping or spinning), incongruent regulation of emotions (reduced positive and increased negative emotion). There is also delay in developmental characteristics such as the language (both receptive and expressive) and motor (particularly in postural control - characteristic is the drop of the head backwards when the infant is held in horizontal position). Studies on various aspects of the endophenotype of certain clinical signs among infants at risk for Autism Spectrum Disorders, such as avoidance of eye contact, delay in verbal communication and increase of the head circumference, may provide useful information and may assist the clinician on follow up in the early diagnosis. Compared to the progress made for an early diagnosis of autism there is relatively no equal progress in the early intervention. However, some methods such as the 'Early Start Denver Model' which focuses in the family, which is the natural environment of the child, may provide a useful framework for effective work with the child and the family.


The influence of sample area on diclofenac sodium quantification by diffuse reflectance IR spectroscopy.
A procedure for the quantitative determination of diclofenac sodium (DS) in commercial capsules and tablets based on Partial Least Squares (PLS) treatment of diffuse reflectance FTIR spectroscopic (DRIFTS) data is described. Two DRIFTS accessories, a Collector II (Spectra-Tech) and a Seagull (Harrick Scientific), were used to collect the spectra. The spectrometer beam area on the surface of the sample was approximately sevenfold smaller for the Collector II accessory compared to the Seagull accessory. Spectra collection using the smaller beam spot resulted in significantly higher quantification errors for the single measurements. To reduce the errors associated with the Collector II accessory spectra were collected seven times while randomly changing the sample position. The mean spectra were used in the analysis. To compare the predictive ability of the constructed models, the relative standard errors of prediction (RSEP) were calculated. The RSEPs were 1.3-2.9% and 2.0-2.6% using the Collector II accessory and 1.0-1.5% and 1.1-1.7% using the Seagull accessory, for calibration and validation data sets, for the different PLS models. Three commercial preparations containing 20.5, 23.2 and 34.5% DS were successfully quantified using the developed models. The proposed procedure can be used as a fast, precise and economic method for DS quantification in tablets and capsules.


On which evidence can we rely when prescribing off-label methotrexate in dermatological practice? - a systematic review with GRADE approach.
If an authorized drug is prescribed for a use that is not described in the Summary of Product Characteristics, this is defined as 'off-label use.' Methotrexate is often used off-label for dermatological indications. Off-label use is permitted if physicians can justify the treatment based on scientific evidence available to them. Our objective here was therefore to summarize the evidence for the effectiveness, efficacy, and safety of the dermatological off-label use of methotrexate in a systematic review. We searched MEDLINE, EMBASE, and CENTRAL for studies for evidence on the effectiveness, efficacy, and safety of the off-label use of methotrexate in dermatological indications up to November 2019. We used the GRADE system to rate the quality of the evidence. The search retrieved 34,583 hits of which 3566 were selected after the title and abstract screening. After the full-text screening, 143 studies were included, which involved 3688 patients in total. We found low-quality evidence for the effectiveness, efficacy, and safety of the off-label use of methotrexate in 31 dermatological diseases. To optimize the quality of evidence to support off-label use, we need high-quality studies in which well-characterized patients are treated with standardized treatments regimens using well-validated outcomes relevant to patients and physicians.


Mentalization-based treatment for patients with borderline personality disorder: an overview.
To describe the concept of mentalization, and its application in understanding the development of psychopathology in patients with borderline personality disorder; to give an account of the main features of mentalization-based treatment; to summarise the evidence supporting its effectiveness. Mentalization is a predominantly preconscious mental activity that enables the individual to understand him/herself and others in terms of subjective states and mental processes. Psychological trauma in childhood is associated with deficits in mentalization and with the development of borderline personality disorder. Mentalization-based treatment is a psychodynamically-oriented manualized psychotherapy for borderline personality disorder that aims to develop a therapeutic process in which the patient's capacity for mentalization becomes the focus of treatment. Randomized controlled trials have demonstrated the effectiveness of this treatment for patients with borderline personality disorder. The development of a psychodynamically-oriented therapeutic intervention that specifically targets the deficits involved in the psychopathology of borderline personality disorder is a crucial step in increasing the effectiveness of treatment. Mental health professionals should be adequately prepared to deliver effective interventions to their patients, such as mentalization-based treatment.


Identification of Cerebral Metal Ion Imbalance in the Brain of Aging Octodon degus.
The accumulation of redox-active transition metals in the brain and metal dyshomeostasis are thought to be associated with the etiology and pathogenesis of several neurodegenerative diseases, and Alzheimer's disease (AD) in particular. As well, distinct biometal imaging and role of metal uptake transporters are central to understanding AD pathogenesis and aging but remain elusive, due inappropriate detection methods. We therefore hypothesized that Octodon degus develop neuropathological abnormalities in the distribution of redox active biometals, and this effect may be due to alterations in the expression of lysosomal protein, major Fe/Cu transporters, and selected Zn transporters (ZnTs and ZIPs). Herein, we report the distribution profile of biometals in the aged brain of the endemic Chilean rodent O. degus-a natural model to investigate the role of metals on the onset and progression of AD. Using laser ablation inductively coupled plasma mass spectrometry, our quantitative images of biometals (Fe, Ca, Zn, Cu, and Al) appear significantly elevated in the aged O. degus and show an age-dependent rise. The metals Fe, Ca, Zn, and Cu were specifically enriched in the cortex and hippocampus, which are the regions where amyloid plaques, tau phosphorylation and glial alterations are most commonly reported, whilst Al was enriched in the hippocampus alone. Using whole brain extracts, age-related deregulation of metal trafficking pathways was also observed in O. degus. More specifically, we observed impaired lysosomal function, demonstrated by increased cathepsin D protein expression. An age-related reduction in the expression of subunit B2 of V-ATPase, and significant increases in amyloid beta peptide 42 (Aβ42), and the metal transporter ATP13a2 were also observed. Although the protein expression levels of the zinc transporters, ZnT (1,3,4,6, and 7), and ZIP7,8 and ZIP14 increased in the brain of aged O. degus, ZnT10, decreased. Although no significant age-related change was observed for the major iron/copper regulator IRP2, we did find a significant increase in the expression of DMT1, a major transporter of divalent metal species, 5'-aminolevulinate synthase 2 (ALAS2), and the proto-oncogene, FOS. Collectively, our data indicate that transition metals may be enriched with age in the brains of O. degus, and metal dyshomeostasis in specific brain regions is age-related.


Child protection and public health: nurses' responsibilities.
Health care workers have been recognized as having a key role in the protection and care of Scotland's children, particularly in respect of identification and detection of child abuse. Nurses, especially health visitors, are often the first professionals to suspect that child abuse has taken place. While previous research has found that health visitors have primarily perceived their role as that of providing support and advice to vulnerable families, there are pressures on them to fulfil a more narrow surveillance role. Concurrent with a lack of clarity about the role of health visitors in child protection, there has been increasing recognition that other nurses can also make an important contribution, including those who do not work directly with children. The aim of the study was to explore nurses' understanding of their professional responsibilities in relation to child protection, and the potential for nurses to be involved in the protection of children from abuse. A qualitative interview-based design was used, and 99 nurses working in an National Health Service trust in a Scottish city were interviewed, either individually or in groups, about their professional involvements in child protection issues. Interview data were subjected to thematic analysis. There was lack of consensus among interviewees about the nursing remit in child protection issues, particularly with respect to the extent to which nurses should actively seek to detect cases of child abuse. An emphasis on identification and detection was not easily accepted by many nurses, and was perceived by some to be a change from their more traditional role of supporting families, as well as being potentially in conflict with some public health responsibilities. In spite of the perception of some nurses that there is a sharp divide between child protection work and public health interventions, many of the child protection roles identified by nurses, such as supporting families, parenting education and service development, are clearly within the ambit of contemporary notions of public health. Furthermore, it is clear that there is a role in child protection for a much wider group of nurses than health visitors.


Serum Neutrophil Gelatinase-Associated Lipocalin: A Diagnostic Marker in Pediatric Sepsis.
Sepsis is a life-threatening condition that arises when the response of the body to infection injures its own tissues and organs. The early prediction of sepsis by current clinical and laboratory methods remains inadequate. Serum neutrophil gelatinase-associated lipocalin level is increased in sepsis irrespective of renal dysfunction. Therefore, we aimed to correlate the serum neutrophil gelatinase-associated lipocalin value determined at admission with clinical progression and severity of disease in critically ill children and to declare its role as a potential diagnostic and prognostic marker for sepsis in critically ill children in the emergency department. A prospective cohort study. The study carried out at the PICU of Menoufia University Hospital. We serially enrolled 120 critically ill children admitted to the PICU at 2 fixed days per week in addition to 40 healthy children served as controls. Clinical examination was performed including calculation of the Pediatric Risk of Mortality and Pediatric Index of Mortality 2. Serum neutrophil gelatinase-associated lipocalin measurement was performed for patients at admission and for the controls. Patients were followed up for 30 days. The discriminatory power of neutrophil gelatinase- associated lipocalin was determined using the receiver-operating characteristic and other predictive likelihood values. Serum neutrophil gelatinase-associated lipocalin level was significantly higher among the total patient cohort and those with sepsis than among the controls (p < 0.001), also in patients with systemic inflammatory response syndrome without sepsis and patients without systemic inflammatory response syndrome (p = 0.04 and <0.001). Furthermore, plasma level of neutrophil gelatinase-associated lipocalin was significantly elevated in nonsurvivors compared with survivors (p < 0. 001). Receiver-operating characteristic curve analysis exhibited an area under the curve of 0.84 for neutrophil gelatinase-associated lipocalin for diagnosis of sepsis, whereas C-reactive protein had an area under the curve of 0.79. Regarding the prognosis, neutrophil gelatinase-associated lipocalin had an area under the curve of 0.74 for prediction of mortality, whereas the area under the curve for Pediatric Risk of Mortality, Pediatric Index of Mortality 2, and C-reactive protein were 0.59, 0.58, and 0.62, respectively. Overall, the data support the view that measurement at admission, serum neutrophil gelatinase-associated lipocalin results in substantial added value for early diagnosis and prognostication of sepsis in critically sick children.


