The effect of flow limitation on the cardiorespiratory response to arousal from sleep under controlled conditions of chemostimulation in healthy older adults.
The influence of flow limitation on the magnitude of the cardiorespiratory response to arousal from sleep is of interest in older people, because they experience considerable flow limitation and frequent arousals from sleep. We studied older flow-limiting subjects, testing the hypothesis that the cardiorespiratory activation response would be larger when arousal occurred during flow limitation, compared to no flow limitation, and chemical stimuli were controlled. In 11 older adults [mean ± standard deviation (SD) age: 68 ± 5 years] ventilation was stabilized using continuous positive airway pressure, and flow limitation was induced by dialling down the pressure. Partial pressure of end-tidal carbon dioxide (PetCO(2)) was maintained by titration of the inspired CO(2) and hyperoxia was maintained using 40% O(2) balanced with nitrogen. Flow limitation at the time of arousal did not augment cardiovascular activation response (heart rate P = 0.7; systolic blood pressure P = 0.6; diastolic blood pressure P = 0.3), whereas ventilation was greater following arousals during flow limitation compared to no flow limitation (P < 0.001). The pre-post-arousal differences in ventilation reflected significant pre-arousal suppression (due to flow limitation) plus post-arousal activation. In summary, the cardiovascular response to arousal from sleep is not influenced by flow limitation at the time of arousal, when chemical stimuli are controlled in older adults. This finding may contribute to the decreased cardiovascular burden associated with sleep-disordered breathing reported in older adults, although our data do not exclude the possibility that flow limitation in the presence of mild hypoxic hypercapnia could increase the cardiovascular response to arousal.


The effectiveness of conically shaped compared with cylindrically shaped interdental brushes - a randomized controlled clinical trial.
The purpose of this study was to compare the effectiveness of conically shaped versus cylindrically shaped interdental brushes (IDB) in patients receiving supportive periodontal therapy. Periodontal maintenance patients volunteered to be enrolled into this randomized controlled examiner-blind parallel study. At baseline and after 3 months, plaque scores, bleeding upon pocket probing scores and probing pocket depth (PPD) were assessed. The type of IDB (conical or cylindrical) was randomly assigned to each patient and individual instruction was provided regarding the method of use and the appropriate size. Only those approximal sites that had sufficient space for the IDB were eligible, and for those sites the data were analysed separately. Analyses were performed for all eligible approximal surfaces and a sub-analysis was performed for vestibular and lingual surfaces. In total, 51 participants attended the baseline and the 3-month clinical appointments. Overall, there was no difference between conical and cylindrical IDBs. However, the conical IDB showed significantly higher plaque and bleeding scores at the lingual approximal sites. The cause of this difference was an increase in plaque and bleeding scores compared with baseline. With respect to the PPD, no difference between the IDBs was observed. Within the limitations of this experiment, the conical IDBs are less effective than cylindrical IDBs with respect to lingual approximal plaque removal. Thus, in patients receiving supportive periodontal therapy, the cylindrical shape should be the first choice of IDB to obtain and maintain gingival health around natural teeth.


Influence of prisoners' mental health on the relational climate of prisons.
Entry into prison involves adapting to a prison culture that is sometimes altered by the effect of imprisonment. Prisons are overcrowded and hold large numbers of inmates suffering from mental disorders and difficulties of adaptation, who affect the delicate equilibrium of the prison environment and can worsen the relational climate. Several bibliographical databases on the influence of the mental health of adult inmates on the prison relationship climate and existing interventions in this regard that have been published in the last 15 years were reviewed. This data was complemented by other information obtained from the online bibliographic indexes of the Ministry of the Interior. There is little literature on the influence of mental health on the relational climate of prisons and existing interventions. However, what literature there is does respond to a current prison reality where inmates with mental health problems have an increased risk of victimization and also a greater predisposition to penitentiary misconduct that causes violations of rules and the consequent application of disciplinary sanctions that lead to segregation. The dysfunctional behaviour of such inmates, as well as the stigmatized treatment they receive, negatively impact the prison social climate, generating pathological relational styles and distorting the prison environment. This creates a need for an adequate number of programs and interventions of sufficient quality to prevent and mitigate their consequences.


[Assessment of health risk in view of changes in labor code and methods used for protecting the environment].
In view of the present and proposed amendments to the Labor Code as well as bearing in mind anticipated harmonization of regulations in this area with those of EEC, the authors emphasize the need for well developed methodology for assessing chemical safety in an occupational environment with special reference to health effects in people exposed to chemicals. Methods for assessing health risk induced by work under conditions of exposure to chemicals were divided into: methods for assessing technological/processing risk, and methods for assessing health risk related to the toxic effect of chemicals. The need for developing means of risk communication in order to secure proper risk perception among people exposed to chemicals and risk managers responsible for prevention against chemical hazards was also stressed. It is suggested to establish a centre for chemical substances in order to settle down all issues pertaining to human exposure to chemicals. The centre would be responsible, under the provisions of the Chemical Substances Act, for the qualitative and quantitative analysis of the present situation and for the development of guidelines on assessment of health risk among persons exposed to chemicals.


Evaluation of surgical therapy by cost-benefit analysis.
Cost-benefit analysis (CBA) is described briefly, emphasizing the contribution this technique can make in justifying health care programs vying with other costly public programs for support. Costs and benefits of two surgical diagnostic techniques are outlined briefly as illustrations of the analysis. The first analyzes routine intraoperative cholangiography indicating the slight benefits that accrue from its use in terms of mortality and hospital expense. A more complicated example is an analysis of a policy of universal annual uterine cervical smears in asymptomatic women for cancer prevention where, granted the assumptions of the model, the costs minus the benefits of the program are estimated to be $565,000/beneficiary/year. The average beneficiary will gain 22 years of life at an average cost/year of about $26,000. The two examples are presented primarily as illustrations of a methodology which is being applied widely in medicine and public policy planning. Second, the marshalling of facts in CBA of a policy of annual uterine smears is of interest to those concerned with the justification of this screening procedure.


Evaluation of the effectiveness of the tuba uterina tubular flap in the peripheral nervous system regeneration in rats.
Nerve conduits could be used to provide a bridge between both nerve endings. In this study, the tuba uterina of female rats were prepared in a vascularized pedicled flap model and it used as a nerve conduit. The aim was to investigate the effectiveness of a vascularized pedicle nerve conduit and its ciliated epithelium in a sciatic nerve defect. The study was conducted between May and August 2018, and used a total of 60, 14-16-week-old female Wistar albino rats. Six groups were created; Cut and Unrepaired Group, Nerve Graft Group, Flap-Forward Group (Tuba uterina tubular flap, forward direction), Flap-Reversed Group (Tuba uterina tubular flap, reverse direction), Graft-Forward Group (Tuba uterina tubular graft, forward direction) and Graft-Reverse Group (Tuba uterina tubuler graft, reverse direction). Nerve regeneration was evaluated 3 months (90 days) after the surgery by the following methods: (1) Sciatic Functional Index (SFI) measurement, (2) Electromyographic (EMG) assessment, (3) Microscopic assessment with the light microscope and (4) Microscopic assessment with the electron microscope. According to the SFI, EMG and microscopic assessments with the light and electron microscope, it was observed that the transfer of tuba uterina tubular conduit as a graft was statistically better in its effect on nerve regeneration than flap transfer, but also indicated that the direction of the ciliated structures had no significant effect. We believe that as this model is improved with future studies, it will shed light on new models, ideas and innovations about nerve conduits.


Capture method affects survival estimates and subsequent interpretation of ecological covariates for a long-lived cervid.
Understanding what variables affect ungulate neonate survival is imperative to successful conservation and management of the species. Predation is commonly cited as a cause-specific source of mortality, and ecological covariates often influence neonate survival. However, variation in survival estimates related to capture methodology has been documented with opportunistically captured neonates generally displaying greater survival than those captured via aid of vaginal implant transmitters (VITs), likely because of increased left truncation observed in the opportunistically captured datasets. Our goal was to assess whether 3- and 6-month survival estimates varied by capture method while simultaneously assessing whether capture method affected model selection and interpretation of ecological covariates for white-tailed deer neonates captured from three study sites from 2014 to 2015 in North Dakota and South Dakota, USA. We found survival varied by capture method for 3-month neonate survival with opportunistically captured neonates displaying up to 26% greater survival than their counterparts captured via VITs; however, this relationship was not present for 6-month survival. We also found model selection and subsequent interpretation of ecological covariates varied when analyzing datasets comprised of neonates captured via VITs, neonates captured opportunistically, and all neonates combined regardless of capture method. When interpreting results from our VIT-only analysis for 3-month survival, we found survival varied by three time intervals and was lowest in the first two weeks of life. Capture method did not affect 6-month survival, which was most influenced by total precipitation occurring during 3 - 8 weeks of a neonate's life and percent canopy cover found at a neonate's capture site. Our results support previous research that capture method must be accounted for when deriving survival estimates for ungulate neonates as it can impact derived estimates and subsequent interpretation of results.


Current diagnosis and management of ovarian cysts.
The epidemiology of ovarian cysts is unclear due to the lack of consistent reporting and a high likelihood of spontaneous resolution. In the USA, postmenopausal women have an ovarian cyst incidence of 18% over a 15-year period. Worldwide, about 7% of women have an ovarian cyst at some point in their lives. In Europe, a large screening trial revealed a 21.2% incidence of ovarian cysts among healthy postmenopausal women. The American College of Obstetricians and Gynecologists (ACOG) stated that simple cysts found on ultrasound may be safely followed without intervention, even in postmenopausal women. These cysts are not likely cancer precursors, nor markers of increased risk, and can be managed conservatively. Simple ovarian cysts appear to be stable or resolve by the next annual examination. These findings support recent recommendations to follow unilocular simple cysts in postmenopausal women without intervention. For those patients, ovarian cancer screening and follow up include a CA-125 blood test and transvaginal ultrasonography (TVU) at baseline, an annual TVU for three additional years, and annual CA-125 tests for five years beyond baseline. The TVU screening examination is considered positive (abnormal and suspicious for ovarian cancer) when findings included: 1) ovarian volume greater than 10 cubic cm; 2) cyst volume greater than ten cubic cm; 3) any solid area or papillary projection extending into the cavity of a cystic ovarian tumor of any size; or, 4) any mixed (solid/cystic) component within a cystic ovarian tumor. Women with positive screening examinations are referred to gynecologic oncology unit for follow-up investigation. Diagnostic consideration and surgical management of ovarian cysts are discussed.


