The application of a simplified system of criterions for audit to improve paediatric cardiologic and paediatric cardiac surgical care: evidence from a regional approach in Hungary.
To evaluate the quality of cardiac and surgical care provided for children with congenital cardiac malformations in the eastern county of Hungary. We used the method of clinical audit based in selection of criterions, developing five such criterions concerning timely diagnosis, access to treatment, and outcome. To examine compliance with these criterions, we analysed retrospectively the routine data relating to children living in Hajdu-Bihar County. The period of observation lasted from January, 1994, until December, 2001, and was divided into two epochs in order to evaluate any changes over time. In the first epoch, 28 infants, representing 0.1% of all newborns, died of congenital cardiac disease, with one of the malformations being recognised post mortem. In the second epoch, 21 infants died, representing 0.09% of newborns. In this group, each malformation was diagnosed before death. In each period, 6 infants died without having undergone cardiac surgery, and having no other non-cardiac disease. The overall postoperative mortality was 9.2% for the first period, and 4.6% for the second. The number of patients waiting for non-urgent repair had reduced significantly by the end of the second epoch. The results show that the timely diagnosis of congenital cardiac malformations is in line with accepted international standards. Conversely, the access to invasive treatment was limited over the period of observation, albeit that the waiting list was reduced significantly by the end of the second epoch. The postoperative mortality for those suffering congenital cardiac malformations is also comparable with international standards, except for mortality during infancy for treatment of complex anomalies. Our audit highlights the need for surgical repair of the more complex malformations during infancy.


Drug network characteristics as a predictor of cessation of drug use among adult injection drug users: a prospective study.
Few studies have examined recovery from opiate and cocaine dependence without treatment, referred to as "natural recovery," "spontaneous recovery," and "spontaneous remission." The present study examined the relationship between network characteristics and cessation of heroin, cocaine, and crack use in a sample of underclass inner-city injection drug users in Baltimore, Maryland. Participants were enrolled in an experimental human immunodeficiency virus (HIV) preventive intervention. Between the baseline and follow-up interviews, which averaged 5.2 months, 24 (7%) of 335 participants reported ceasing to use heroin, cocaine, and crack. Individuals who had reported cessation of drug use at follow-up had reported at baseline a smaller proportion of their network members with whom they used drugs (p < .02). Using multiple logistic regression analyses and adjusting for baseline drug use, enrollment in drug treatment, and demographic and background variables, cessation of drug use was associated with a lower proportion of personal network members in one's drug network (odds ratio [OR] = 25.4, p < .05). The data from this study suggest that network members have potential for social influence on the cessation of drug use.


Glycogen synthase kinase-3beta heterozygote knockout mice as a model of findings in postmortem schizophrenia brain or as a model of behaviors mimicking lithium action: negative results.
In mice glycogen synthase kinase (GSK)-3beta heterozygote knockout status was reported to cause reduced immobility in the Porsolt forced swim test and reduced amphetamine-induced hyperactivity, behaviors that mimic the effects of lithium. GSK-3beta protein and mRNA level and activity have been reported to be reduced in the postmortem brain of schizophrenia patients and this could suggest the involvement of GSK-3beta in the etiology of schizophrenia. However, apomorphine-induced stereotyping was reported to be unchanged in GSK-3beta heterozygote (HZ) knockout (KO) mice. As such behaviors are not always robust, study in another laboratory seemed indicated. Motor activity and coordination were assessed in the rotarod test. Behavior was studied in the following tests: pilocarpine-induced seizures model for lithium action, Porsolt forced swim test, tail suspension test, elevated plus-maze, large open field, startle response and prepulse inhibition of acoustic startle response, amphetamine-induced hyperactivity, and apomorphine-induced stereotypic climbing. We could not confirm the report that GSK-3beta HZ KO mice exhibit reduced immobility in the Porsolt forced swim or reduced amphetamine-induced hyperactivity in a manner mimicking the behavioral effects of lithium. We did not find increased apomorphine-induced stereotypic climbing or disruption of prepulse inhibition, suggesting that human postmortem findings regarding GSK-3beta in schizophrenia are not mediated by changes in dopamine receptors and are not the cause of prepulse inhibition deficits in schizophrenia. These data do not support the role of GSK-3beta in schizophrenia or in the mechanism of therapeutic action of lithium. Although differences in the genetic background of the GSK-3beta HZ KOs used in the present study compared with that of the previous study could be responsible, such results could suggest that the previously reported effects of GSK-3beta knockout on behavior are not robust.