Human embryonic and mesenchymal stem cells express different nuclear proteomes.
Human embryonic stem cells (hESCs) are characterized by their immortality and pluripotency. Human mesenchymal stem cells (hMSC), on the other hand, have limited self-renewal and differentiation capabilities. The underlying molecular differences that account for this characteristic self-renewal and plasticity are, however, poorly understood. This study reports a nuclear proteomic analysis of human embryonic and bone marrow-derived mesenchymal stem cells. Our proteomic screen highlighted a 5-fold difference in the expression of Reptin52. We show, using two-dimensional difference gel electrophoresis (2-DIGE), western analysis, and quantitative reverse transcriptase polymerase chain reaction, that Reptin52 is more abundantly expressed in hESC than hMSC. Moreover, we observed differential expression of Pontin52 and beta-catenin-proteins known to interact with Reptin52. This difference in the expression of Reptin52 and Pontin52 (known regulators of beta-catenin) further supports a role for Wnt signaling in stem cell self-renewal and proliferation.


Four routes of cognitive evolution.
Four routes of cognitive evolution are distinguished: phylogenetic construction, in which natural selection produces qualitative change to the way a cognitive mechanism operates (language); phylogenetic inflection, in which natural selection biases the input to a cognitive mechanism (imprinting and spatial memory); ontogenetic construction, in which developmental selection alters the way a cognitive mechanism operates (face recognition and theory of mind); and ontogenetic inflection, in which developmental selection changes the input to a cognitive mechanism (imitation). This framework integrates findings from evolutionary psychology (i.e., all research on the evolution of mentality and behavior). In contrast with human nativist evolutionary psychology, it recognizes the adaptive significance of developmental processes, conserves the distinction between cognitive and noncognitive mechanisms, and encompasses research on human and nonhuman animals.


Protective role of epigallocatechin-3-gallate on arsenic induced testicular toxicity in Swiss albino mice.
Arsenic, often referred to as the king of poisons is carcinogenic in humans and animals. It affects multiorgan systems including reproduction. The present study was undertaken to explore the protective role of green tea compound, epigallocatechin-3-gallate (EGCG) on arsenic induced testicular toxicity in Swiss albino mice. Thirty two adult male mice were randomly assigned to four groups (n=8). Group I served as control without test chemical. The group II received arsenic (200ppm) through drinking water, group III received only EGCG (20mg/kgb.wt., intraperitoneally, alternate days) and group IV was administered arsenic+EGCG for 40days. Factorial experimental design was employed to assess the treatment effect. The EGCG restored arsenic induced decrements in epididymal sperm concentration, kinematic attributes (total motility, rapid, progressive motile, fast progressive, VSL, VAP, VCL, BCF, LIN, WOB, STR and Type A), structutal membrane integrity, functional membrane integrity and mitochondrial membrane potential. As evidenced by the histoarchitectural studies, the EGCG reversed the deleterious effects of arsenic on testicular malondialdehyde (p<0.05) levels, reduced glutathione, antioxidative enzymes and spermatogenesis. Overall, the results suggest that EGCG reduces the testicular oxidative stress induced by arsenic poisoning and thereby protect the reproductive system.


Role of CYP3A5 in abnormal clearance of methadone.
To report a case of unusually low concentrations of methadone in a polydrug abuser during maintenance treatment with methadone. A 25-year-old man (weight 55 kg, height 165 cm) with a 12-year history of polydrug abuse was admitted to an opiates withdrawal methadone program. At the time of our observation, he was using both cannabinoids and heroin; no other medical conditions were discovered. Within the opiates withdrawal methadone program, under medical supervision, the patient started methadone therapy (20 mg/day). Two weeks later, an Abuscreen assay for methadone screening in the urine was negative and, to prevent the development of withdrawal symptoms, the dose of methadone was increased to 60 mg/day. One day later, the patient was asked to collect another urine sample in the presence of a nurse. The Abuscreen for methadone in urine remained negative. Evaluation of urinary samples collected over 24 hours documented low concentrations of methadone and high levels of 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (the primary metabolite of methadone). Evaluation for the presence of the most common polymorphisms in the cytochrome P450 and P-glycoprotein genes showed that the patient was heterozygous for the CYP3A5(*)1 allele and for 2 single nucleotide polymorphisms in the P-glycoprotein gene (1236C/T and 3435C/T). In this patient, poor methadone adherence was ruled out because of the presence of physicians and nurses during both methadone maintenance treatment and Abuscreen screening. Moreover, because the patient reported only heroin and cannabis at the time of evaluation, drug interactions were ruled out as possible causes for the rapid clearance of methadone. In this case, CYP3A5 polymorphism may have played a role in the rapid methadone metabolism.


Does perceived trainability of dog (Canis lupus familiaris) breeds reflect differences in learning or differences in physical ability?
Researchers have reported perceived differences in trainability between different dog breeds. These reports could either be the result of underlying differences in learning or differences in physical capabilities. Four studies were conducted to investigate this issue. In Study 1 the level of agility metal-winners amongst those breeds perceived to be high and low in trainability did not deviate significantly from their respective levels of participation in the sport. In Study 2 the level of precision amongst those dogs perceived to be high and low in trainability did not deviate significantly in a real agility competition (P>0.05), but these dogs did differ in speed (P<0.05). In Study 3 the amount of training time necessary to achieve agility precision mastery did not significantly differ amongst dogs from breeds perceived to be high and low in trainability (P>0.05), but there was a significant difference in speed. Finally, in Study 4 breeds considered to be high in trainability were found to be relatively physically homogenous in respects to height, in comparison to breeds considered to be low in trainability. Overall, the results of these studies are more supportive of a physical capability interpretation of perceived breed differences in trainability, than a more cognitive interpretation.


Analysis of Clinically Relevant Factors for Pulmonary Hypertension in Maintenance Hemodialysis Patients.
BACKGROUND Pulmonary hypertension (PH) is common in patients with maintenance hemodialysis (MHD) and is associated with high mortality. This study analyzed clinically relevant factors for pulmonary hypertension in MHD patients and the effect of serum pentraxin3 (PTX3) in the pathogenesis of PH to provide the basis for early diagnosis and treatment of MHD patients with PH. MATERIAL AND METHODS This study included 60 MHD patients (group A) and 30 healthy controls (group B). Group A was further divided into PH and non-PH groups. Clinical characteristics, auxiliary examination results and serum PTX3 level of the PH and non-PH groups were compared. Binary logistic regression was used to assess the risk factors for PH in MHD patients. ROC curve was applied to evaluate the diagnostic value of PTX3 in PH. RESULTS The incidence rate of PH in MHD patients was 50%, and most presented as mild to moderate. Compared with the non-PH group, patients in PH group presented significantly longer atrial diameter, right ventricular diameter and main pulmonary artery diameter (P<0.05), as well as higher PTX3 and NT-proBNP level. Atrial diameter and PTX3 level were the risk factors for PH in MHD patients. AUC of PTX3 was 0.721 (95%CI: 0.590-0.851, P=0.003). CONCLUSIONS The prevalence of PH was higher in MHD patients and mostly presented as mild to moderate. Such patients often developed heart structural changes and cardiac ultrasound was highly recommended. Serum PTX3 level was significantly elevated and could be used as a marker of PH in MHD patients.


Seasonal variation in tsetse fly apparent density and Trypanosoma spp. infection rate and occurrence of drug-resistant trypanosomes in Lambwe, Kenya.
Tsetse flies are major arthropod vectors of trypanosomes that cause debilitating African animal trypanosomiasis. The emergence of drug-resistant trypanosomes is a common problem in sub-Saharan Africa. This study aimed to identify tsetse flies' seasonal variation in apparent densities and their infection rates and the occurrence of drug-resistant trypanosomes. Tsetse flies were collected from Lambwe, Kenya, during May and September 2021. Genomic DNA was extracted from them, and the ITS1 gene was amplified to detect Trypanosoma infection with subsequent species determination. Transporter genes DMT, E6M6, TbAT/P2, and TcoAde2 were targeted to detect polymorphisms associated with drug-resistance, using sequencing and comparison to drug-sensitive trypanosome species referenced in Genbank. A total of 498 tsetse flies and 29 non-tsetse flies were collected. The apparent density of flies was higher in wet season 6.2 fly per trap per density (FTD) than in the dry season 2.3 FTD (P = 0.001), with n = 386 and n = 141 flies caught in each season, respectively. Male tsetse flies (n = 311) were more numerous than females (n = 187) (P = 0.001). Non-tsetse flies included Tabanids and Stomoxys spp. Overall, Trypanosoma infection rate in tsetse was 5% (25/498) whereby Trypanosoma vivax was 4% (11/25), Trypanosoma congolense 36% (9/25), and Trypanosoma brucei 20% (5/25) (P = 0.186 for the distribution of the species), with infections being higher in females (P = 0.019) and during the wet season (P < 0.001). Numerous polymorphisms and insertions associated with drug resistance were detected in DMT and E6M6 genes in two T. congolense isolates while some isolates lacked these genes. T. brucei lacked TbAT/P2 genes. TcoAde2 sequences in three T. congolense isolates were related to those observed in trypanosomes from cattle blood in our previous study, supporting tsetse fly involvement in transmission in the region. We report Trypanosoma associated with trypanocidal drug-resistance in tsetse flies from Lambwe, Kenya. Female tsetse flies harbored more Trypanosoma infections than males. Tsetse transmission of trypanosomes is common in Lambwe. Risk of trypanosome infection would seem higher in the wet season, when tsetse flies and Trypanosoma infections are more prevalent than during the dry season. More efforts to control animal trypanosome vectors in the region are needed, with particular focus on wet seasons.