Structure and phase equilibria of the soybean lecithin/PEG 40 monostearate/water system.
PEG stearates are extensively used as emulsifiers in many lipid-based formulations. However, the scheme of the principles of the lipid-surfactant polymer interactions are still poorly understood and need more studies. A new phase diagram of a lecithin/PEG 40 monostearate/water system at 30 °C is reported. First, we have characterized the binary PEG 40 monostearate/water system by the determination of the critical micelle concentration value and the viscous properties. Then, the ternary phase behavior and the influence of phase structure on their macroscopic properties are studied by a combination of different techniques, namely, optical microscopy, small-angle X-ray scattering, differential scanning calorimetry, and rheology. The phase behavior is complex, and some samples evolve even at long times. The single monophasic regions correspond to micellar, swollen lamellar, and lamellar gel phases. The existence of extended areas of phase coexistence (hexagonal, cubic, and lamellar liquid crystalline phases) may be a consequence of the low miscibility of S40P in the lecithin bilayer as well as of the segregation of the phospholipid polydisperse hydrophobic chains. The presence of the PEG 40 monostearate has less effect in the transformation to the cubic phase for lecithin than that found in other systems with simple glycerol-based lipids.


DPD Testing Before Treatment With Fluoropyrimidines in the Amsterdam UMCs: An Evaluation of Current Pharmacogenetic Practice.
The fluoropyrimidines (FP) (5-Fluorouracil, capecitabine, and tegafur) are commonly used anti-cancer drugs, but lead to moderate to severe toxicity in about 10-40% of patients. DPD testing [either the enzyme activity of dihydropyrimidine dehydrogenase (DPD) or the DPYD genotype] identifies patients at higher risk for toxicity who may be treated more safely with a lower drug dose. The Netherland's National guideline for colon carcinoma was updated in 2017 to recommend DPYD genotyping before treatment with FP. Pretreatment DPYD genotyping identifies approximately 50% of the patients that will develop severe FP toxicity. The aim of the study was to assess the uptake of DPD testing in the Amsterdam University Medical Centers over time and to evaluate stakeholder experiences to indicate barriers and facilitators of implementation in routine clinical care. We used a mixed-method approach involving electronic patient records of 753 unique patients and pharmacy information systems analyses and fifteen semi-structured interviews with oncologists, pharmacists, and patients. The constellation perspective was used to identify barriers and facilitators at the level of practice, culture and structure. The proportion of FP users who were DPD tested pretreatment showed an increase from 1% (1/86) in Q2-2017 up to 87% (73/84) in Q4-2018. Unlike a landmark paper published in 2015, the National guideline for colorectal carcinoma followed by meetings to achieve local consensus led to this steep increase in the proportion of patients tested. Facilitating factors for stakeholders to implement testing included the existence of clear protocols, (anecdotal) evidence of the utility, being aware that peers are adhering to standard practice and clear and simple procedures for ordering and reporting. Main barriers included the lack of clear divisions of responsibilities, the lack of consensus on a test approach, long turn-around times and non-user-friendly IT-infrastructures. More professional education on the utility and limitations of pharmacogenetic testing was desired by most stakeholders. While the evidence for DPD testing was sufficient, only after the update of a National guideline and local consensus meetings the proportion of FP users that were DPD tested pretreatment rose to 87%. The implementation of personalized medicine requires stakeholders involved to attune practice, culture and structure.


Extracellular thioredoxin levels are increased in patients with acute lung injury.
Acute lung injury (ALI) and its extreme manifestation the acute respiratory distress syndrome (ARDS) complicate a wide variety of serious medical and surgical conditions. Thioredoxin is a small ubiquitous thiol protein with redox/inflammation modulatory properties relevant to the pathogenesis of ALI. We therefore investigated whether thioredoxin is raised extracellulary in patients with ALI and whether the extent of any increase is dependent upon the nature of the precipitating insult. Bronchoalveolar lavage (BAL) fluid and plasma samples were collected from patients with ALI (n=30) and healthy controls (n=18, plasma; n=14, BAL fluid). Lung tissue was harvested from a separate group of patients and controls (n=10). Thioredoxin was measured by ELISA in fluids and by immunohistochemistry in tissue. Interleukin (IL)-8 levels were determined by ELISA. Disease severity was assessed as APACHE II and SOFA scores. BAL fluid levels of thioredoxin were higher in patients with ALI than in controls (median 61.6 ng/ml (IQR 34.9-132.9) v 16.0 ng/ml (IQR 8.9-25.1), p<0.0001); plasma levels were also significantly higher. When compared with controls, sections of wax embedded lung tissue from patients with ALI showed greater positive staining for thioredoxin in alveolar macrophages and type II epithelial cells. BAL fluid levels of thioredoxin correlated with IL-8 levels in BAL fluid but not with severity of illness scores or mortality. BAL fluid levels of thioredoxin, IL-8, and neutrophils were significantly greater in patients with ALI of pulmonary origin. Extracellular thioredoxin levels are raised in patients with ALI, particularly of pulmonary origin, and have a significant positive association with IL-8. Extracellular thioredoxin levels could provide a useful indication of inflammation in ALI.


Health-related quality of life in patients with adolescent idiopathic scoliosis: a matched follow-up at least 20 years after treatment with brace or surgery.
No results on long-term outcome in terms of health-related quality of life (HRQL) have previously been presented for patients treated for adolescent idiopathic scoliosis. A consecutive series of patients with adolescent idiopathic scoliosis, treated between 1968 and 1977 before the age of 21, either with distraction and fusion using Harrington rods [surgical treatment group (ST), n=156; 145 females and 11 males] or with a brace [brace treatment group (BT), n=127; 122 females and 5 males] were followed at least 20 years after completion of the treatment. Ninety-four percent of ST and 91% of BT patients filled in a questionnaire comprising the SF-36, Psychological General Well-Being Index (PGWB), Oswestry Disability Back Pain Questionnaire, parts of SRS/MODEM'S questionnaire and study-specific questions concerning the treatment, as a part of an unbiased personal follow-up examination including radiography and clinical examination. An age- and sex-matched control group of 100 persons was randomly selected and subjected to the same examinations. The results showed no differences in terms of sociodemographic data between the groups. Both ST and BT patients had a slightly, but significantly, reduced physical function using the SF-36 subscales, SF-36/Physical Component Summary (PCS) score as well as the Oswestry Disability Back Pain Questionnaire compared to the controls. Neither the mental subscales and the Mental Component Summary (MCS) score of SF-36 nor the PGWB index showed any significant difference between the groups. Forty-nine percent of ST, 34% of BT and 15% of controls admitted limitation of social activities due to their back [P<0.001 ST vs controls, P=0.0010 BT vs controls, and n.s. (P=0.024) ST vs BT], mostly due to difficulties with physical participation in activities or self-consciousness about appearance. Pain was a minor reason for limitation. No correlation was found between the outcome scores and curve size after treatment, curve type, total treatment time or age at completed treatment. Patients treated for adolescent idiopathic scoliosis were found to have approximately the same HRQL as the general population. A minority of the patients (4%) had a severely decreased psychological well-being, and a few (1.5%) were severely physically disabled due to the back.


Agents and robots for collaborating and supporting physicians in healthcare scenarios.
Monitoring patients through robotics telehealth systems is an interesting scenario where patients' conditions, and their environment, are dynamic and unknown variables. We propose to improve telehealth systems' features to include the ability to serve patients with their needs, operating as human caregivers. The objective is to support the independent living of patients at home without losing the opportunity to monitor their health status. Application scenarios are several, and they spread from simple clinical assisting scenarios to an emergency one. For instance, in the case of a nursing home, the system would support in continuously monitoring the elderly patients. In contrast, in the case of an epidemic diffusion, such as COVID-19 pandemic, the system may help in all the early triage phases, significantly reducing the risk of contagion. However, the system has to let medical assistants perform actions remotely such as changing therapies or interacting with patients that need support. The paper proposes and describes a multi-agent architecture for intelligent medical care. We propose to use the beliefs-desires-intentions agent architecture, part of it is devised to be deployed in a robot. The result is an intelligent system that may allow robots the ability to select the most useful plan for unhandled situations and to communicate the choice to the physician for his validation and permission.


TNFα-mediated activation of NF-κB downregulates sodium-iodide symporter expression in thyroid cells.
The sodium-iodide symporter (NIS) mediates transport of iodide across the basolateral membrane of thyroid cells. NIS expression in thyroid cancer (TC) cells allows the use of radioactive iodine (RAI) as a diagnostic and therapeutic tool, being RAI therapy the systemic treatment of choice for metastatic disease. Still, a significant proportion of patients with advanced TC lose the ability to respond to RAI therapy and no effective alternative therapies are available. Defective NIS expression is the main reason for impaired iodide uptake in TC and NIS downregulation has been associated with several pathways linked to malignant transformation. NF-κB signaling is one of the pathways associated with TC. Interestingly, NIS expression can be negatively regulated by TNF-α, a bona fide activator of NF-κB with a central role in thyroid autoimmunity. This prompted us to clarify NF-kB's role in this process. We confirmed that TNF-α leads to downregulation of TSH-induced NIS expression in non-neoplastic thyroid follicular cell-derived models. Notably, a similar effect was observed when NF-κB activation was triggered independently of ligand-receptor specificity, using phorbol-myristate-acetate (PMA). TNF-α and PMA downregulation of NIS expression was reverted when NF-κB-dependent transcription was blocked, demonstrating the requirement for NF-kB activity. Additionally, TNF-α and PMA were shown to have a negative impact on TSH-induced iodide uptake, consistent with the observed transcriptional downregulation of NIS. Our data support the involvement of NF-κB-directed transcription in the modulation of NIS expression, where up- or down-regulation of NIS depends on the combined output to NF-κB of several converging pathways. A better understanding of the mechanisms underlying NIS expression in the context of normal thyroid physiology may guide the development of pharmacological strategies to increase the efficiency of iodide uptake. Such strategies would be extremely useful in improving the response to RAI therapy in refractory-TC.


The systemic use of procaine in the treatment of the elderly: a review.
This article is a review and evaluation of the world literature on the systemic use of procaine in the treatment of the aging process and the common chronic diseases of later life. Included are data from 285 articles and books, describing treatment in more than 100,000 patients in the past 25 years. Except for a possible antidepressant effect, there is no convincing evidence that procaine (or Gerovital, of which procaine is the major component) has any value in the treatment of disease in older patients. If procaine has an antidepressant effect, there is some likelihood that this accounts for the reports of decreased complaints referable to the musculoskeletal, cardiovascular, endocrine sexual, gastrointestinal and respiratory systems.