Sex-specific morphological and physiological differences in the moss Ceratodon purpureus (Dicranales).
Dioecy and sexual dimorphism occur in many terrestrial plant species but are especially widespread among the bryophytes. Despite the prevalence of dioecy in non-vascular plants, surprisingly little is known about how fine-scale sex-specific cell and leaf morphological traits are correlated with sex-specific physiology and population sex ratios. Such data are critical to understanding the inter-relationship between sex-specific morphological and physiological characters and how their relationship influences population structure. In this study, these data types were assessed to determine how they vary across three populations within one moss species and whether fine-scale morphological traits scale up to physiological and sex ratio characteristics. Twenty cell-, leaf- and canopy-level traits and two photochemical measurements were compared between sexes and populations of the dioecious moss Ceratodon purpureus . Field population-expressed sex ratios were obtained for the same populations. Male and female plants differed in cell, leaf and photochemical measures. These sexual dimorphisms were female biased, with females having larger and thicker leaves and greater values for chlorophyll fluorescence-based, leaf photochemistry measurements than males. Female traits were also more variable than male traits. Interestingly, field population sex ratios were significantly male biased in two study populations and female biased in the third study population. The results demonstrate that the larger morphology and the greater physiological output of female C. purpureus gametophytes compared with males occurs across populations and is likely to have significant effects on resource allocation and biotic interactions. However, this high level of dimorphism does not explain population sex ratio variation in the three study populations tested. This research lays the groundwork for future studies on how differential sex-specific variation in cell and leaf traits influences bryophyte plant fitness.


From which level of competition in clubs are adolescents at greater risk of injury compared with outside-of-clubs athletes? A school-based study.
Sport practice is a key factor in a person's physical and mental health but, for adolescent athletes, some injuries lead to health problems in the long term. The literature provides multiple factors for understanding injury but does not give information about injury risk related to each level of play in a large sample of multisport athletes. This study investigates this relationship in 14- to 19-year-old adolescents. The survey on adolescents and health was conducted in classrooms of France, from February to March 2015. Only sports players were included in the analyses (n = 986). The levels of play were divided into five categories: outside of a club/no competition, club player/no competition, club player/local level, club player/state level and club player/national and higher level. A three-step binary logistic regression analysis with age, sex, type of sport, weekly hours of exposure, and level of play was used. During the past year, 48.1% of the adolescents were injured. Age and sex were not risk factors. The injury risk associated with the increases in level of play is higher than those related to the hours of exposure per week or the type of sport. In clubs, adolescents who do not compete or play at a local level showed no evidence of greater injury risk whereas state-level and national- and higher-level athletes were at greater risk than outside-of-club players (OR = 2.18, 95%CI = 1.13-3.94 and OR = 3.89, 95%CI = 2.07-7.31, respectively). Adolescents who play sports in clubs are clearly more exposed to injury than those who play outside of a club, mainly from state level. Age and sex are not related to injury. Future epidemiological studies should control adolescents' level of play. Special attention should be accorded to the injury risk of athletes playing at these levels of competition.