Molecular analysis of the hot spot region related to length mutations in wheat chloroplast DNAs. I. Nucleotide divergence of genes and intergenic spacer regions located in the hot spot region.
The nucleotide divergence of chloroplast DNAs around the hot spot region related to length mutation in Triticum (wheat) and Aegilops was analyzed. DNA sequences (ca. 4.5 kbp) of three chloroplast genome types of wheat complex were compared with one another and with the corresponding region of other grasses. The sequences region contained rbcL and psaI, two open reading frames, and a pseudogene, rpl23' (pseudogene for ribosomal protein L23) disrupted by AT-rich intergic spacer regions. The evolution of these genes in the closely related wheat complex is characterized by nonbiased nucleotide substitutions in terms of being synonymous/nonsynonymous, having A-T pressure transitions over transversions, and frequent changes at the third codon position, in contrast with the gene evolution among more distant plant groups where biased nucleotide substitutions have frequently occurred. The sequences of these genes had diverged almost in proportion to taxonomic distance. The sequence of the pseudogene rpl23' changed approximately two times faster than that of the coding region. Sequence comparison between the pseudogene and its protein-coding counterpart revealed different degrees of nucleotide homology in wheat, rice and maize, suggesting that the transposition timing of the pseudogene differed and/or that different rates of gene conversion operated on the pseudogene in the cpDNA of the three plant groups in Gramineae. The intergenic spacer regions diverged approximately ten times faster than the genes. The divergence of wheat from barley, and that from rice are estimated based on the nucleotide similarity to be 1.5, 10 and 40 million years, respectively.


Nonredundant roles for cytochrome c2 and two high-potential iron-sulfur proteins in the photoferrotroph Rhodopseudomonas palustris TIE-1.
The purple bacterium Rhodopseudomonas palustris TIE-1 expresses multiple small high-potential redox proteins during photoautotrophic growth, including two high-potential iron-sulfur proteins (HiPIPs) (PioC and Rpal_4085) and a cytochrome c2. We evaluated the role of these proteins in TIE-1 through genetic, physiological, and biochemical analyses. Deleting the gene encoding cytochrome c2 resulted in a loss of photosynthetic ability by TIE-1, indicating that this protein cannot be replaced by either HiPIP in cyclic electron flow. PioC was previously implicated in photoferrotrophy, an unusual form of photosynthesis in which reducing power is provided through ferrous iron oxidation. Using cyclic voltammetry (CV), electron paramagnetic resonance (EPR) spectroscopy, and flash-induced spectrometry, we show that PioC has a midpoint potential of 450 mV, contains all the typical features of a HiPIP, and can reduce the reaction centers of membrane suspensions in a light-dependent manner at a much lower rate than cytochrome c2. These data support the hypothesis that PioC linearly transfers electrons from iron, while cytochrome c2 is required for cyclic electron flow. Rpal_4085, despite having spectroscopic characteristics and a reduction potential similar to those of PioC, is unable to reduce the reaction center. Rpal_4085 is upregulated by the divalent metals Fe(II), Ni(II), and Co(II), suggesting that it might play a role in sensing or oxidizing metals in the periplasm. Taken together, our results suggest that these three small electron transfer proteins perform different functions in the cell.


A simple mathematical model for manganese oxide-coated montmorillonite as a catalyst for water oxidation: from nano to macro sized manganese oxide.
Here, we propose a mathematical model that gives a good fit to the experimental data for water oxidation by Mn oxide-coated montmorillonite with different Mn content. Our data show that the water oxidation may progress by cooperation among only neighbor Mn ions on montmorillonite. It is a promising model for finding more about the mechanism of multi-electron reactions.


Resilience profiles across context: A latent profile analysis in a German, Greek, and Swiss sample of adolescents.
The present study investigated resilience profiles (based on levels of symptoms of anxiety and depression and five dimensions of protective factors) of 1,160 students from Germany (n = 346, 46.0% females, Mage = 12.77, SDage = 0.78), Greece (n = 439, 54.5% females, Mage = 12.68, SDage = 0.69), and Switzerland (n = 375, 44.5% females, Mage = 12.29, SDage = 0.88) using latent profile analyses. We also checked for measurement invariance and investigated the influence of gender and migration on class membership. A three-profile-solution was found for Switzerland (nonresilient 22.1%, moderately resilient 42.9%, untroubled 34.9%), and a four-profile-solution was the best fitting model for Germany (nonresilient 15.7%, moderately resilient 44.2%, untroubled 27.3%, resilient 12.7%) and Greece (nonresilient 21.0%, moderately resilient 30.8%, untroubled 24.9%, resilient 23.3%). Measurement invariance did not hold across the three countries. Profile differences regarding class membership predictions were detected for Germany and Greece, but none for Switzerland. Results implicate that resilience profiles are highly contextually sensitive, and resilience research findings should not be generalized considering the particularity of contexts, people, and outcomes.


A retrospective study on use of palliative care for patients with alcohol related end stage liver disease in United States.
Palliative care (PC) has been shown to be beneficial in end stage liver disease (ESLD), yet the hospitalization data for PC utilization is unknown. To identify the trend of PC utilization for the special population of alcohol-associated ESLD patients, factors affecting its use and ascertain its impact on healthcare utilization. We analyzed around 78 million discharges from the 2007-2014 national inpatient sample and 2010-2014 national readmission database including adult patients admitted for decompensated alcohol-associated cirrhosis. We identified patients with PC consultation as a secondary diagnosis. Odds ratios (OR) and means were adjusted for confounders using multivariate regression analysis models. Out of the total 1421849 hospitalizations for decompensated liver cirrhosis, 62782 (4.4%) hospitalizations had a PC consult, which increased from 0.8% (1258) of all alcohol-associated ESLD hospitalizations in 2007 to 6.6% in 2014 (P < 0.01). Patient and hospital characteristics associated with increased odds of PC utilization were advanced age, lower income, Medicaid coverage, teaching institution, urban location, length of stay > 3 d, prolonged ventilation, and administration of total parenteral nutrition (all P < 0.01). Palliative encounters in alcohol-associated ESLD and acute-on-chronic liver failure (ACLF) score were associated with increased odds of discharge to a rehabilitation facility, but significantly lower odds of 30-d readmissions (aOR: 0.35, 95%CI: 0.31-0.41), lower total hospitalization charges and lower mean hospitalization days (all P < 0.01). Inpatient PC is sparingly used for patients with decompensated alcohol related liver disease, however it has increased over the past decade. PC consultation is associated with lower 30-d readmission rates on multivariate analysis, and lower hospitalization cost and length of stay in patients with ACLF score ≥ 2.


Regenerative endodontics: a state of the art.
Scientific advances in the creation of restorative biomaterials, in vitro cell culture technology, tissue grafting, tissue engineering, molecular biology and the human genome project provide the basis for the introduction of new technologies into dentistry. Non-vital infected teeth have long been treated with root canal therapy (for mature root apex) and apexification (for immature root apex), or doomed to extraction. Although successful, current treatments fail to re-establish healthy pulp tissue in these teeth. But, what if the non-vital tooth could be made vital once again? That is the hope offered by regenerative endodontics, an emerging field focused on replacing traumatized and diseased pulp with functional pulp tissue. Restoration of vitality of non-vital tooth is based on tissue engineering and revascularization procedures. The purpose of this article is to review these biological procedures and the hurdles that must be overcome to develop regenerative endodontic procedures.


Predictors for the white coat effect in general practice patients with suspected and treated hypertension.
Ambulatory blood pressure monitoring was introduced more than 40 years ago and is accepted as a clinically useful method to evaluate the white coat effect in patients with suspected and established hypertension. To study the differences between blood pressure readings taken in the physician's office in the primary healthcare setting, and ambulatory readings, and to find possible predictors. Prospective study. Two primary healthcare centres in Norway. The study included 221 patients, 107 of whom were on antihypertensive treatment, and 114 of whom were under investigation for possible hypertension. Differences between blood pressure readings taken in the physician's office and ambulatory readings were calculated. Independent predictors for the white coat effect were calculated using linear regression analysis. The difference between blood pressure readings taken in the office and ambulatory readings was 27 mmHg systolic and 11 mmHg diastolic. For the systolic readings, the following factors were independent predictors of the amount of the white coat effect: mean blood pressure, age, history of smoking, family history of cardiovascular disease, and antihypertensive treatment. For the diastolic readings, they were: mean blood pressure, history of smoking, and sex of the patient (with this being most significant for women). Ambulatory blood pressure measurement is of significant value in identifying patients with white coat hypertension. It can be an important supplement for use in the diagnosis and follow-up of patients with hypertension in general practice.