Spontaneous resolution of hemophagocytic syndrome and disseminated intravascular coagulation associated with parvovirus b19 infection in a previously healthy child.
A 10-year-old male with a brain abscess developed pancytopenia, liver dysfunction, disseminated intravascular coagulation (DIC) and decrease of immunoglobulin A (IgA) level during postoperative antibiotic and anticonvulsant therapy. A bone marrow examination revealed hemophagocytosis. Real-time PCR revealed parvovirus B19 infection. The hemophagocytic syndrome resolved without specific treatment. To our knowledge, this is the first report of a spontaneous resolution of parvovirus B19-associated hemophagocytic syndrome and DIC.


Comparison of conventional and high-sensitivity troponin in patients with chest pain: a collaborative meta-analysis.
Multiple studies have evaluated the diagnostic and prognostic performance of conventional troponin (cTn) and high-sensitivity troponin (hs-cTn). We performed a collaborative meta-analysis comparing cTn and hs-cTn for diagnosis of acute myocardial infarction (AMI) and assessment of prognosis in patients with chest pain. MEDLINE/PubMed, Cochrane CENTRAL, and EMBASE were searched for studies assessing both cTn and hs-cTn in patients with chest pain. Study authors were contacted and many provided previously unpublished data. From 17 included studies, there were 8,644 patients. Compared with baseline cTn, baseline hs-cTn had significantly greater sensitivity (0.884 vs 0.749, P < .001) and negative predictive value (NPV; 0.964 vs 0.935, P < .001), whereas specificity (0.816 vs 0.938, P < .001) and positive predictive value (0.558 vs 0.759, P < .001) were significantly reduced. Based on summary receiver operating characteristic curves, test performance for the diagnosis of AMI was not significantly different between baseline cTn and hs-cTn (0.90 [95% CI 0.85-0.95] vs 0.92 [95% CI 0.90-0.94]). In a subanalysis of 6 studies that alternatively defined AMI based on hs-cTn, cTn had lower sensitivity (0.666, P < .001) and NPV (0.906, P < .001). Elevation of baseline hs-cTn, but negative baseline cTn, was associated with increased risk of death or nonfatal myocardial infarction during follow-up (P < .001) compared with both negative. High-sensitivity troponin has significantly greater early sensitivity and NPV for the diagnosis of AMI at the cost of specificity and positive predictive value, which may enable early rule in/out of AMI in patients with chest pain. Baseline hs-cTn elevation in the setting of negative cTn is also associated with increased nonfatal myocardial infarction or death during follow-up.


Chromosomal microarray testing identifies a 4p terminal region associated with seizures in Wolf-Hirschhorn syndrome.
Wolf-Hirschhorn syndrome (WHS) is a contiguous gene deletion syndrome involving variable size deletions of the 4p16.3 region. Seizures are frequently, but not always, associated with WHS. We hypothesised that the size and location of the deleted region may correlate with seizure presentation. Using chromosomal microarray analysis, we finely mapped the breakpoints of copy number variants (CNVs) in 48 individuals with WHS. Seizure phenotype data were collected through parent-reported answers to a comprehensive questionnaire and supplemented with available medical records. We observed a significant correlation between the presence of an interstitial 4p deletion and lack of a seizure phenotype (Fisher's exact test p=3.59e-6). In our cohort, there were five individuals with interstitial deletions with a distal breakpoint at least 751 kbp proximal to the 4p terminus. Four of these individuals have never had an observable seizure, and the fifth individual had a single febrile seizure at the age of 1.5 years. All other individuals in our cohort whose deletions encompass the terminal 751 kbp region report having seizures typical of WHS. Additional examples from the literature corroborate these observations and further refine the candidate seizure susceptibility region to a region 197 kbp in size, starting 368 kbp from the terminus of chromosome 4. We identify a small terminal region of chromosome 4p that represents a seizure susceptibility region. Deletion of this region in the context of WHS is sufficient for seizure occurrence.


Exercise-induced hyperkalaemia: effects of beta-adrenoceptor blocker vs diuretic.
Four groups of eight normotensive male volunteers performed a 60 min bicycle exercise test before and after 2 weeks of either placebo, hydrochlorothiazide (HCTZ, 25 mg day-1), pindolol (PIND, 10 mg day-1) or both drugs in combination using a double-blind, randomized design. During exercise on placebo serum potassium increased by 0.8 mmol l-1. HCTZ significantly decreased potassium levels at rest and during exercise by 0.2 mmol l-1. PIND did not affect resting potassium levels but potentiated the increase by 0.4 mmol l-1 at the end of exercise, and delayed the return to normal of serum potassium after exercise. The addition of HCTZ to PIND offset the potentiating effect of PIND on exercise-induced hyperkalaemia (only after prolonged exercise) and accelerated the return to baseline after exercise. The results indicate that the hypokalaemic effect of HCTZ can oppose the hyperkalaemic effect of PIND during prolonged physical exercise and particularly during recovery.


Factors regulating trophoblast invasion.
Implantation in the human is unique. This uniqueness is characterized on the maternal side by a spontaneous and massive decidualization of the endometrium and on the embryonic side by an almost unlimited invasive potential. Human embryos express an intrinsic invasive potential, which allows them to implant almost anywhere except in the endometrium because it protects itself from implantation. Human implantation is thus only possible during a limited period of time known as the implantation window. This mini review stresses the importance of studying trophoblast invasion into the endometrium as a model for human implantation. Cytotrophoblastic cells (CTB) can easily be isolated from first-trimester legal abortions and retain their invasive behavior when cultured in vitro. This model shows that matrix metalloproteinases (MMPs) are produced by CTB and are instrumental to their invasive behavior. Embryo implantation and tumor invasion use these same biochemical mediators for invasion. However, in contrast to tumor invasion, trophoblast invasion is limited both in time and space: it occurs during the first trimester of pregnancy and invasion does not go beyond the proximal third of the myometrium. Factors regulating MMP expression are of maternal and fetal origin.


Immunocompetence analysis of the aquatic snail Lymnaea stagnalis exposed to urban wastewaters.
Wastewater treatment plant effluents from urban area are a well-known source of chronic multiple micropollution to the downstream living organisms. In this study, ecologically relevant laboratory-bred freshwater gastropods, Lymnaea stagnalis, were exposed for 29 days to raw effluents of a wastewater treatment plant in Lyon area (France). A time-course analysis of individual markers of immunocompetence (hemocyte density and viability, hemocyte NADPH activity, phenol oxidase activity, and capacity of phagocytosis) has shown slight trends of inflammatory-like responses induced by the 100% effluents. So far, no short-term hazard for L. stagnalis can be revealed. However, over the long term, such environmental stress-stimulating immune responses could provoke deleterious life history trade-offs because the immune system is known to be highly energy-consuming.


Histologic study of intravelar structures in normal human adult specimens.
To describe more clearly the tissue composition and structure of the human soft palate with particular emphasis on the central portion between the faucial pillars. Nine female and three male normal adult human soft palate cadaver specimens. The anterior soft palate consists of fairly uniform layers. The anterior one fourth contains a substantial investment of mucous-secreting glandular tissue, as well as an abundance of adipose tissue. The tensor veli palatini tendon is prominent in the most anterior region just posterior to the hard palate and close to the nasal surface. The middle one third of the soft palate is largely invested with muscle tissue consisting of (1) levator veli palatini fibers coursing transversely across the midline without a septal interruption, (2) musculus uvulae fibers encapsulated in a sheath and coursing longitudinally, perpendicular to and cradled by the levator sling, and (3) palatopharyngeus fibers located laterally and not approaching the midline. Musculus uvulae is variable across and within specimens in terms of its paired versus unpaired nature. The anatomy of the soft palate from the posterior border of the hard palate to the levator veli palatini sling is consistent among specimens, suggesting that structures in this region have a uniform function across subjects. The paired versus unpaired nature of musculus uvulae is variable both within and between specimens. The posterior one third of the soft palate is variable across specimens with regard to the relative amount and distribution of different tissue types.


Detecting Pneumoperitoneum via Point-of-Care Abdominal Ultrasound: To See Beyond Touch.
Point-of-care ultrasound is increasingly important in the management of acute medical emergencies. An elderly man was brought to the emergency department after 2 days of fever and urinary retention. He was drowsy and had peri-arrest arrhythmia. He was hypoperfused peripherally with a systolic blood pressure of 45 mmHg and so was managed as for septic shock with no obvious aetiology. Chest and abdominal physical examinations were unremarkable. The source of sepsis was unclear. A point-of-care abdominal ultrasound was performed by the reviewing internist which detected a pneumoperitoneum, leading to a change in diagnosis to a perforated viscus which was confirmed later by a CT of the abdomen and pelvis. Pneumoperitoneum leading to septic shock is a medical emergency, but diagnosis is frequently delayed as the physical examination and chest and abdominal x-rays may fail to detect the condition in the critically ill patient, leading to delayed medical and surgical intervention.There is a role for point-of-care abdominal ultrasound for the early diagnosis of pneumoperitoneum with the air reverberation artefact.Point-of-care ultrasound of the abdomen to identify intra-abdominal air is an important physical examination adjunct in undifferentiated septic shock and should be incorporated into the routine care of patients and included in the acute medicine training syllabus.


Canadian families caring for members with mental illness: a vicious cycle.
The purpose of this qualitative, descriptive study was to explore the perspectives of individuals who were caring for a family member with a mental illness, with particular attention to housing, quality of supports, and formal care services. Eleven focus groups with family caregivers (N = 75) were conducted. Both individual and team thematic analyses were undertaken until interpretations of the experiences of the participants were inductively developed and conceptualized into a holistic interpretation. Findings revealed that family caregivers were part of a "circle of care," supporting the independence of the individual with mental illness while attempting to protect their family member with mental illness. However, findings suggested that this circle of care led to a "vicious cycle" of caregiving. Three major themes were identified: witnessing inadequacies, working behind the scenes, and creating a better world. Health-promoting family nursing practice and policy implications, as well as areas for further research, are discussed.