Targeting MerTK Enhances Adaptive Immune Responses After Radiation Therapy.
The role of MerTK, a member of the Tyro3-Axl-MerTK family of receptor tyrosine kinase, in the immune response to radiation therapy (RT) is unclear. We investigated immune-mediated tumor control after RT in murine models of colorectal and pancreatic adenocarcinoma using MerTK wild-type and knock-out hosts and whether inhibition of MerTK signaling with warfarin could replicate MerTK knock-out phenotypes. Wild-type and MerTK-/- BALB/c mice were grafted in the flanks with CT26 tumors and treated with computed tomography guided RT. The role of macrophages and CD8 T cells in the response to radiation were demonstrated with cell depletion studies. The role of MerTK in priming immune responses after RT alone and with agonist antibodies to the T cell costimulatory molecule OX40 was evaluated in a Panc02-SIY model antigen system. The effect of warfarin therapy on the in-field and abscopal response to RT was demonstrated in murine models of colorectal adenocarcinoma. The association between warfarin and progression-free survival for patients treated with SABR for early-stage non-small cell lung cancer was evaluated in a multi-institutional retrospective study. MerTK-/- hosts had better tumor control after RT compared with wild-type mice in a macrophage and CD8 T cell-dependent manner. MerTK-/- mice showed increased counts of tumor antigen-specific CD8 T cells in the peripheral blood after tumor-directed RT alone and in combination with agonist anti-OX40. Warfarin therapy phenocopied MerTK-/- for single-flank tumors treated with RT and improved abscopal responses for RT combined with anti-CTLA4. Patients on warfarin therapy when treated with SABR for non-small cell lung cancer had higher progression-free survival rates compared with non-warfarin users. MerTK inhibits adaptive immune responses after SABR. Because warfarin inhibits MerTK signaling and phenocopies genetic deletion of MerTK in mice, warfarin therapy may have beneficial effects in combination with SABR and immune therapy in patients with cancer.


Population-specific association between a polymorphic variant in ST18, encoding a pro-apoptotic molecule, and pemphigus vulgaris.
Pemphigus vulgaris (PV) is a severe autoimmune blistering disease caused by anti-epithelial antibodies, leading to disruption of cell-cell adhesion. Although the disease is exceedingly rare worldwide, it is known to be relatively prevalent in Jewish populations. The low prevalence of the disease represents a significant obstacle to a genome-wide approach to the mapping of susceptibility genes. We reasoned that the study of a genetically homogeneous cohort characterized by a high prevalence of PV may help exposing associated signals while reducing spurious results due to population sub-structure. We performed a genome-wide association study using 300K single-nucleotide polymorphisms (SNPs) in a case-control study of 100 PV patients of Jewish descent and 397 matched control individuals, followed by replication of significantly associated SNPs in three additional cohorts of Jewish, Egyptian, and German origin. In addition to the major histocompatibility complex locus, a genomic segment on 8q11.23 that spans the ST18 gene was also found to be significantly associated with PV. This association was confirmed in the Jewish and Egyptian replication sets but not in the German sample, suggesting that ST18-associated variants may predispose to PV in a population-specific manner. ST18 regulates apoptosis and inflammation, two processes of direct relevance to the pathogenesis of PV. Further supporting the relevance of ST18 to PV, we found this gene to be overexpressed in the skin of PV patients as compared with healthy individuals.


Mainstreaming Caenorhabditis elegans in experimental evolution.
Experimental evolution provides a powerful manipulative tool for probing evolutionary process and mechanism. As this approach to hypothesis testing has taken purchase in biology, so too has the number of experimental systems that use it, each with its own unique strengths and weaknesses. The depth of biological knowledge about Caenorhabditis nematodes, combined with their laboratory tractability, positions them well for exploiting experimental evolution in animal systems to understand deep questions in evolution and ecology, as well as in molecular genetics and systems biology. To date, Caenorhabditis elegans and related species have proved themselves in experimental evolution studies of the process of mutation, host-pathogen coevolution, mating system evolution and life-history theory. Yet these organisms are not broadly recognized for their utility for evolution experiments and remain underexploited. Here, we outline this experimental evolution work undertaken so far in Caenorhabditis, detail simple methodological tricks that can be exploited and identify research areas that are ripe for future discovery.