[Option for different surgeries in treating small gastrointestinal stromal tumors].
To explore the differences in biological characteristics for the small gastrointestinal stromal tumors and the incidence of complications and recurrence between the traditional surgical treatment and endoscopic treatment. Methods: We collected the relevant clinical and pathological data from patients who were diagnosed as gastrointestinal stromal tumors with the diameter less than 2 cm by the Department of Pathology of Xiangya Hospital from January 2009 to December 2015. The complications and recurrence after the surgical treatment were analyzed. Results: In patients with small gastrointestinal stromal tumors, the proportion of female was higher than that of male (male:female=1:1.69). The median age for patient with this disease was 49 years old and it was more common in middle-aged and elderly. Most lesions were found in the stomach, followed by the esophagus and the small intestine. The small gastrointestinal stromal tumors occurred in the colon and rectum were rare. There was 60.3% (47/78) patients with abdominal pain, 7.7% (6/78) patients with hematochezia or melena, and 98.7% (78/79) with small gastrointestinal stromal tumors' mitotic count ≤5/50 HPF. The positive rates for CD, CD34, DOG-1, actin-smooth, and S-100 were 98.7%, 86.1%, 82.3%, 31.6%, and 24.1%, respectively. Three patients occurred surgical complications, 2 suffered recurrence during the follow-up. There was no significant difference in the incidence of complications and recurrence between the traditional surgical treatment and endoscopic treatment (P>0.05). Conclusion: Small gastrointestinal stromal tumors' malignant potential is low, and the recurrence and metastasis rate is low. Its biological behavior tends to be benign. The traditional surgical treatment and endoscopic treatment are both safe and effective for small gastrointestinal stromal tumor. Endoscopic treatment has the advantages in lower cost, shorter hospitalization time, and small trauma. Therefore, endoscopic treatment could be the first choice for small GIST resection under the condition of mature endoscopic technology.


Incidence of SARS-CoV-2 Infection, Emergency Department Visits, and Hospitalizations Because of COVID-19 Among Persons Aged ≥12 Years, by COVID-19 Vaccination Status - Oregon and Washington, July 4-September 25, 2021.
Population-based rates of infection with SARS-CoV-2 (the virus that causes COVID-19) and related health care utilization help determine estimates of COVID-19 vaccine effectiveness and averted illnesses, especially since the SARS-CoV-2 B.1.617.2 (Delta) variant began circulating in June 2021. Among members aged ≥12 years of a large integrated health care delivery system in Oregon and Washington, incidence of laboratory-confirmed SARS-CoV-2 infection, emergency department (ED) visits, and hospitalizations were calculated by COVID-19 vaccination status, vaccine product, age, race, and ethnicity. Infection after full vaccination was defined as a positive SARS-CoV-2 molecular test result ≥14 days after completion of an authorized COVID-19 vaccination series.* During the July-September 2021 surveillance period, SARS-CoV-2 infection occurred among 4,146 of 137,616 unvaccinated persons (30.1 per 1,000 persons) and 3,009 of 344,848 fully vaccinated persons (8.7 per 1,000). Incidence was higher among unvaccinated persons than among vaccinated persons across all demographic strata. Unvaccinated persons with SARS-CoV-2 infection were more than twice as likely to receive ED care (18.5%) or to be hospitalized (9.0%) than were vaccinated persons with COVID-19 (8.1% and 3.9%, respectively). The crude mortality rate was also higher among unvaccinated patients (0.43 per 1,000) than in fully vaccinated patients (0.06 per 1,000). These data support CDC recommendations for COVID-19 vaccination, including additional and booster doses, to protect individual persons and communities against COVID-19, including illness and hospitalization caused by the Delta variant (1).


Evaluation of preoperative duloxetine use for postoperative analgesia following laparoscopic cholecystectomy: A randomized controlled trial.
Background: The pain pattern after laparoscopic cholecystectomy (LC) is complex and distinct from postoperative pain after other laparoscopic procedures, suggesting that procedure-specific optimal analgesic management plans should be proposed. Duloxetine, a non-opioid neuromodulator, has been widely used to manage pain with dual central and peripheral analgesic properties. Aims: To assess the effect of preoperative administration of duloxetine compared to placebo on postoperative pain control in patients undergoing LC. Patients and Methods: This study was a randomized, parallel-group, placebo-controlled, double-blinded study performed on patients undergoing LC. Patients were randomly divided into two groups of 30 each on the day of surgery in the preoperative holding area, using a computer-generated random number to receive 60 mg duloxetine as a single oral dose 2 h before the procedure or placebo. The primary outcome was the difference in the mean of visual analogue scale (VAS) scores between the two studied groups, as measured by the area under the curve (AUC) of the VAS scores. Results: The derived AUC of VAS scores in the duloxetine group (757.89 ± 326.01 mm × h) was significantly lower than that calculated for the control group (1005.1 ± 432.5 mm × h). The mean postoperative VAS scores recorded at 4 and 24 h were statistically different between the study groups (p = 0.041 and 0.003, respectively). As observed in the survival curve analysis, there was no significant difference (p = 0.665) for the time until the patient's first request for rescue medications in the two groups. The frequency of postoperative nausea and vomiting (PONV) was lower in patients of the duloxetine group than that recorded in those allocated to the control group at 8 and 24-h time intervals (p = 0.734 and 0.572, respectively). Conclusion: Preoperative use of duloxetine reduces postoperative pain significantly compared with placebo. In addition, its use is associated with a reduction in PONV. These preliminary findings suggest that duloxetine could play a role in the acute preoperative period for patients undergoing LC. Clinical Trial Registration: [https://clinicaltrials.gov/ct2/show/NCT05115123, identifier NCT05115123].


Factors associated with developing vaginal dryness symptoms in women transitioning through menopause: a longitudinal study.
To evaluate factors associated with incident self-reported vaginal dryness and the consequences of this symptom across the menopausal transition in a multiracial/ethnic cohort of community-dwelling women. We analyzed questionnaire and biomarker data from baseline and 13 approximately annual visits over 17 years (1996-2013) from 2,435 participants in the Study of Women's Health Across the Nation, a prospective cohort study. We used discrete-time Cox proportional-hazards regression to identify predictors of incident vaginal dryness and to evaluate vaginal dryness as a predictor of pain during intercourse and changes in sexual intercourse frequency. The prevalence of vaginal dryness increased from 19.4% among all women at baseline (ages 42-53 years) to 34.0% at the 13th visit (ages 57-69 years). Advancing menopausal stage, surgical menopause, anxiety, and being married were positively associated with developing vaginal dryness, regardless of partnered sexual activity. For women not using hormone therapy, higher concurrent levels of endogenous estradiol were inversely associated (multivariable-adjusted hazard ratio: 0.94 per 0.5 standard deviation increase, 95% confidence interval: 0.91-0.98). Concurrent testosterone levels, concurrent dehydroepiandrosterone sulfate levels, and longitudinal change in any reproductive hormone were not associated with developing vaginal dryness. Both vaginal dryness and lubricant use were associated with subsequent reporting of pain during intercourse, but not with a decline in intercourse frequency. In these longitudinal analyses, our data support many clinical observations about the relationship between vaginal dryness, menopause, and pain during intercourse, and suggest that reporting of vaginal dryness is not related to androgen level or sexual intercourse frequency.


Peroxisome Proliferator-Activated Receptor-δ Acts within Peripheral Myeloid Cells to Limit Th Cell Priming during Experimental Autoimmune Encephalomyelitis.
Peroxisome proliferator-activated receptor (PPAR)-δ is a fatty acid-activated transcription factor that regulates metabolic homeostasis, cell growth, and differentiation. Previously, we reported that mice with a global deficiency of PPAR-δ develop an exacerbated course of experimental autoimmune encephalomyelitis (EAE), highlighting a role for this nuclear receptor in limiting the development of CNS inflammation. However, the cell-specific contribution of PPAR-δ to the more severe CNS inflammatory response remained unclear. In this study, we studied the specific involvement of PPAR-δ in myeloid cells during EAE using mice that had Cre-mediated excision of floxed Ppard driven by the lysozyme M (LysM) promoter (LysM Cre :Ppard fl/fl). We observed that LysM Cre :Ppard fl/fl mice were more susceptible to EAE and developed a more severe course of this disease compared with Ppard fl/fl controls. The more severe EAE in LysM Cre :Ppard fl/fl mice was associated with an increased accumulation of pathogenic CD4+ T cells in the CNS and enhanced myelin-specific Th1 and Th17 responses in the periphery. Adoptive transfer EAE studies linked this EAE phenotype in LysM Cre :Ppard fl/fl mice to heightened Th responses. Furthermore, studies using an in vitro CD11b+ cell:Th cell coculture system revealed that CD11b+CD11c+ dendritic cells (DC) from LysM Cre :Ppard fl/fl mice had a heightened capacity to prime myelin oligodendrocyte glycoprotein (MOG)-specific Th cells compared with Ppard fl/fl counterparts; the effects of DC on Th1 cytokine production were mediated through production of the IL-12p40 homodimer. These studies revealed a role for PPAR-δ in DC in limiting Th cell priming during EAE.