Adherence to a cardiac rehabilitation home program model of care: a comparison to a well-established traditional on-site supervised program.
Despite the proven benefits of cardiac rehabilitation (CR), adherence to programs remains suboptimal. To improve adherence, alternative models of care, such as using home programs (HP), have been recommended. Little information exists, however, about its effect on adherence in real-world settings. Therefore, this study's primary objective was to compare adherence of patients in an HP and traditional on-site program (TP) model of CR. The secondary objective was to compare their clinical and demographic profiles and changes in cardiovascular fitness. We implemented a retrospective review of 200 consecutively enrolled patients who chose either a TP or HP model. Profile data was collected at intake assessment. Adherence, defined as attendance to prescheduled contacts or on-site visits in each respective cohort, served as a primary outcome measure. Secondary outcomes included completion of program and cardiopulmonary fitness levels at 6 months. We found that the HP cohort had patients who were significantly younger, male (significantly moreso than female), were more geographically removed from the on-site centre, were employed, and (or) had greater cardiopulmonary fitness at initiation. Similar mean attendance (p = 0.21) and completion rates (p = 0.22) were seen between models. Both groups attained similar gains in cardiovascular fitness (p = 0.79). Analysis of adherence shows the HP to be a suitable option for patients who face barriers for TP-CR participation.


Education and training need to be brought up-to-date.
Training of hospital employees consumes a large segment of total hospital dollars, so it is important to document clear proof of value. If training programs are to be successful, they must be relevant to the measured needs of the institution. In addition to this essential characteristic of the training process, the author discusses six others that affect the outcome of such programs.


Bayesian adaptive randomized trial design for patients with recurrent glioblastoma.
To evaluate whether the use of Bayesian adaptive randomized (AR) designs in clinical trials for glioblastoma is feasible and would allow for more efficient trials. We generated an adaptive randomization procedure that was retrospectively applied to primary patient data from four separate phase II clinical trials in patients with recurrent glioblastoma. We then compared AR designs with more conventional trial designs by using realistic hypothetical scenarios consistent with survival data reported in the literature. Our primary end point was the number of patients needed to achieve a desired statistical power. If our phase II trials had been a single, multiarm trial using AR design, 30 fewer patients would have been needed compared with a multiarm balanced randomized (BR) design to attain the same power level. More generally, Bayesian AR trial design for patients with glioblastoma would result in trials with fewer overall patients with no loss in statistical power and in more patients being randomly assigned to effective treatment arms. For a 140-patient trial with a control arm, two ineffective arms, and one effective arm with a hazard ratio of 0.6, a median of 47 patients would be randomly assigned to the effective arm compared with 35 in a BR trial design. Given the desire for control arms in phase II trials, an increasing number of experimental therapeutics, and a relatively short time for events, Bayesian AR designs are attractive for clinical trials in glioblastoma.


Discovery of fully human anti-MET monoclonal antibodies with antitumor activity against colon cancer tumor models in vivo.
The receptor tyrosine kinase MET is a major component controlling the invasive growth program in embryonic development and in invasive malignancies. The discovery of therapeutic antibodies against MET has been difficult, and antibodies that compete with hepatocyte growth factor (HGF) act as agonists. By applying phage technology and cell-based panning strategies, we discovered two fully human antibodies against MET (R13 and R28), which synergistically inhibit HGF binding to MET and elicit antibody-dependent cellular cytotoxicity. Cell-based phosphorylation assays demonstrate that R13 and R28 abrogate HGF-induced activation of MET, AKT1, ERK1/2, and HGF-induced migration and proliferation. FACS experiments suggest that the inhibitory effect is mediated by "locking" MET receptor in a state with R13, which then increases avidity of R28 for the extracellular domain of MET, thus blocking HGF binding without activating the receptor. In vivo studies demonstrate that the combination of R13/28 significantly inhibited tumor growth in various colon tumor xenograft models. Inhibition of tumor growth was associated with induction of hypoxia. Global gene expression analysis shows that inhibition of HGF/MET pathway significantly upregulated the tumor suppressors KLF6, CEACAM1, and BMP2, the negative regulator of phosphatidylinositol-3-OH-kinase PIK3IP1, and significantly suppressed SCF and SERPINE2, both enhancers of proliferation and invasiveness. Moreover, in an experimental metastasis model, R13/28 increased survival by preventing the recurrence of otherwise lethal lung metastases. Taken together, these results underscore the utility of a dual-antibody approach for targeting MET and possibly other receptor tyrosine kinases. Our approach could be expanded to drug discovery efforts against other cell surface proteins.


Extreme-scale Dynamic Exploration of a Distributed Agent-based Model with the EMEWS Framework.
Agent-based models (ABMs) integrate multiple scales of behavior and data to produce higher-order dynamic phenomena and are increasingly used in the study of important social complex systems in biomedicine, socio-economics and ecology/resource management. However, the development, validation and use of ABMs is hampered by the need to execute very large numbers of simulations in order to identify their behavioral properties, a challenge accentuated by the computational cost of running realistic, large-scale, potentially distributed ABM simulations. In this paper we describe the Extreme-scale Model Exploration with Swift (EMEWS) framework, which is capable of efficiently composing and executing large ensembles of simulations and other "black box" scientific applications while integrating model exploration (ME) algorithms developed with the use of widely available 3rd-party libraries written in popular languages such as R and Python. EMEWS combines novel stateful tasks with traditional run-to-completion many task computing (MTC) and solves many problems relevant to high-performance workflows, including scaling to very large numbers (millions) of tasks, maintaining state and locality information, and enabling effective multiple-language problem solving. We present the high-level programming model of the EMEWS framework and demonstrate how it is used to integrate an active learning ME algorithm to dynamically and efficiently characterize the parameter space of a large and complex, distributed Message Passing Interface (MPI) agent-based infectious disease model.


Sources, uses, strengths and limitations of data collected in primary care in England.
In the UK, general practitioners are responsible for both providing primary care services and for organising referrals for specialist care. Consequently, the medical records held by general practitioners include details of all diagnoses and prescribed drugs, in many cases extending from birth. In an increasing number of general practices, these records are held in electronic format and are potentially available for extraction and analysis. Electronic data from general practice therefore offer unique opportunities to plan and monitor health services, measure the quality of care provided by the National Health Service, and undertake population-based research.


Analysis of enoxaparin dose titration at a large, tertiary teaching facility.
Therapeutic drug monitoring of enoxaparin with antifactor Xa levels (AXALs) is recommended in some populations; however, the approach to dose titration is poorly described. Our study at a large, tertiary teaching facility examined the dose response to titration of enoxaparin based on AXAL. Patients from 2008 to 2012 receiving enoxaparin were included, provided 2 or more steady state AXAL were obtained within 30 days and that the enoxaparin was prescribed for treatment rather than prophylaxis. The primary outcome was the percentage of dose change required to obtain goal range AXAL following dose titration. Eighty-seven patients were available for analysis with the following key characteristics: renal dysfunction during treatment 72%, obesity 8%, and solid organ transplant 26%. Initial goal AXAL was attained in 27 (31%) patients, and ultimately 54 (62%) patients achieved goal AXAL. Of the 31 patients who had initial AXAL above goal, 13 (42%) patients reached goal with a median dose decrease of 24%. In the 29 patients who had an initial AXAL below goal, 11 (38%) achieved therapeutic AXAL with a median dose increase of 16%. The AXAL monitoring can guide enoxaparin titration with subtherapeutic or supratherapeutic AXAL and an increase or decrease of roughly 20% is suggested as an initial change.


Complete chloroplast genome of the orchid Cattleya crispata (Orchidaceae:Laeliinae), a Neotropical rupiculous species.
A partial genome dataset was sequenced for the orchid Cattleya crispata using both Illumina and 454 technologies. The chloroplast genome was assembled using iterative runs of MIRA software that yielded a circular molecule with 148,343 bp in length and deposited in GenBank database (Accession Number KP168671). The plastid genome conserved the quadripartite structure present in most Orchidaceae chloroplasts and was composed by 79 protein-coding genes, 39 tRNAs and 8 rRNAs. Genome structure, gene order and orientation were similar to previously described chloroplasts for Cymbidium orchids, differing in gene order for petN and psbM genes. Data described here contain the first report of a complete chloroplast for the Neotropical subtribe Laeliinae and may contribute to improve the phylogenetic resolution and allow the development of new molecular markers for population genetic studies of orchids.


Effects of differentiation of embryonal carcinoma cells (P19) on mitochondrial DNA content in vitro.
The embryonal carcinoma cell line P19 is derived from mouse teratocarcinomas. These pluripotent cells can be induced to differentiate into a variety of cell types by exposure to various drugs. We used retinoic acid to induce embryonal carcinoma cells to differentiate into neuronlike cells. In this study, we show that changes occur in mitochondria during differentiation of embryonal carcinoma cells to neuronlike cells. We found that various morphologic parameters such as mitochondrial fractional area and mitochondrial size decrease as embryonal carcinoma cells differentiate into neuronlike cells. Similar changes were also observed in mitochondrial DNA content. Stereologic analysis of cell preparations provided a measure of mitochondrial fractional area per cell and mtDNA content was assessed by radiolabeled mtDNA probe. This study establishes that mitochondria are regulated as cells differentiate.


Support for Relatives Bereaved by Psychiatric Patient Suicide: National Confidential Inquiry Into Suicide and Homicide Findings.
International suicide prevention strategies recommend providing support to families bereaved by suicide. The study objectives were to measure the proportion of cases in which psychiatric professionals contact next of kin after a patient's suicide and to investigate whether specific, potentially stigmatizing patient characteristics influence whether the family is contacted. Annual survey data from England and Wales (2003-2012) were used to identify 11,572 suicide cases among psychiatric patients. Multivariate regression analysis was used to describe the association between specific covariates (chosen on the basis of clinical judgment and the published literature) and the probability that psychiatric staff would contact bereaved relatives of the deceased. Relatives were not contacted after the death in 33% of cases. Contrary to the hypothesis, a violent method of suicide was independently associated with greater likelihood of contact with relatives (adjusted odds ratio=1.67). Four patient factors (forensic history, unemployment, and primary diagnosis of alcohol or drug dependence or misuse) were independently associated with less likelihood of contact with relatives. Patients' race-ethnicity and recent alcohol or drug misuse were not associated with contact with relatives. Four stigmatizing patient-related factors reduced the likelihood of contacting next of kin after patient suicide, suggesting inequitable access to support after a potentially traumatic bereavement. Given the association of suicide bereavement with suicide attempt, and the possibility of relatives' shared risk factors for suicide, British psychiatric services should provide more support to relatives after patient suicide.