Comparison of heparin dosing based on actual body weight in non-obese, obese and morbidly obese critically ill patients.
Obesity is endemic in the United States and obese patients are at increased risk of thromboembolism but little data are available for dosing unfractionated heparin (UFH). We evaluated the relationship between obesity and UFH efficacy during critical illness by examining UFH infusions in non-obese, obese, and morbidly obese critically ill patients. Retrospective review of UFH infusions in non-obese, obese, and morbidly obese critically ill patients. Heparin was initiated without a bolus at 16 units/kg/h or 12 units/kg/h in obese and morbidly obese patients. Demographics, UFH dosage/therapy duration, laboratory values, and bleeding events were reviewed for patients receiving UFH for >24 h. Steady state (SS) was defined as the dosage that resulted in three consecutive activated partial thromboplastin times (aPTT) within target range. Sixty-two patients were analyzed including 21 non-obese (mean body mass index (BMI) 24.2 ± 2.3); 21 obese (BMI 34.1 ± 3.1); and 20 morbidly obese (mean BMI 55.3 ± 13.7). Patients had otherwise similar demographics. Although 92% had at least one therapeutic aPTT, only 55% of patients reached SS. Six patients developed minor bleeding, but no major hemorrhagic complications. The dosing of heparin based on actual body weight (units/kg/h) and time to first therapeutic aPTT was similar between groups, but dose was statistically higher at steady state in the non-obese (16.3 ± 5.3 non-obese, 11.6 ± 5.5 obese and 11.1 ± 1.2 obese, P = 0.01) with similar times to steady state. Dosing of UFH in morbidly obese and obese critically ill patients based on actual body weight and a reduced initial dose was associated with similar time to first therapeutic aPTT and steady state.


A plant-derived VLP influenza vaccine elicits a balanced immune response even in very old mice with co-morbidities.
The elderly are at high risk from influenza, in part because immunity wanes with age and through the accumulation of comorbidities. A novel plant-derived virus-like-particle (VLP) vaccine bearing influenza hemagglutinin can induce a balanced humoral and cellular response in old mice (16-18 months) while split virion vaccines elicit mostly antibodies. Because mice also collect comorbidities and lose immune competence as they age, we wished to determine how the plant-derived VLP vaccine would perform in animals approaching the end of their life-span. Old (24-26 months) female BALB/c mice received two intramuscular doses of H1-VLP vaccine, an inactivated H1N1 vaccine (IIV) (both based on A/H1N1/California/07/09) (3μg each) or PBS. Serum was collected on day 42 and humoral responses were measured by enzyme-linked immunosorbent assay (ELISA), microneutralization (MN) and hemagglutination inhibition (HI) assays. Influenza-specific splenocyte CD4+ & CD8+ T cell responses were measured by flow cytometry. Full body computed tomography (CT) and structured necropsies were performed on day 42. Comorbidities including reduced lung volume (kyphosis), masses, abscesses, etc. were assessed using a standard scoring system (1-21) and mice with scores ≥5 were considered to have important comorbidities. Overall, 53.3% of the animals had significant comorbidities. Three weeks post-boost, HI and MN titres were mostly undetectable but ELISA titres were significantly higher in the H1-VLP animals compared to the IIV group (GMT (95% CI): 961 (427, 2163) vs 425 (200, 903): p = 0.03). Both CD4+(TNFα, IFNγ) and CD8+ (IFNγ) T cell responses were also greater in the H1-VLP group than the IIV. Even in very old mice with comorbidities, the plant-made H1-VLP vaccine elicited a stronger and more balanced immune response than IIV. Animals with fewer comorbidities tended to have the better composite (humoral and cellular) responses. These novel vaccines have the potential to address some of the limitations of current vaccines in the elderly.