Evolving Concepts in Cardiac Physiologic Pacing in the Era of Conduction System Pacing.
Cardiac physiologic pacing (CPP) has become a well-established therapy for patients with cardiomyopathy (left ventricular ejection fraction <35%) in the presence of a left bundle branch block. In addition, CPP can be highly beneficial in patients with pacing-induced cardiomyopathy and patients with existing cardiomyopathy expected to have a right ventricular pacing burden of >40%. The benefits of CPP with traditional biventricular pacing are only realized if adequate resynchronization can be achieved. However, left ventricular lead implantation can be limited by individual anatomic variation within the coronary venous system and can be adversely affected by underlying abnormal myocardial substrate (i.e., scar tissue), especially if located within the basal lateral wall. In the last 7 years the investigation of conduction system pacing (CSP) and its potential salutary benefits are being realized and have led to a rapid evolution in the field of cardiac resynchronization pacing. However, supportive evidence for CSP for patients eligible for cardiac resynchronization remains limited compared with data available for biventricular cardiac resynchronization, mostly derived from leading CSP investigative centers. In this review, we perform an up-to-date comprehensive review of the available literature on CPP.


PSTPIP2 dysregulation contributes to aberrant terminal differentiation in GATA-1-deficient megakaryocytes by activating LYN.
GATA1 mutations are tightly associated with transient myeloproliferative disorder (TMD) and acute megakaryoblstic leukemia (AMKL) in children with Down syndrome. Numerous genes are altered in GATA-1-deficient megakaryocytes, which may contribute to the hyperproliferation and abnormal terminal differentiation of these malignant cells. In this study, we demonstrate that Pstpip2 is a GATA-1-repressed gene that controls megakaryopoiesis. Ectopic expression of PSTPIP2 impaired megakaryocytic differentiation as evidenced by a decrease of CD41 expression and reduced DNA content in K562 cells. PSTPIP2 overexpression also caused enhanced activation of Src family kinases and subsequently reduced ERK phosphorylation. Consistently, PSTPIP2 knockdown showed the opposite effect on differentiation and signaling. Moreover, the W232A mutant of PSTPIP2, defective in its interaction with PEST family phosphatases that recruit c-Src terminal kinase (CSK) to suppress Src family kinases, failed to inhibit differentiation and lost its ability to enhance Src family kinases or reduce ERK phosphorylation. In fact, the W232A mutant of PSTPIP2 promoted megakaryocyte differentiation. These observations suggest that PSTPIP2 recruiting PEST phosphatases somehow blocked CSK activity and led to enhanced activation of Src family kinases and reduced ERK phosphorylation, which ultimately repressed megakaryocyte differentiation. Supporting this idea, PSTPIP2 interacted with LYN and the expression of a dominant negative LYN (LYN DN) overwhelmed the inhibitory effect of PSTPIP2 on differentiation and ERK signaling. In addition, a constitutively active LYN (LYN CA) normalized the enhanced megakaryocyte differentiation and repressed ERK signaling in PSTPIP2 knockdown cells. Finally, we found that PSTPIP2 repressed ERK signaling, differentiation, and proliferation and verified that PSTPIP2 upregulation repressed megakaryocyte development in primary mouse bone marrow cells. Our study thus reveals a novel mechanism by which dysregulation of PSTPIP2 due to GATA-1 deficiency may contribute to abnormal megakaryocyte proliferation and differentiation in pathogenesis of related diseases.


Assessment of Body Composition and Sport Performance of Brazilian Paralympic Swim Team Athletes.
Body-composition assessments of high-performance athletes are very important for identifying physical performance potential. Although the relationship between the kinanthropometric characteristics and performance abilities of Olympic swimmers is extremely important, this subject is not completely understood for Paralympic swimmers. To investigate the relationship between body composition and sport performance in Brazilian Paralympic swimmers 6 mo after training. Experimental pre/posttest design. Research laboratory and field evaluations of swimming were conducted to verify the 50-m freestyle time of each athlete. 17 Brazilian Paralympic swim team athletes (12 men, 5 women). Body-composition assessments were performed using a BOD POD, and swimming performance was assessed using the 50-m freestyle, which was performed twice: before and after 6 mo of training. Increased lean mass and significantly reduced relative fat mass and swimming time (P < .05) were observed 6 mo after training. Furthermore, a positive correlation between body-fat percentage and performance (r = .66, P < .05) was observed, but there was no significant correlation between body density and performance (r = -.14, P > .05). After a 6-mo training period, Paralympic swimmers presented reduced fat mass and increased lean body mass associated with performance, as measured by 50-m freestyle time. These data suggest that reduced fat-mass percentage was significantly correlated with improved swimming performance in Paralympic athletes.


Postpartal recurrent non-ST elevation myocardial infarction in essential thrombocythaemia: case report and review of the literature.
Normal pregnancy corresponds to a procoagulant state. Acute myocardial infarction during pregnancy is rare, yet considering the low non-pregnant risk score of childbearing women it is still surprisingly frequent. We report a case of postpartum recurrent non-ST elevation myocardial infarction in a 40-year-old caucasian woman with essential thrombocythaemia in the presence of a positive JAK-2 mutation and an elevated anti-cardiolipin IgM antibody titer. In the majority of cases of myocardial infarction in pregnancy or in the peripartal period, atherosclerosis, a thrombus or coronary artery dissection is observed. The combination of essential thrombocythaemia and elevated anti-cardiolipin IgM antibody titer in the presence of several cardiovascular risk factors seems to be causative in our case. In conclusion, with the continuing trend of childbearing at older ages, rare or unlikely conditions leading to severe events such as myocardial infarction must be considered in pregnant women.


Nitric oxide as a putative retinal axon pathfinding and target recognition cue in Xenopus laevis.
Nitric oxide (NO) is an atypical neurotransmitter synthesized by the enzyme nitric oxide synthase (NOS) during many stages of the Xenopus laevis life cycle. This research investigates whether the gas NO is involved in axon guidance, the neurodevelopmental process in which axons travel through the brain to their appropriate target locations to form functional neural circuitry. Through immunocytochemistry and direct labeling of the NO gas with a fluorescent dye, we have found that NOS expression corresponds spatiotemporally with the beginning of retinal axon innervation of the optic tectum in X. laevis. Our function-blocking studies in which NO is chemically inhibited suggest that NO may be necessary for correct pathfinding and targeting, evidenced by qualitative widening of the optic tract and aberrant target innervation.


Prevalence and correlates of nocturnal desaturations in a sample of elderly people.
We designed an epidemiological study to estimate the prevalence and correlates of nocturnal desaturations in a sample of elderly subjects from the general population. Sleep-related respiratory disturbances were assessed by questionnaire and MESAM IV (MADAUS electronic sleep apnoea monitor) ambulatory monitoring. An oxygen desaturation index (ODI), oxygen desaturation being defined as a decrease in SaO2 of or exceeding 4%, was then computed from these data. An oxygen desaturation index > or = 10 was observed in 27.0% of the 293 subjects studied (mean age 76.6+/-5.7 y, median=75 y, min.=69, max.=99), an oxygen desaturation index > or = 30in 4.4%. Multivariate analysis identified as correlates to an oxygen desaturation index > or = 10: male gender (OR=1.80; P=0.04), a high body mass index (BMI) in men (OR=1.20 per kg m2; P=0.0009), and advanced age in women (OR=1.09 per y; P=0.02). A positive association was found between loud snoring (OR=1.75; P=0.06) and an oxygen desaturation index > or = 10. However, there was no statistically significant relationship between an oxygen desaturation index > or = 10 and either daytime somnolence (OR=1.50; P=0.19) or trouble getting to sleep (OR=0.59; P=0.09). We found no significant relationship in our sample between oxygen desaturation index and arterial hypertension or cardiac dysrhythmia. Previous studies on younger populations have reported different results. It may well be the advanced age of our sample that explains these inconsistencies. For the elderly persons we studied our results underline the relatively high prevalence of sleep-related respiratory disturbances. However, these may be of less consequence than in younger populations.


Efficiency of a turbidity-based, real-time control strategy applied to a retention tank: a simulation study.
This paper compares several real-time control (RTC) strategies for a generic configuration consisting of a storage tank with two overflow facilities. Two of the strategies only make use of flow rate data, while the third also introduces turbidity data in order to exercise dynamic control between two overflow locations. The efficiency of each strategy is compared over a wide range of system setups, described by two parameters. This assessment is performed by simulating the application of control strategies to actual measurements time series recorded on two sites. Adding turbidity measurements into an RTC strategy leads to a significant reduction in the annual overflow pollutant load. The pollutant spills spared by such a control strategy strongly depend on the site and on the flow rate based strategy considered as a reference. With the datasets used in this study, values ranging from 5 to 50% were obtained.