Impact of Grazing Management on Soil Nitrogen, Phosphorus, Potassium, and Sulfur Distribution.
Little information is available directly comparing soil nutrient distribution under different defoliation managements. During 1990 (116 d) and 1991 (141 d), 'Callie' bermudagrass (Cynodon dactylon var. aridus Harlan et de Wet) pastures grazed by Holstein heifers (Bos taurus) were used to determine the effects of two rotational stocking methods and continuous stocking on lateral and vertical distribution of extractable N, P, K, and S. A hay management also was included to compare soil responses under grazing and clipping. Nutrient distribution and concentration in the Ap1 horizon (0- to 15-cm soil depth) did not differ among grazing methods, but N, P, and K accumulated in the third of the pastures closest to shade, water sources, and supplement feeders (lounging areas where cattle tend to congregate or rest). Similar observations were made with K in the Ap2 horizon (15- to 30-cm soil depth). Nutrient concentrations were lower or tended to be lower in the Ap1 horizon of the hay management than in grazed pastures because of nutrient removal in harvested herbage. Across defoliation managements, greater extractable N, P, and K concentrations were observed in the Ap1 horizon in 1991 than in 1990. For N and K, this was attributed to fertilizer inputs in all managements and partially to supplemental feed inputs in grazed pastures. Increases in extractable P appeared to be associated primarily with flooding of the experimental site in late 1991. This study suggests that grazing method of well-managed pastures may have little effect on short-term (2 yr) soil nutrient distribution, especially when grazing occurs during months when temperatures are high.


Stability of amoxicillin-clavulanate in BACTEC medium determined by high-performance liquid chromatography and bioassay.
The stabilities of amoxicillin (16 micrograms/ml) and clavulanate (8 micrograms/ml), alone and in combination in BACTEC medium (Middlebrook 7H12B medium), were determined by high-performance liquid chromatography (HPLC) and bioassay. By HPLC, the half-life of amoxicillin (trihydrate and sodium) in combination with clavulanate in nonradiolabelled 7H12B medium was 6.7 days, whereas the half-life of clavulanate in combination with amoxicillin was 2.0 days. By bioassay, the half-lives of amoxicillin trihydrate and clavulanate in radiolabelled 7H12B medium were comparable (7 and 2 days, respectively) to those determined by HPLC. When clavulanate was tested alone, the half-life was determined to be 1.88 days by HPLC and 1.87 days by bioassay. The relatively short half-life of clavulanate can be adjusted by a procedure of "topping up," or adding one-half the concentration of clavulanate every second day, in order to allow accurate amoxicillin-clavulanate MIC testing with the BACTEC mycobacterial susceptibility system.


[Anxiety and hemodynamics: correlations in hypertensive patients].
Syndrome analysis of functional relationships between personality disorders, autonomic regulation and psychic performance in relation to hemodynamic condition of hypertensive patients. MMP1, Tailor's anxiety scale, Burdon and Schulte tables, variation pulsometry, tetrapolar chest rheography were used in examination of 86 patients with stage I and II hypertension. In early hypertension, weak anxiety and subclinical emotional disturbances, dominant was a parasympathetic trend of the vegetative tone without central control of the cardiac rhythm in hyperkinetic circulation. Changes in cortical neurodynamics manifested with minor attenuation of the ability to focus attention. Progression of angiospasticity and anxiety gave rise to psychic maladjustment in the form of anxiety-depression disorders with autonomic dysfunction in hypokinetic circulation, overcentralization of cardiac rhythm control. This was associated with a decline in mental performance. Systemic investigations of the emotional sphere, central hemodynamics and autonomic regulation of cardiac rhythm enable assessment of multilevel structure and severity of psychocardiac syndrome in hypertensive patients which is important both for therapeutic and expert practice.


Extracellular miR-146a-5p Induces Cardiac Innate Immune Response and Cardiomyocyte Dysfunction.
Previous studies have demonstrated that transient myocardial ischemia leads to release of cellular nucleic acids such as RNA. Extracellular RNA reportedly plays a pivotal role in myocardial inflammation and ischemic injury in animals. RNA profiling has identified that numerous microRNA (miRNAs), such as ss-miR-146a-5p, are upregulated in plasma following myocardial ischemia, and certain uridine-rich miRNAs exhibit strong proinflammatory effects in immune cells via ssRNA-sensing mechanism. However, the effect of extracellular miRNAs on myocardial inflammation and cardiac cell function remains unknown. In this study, we treated adult mouse cardiomyocytes with miR-146a-5p loaded in extracellular vesicles and observed a dose- and TLR7-dependent production of CXCL-2, IL-6, and TNF-α. In vivo, a single dose of myocardial injection of miR-146a-5p induced both cytokine expression (CXCL2, IL-6, and TNF-α) and innate immune cell activation (CD45+ leukocytes, Ly6Cmid+ monocytes, Ly6G+ neutrophils), which was significantly attenuated in the hearts of TLR7 KO mice. We discovered that conditioned media from miR-146a-treated macrophages stimulated proinflammatory cytokine production in adult cardiomyocytes and significantly inhibited their sarcomere shortening. Finally, using an electric cell impedance-sensing assay, we found that the conditioned media from miR-146a-treated cardiac fibroblasts or cardiomyocytes impaired the barrier function of coronary artery endothelial cells. Taken together, these data demonstrate that extracellular miR-146a-5p activates multiple cardiac cells and induces myocardial inflammation and cardiomyocyte dysfunction via intercellular interaction and innate immune TLR7 nucleic acid sensing.


Identification of a core set of 58 gene transcripts with broad and specific expression in the microvasculature.
Pathological angiogenesis is an integral component of many diseases. Antiangiogenesis and vascular targeting are therefore promising new therapeutic principles. However, few endothelial-specific putative drug targets have been identified, and information is still limited about endothelial-specific molecular processes. Here we aimed at determining the endothelial cell-specific core transcriptome in vivo. Analysis of publicly available microarray data identified a mixed vascular/lung cluster of 132 genes that correlated with known endothelial markers. Filtering against kidney glomerular/nonglomerular and brain vascular/nonvascular microarray profiles separated contaminating lung markers, leaving 58 genes with broad and specific microvascular expression. More than half of these have not previously been linked to endothelial functions or studied in detail before. The endothelial cell-specific expression of a selected subset of these, Eltd1, Gpr116, Ramp2, Slc9a3r2, Slc43a3, Rasip1, and NM_023516, was confirmed by real-time quantitative polymerase chain reaction and/or immunohistochemistry. We have used a combination of publicly available and own microarray data to identify 58 gene transcripts with broad yet specific expression in microvascular endothelium. Most of these have unknown functions, but many of them are predicted to be cell surface expressed or implicated in cell signaling processes and should therefore be explored as putative microvascular drug targets.


Hydration and ion association of La3+ and Eu3+ salts in aqueous solution.
Aqueous solutions of five lanthanide salts: LaCl3, La(NO3)3, La2(SO4)3, Eu(NO3)3 and Eu2(SO4)3 have been studied at 25 °C by dielectric relaxation spectroscopy over the frequency range 0.05 ≤ ν/GHz ≤ 89. Detailed analysis of the solvent-related modes located at higher frequencies showed that both La3+ and Eu3+ are strongly hydrated, even including partial formation of a third hydration shell similar to that of Al3+(aq). Up to two solute-related modes could be detected at lower frequencies, due to the formation of various types of 1 : 1 ion pairs (IPs). All five salts showed modest levels of association in the order Cl- < NO3- ≪ SO42-, mostly in the form of double-solvent-separated IPs with small amounts of solvent-shared IPs. Overall association constants, , calculated from the stepwise IP formation constants were consistent with literature values.


Sensitivity-enhanced three-dimensional and carbon-detected two-dimensional NMR of proteins using hyperpolarized water.
Signal enhancements of up to two orders of magnitude in protein NMR can be achieved by employing HDO as a vector to introduce hyperpolarization into folded or intrinsically disordered proteins. In this approach, hyperpolarized HDO produced by dissolution-dynamic nuclear polarization (D-DNP) is mixed with a protein solution waiting in a high-field NMR spectrometer, whereupon amide proton exchange and nuclear Overhauser effects (NOE) transfer hyperpolarization to the protein and enable acquisition of a signal-enhanced high-resolution spectrum. To date, the use of this strategy has been limited to 1D and 1H-15N 2D correlation experiments. Here we introduce 2D 13C-detected D-DNP, to reduce exchange-induced broadening and other relaxation penalties that can adversely affect proton-detected D-DNP experiments. We also introduce hyperpolarized 3D spectroscopy, opening the possibility of D-DNP studies of larger proteins and IDPs, where assignment and residue-specific investigation may be impeded by spectral crowding. The signal enhancements obtained depend in particular on the rates of chemical and magnetic exchange of the observed residues, thus resulting in non-uniform 'hyperpolarization-selective' signal enhancements. The resulting spectral sparsity, however, makes it possible to resolve and monitor individual amino acids in IDPs of over 200 residues at acquisition times of just over a minute. We apply the proposed experiments to two model systems: the compactly folded protein ubiquitin, and the intrinsically disordered protein (IDP) osteopontin (OPN).


Evolving Business Models in Orthotics.
This submission provides an important historical context for understanding the current challenge facing the Orthotic and Prosthetic community in Alberta including Alberta Aids to Daily Living (AADL), Suppliers, and Providers: maintaining sustainable access to Orthotic care for people with mobility disorders in the face of declining real rates of reimbursement combined with increasing costs and a shortage of skilled Clinicians. Under the Canada Health Act, the federal government delegates responsibility for providing health care to the provinces. This delegation of responsibility to the provinces results in a degree of variability of funding of Orthotics and Prosthetics between provinces across the country. Funding of Orthotics and Prosthetics in Alberta is characterized by structural inequities that favour Prosthetics at the expense of Orthotics. To the extent that the structural inequities that exist in Alberta are related to governance by volunteer-run, non-profit organizations, they may be generalized to the Canadian experience. Finally, in a Call to Action a number of recommendations are made to address the challenge of sustainable access to Orthotic care in Alberta serving as a model for other provinces across Canada.