ETV5 regulates proliferation and cell cycle genes in the INS-1 (832/13) cell line independently of the concentration of secreted insulin.
The transcription factor E-twenty-six variant 5 (ETV5) regulates acute insulin secretion. Adequate insulin secretion is dependent on pancreatic β-cell size and cell proliferation, but the effects of ETV5 on proliferation, cell number, and viability, as well as its relationship with insulin secretion, have not been established yet. Here, we partially silenced ETV5 in the INS-1 (832/13) cell line by siRNA transfection and then measured secreted insulin concentration at different time points, observing similar levels to control cells. After 72 h of ETV5 silencing, we observed decreased cell number and proliferation, without any change in viability or apoptosis. Thus, partial silencing of ETV5 modulates cell proliferation in INS-1 (832/13) independently of secreted insulin levels via upregulation of E2F1 and of inhibitors of the cyclin/CDKs complexes (p21Cdkn1a , p27Cdkn1b , and p57Cdkn1c ) and downregulation of cell cycle activators (PAK3 and FOS).


Low Serum Angiopoietin-1, High Serum Angiopoietin-2, and High Ang-2/Ang-1 Protein Ratio are Associated with Early Onset Sepsis in Surinamese Newborns.
Vascular inflammation and leakage in sepsis is mediated by Angiopoietin-1 (Ang-1) and Angiopoietin-2 (Ang-2) and their phosphorylation of the endothelial Tie-2 receptor. This study investigates levels of Ang-1 and Ang-2 in newborns to gain insight in the vascular pathophysiology of early onset sepsis (EOS) within 72 h after birth. A prospective cohort study was performed among 71 Surinamese newborns treated with antibiotics for suspected EOS and 20 control newborns. Newborns with suspected EOS were divided in two groups: blood culture negative and positive EOS. Ang-1 and Ang-2 levels were measured in serum obtained at the start of antibiotic treatment and at re-evaluation after 48 to 72 h. In this cohort 8.5% of newborns had a positive blood culture. At the start of antibiotic treatment Ang-1 serum levels were lower (P < 0.01), and Ang-2 and Ang-2/Ang-1 serum protein ratios were higher (P < 0.01 and P < 0.01, respectively) in newborns with blood culture positive EOS than in controls. These levels were not dependent on timing of first blood draw after birth. After 48 to 72 h levels of Ang-1 further decreased in blood culture positive EOS, while in the other groups no change was observed. Our findings support the hypothesis that a disbalance in the Angiopoietins plays a role in the vascular pathophysiology of EOS.


Adjustable trapping position for single cells using voltage phase-controlled method.
We present an advanced technique improving upon the micron-sized particle trap integrated in biochip systems using a planar structure to generate an adjustable trapping position by utilizing voltage phase-controlled (VPC) method and negative dielectrophoresis (nDEP) theory in high conductivity physiological media. The designed planar and split structure is composed of independent components of measuring and trapping micro-electrodes. Through different voltage configurations on the device, the trapped position of single particles/cells was selected and adjusted in vertical and horizontal directions. The numerical simulations verify our theoretical predictions of the effects at the various voltages. It shows that the trapped position can be adjusted in the vertical (0 to 26 μm) and horizontal (0 to 74 μm) directions. In experiments, the single particles/cells is captured, measured, and then released, with the same process being repeated twice to demonstrate the precision of the positioning. The measurement results determined that particles at various heights result in different magnitude values, while the impedance error is less than 5% for the proposed electrode layout. Finally, the experiments are performed to verify that a particle/cell can be precisely trapped on the selected site in both the vertical and horizontal directions.