Herpangina surveillance in Japan, 1982-1989. A report of the national epidemiological surveillance of infectious agents in Japan.
In Japan, herpangina cases increase every summer with an incidence peak in July. During the past eight years from January, 1982 to December, 1989, a total of 3,974 viruses from herpangina cases were reported from 47 participating laboratories, and coxsackie A (CA) viruses accounted for most of them, 3,055 (76.9%). The major types associated with herpangina were, in order of frequency, CA4, 10, 5, 6, 2 and 3, representing 87.2%-80.8% of total isolations reported for each respective type in this period. Eighty-four point four percent of the total virus isolations were from children at four years of age or younger. More than 80% of total CA virus isolations were from nasopharyngeal specimens and nearly 90% of the viruses were isolated in mice.


Chemoprevention of prostate cancer.
Due to the public health impact of prostate cancer including the burden of screening and treatment, there is significant public interest in the potential prevention of this disease. We review the most recent results of large scale randomized clinical trials. We review the potential agents, their hypothesized mechanisms of action and challenges for the design of chemoprevention trials, including the 3 large scale trials SELECT (testing selenium and vitamin E), PCPT (testing finasteride) and REDUCE (testing dutasteride). The initial results of SELECT have now been reported and demonstrate no impact of selenium or vitamin E on the risk of prostate cancer. The REDUCE trial results should be available within the year. The results of the PCPT demonstrate a significant (measured relative risk reduction of 24.8%) reduction in the risk of prostate cancer. The initial observation of an excess risk of high grade disease appears to be related to improved detection of cancer and high grade cancer related to the improved sensitivity of prostate specific antigen, digital rectal examination and prostate biopsy for cancer and high grade cancer detection. Modeling studies suggest that with finasteride the risk of high grade cancer is unchanged or reduced. Sexual dysfunction and gynecomastia were observed but the rates were low. Recommendations for the prevention of prostate cancer must be based on outcomes of well designed randomized trials. Men undergoing prostate cancer screening should be informed of the potential for the reduction in risk with finasteride.


A RNA-Seq Analysis to Describe the Boar Sperm Transcriptome and Its Seasonal Changes.
Understanding the molecular basis of cell function and ultimate phenotypes is crucial for the development of biological markers. With this aim, several RNA-seq studies have been devoted to the characterization of the transcriptome of ejaculated spermatozoa in relation to sperm quality and fertility. Semen quality follows a seasonal pattern and decays in the summer months in several animal species. The aim of this study was to deeply profile the transcriptome of the boar sperm and to evaluate its seasonal changes. We sequenced the total and the short fractions of the sperm RNA from 10 Pietrain boars, 5 collected in summer and 5 five sampled in winter, and identified a complex and rich transcriptome with 4,436 coding genes of moderate to high abundance. Transcript fragmentation was high but less obvious in genes related to spermatogenesis, chromatin compaction and fertility. Short non-coding RNAs mostly included piwi-interacting RNAs, transfer RNAs and microRNAs. We also compared the transcriptome of the summer and the winter ejaculates and identified 34 coding genes and 7 microRNAs with a significantly distinct distribution. These genes were mostly related to oxidative stress, DNA damage and autophagy. This is the deepest characterization of the boar sperm transcriptome and the first study linking the transcriptome and the seasonal variability of semen quality in animals. The annotation described here can be used as a reference for the identification of markers of sperm quality in pigs.


Estrogen receptor alpha mediates epithelial to mesenchymal transition, expression of specific matrix effectors and functional properties of breast cancer cells.
The 17β-estradiol (E2)/estrogen receptor alpha (ERα) signaling pathway is one of the most important pathways in hormone-dependent breast cancer. E2 plays pivotal roles in cancer cell growth, survival, and architecture as well as in gene expression regulatory mechanisms. In this study, we established stably transfected MCF-7 cells by knocking down the ERα gene (designated as MCF-7/SP10+ cells), using specific shRNA lentiviral particles, and compared them with the control cells (MCF-7/c). Interestingly, ERα silencing in MCF-7 cells strongly induced cellular phenotypic changes accompanied by significant changes in gene and protein expression of several markers typical of epithelial to mesenchymal transition (EMT). Notably, these cells exhibited enhanced cell proliferation, migration and invasion. Moreover, ERα suppression strongly affected the gene and protein expression of EGFR and HER2 receptor tyrosine kinases, and various extracellular matrix (ECM) effectors, including matrix metalloproteinases and their endogenous inhibitors (MMPs/TIMPs) and components of the plasminogen activation system. The action caused by E2 in MCF-7/c cells in the expression of HER2, MT1-MMP, MMP1, MMP9, uPA, tPA, and PAI-1 was abolished in MCF-7/SP10+ cells lacking ERα. These data suggested a regulatory role for the E2/ERα pathway in respect to the composition and activity of the extracellular proteolytic molecular network. Notably, loss of ERα promoted breast cancer cell migration and invasion by inducing changes in the expression levels of certain matrix macromolecules (especially uPA, tPA, PAI-1) through the EGFR-ERK signaling pathway. In conclusion, loss of ERα in breast cancer cells results in a potent EMT characterized by striking changes in the expression profile of specific matrix macromolecules highlighting the potential nodal role of matrix effectors in breast cancer endocrine resistance.


Target size of neurotoxic esterase and acetylcholinesterase as determined by radiation inactivation.
The target size of neurotoxic esterase (NTE), the putative target site for the initiation of organophosphorus-compound-induced delayed neurotoxicity, and acetylcholinesterase (AChE) from hen brain were examined by determining the rate at which the activities of the esterases were destroyed by ionizing irradiation. Samples of hen brain were prepared by slowly drying a microsomal preparation under vacuum. The dried samples were then irradiated with electrons from a 1 MeV Van de Graaff generator. The doses ranged from 0 to 28 Mrad. The radiation doses were calibrated by the rate of inactivation of T1-bacteriophage plaque induction. Following the irradiation procedure, the samples were resuspended in buffer and enzymic activity was measured. The target size of NTE from hen brain was determined to be about 105 kDa, whereas hen brain AChE was found to have a target size of about 53 kDa. The target size of NTE was found to be similar in experiments with rat brain and cat brain. In addition, commercial preparations of electric-eel electric-organ AChE and horse serum butyrylcholinesterase were found to have target sizes that were identical with each other, and also were very similar to that of AChE from hen brain.


A Cone-beam Computed Tomographic Study of Apical Surgery-related Morphological Characteristics of the Distolingual Root in 3-rooted Mandibular First Molars in a Chinese Population.
Apical surgery on the separate distolingual (DL) root of a 3-rooted mandibular first molar is thought to be difficult because DL roots are always severely curved, small, and shorter than distobuccal (DB) roots, and they are located far from the buccal cortical bone. The purpose of this study was to use cone-beam computed tomographic images to investigate the apical surgery-related morphological characteristics of DL roots in a Chinese population. The screening process identified 83 subjects with 128 mandibular first molars with separate DL roots. The degree of root canal curvature was measured in the mesiodistal (MD) and buccolingual (BL) planes using the Schneider method. Distances were measured from the DL root apex to the buccal cortical bone, to the buccal cortical bone opposite the DB root apex, and to the buccal cortical bone opposite the mesiobuccal (MB) root apex. The lengths of the remaining DB, MB, and DL roots were measured after resection of 3 mm of the DL apical root. Angulations of the DL root canal were determined before and after DL apical root resection. The teeth were classified into 3 types: type I, straight in the MD and BL planes; type II, straight in the MD plane and curved in the BL plane; and type III, curved in the MD and BL planes. Only 5 molars (3.9%) were classified as type I, whereas 52 (40.6%) molars were type II and 71 (55.5%) molars were type III. The type I DL root has a significantly shorter length and longer distance from the DL root apex to the buccal cortical bone than type II and III roots (P < .05). The distance from the DL root apex to the buccal cortical bone opposite the MB root apex is significantly longer than the distance to the buccal cortical bone opposite the DB root apex in types I and II (P < .05), whereas the distance is nearly equal for type III. The angulations for root-end preparation of types II and III DL roots vary from 57.5° to 129.1° and from 55.8° to 128.1°, respectively. A new classification was proposed for DL roots in 3-rooted mandibular first molars based on the root canal curvature. Type I is unsuitable for apical surgery. Access to the type II DL root apex should be through the DB root apex, whereas access to the type III DL root apex through the MB root apex is more feasible. Apical surgery on types II and III DL roots may be accomplished when the depth of the root-end preparation is reasonably reduced, and fine and personalized angulated ultrasonic retro tips are used.


The Bacillus subtilis GntR family repressor YtrA responds to cell wall antibiotics.
The transglycosylation step of cell wall synthesis is a prime antibiotic target because it is essential and specific to bacteria. Two antibiotics, ramoplanin and moenomycin, target this step by binding to the substrate lipid II and the transglycosylase enzyme, respectively. Here, we compare the ramoplanin and moenomycin stimulons in the Gram-positive model organism Bacillus subtilis. Ramoplanin strongly induces the LiaRS two-component regulatory system, while moenomycin almost exclusively induces genes that are part of the regulon of the extracytoplasmic function (ECF) σ factor σ(M). Ramoplanin additionally induces the ytrABCDEF and ywoBCD operons, which are not part of a previously characterized antibiotic-responsive regulon. Cluster analysis reveals that these two operons are selectively induced by a subset of cell wall antibiotics that inhibit lipid II function or recycling. Repression of both operons requires YtrA, which recognizes an inverted repeat in front of its own operon and in front of ywoB. These results suggest that YtrA is an additional regulator of cell envelope stress responses.