Bacteriophage T4 helicase loader protein gp59 functions as gatekeeper in origin-dependent replication in vivo.
Bacteriophage T4 initiates origin-dependent replication via an R-loop mechanism in vivo. During in vitro reactions, the phage-encoded gp59 stimulates loading of the replicative helicase, gp41, onto branched intermediates, including origin R-loops. However, although gp59 is essential for recombination-dependent replication from D-loops, it does not appear to be required for origin-dependent replication in vivo. In this study, we have analyzed the origin-replicative intermediates formed during infections that are deficient in gp59 and other phage replication proteins. During infections lacking gp59, the initial replication forks from two different T4 origins actively replicated both leading- and lagging-strands. However, the retrograde replication forks from both origins were abnormal in the gp59-deficient infections. The lagging-strand from the initial fork was elongated as a new leading-strand in the retrograde direction without lagging-strand synthesis, whereas in the wild-type, leading- and lagging-strand synthesis appeared to be coupled. These results imply that gp59 inhibits the polymerase holoenzyme in vivo until the helicase-primase (gp41-gp61) complex is loaded, and we thereby refer to gp59 as a gatekeeper. We also found that all origin-replicative intermediates were absent in infections deficient in the helicase gp41 or the single-strand-binding protein gp32, regardless of whether gp59 was present or absent. These results argue that replication from the origin in vivo is dependent on both the helicase and single-strand-binding protein and demonstrate that the strong replication defect of gene 41 and 32 single mutants is not caused by gp59 inhibition of the polymerase.


AMPK as a Therapeutic Target for Treating Metabolic Diseases.
The AMP-activated protein kinase (AMPK) is a central regulator of multiple metabolic pathways and may have therapeutic importance for treating obesity, insulin resistance, type 2 diabetes (T2D), non-alcoholic fatty liver disease (NAFLD), and cardiovascular disease (CVD). Given the ubiquitous expression of AMPK, it has been a challenge to evaluate which tissue types may be most beneficially poised for mediating the positive metabolic effects of AMPK-centered treatments. In this review we evaluate the metabolic phenotypes of transgenic mouse models in which AMPK expression and function have been manipulated, and the impact this has on controlling lipid metabolism, glucose homeostasis, and inflammation. This information may be useful for guiding the development of AMPK-targeted therapeutics to treat chronic metabolic diseases.


Formation and stability of oxygen-rich bubbles that shape photosynthetic mats.
Gas release in photic-zone microbialites can lead to preservable morphological biosignatures. Here, we investigate the formation and stability of oxygen-rich bubbles enmeshed by filamentous cyanobacteria. Sub-millimetric and millimetric bubbles can be stable for weeks and even months. During this time, lithifying organic-rich laminae surrounding the bubbles can preserve the shape of bubbles. Cm-scale unstable bubbles support the growth of centimetric tubular towers with distinctly laminated mineralized walls. In environments that enable high photosynthetic rates, only small stable bubbles will be enclosed by a dense microbial mesh, while in deep waters extensive microbial mesh will cover even larger photosynthetic bubbles, increasing their preservation potential. Stable photosynthetic bubbles may be preserved as sub-millimeter and millimeter-diameter features with nearly circular cross-sections in the crests of some Proterozoic conical stromatolites, while centrimetric tubes formed around unstable bubbles provide a model for the formation of tubular carbonate microbialites that are not markedly depleted in (13)C.


The value of supplementary anatomy workshops for improving undergraduate performance.
With many anatomy courses within the medical curriculum suffering reductions in lecture and dissection time, supplementary programs in anatomy have shown to improve performance of medical students. This study was conducted to evaluate the efficacy of anatomy workshops on undergraduate anatomy grade performance. From 2012 to 2014, mean anatomy and physiology scores of medical students who attended anatomy workshops were compared to those who did not attend. Furthermore, mean scores in anatomy were also compared between students who attended one workshop to those who attended multiple workshops. Data analyses included comparing mean test scores using Student's t-test for normally distributed data. A total of 55 medical students were included in this study. Significant differences in both anatomy (p < 0.0001) and physiology (p = 0.0113) scores were found between medical students who attended the anatomy workshops versus those who did not. Similarly, medical students who attended multiple workshops, compared to those that attended a single workshop, obtained significantly higher anatomy grade performance (p = 0.0043). The results of this study are supported by comparable studies that have shown similar improvements in grade performance. In a time where reduced undergraduate medical teaching time is prevalent, supplementary education in the basic sciences, with an emphasis on anatomy, may prove a successful adjunct to the medical curriculum.


The association between pre-tonsillectomy education and postoperative emergency department returns: A retrospective cohort pilot study.
Tonsillectomy is the most common pediatric surgery in Canada. Post-tonsillectomy 30-day Emergency Department (ED) visit rates are higher than other pediatric day surgeries. To date, there have been no studies assessing whether additional preoperative education directed by Child Life Specialists impacts preventable ED visits. The primary aim of this study was to evaluate whether additional preoperative tonsillectomy education is feasible and is associated with fewer ED visits and admissions in the immediate postoperative period compared to standard care. The secondary aim of this study was to assess whether this education was associated with a lower ED visit rate for preventable causes. This is a retrospective chart review conducted in an academic tertiary pediatric hospital. Patients undergoing tonsillectomy surgery (from 2014 to 2019) were divided into three groups: consultation with a Child Life Specialist plus educational Booklet plus traditional surgeon-led education (CLS), educational Booklet plus surgeon-led education (Booklet), and traditional surgeon-led education (Traditional). The feasibility of the CLS education was assessed and the 30-day ED visit and admission rates were compared between groups. Visits included patients who returned to ED post-tonsillectomy and were not admitted, whereas admission included those who returned to ED and were admitted. 2081 patients undergoing tonsillectomy were included. 329 (15.8%) presented to the ED (within a median of 5 days), and 92 (4.4%) were admitted. ED visit/admission rates by group were: 14.7%/4.4% (CLS), 15.8%/4.1% (Booklet), and 16.2%/4.7% (Traditional) (p = 0.81/p = 0.84). The most common reason for return to ED was Hemorrhage (4.9%). Patients also returned to the ED for preventable reasons such as dehydration, pain, nausea/vomitting and fevers. Additional preoperative tonsillectomy education is feasible but is not associated with fewer ED visits and admissions, or fewer ED visits for preventable causes. Further research is needed to identify the optimal intervention to address the high post-tonsillectomy ED visit rate.


Endocrine cells in the gastrointestinal tracts of 3 teleostean species.
Endocrine cells of the gastrointestinal tracts of 3 teleostean species (Lepomis gibbosus, Morone americana and Notemigonus crysoleucas) were studied using several staining techniques: Masson-Hamperl argentaffin reaction, Singh's argyrophil reaction, Schmorl's ferricyanide test and MacConnail's lead hematoxylin; only the argyrophil method showed a positive reaction. In L. gibbosus and M. americana, endocrine cells of proximal (corporic) stomach were seen singly among the surface epithelial cells while those of distal (pyloric) stomach were more numerous and located in irregular patches. A small number of positive cells were found in the gastric glands of corporic stomach. A few argyrophil cells were seen among the surface intestinal epithelial cells of N. crysoleucas but none were seen in the intestines of L. gibbosus and M. americana.


FGF signaling network in the gastrointestinal tract (review).
Fibroblast growth factor (FGF) signals are transduced through FGF receptors (FGFRs) and FRS2/FRS3- SHP2 (PTPN11)-GRB2 docking protein complex to SOS-RAS-RAF-MAPKK-MAPK signaling cascade and GAB1/GAB2-PI3K-PDK-AKT/aPKC signaling cascade. The RAS approximately MAPK signaling cascade is implicated in cell growth and differentiation, the PI3K approximately AKT signaling cascade in cell survival and cell fate determination, and the PI3K approximately aPKC signaling cascade in cell polarity control. FGF18, FGF20 and SPRY4 are potent targets of the canonical WNT signaling pathway in the gastrointestinal tract. SPRY4 is the FGF signaling inhibitor functioning as negative feedback apparatus for the WNT/FGF-dependent epithelial proliferation. Recombinant FGF7 and FGF20 proteins are applicable for treatment of chemotherapy/radiation-induced mucosal injury, while recombinant FGF2 protein and FGF4 expression vector are applicable for therapeutic angiogenesis. Helicobacter pylori, a causative pathogen for peptic ulcer diseases, chronic atrophic gastritis and gastric cancer, injects bacterial proteins into gastric epithelial cells by using Type IV secretion system, which leads to FGF signaling activation through FGF2 upregulation as well as CagA-dependent SHP2 activation. FGFR2 gene is preferentially amplified and overexpressed in diffuse-type gastric cancer. PD173074 is a small-molecule inhibitor for FGFR, while RO4396686 and SU6668 are small-molecule inhibitors for FGFR and other tyrosine kinases. Cocktail therapy using multiple protein kinase inhibitors could enhance the therapeutic effects for gastrointestinal cancer through the reduction of recurrence associated with somatic mutations of drug-target genes. Single nucleotide polymorphism (SNP) and copy number polymorphism (CNP) of genes encoding FGF signaling molecules will be identified as novel risk factors of gastrointestinal cancer. Personalized prevention and personalized medicine based on the combination of genetic screening and novel therapeutic agents could dramatically improve the prognosis of cancer patients.


Quantification of bleeding volume using computed tomography and clinical complications after percutaneous renal biopsy.
Background: The aim of this study was to investigate specific bleeding volume after percutaneous renal biopsy (PRB) and the correlation between bleeding volume and clinical parameters. Methods: A retrospective study of 252 consecutive patients (153 male patients and 99 female patients) who underwent PRB at the Department of Nephrology, Japanese Red Cross Ishinomaki Hospital, between July 2013 and January 2016 was conducted. PRB was performed under ultrasound guidance using an automated spring-loaded biopsy device and a 16-cm, 16-gauge needle. Patients underwent computed tomography (CT) the day after PRB. Bleeding volume after PRB was evaluated using reconstructed CT data. Results: The median bleeding volume after PRB was 38 mL (25th-75th percentile, 18-85 mL), with ≥4 punctures identified as a risk factor for massive bleeding. The incidence rates of macrohematuria, transient hypotension and bladder obstruction were 14.3, 8.7 and 4.7%, respectively. Post-PRB blood transfusion and intervention were required in 4.7 and 0.8% of patients, respectively. Conclusion: Although it is difficult to assess the risk for massive bleeding prior to PRB, we do provide evidence of a specific increased risk with ≥4 puncture attempts, and recommend careful follow-up of these patients.


[Treatment of urogenital neoplasms in males].
The carcinomas of the male genitourinary tract--prostate, testicle, bladder, kidney and penis--are discussed with regard to incidence, prognosis and therapeutic possibilities. The results of drug treatment, especially for prostatic and non-seminoma testicular cancers, are presented in detail.