Detection and localization of proteinuria by dynamic contrast-enhanced magnetic resonance imaging using MS325.
After renal transplantation, persistent glomerular disease affecting the native kidneys typically causes albuminuria, at least for a period of time, making it difficult to determine in a noninvasive fashion whether proteinuria originates in the native kidneys or the renal allograft. To address this problem, dynamic contrast-enhanced magnetic resonance imaging (MRI) using gadolinium (Gd)-based albumin-bound blood pool contrast agent (MS325) to localize proteinuria was investigated. Glomerular proteinuria was induced in Sprague-Dawley rats by intravenous injection of puromycin aminonucleoside (PAN), whereas control rats received physiologic saline vehicle. Both groups of animals underwent a 40-min dynamic contrast-enhanced MRI using radio frequency spoiled gradient echo imaging sequence after injection of Gd-labeled MS325. Contrast uptake and clearance curves for cortex and medulla were determined from acquired MR images. Compared with controls, proteinuric rats exhibited significantly lower elimination rate constants. The use of gadopentetate dimeglumine (Gd-DTPA) as a contrast agent showed smaller and less specific differences between proteinuric and control groups. In rats with one proteinuric kidney (PAN-treated) and one normal kidney (transplanted from a normal rat), MRI using MS325 was able to differentiate between the two kidneys. The results suggest that MRI with an albumin-bound blood pool contrast agent may be a useful noninvasive way to localize proteinuria. If this technique can be successfully applied in human patients, it may allow for the localization of proteinuria after kidney transplant and thereby provide a noninvasive way to detect disease affecting the renal allograft.


Understanding the use of contextual cues: design implications for medication adherence technologies that support remembering.
Forgetfulness is one of the main reasons of unintentional medication non-adherence. Adherence technologies that help people remember to take their medications on time often do not take into account the context of people's everyday lives. Existing evidence that highlights the effectiveness of remembering strategies that rely on contextual cues is largely based on research with older adults, and thus it is not clear whether it can be generalized to other populations or used to inform the design of wider adherence technologies that support medication self-management. Understanding how younger populations currently remember medications can inform the design of future adherence technologies that take advantage of existing contextual cues to support remembering. We conducted three surveys with a total of over a thousand participants to investigate remembering strategies used by different populations: women who take oral contraception, parents and carers who give antibiotics to their children, and older adults who take medications for chronic conditions. Regardless of the population or the type of regimen, relying on contextual cues-routine events, locations, and meaningful objects-is a common and often effective strategy; combinations of two or more types of cues are more effective than relying on a single cue. To effectively support remembering, adherence technologies should help users recognize contextual cues they already have at their disposal and reinforce relevant cues available in their environment. We show that, given the latest developments in technology, such support is already feasible.


A tree-based conservation scoring method for short linear motifs in multiple alignments of protein sequences.
The structure of many eukaryotic cell regulatory proteins is highly modular. They are assembled from globular domains, segments of natively disordered polypeptides and short linear motifs. The latter are involved in protein interactions and formation of regulatory complexes. The function of such proteins, which may be difficult to define, is the aggregate of the subfunctions of the modules. It is therefore desirable to efficiently predict linear motifs with some degree of accuracy, yet sequence database searches return results that are not significant. We have developed a method for scoring the conservation of linear motif instances. It requires only primary sequence-derived information (e.g. multiple alignment and sequence tree) and takes into account the degenerate nature of linear motif patterns. On our benchmarking, the method accurately scores 86% of the known positive instances, while distinguishing them from random matches in 78% of the cases. The conservation score is implemented as a real time application designed to be integrated into other tools. It is currently accessible via a Web Service or through a graphical interface. The conservation score improves the prediction of linear motifs, by discarding those matches that are unlikely to be functional because they have not been conserved during the evolution of the protein sequences. It is especially useful for instances in non-structured regions of the proteins, where a domain masking filtering strategy is not applicable.


What is the most relevant standard of success in assisted reproduction? Is there a single 'parameter of excellence'?
An optimal standard of success reflects all risk and safety aspects of the treatment-related procedures. In this debate, we question whether one single standard of success can cover the complexity of assisted reproduction technology (ART). To emphasize the use of different parameters as standards of success, we calculated data from all IVF and ICSI cycles initiated at the Fertility Clinic, Rigshospitalet, Copenhagen, Denmark between 1999 and 2001. Several already established parameters were computed and, additionally, new suggestions for end-points were calculated. Three parameters were proposed as the best standards of success in ART: (i) number of oocytes per aspiration (8.7); (ii) number of ongoing implantations per embryo transferred (20%); and (iii) number of deliveries per embryo transferred (14%). These parameters cover aspects of all steps in ART, i.e. the stimulation, laboratory and embryo transfer/outcome phase, also termed the pre-in vitro, in vitro and post-in vitro phase. We suggest that the final description of an ART programme should be a balanced choice of the three standards of success proposed herein. If these standards are implemented as national gold standards, the future goal of homogeneous reporting of success rates in ART is within reach.


Decade of progress in motor functional neurological disorder: continuing the momentum.
Functional neurological disorder (FND) is a prevalent, disabling and costly condition at the neurology-psychiatry intersection. After being marginalised in the late 20th century, there has been renewed interest in this field. In this article, we review advances that have occurred over the past decade (2011-2020) across diagnosis, mechanisms, aetiologies, treatments and stigma in patients with motor FND (mFND, that is, functional movement disorder and functional limb weakness). In each content area, we also discuss the implications of recent advances and suggest future directions that will help continue the momentum of the past decade. In diagnosis, a major advance has been the emphasis on rule-in physical signs that are specific for hyperkinetic and hypokinetic functional motor symptoms. Mechanistically, greater importance has been given to determining 'how' functional neurological symptoms develop, highlighting roles for misdirected attention, expectation and self-agency, as well as abnormal influences of emotion/threat processing brain areas on motor control circuits. Aetiologically, while roles for adverse life experiences remain of interest in mFND, there is recognition of other aetiologic contributors, and efforts are needed to investigate links between aetiological factors and mechanisms. This decade has seen the first randomised controlled trials for physiotherapy, multidisciplinary rehabilitation and psychotherapy performed in the field, with consensus recommendations for physiotherapy, occupational therapy and outcome measures also published. Across patients, clinicians, healthcare systems and society, stigma remains a major concern. While challenges persist, a patient-centred integrated clinical neuroscience approach is primed to carry forward the momentum of the past decade into the future.


Clinical pharmacokinetics of depot leuprorelin.
Leuprorelin acetate is a synthetic agonist analogue of gonadotropin-releasing hormone. Continued leuprorelin administration results in suppression of gonadal steroid synthesis, resulting in pharmacological castration. Since leuprorelin is a peptide, it is orally inactive and generally given subcutaneously or intramuscularly. Sustained release parenteral depot formulations, in which the hydrophilic leuprorelin is entrapped in biodegradable highly lipophilic synthetic polymer microspheres, have been developed to avoid daily injections. The peptide drug is released from these depot formulations at a functionally constant daily rate for 1, 3 or 4 months, depending on the polymer type [polylactic/glycolic acid (PLGA) for a 1-month depot and polylactic acid (PLA) for depot of >2 months], with doses ranging between 3.75 and 30mg. Mean peak plasma leuprorelin concentrations (C(max)) of 13.1, 20.8 to 21.8, 47.4, 54.5 and 53 microg/L occur within 1 to 3 hours of depot subcutaneous administration of 3.75, 7.5, 11.25, 15 and 30 mg, respectively, compared with 32 to 35 microg/L at 36 to 60 min after a subcutaneous injection of 1mg of a non-depot formulation. Sustained drug release from the PLGA microspheres maintains plasma concentrations between 0.4 and 1.4 microg/L over 28 days after single 3.75, 7.5 or 15mg depot injections. Mean areas under the concentration-time curve (AUCs) are similar for subcutaneous or intravenous injection of short-acting leuprorelin 1mg; a significant dose-related increase in the AUC from 0 to 35 days is noted after depot injection of leuprorelin 3.75, 7.5 and 15mg. Mean volume of distribution of leuprorelin is 37L after a single subcutaneous injection of 1mg, and 36, 33 and 27L after depot administration of 3.75, 7.5 and 15mg, respectively. Total body clearance is 9.1 L/h and elimination half-life 3.6 hours after a subcutaneous 1mg injection; corresponding values after intravenous injection are 8.3 L/h and 2.9 hours. A 3-month depot PLA formulation of leuprorelin acetate 11.25mg ensures a C(max) of around 20 microg/L at 3 hours after subcutaneous injection, and continuous drug concentrations of 0.43 to 0.19 microg/L from day 7 until before the next injection. Recently, an implant that delivers leuprorelin for 1 year has been evaluated. Serum leuprorelin concentrations remained at a steady mean of 0.93 microg/L until week 52, suggesting zero-order drug release from the implant. In general, regular or depot leuprorelin treatment is well tolerated. Local reactions are more common after application of the 3- or 4-month depot in comparison with the 1-month depot.


Ubiquitylation of the chemokine receptor CCR7 enables efficient receptor recycling and cell migration.
The chemokine receptor CCR7 is essential for lymphocyte and dendritic cell homing to secondary lymphoid organs. Owing to the ability to induce directional migration, CCR7 and its ligands CCL19 and CCL21 are pivotal for the regulation of the immune system. Here, we identify a novel function for receptor ubiquitylation in the regulation of the trafficking process of this G-protein-coupled seven transmembrane receptor. We discovered that CCR7 is ubiquitylated in a constitutive, ligand-independent manner and that receptor ubiquitylation regulates the basal trafficking of CCR7 in the absence of chemokine. Upon CCL19 binding, we show that internalized CCR7 recycles back to the plasma membrane via the trans-Golgi network. An ubiquitylation-deficient CCR7 mutant internalized normally after ligand binding, but inefficiently recycled in immune cells and was transiently retarded in the trans-Golgi network compartment of HEK293 transfectants. Finally, we demonstrate that the lack of CCR7 ubiquitylation profoundly impairs immune cell migration. Our results provide evidence for a novel function of receptor ubiquitylation in the regulation of CCR7 recycling and immune cell migration.


The association between caffeine and cognitive decline: examining alternative causal hypotheses.
Numerous studies suggest that higher coffee consumption may reduce the rate of aging-related cognitive decline in women. It is thus potentially a cheap and widely available candidate for prevention programs provided its mechanism may be adequately understood. The assumed effect is that of reduced amyloid deposition, however, alternative pathways notably by reducing depression and diabetes type 2 risk have not been considered. A population study of 1,193 elderly persons examining depressive symptomatology, caffeine consumption, fasting glucose levels, type 2 diabetes onset, serum amyloid, and factors known to affect cognitive performance was used to explore alternative causal models. Higher caffeine consumption was found to be associated with decreased risk of incident diabetes in men (HR = 0.64; 95% CI 0.42-0.97) and increased risk in women (HR = 1.51; 95% CI 1.08-2.11). No association was found with incident depression. While in the total sample lower ratio Aβ42/Aβ40 levels (OR = 1.36, 95% CI 1.05-1.77, p = 0.02) were found in high caffeine consumers, this failed to reach significance when the analyses were stratified by gender. We found no evidence that reduced risk of cognitive decline in women with high caffeine consumption is moderated or confounded by diabetes or depression. The evidence of an association with plasma beta amyloid could not be clearly demonstrated. Insufficient proof of causal mechanisms currently precludes the recommendation of coffee consumption as a public health measure. Further research should focus on the high estrogen content of coffee as a plausible alternative explanation.


River water analysis using a multiparametric approach: Portuguese river as a case study.
The Ave River in northern Portugal has a history of riverbanks and water quality degradation. The river water quality was assessed by physicochemical, biological (macroinvertebrates) and microbiological (Enterococcus spp. and Escherichia coli) parameters in six locations (A-F, point A being the nearest to the source) throughout its course during a year. Epilithic biofilms were studied through polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Antimicrobial susceptibility testing helped with selecting isolates (n = 149 E. coli and n = 86 enterococci) for further genetic characterization. Pursuant to physicochemical and macroinvertebrates-based parameters, the river water was of reasonable quality according to European legislation (Directive 2000/60/EC). However, the microbiological analysis showed increased fecal contamination downstream from point C. At point D, four carbapenem-resistant E. coli isolates were recovered. Paradoxically, point D was classified as a point of 'Good Water Quality' according to macroinvertebrates results. Point F presented the highest contamination level and incidence of multidrug-resistant (MDR) isolates in the water column (13 MDR enterococci out of 39 and 33 MDR E. coli out of 97). Epilithic biofilms showed higher diversity in pristine points (A and B). Thus, biological and microbiological parameters used to assess the water quality led to divergent results; an outcome that reinforces the need for a holistic evaluation.


Building a viral capsid in the presence of genomic RNA.
Virus capsid assembly has traditionally been considered as a process that can be described primarily via self-assembly of the capsid proteins, neglecting interactions with other viral or cellular components. Our recent work on several ssRNA viruses, a major class of viral pathogens containing important human, animal, and plant viruses, has shown that this protein-centric view is too simplistic. Capsid assembly for these viruses relies strongly on a number of cooperative roles played by the genomic RNA. This realization requires a new theoretical framework for the modeling and prediction of the assembly behavior of these viruses. In a seminal paper Zlotnick [J. Mol. Biol. 241, 59 (1994)] laid the foundations for the modeling of capsid assembly as a protein-only self-assembly process, illustrating his approach using the example of a dodecahedral study system. We describe here a generalized framework for modeling assembly that incorporates the regulatory functions provided by cognate protein-nucleic-acid interactions between capsid proteins and segments of the genomic RNA, called packaging signals, into the model. Using the same dodecahedron system we demonstrate, using a Gillespie-type algorithm to deal with the enhanced complexity of the problem instead of a master equation approach, that assembly kinetics and yield strongly depend on the distribution and nature of the packaging signals, highlighting the importance of the crucial roles of the RNA in this process.


Hospital case volume and appropriate prescriptions at hospital discharge after acute myocardial infarction: a nationwide assessment.
In acute myocardial infarction, the relationship between volume and quality indicators (QIs) is poorly documented. Through a nationwide assessment of QIs at discharge repeated for 3 years, we aimed to quantify the relationship between volume and QIs in survivors after acute myocardial infarction. Almost all healthcare centers in France participated. Medical records were randomly selected. Data collection was performed by an independent group. QIs for acute myocardial infarction were defined by an expert consensus group as appropriate prescription at discharge of aspirin, clopidogrel, β-blocker, statin, and an angiotensin-converting enzyme inhibitor in patients with left ventricular ejection fraction <0.40. A composite QI was calculated through the use of the all-or-none method. Volume was classified into 7 categories based on the number of admissions for acute myocardial infarctions in 2008 (centers with <10 acute myocardial infarctions were excluded). Odds ratios adjusted for age and sex with 95% confidence interval for volume categories were calculated by use of logistic regression for each QI. Temporal changes were tested in centers that participated in all 3 campaigns. A total of 46 390 records were examined: 18 159 in 2008, 12 837 in 2009, and 15 394 in 2010. Two hundred ninety-one centers were eligible for the temporal analysis. There was a significant increase between 2008 and 2009 in appropriate prescription of antiplatelet agents, β-blockers, angiotensin-converting enzyme inhibitor, statins at discharge, and the composite indicator. Similarly, a significant increase was observed between 2009 and 2010 in appropriate prescription of angiotensin-converting enzyme inhibitor and β-blockers and in the composite QI. Compared with a volume of >300, a significantly lower rate of all QIs was observed in centers with the lowest volume. Odds ratios progressively decreased with increasing volume. Despite a significant increase in the composite QI over the 3 years, a significant relationship persisted between volume and quality of care. Analysis of QIs at discharge demonstrates the existence of a relationship between volume and appropriate prescriptions at discharge. Centers with the highest volume perform better on quality measures than centers with lower volumes. Temporal analysis over 3 consecutive years confirms this relationship and shows that it persists despite improvement in QIs between 2008 and 2010.


Electrically evoked auditory brain stem responses for lateral and medial placement of the Clarion HiFocus electrode.
The purpose of this study was to compare the electrically evoked auditory brain stem response (EABR) for lateral and medial placement of the Clarion HiFocus cochlear implant electrode array via the electrode positioning system (EPS). Twenty-five adult and pediatric cochlear implant recipients participated in the study. Intraoperatively recorded EABRs were evoked by stimuli via three intracochlear electrodes representing apical, medial, and basal locations, and responses were elicited before and after positioner insertion. Evoked potential measures of wave V amplitude and threshold were examined for statistical significance using ANOVA for repeated measures and Chi-Square methods. For a given supra-threshold stimulus level, the increase in EABR wave V amplitude was significantly larger after EPS placement compared to before EPS placement for electrodes 1 (apical) and 13 (basal). Likewise, when the stimulus was decreased to obtain a minimal amplitude, the wave V threshold was significantly lower after EPS placement for electrodes 7 (medial) and 13. The number of measurements that showed decreased wave V threshold after EPS insertion was significantly dependent on intracochlear electrode location. Placement of the Clarion Electrode Positioning System following HiFocus electrode insertion resulted in a reduction in the electrical current required to activate the auditory system. The effect of the EPS was greatest for the basal location, demonstrated by lower wave V thresholds and a larger percentage increase in wave V amplitude. The EABR reflected electrophysiologic changes relative to lateral-to-medial changes in intracochlear electrode position due to the EPS.


A Comment on Frontal Fibrosing Alopecia (Axel Munthe's Syndrome).
Frontal fibrosing alopecia (FFA) represents a peculiar condition with a quasi-symmetrical, marginal alopecia along the frontal and temporal hairline with scarring. Steven Kossard is credited with the original description of the condition in 1994. Since its first description, FFA has become increasingly common while its etiology has remained obscure. While FFA has been related to lichen planopilaris, in fact, the pattern of clinical disease presentation might be more specific for the condition than the underlying inflammatory autoimmune reaction. It has been speculated as to whether FFA existed before Kossard's original report since artistic depictions dating from the 15th to 16th century often show a high frontotemporal hairline. Rather, these represent fashionable forms of frontal pseudo-alopecia than FFA. Nevertheless, there is compelling evidence that FFA existed well before 1994 from the Swedish Physician Axel Munthe's (1857-1948) account of the distinctive features of the condition in his book of memoirs "The Story of San Michele." Therefore, Axel Munthe is to be acknowledged the first description of FFA in 1929.