Van der Woude Syndrome With a Novel Mutation in the IRF6 Gene.
Van der Woude syndrome (VWS) is a rare autosomal dominant disease, first reported in the literature in 1845 by Demarquay and subsequently thoroughly described in 1954 by Van der Woude. Van der Woude Syndrome is the most common form of syndromic orofacial clefting and individuals with this syndrome account for 2% of all cleft cases. Van der Woude syndrome clinically presents with congenital lip pits. These lip pits occur on paramedian portion of the vermillion border of the lip. In VWS, congenital lip pits occur in concurrence with cleft lip and/or cleft palate and represent the most common clinical problem occurring in 80% of the patients. Lip pits result due to notching of the lips at an early stage of development with fixation of tissues at the base of the notch or they may result from a failure of complete union of embryonic lateral sulci of lip. Single lip sinuses without any cleft syndrome are rare; lower lip fistulas in VWS are generally asymptomatic, and surgical management is usually accomplished because of aesthetic concerns. However, in some cases, patients may complain of watery drainage or hypotonia of the lower lip. Herein, the authors report a novel frameshift mutation in IRF6 gene which may contribute to better understanding the genetic aspect of VWS.


Back to the Future: Lessons Learned From the 1918 Influenza Pandemic.
2018 marks the 100-year anniversary of the 1918 influenza pandemic, which killed ~50 million people worldwide. The severity of this pandemic resulted from a complex interplay between viral, host, and societal factors. Here, we review the viral, genetic and immune factors that contributed to the severity of the 1918 pandemic and discuss the implications for modern pandemic preparedness. We address unresolved questions of why the 1918 influenza H1N1 virus was more virulent than other influenza pandemics and why some people survived the 1918 pandemic and others succumbed to the infection. While current studies suggest that viral factors such as haemagglutinin and polymerase gene segments most likely contributed to a potent, dysregulated pro-inflammatory cytokine storm in victims of the pandemic, a shift in case-fatality for the 1918 pandemic toward young adults was most likely associated with the host's immune status. Lack of pre-existing virus-specific and/or cross-reactive antibodies and cellular immunity in children and young adults likely contributed to the high attack rate and rapid spread of the 1918 H1N1 virus. In contrast, lower mortality rate in in the older (>30 years) adult population points toward the beneficial effects of pre-existing cross-reactive immunity. In addition to the role of humoral and cellular immunity, there is a growing body of evidence to suggest that individual genetic differences, especially involving single-nucleotide polymorphisms (SNPs), contribute to differences in the severity of influenza virus infections. Co-infections with bacterial pathogens, and possibly measles and malaria, co-morbidities, malnutrition or obesity are also known to affect the severity of influenza disease, and likely influenced 1918 H1N1 disease severity and outcomes. Additionally, we also discuss the new challenges, such as changing population demographics, antibiotic resistance and climate change, which we will face in the context of any future influenza virus pandemic. In the last decade there has been a dramatic increase in the number of severe influenza virus strains entering the human population from animal reservoirs (including highly pathogenic H7N9 and H5N1 viruses). An understanding of past influenza virus pandemics and the lessons that we have learnt from them has therefore never been more pertinent.


Phat Queens Emerge Fashionably Late: Body Size and Condition Predict Timing of Spring Emergence for Queen Bumble Bees.
For insects, the timing of many life history events (phenology) depends on temperature cues. Body size is a critical mediator of insect responses to temperature, so may also influence phenology. The determinants of spring emergence of bumble bee queens are not well understood, but body size is likely important for several reasons. In fall, queens accumulate energy stores to fuel overwinter survival. Accumulation of fat stores prior to and depletion of fat stores during overwintering are likely size-dependent: larger queens can accumulate more lipids and have lower mass-specific metabolic rates. Therefore, larger queens and queens in relatively better condition may have delayed depletion of energy stores, allowing for later spring emergence. To test whether timing of spring emergence is associated with body size and condition, we captured 295 Bombus huntii queens in Laramie, WY, during the 2020 and 2021 growing seasons, weighed them, and measured intertegular width (a size metric unaffected by variation in feeding and hydration state). Early emerging queens were smaller than later emerging queens across years. Mass relative to intertegular width increased as the season progressed suggesting, as predicted, that body condition influences the timing of spring emergence for these crucial pollinators.


TAPAS: A Thresholding Approach for Probability Map Automatic Segmentation in Multiple Sclerosis.
Total brain white matter lesion (WML) volume is the most widely established magnetic resonance imaging (MRI) outcome measure in studies of multiple sclerosis (MS). To estimate WML volume, there are a number of automatic segmentation methods available, yet manual delineation remains the gold standard approach. Automatic approaches often yield a probability map to which a threshold is applied to create lesion segmentation masks. Unfortunately, few approaches systematically determine the threshold employed; many methods use a manually selected threshold, thus introducing human error and bias into the automated procedure. In this study, we propose and validate an automatic thresholding algorithm, Thresholding Approach for Probability Map Automatic Segmentation in Multiple Sclerosis (TAPAS), to obtain subject-specific threshold estimates for probability map automatic segmentation of T2-weighted (T2) hyperintense WMLs. Using multimodal MRI, the proposed method applies an automatic segmentation algorithm to obtain probability maps. We obtain the true subject-specific threshold that maximizes the Sørensen-Dice similarity coefficient (DSC). Then the subject-specific thresholds are modeled on a naive estimate of volume using a generalized additive model. Applying this model, we predict a subject-specific threshold in data not used for training. We ran a Monte Carlo-resampled split-sample cross-validation (100 validation sets) using two data sets: the first obtained from the Johns Hopkins Hospital (JHH) on a Philips 3 Tesla (3T) scanner (n = 94) and a second collected at the Brigham and Women's Hospital (BWH) using a Siemens 3T scanner (n = 40). By means of the proposed automated technique, in the JHH data we found an average reduction in subject-level absolute error of 0.1 mL per one mL increase in manual volume. Using Bland-Altman analysis, we found that volumetric bias associated with group-level thresholding was mitigated when applying TAPAS. The BWH data showed similar absolute error estimates using group-level thresholding or TAPAS likely since Bland-Altman analyses indicated no systematic biases associated with group or TAPAS volume estimates. The current study presents the first validated fully automated method for subject-specific threshold prediction to segment brain lesions.


Design of a 9K illumina BeadChip for polar bears (Ursus maritimus) from RAD and transcriptome sequencing.
Single-nucleotide polymorphisms (SNPs) offer numerous advantages over anonymous markers such as microsatellites, including improved estimation of population parameters, finer-scale resolution of population structure and more precise genomic dissection of quantitative traits. However, many SNPs are needed to equal the resolution of a single microsatellite, and reliable large-scale genotyping of SNPs remains a challenge in nonmodel species. Here, we document the creation of a 9K Illumina Infinium BeadChip for polar bears (Ursus maritimus), which will be used to investigate: (i) the fine-scale population structure among Canadian polar bears and (ii) the genomic architecture of phenotypic traits in the Western Hudson Bay subpopulation. To this end, we used restriction-site associated DNA (RAD) sequencing from 38 bears across their circumpolar range, as well as blood/fat transcriptome sequencing of 10 individuals from Western Hudson Bay. Six-thousand RAD SNPs and 3000 transcriptomic SNPs were selected for the chip, based primarily on genomic spacing and gene function respectively. Of the 9000 SNPs ordered from Illumina, 8042 were successfully printed, and - after genotyping 1450 polar bears - 5441 of these SNPs were found to be well clustered and polymorphic. Using this array, we show rapid linkage disequilibrium decay among polar bears, we demonstrate that in a subsample of 78 individuals, our SNPs detect known genetic structure more clearly than 24 microsatellites genotyped for the same individuals and that these results are not driven by the SNP ascertainment scheme. Here, we present one of the first large-scale genotyping resources designed for a threatened species.


Tracheobronchomegaly--the Mounier-Kuhn syndrome: report of two cases and review of the literature.
Tracheobronchomegaly (TBM) is a rare disorder of uncertain aetiology, characterized by marked dilatation of the trachea and main bronchi, bronchiectasis and recurrent lower respiratory tract infections. Two patients with TBM are presented. In one case, a marked decrease of elastic and smooth muscle tissue was present in the bronchial biopsy specimens, obtained by rigid bronchoscopy. The airways of the second patient were visualized using computed tomography. The dimensions of the airways of our patients are compared with the normal values supplied in the literature.


Risk factors for intraocular pressure elevation after descemet stripping automated endothelial keratoplasty.
To identify the incidence of and risk factors for intraocular pressure (IOP) elevation after Descemet stripping automated endothelial keratoplasty (DSAEK). Retrospective review was conducted of 68 consecutive DSAEK procedures alone, or in combination with phacoemulsification with intraocular lens implantation or exchange, performed by two surgeons at the University of Texas Southwestern Medical Center between 2005 and 2009. Eyes that developed IOP elevation above 21 mm Hg after DSAEK and requiring initiation or escalation of glaucoma therapy were evaluated. Thirty-seven (54%) eyes showed IOP elevation responsive to medical treatment by a mean follow-up of 11.38 +/- 7.81 months. Six (8.8%) eyes required glaucoma surgery. In the eyes, which developed elevated IOP, gonioscopy did not reveal any new peripheral anterior synechiae formation. Prolonged topical steroid usage, rebubbling, combined DSAEK/cataract surgery, or repeat DSAEK were not significant factors (P>0.05) for development of elevated IOP, but history of previous glaucoma or ocular hypertension (OHTN) was significant (P=0.007). Intraocular pressure elevation is not uncommon in eyes after DSAEK, but most cases can be controlled with conservative management. Intraocular pressure elevation post-DSAEK occurred by mechanisms other than peripheral anterior synechial angle closure. The only significant risk factor for development of elevated IOP in our series was a previous history of glaucoma or OHTN.


Pharmacological characterization of the novel nonpeptide orphanin FQ/nociceptin receptor agonist Ro 64-6198: rapid and reversible desensitization of the ORL1 receptor in vitro and lack of tolerance in vivo.
The novel nonpeptide orphanin FQ/nociceptin (OFQ/N) ligand [(1S,3aS)-8-(2,3,3a,4,5,6-hexahydro-1H-phenalen-1-yl)-1-phenyl-1,3,8-triaza-spiro[4.5]decan-4-one] (Ro 64-6198) was characterized in vitro and in vivo for its agonistic potential. Ro 64-6198 was 130- to 3500-fold selective for the OFQ/N receptor (ORL1) compared with opiate receptors. In the cAMP inhibition assay, Ro 64-6198 was a full agonist at the ORL1 and a partial agonist at the mu opiate receptor. When human embryonic kidney 293 cells stably expressing the human ORL1 receptor were pre-exposed (30 min) to either OFQ/N or Ro 64-6198, the ability of both agonists to inhibit forskolin-mediated cAMP accumulation was strongly reduced, indicating a functional desensitization of the second messenger cascade. However, acidic washes of OFQ/N-exposed cells fully restored the sensitivity of the ORL1 receptor for agonists. In contrast, the cAMP response in Ro 64-6198-exposed cells remained impaired after acidic washes, suggesting sustained receptor internalization at 30 min. In agreement with this finding, the number of cell-surface ORL1 receptors was significantly reduced after Ro 64-6198 pre-exposure, and this effect could be blocked with high sucrose concentrations. When Ro 64-6198 was chronically administered to rats, no signs of tolerance to its anxiolytic-like effects were detected following 15 days of daily drug exposure. In agreement with the behavioral results, Ro 64-6198 was able to reduce brain ORL1 binding sites in both acutely and chronically treated rats. Full recovery of ORL1 binding sites was observed 24 h after Ro 64-6198 administration with a t1/2 of approximately 5.5 h. These data show that nonpeptide agonists at the ORL1 receptor have a good clinical potential as anxiolytics without causing tolerance.


Respiratory disorders and allergies in tea packers.
The aim of this study was to determine whether respiratory symptoms or cross-shift declines in lung function were related to occupational exposure to tea dust. A cross-sectional epidemiological study was conducted at a tea-packing plant. Subjects completed a questionnaire, spirometry before and after a full work shift, skin prick testing and venipuncture. Among the 83% of the workers at the site who participated, the prevalences of asthma, wheezing, hay fever and atopy were similar to the general population. Work-related nasal symptoms were more commonly reported by blenders and operators. There were six (3.2%) subjects with a cross-shift decline in the forced expiratory volume in 1 s of > 10%. Specific immunoglobulin E antibodies to black or chamomile tea were observed in 10 (5.6%) employees. As there was little evidence of specific allergic sensitization to the tea varieties tested, the excess of work-related respiratory and nasal symptoms probably represented non-specific irritation.


Barbadin selectively modulates FPR2-mediated neutrophil functions independent of receptor endocytosis.
FPR2, a member of the family of G protein-coupled receptors (GPCRs), mediates neutrophil migration, a response that has been linked to β-arrestin recruitment. β-Arrestin regulates GPCR endocytosis and can also elicit non-canonical receptor signaling. To determine the poorly understood role of β-arrestin in FPR2 endocytosis and in NADPH-oxidase activation in neutrophils, Barbadin was used as a research tool in this study. Barbadin has been shown to bind the clathrin adaptor protein (AP2) and thereby prevent β-arrestin/AP2 interaction and β-arrestin-mediated GPCR endocytosis. In agreement with this, AP2/β-arrestin interaction induced by an FPR2-specific agonist was inhibited by Barbadin. Unexpectedly, however, Barbadin did not inhibit FPR2 endocytosis, indicating that a mechanism independent of β-arrestin/AP2 interaction may sustain FPR2 endocytosis. This was confirmed by the fact, that FPR2 also underwent agonist-promoted endocytosis in β-arrestin deficient cells, albeit at a diminished level as compared to wild type cells. Dissection of the Barbadin effects on FPR2-mediated neutrophil functions including NADPH-oxidase activation mediated release of reactive oxygen species (ROS) and chemotaxis revealed that Barbadin had no effect on chemotactic migration whereas the release of ROS was potentiated/primed. The effect of Barbadin on ROS production was reversible, independent of β-arrestin recruitment, and similar to that induced by latrunculin A. Taken together, our data demonstrate that endocytic uptake of FPR2 occurs independently of β-arrestin, while Barbadin selectively augments FPR2-mediated ROS production independently of receptor endocytosis. Given that Barbadin binds to AP2 and prevents the AP2/β-arrestin interaction, our results indicate a role for AP2 in FPR2-mediated ROS release from neutrophils.


Genetic Structure Analysis of a Collection of Tunisian Durum Wheat Germplasm.
The Tunisian durum wheat germplasm includes modern cultivars and traditional varieties that are still cultivated in areas where elite cultivars or intensive cultivation systems are not suitable. Within the frame of a collection program of the National Gene Bank of Tunisia (NGBT), durum wheat germplasm was collected from different Tunisian agro-ecological zones. The collected samples were studied using simple sequence repeats (SSRs) markers to explore the genetic diversity and evaluate the genetic structure in Tunisian germplasm. The results demonstrated significant diversity in the Tunisian durum wheat germplasm, with clear differentiation between traditional varieties and modern cultivars. The population structure analysis allowed the identification of five subpopulations, two of which appear to be more strongly represented in germplasm collected in central and southern Tunisia, where environmental conditions at critical development phases of the plant are harsher. Moreover these subpopulations are underrepresented in modern varieties, suggesting that traits of adaptation useful for breeding more resilient varieties might be present in central and southern germplasm. Moreover, our results will support, the activity of in situ on farm conservation of Tunisian durum wheat germplasm started by the National Gene Bank of Tunisia along with the ex situ approach.


Q Fever Knowledge, Attitudes and Vaccination Status of Australia's Veterinary Workforce in 2014.
Q fever, caused by Coxiella burnetii, is a serious zoonotic disease in humans with a worldwide distribution. Many species of animals are capable of transmitting C. burnetii, and consequently all veterinary workers are at risk for this disease. An effective Q fever vaccine has been readily available and used in Australia for many years in at-risk groups, and the European Centre for Disease Prevention and Control has recently also called for the use of this vaccine among at-risk groups in Europe. Little is known about attitudes towards this vaccine and vaccine uptake in veterinary workers. This study aimed to determine the Q fever vaccination status of veterinarians and veterinary nurses in Australia and to assess and compare the knowledge and attitudes towards Q fever disease and vaccination of each cohort. An online cross-sectional survey performed in 2014 targeted all veterinarians and veterinary nurses in Australia. Responses from 890 veterinarians and 852 veterinary nurses were obtained. Binary, ordinal and multinomial logistic regression were used to make comparisons between the two cohorts. The results showed that 74% of veterinarians had sought vaccination compared to only 29% of veterinary nurses. Barriers to vaccination among those not vaccinated did not differ between cohorts, and included a lack of perceived risk, financial expense, time constraints, and difficulty in finding a vaccine provider. Poor knowledge and awareness of Q fever disease and vaccination were additional and notable barriers for the veterinary nursing cohort, suggesting veterinary clinics and veterinarians may not be meeting their legal responsibility to educate staff about risks and risk prevention. Further evaluation is needed to identify the drivers behind seeking and recommending vaccination so that recommendations can be made to improve vaccine uptake.


Endotoxin-induced early gene expression in C3H/HeJ (Lpsd) macrophages.
C3H/HeJ (Lpsd) macrophages have been shown to respond to certain LPSs, especially from rough mutant bacteria. C3H/OuJ (Lpsn) macrophages are induced by wild-type LPS, rough LPS, or lipid A to express many genes, including TNF-alpha, TNFR-2, IL-1 beta, IP-10, D3, and D8. C3H/HeJ macrophages failed to induce any of these genes when cultured with wild-type LPS or synthetic lipid A, even when pretreated with IFN-gamma. However, rough mutant Salmonella minnesota Ra, Rc, and Rd LPS, and Escherichia coli D31 m3 Rd LPS induced Lpsd macrophages to express a subset of genes within the gene panel. Because bioactive preparations contained trace quantities of endotoxin protein(s), a deoxycholate-modified, phenol-water method was used to repurify rough LPS into an aqueous phase, and extract endotoxin proteins into a phenol phase. Repurified LPS failed to stimulate Lpsd macrophages; however, phenol fractions were approximately 10% as potent in Lpsd macrophages as crude rough LPS. Full potency was restored in C3H/HeJ macrophages when aqueous phase LPS and phenol-phase proteins were co-precipitated, suggesting that LPS and endotoxin proteins interact synergistically. Endotoxin proteins alone induced TNF-alpha, TNFR-2, and IL-1 beta, but not IP-10, D3, and D8 genes in both Lpsd and Lpsn macrophages. Tyrosine phosphorylation of three 41- to 47-kDa proteins was induced by endotoxin proteins, but not by LPS, in Lpsd macrophages. Thus, endotoxin proteins seem to activate a signaling pathway(s) that converges (distal to the Lps gene product) with a subset of LPS-signaling pathways.


Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
We demonstrate that cell-free extracts prepared from activated eggs of X. laevis by a method similar to that of Lohka and Masui initiate and complete semiconservative DNA replication of sperm nuclei and plasmid DNA. The efficiency of replication is comparable to that in the intact egg. Under optimal conditions 70%-100% of nuclei, and up to 38% of naked DNA molecules replicate completely. Genuine initiation of replication occurs rather than elongation of preformed primers or priming of irreversibly denatured templates. Rereplication of templates is observed under certain conditions. In addition to replicating DNA, these extracts also assemble nucleus-like structures from naked DNA.


Patients with Initial Negative RT-PCR and Typical Imaging of COVID-19: Clinical Implications.
The sensitivity of reverse transcriptase polymerase chain reaction (RT-PCR) has been questioned due to negative results in some patients who were strongly suspected of having coronavirus disease 2019 (COVID-19). The aim of our study was to analyze the prognosis of infected patients with initial negative RT-PCR in the emergency department (ED) during the COVID-19 outbreak. This study included two cohorts of adult inpatients admitted into the ED. All patients who were suspected to be infected with SARS-CoV-2 and who underwent a typical chest CT imaging were included. Thus, we studied two distinct cohorts: patients with positive RT-PCR (PCR+) and those with negative initial RT-PCR (PCR-). The data were analyzed using Bayesian methods. We included 66 patients in the PCR- group and 198 in the PCR+ group. The baseline characteristics did not differ except in terms of a proportion of lower chronic respiratory disease in the PCR- group. We noted a less severe clinical presentation in the PCR- group (lower respiratory rate, lower oxygen need and mechanical ventilation requirement). Hospital mortality (9.1% vs. 9.6%) did not differ between the two groups. Despite an initially less serious clinical presentation, the mortality of patients infected by SARS-CoV-2 with a negative RT-PCR did not differ from those with positive RT-PCR.