Impaired processing speed in categorical perception: Speech perception of children who stutter.
There have been controversial debates across multiple disciplines regarding the underlying mechanism of developmental stuttering. Stuttering is often related to issues in the speech production system; however, the presence and extent of a speech perception deficit is less clear. This study aimed to investigate the speech perception of children who stutter (CWS) using the categorical perception paradigm to examine their ability to categorize different acoustic variations of speech sounds into the same or different phonemic categories. In this study, 15 CWS and 16 children who do not stutter (CWNS) completed identification and discrimination tasks involving acoustic variations of Cantonese speech sounds in three stimulus contexts: consonants (voice onset times, VOTs), lexical tones, and vowels. The results showed similar categorical perception performance in boundary position and width in the identification task and similar d' scores in the discrimination task between the CWS and CWNS groups. However, the reaction times (RTs) were slower in the CWS group compared with the CWNS group in both tasks. Moreover, the CWS group had slower RTs in identifying stimuli located across categorical boundaries compared with stimuli located away from categorical boundaries. Overall, the data implied that the phoneme representation evaluated in speech perception might be intact in CWS as revealed by similar patterns in categorical perception as those in CWNS. However, the CWS group had slower processing speeds during categorical perception, which may indicate an insufficiency in accessing the phonemic representations in a timely manner, especially when the acoustic stimuli were ambiguous.


Verrucous cyst.
Human papillomavirus has been detected in a variety of cutaneous premalignant and malignant conditions, as well as in benign cutaneous warts. Changes consistent with epidermis papilloma virus infection occurring in epidermal cysts have recently been reported in the dermatologic literature only in plantar epidermal cysts. We report five examples of a form of cutaneous cyst that is histologically distinctive and that has not been previously reported, to our knowledge. We describe five cases of epidermal cysts showing marked verrucous changes of their stratified squamous inner linings, some of which also show proliferative changes. Human papillomavirus genomes were detected via polymerase chain reaction. None of the DNA probes for Southern blot hybridization demonstrated homology with the human papillomavirus types usually associated with cutaneous warts or carcinoma. We believe this cyst to represent a newly described manifestation of human papillomavirus infection, and propose the name verrucous cyst for this entity. Further studies may reveal the specific human papillomavirus type(s) associated with the verrucous cyst.


Falling through the cracks: challenges and opportunities for improving transitional care for persons with continuous complex care needs.
Persons with continuous complex care needs frequently require care in multiple settings. During transitions between settings, this population is particularly vulnerable to experiencing poor care quality and problems of care fragmentation. Despite how common these transitions have become, the challenges of improving care transitions have received little attention from policy makers, clinicians, and quality improvement entities. This article begins with a definition of transitional care and then discusses the nature of the problem, its prevalence, manifestations of poorly executed transitions, and potentially remediable barriers. Necessary elements for effective transitions are then presented, followed by promising new directions for quality improvement at the level of the delivery system, information technology, and national health policy. The article concludes with a proposed research agenda designed to advance the science of high-quality transitional care.


Prenatal Diagnosis of Vasa Previa by Routine Transvaginal Color Doppler.
The prenatal diagnosis of vasa previa is essential to achieving a safe delivery in patients who suffer from the condition. Transvaginal ultrasound with color Doppler performed at the time of a routine mid-trimester ultrasound is a valuable tool in terms of achieving a timely and accurate diagnosis of vasa previa.


Distal Versus Proximal Radial Artery Access for Cardiac Catheterization: 30-Day Outcomes of the DIPRA Study.
Background Proximal radial artery (pRA) access for cardiac catheterization is safe but can jeopardize subsequent use of the artery because of occlusion. Distal radial artery (dRA) access in the anatomical snuffbox preserves the radial artery, but safety and potential detrimental effects on hand function are unknown. Methods and Results In the DIPRA (Distal Versus Proximal Radial Artery Access for Cardiac Catheterization and Intervention) study, a single-center trial, 300 patients were randomized 1:1 to cardiac catheterization through dRA or pRA. The primary end point of change in hand function from baseline to 30 days was a composite of the QuickDASH (Quick Disabilities of the Arm, Shoulder and Hand) questionnaire, hand-grip test, and thumb forefinger pinch test. Secondary end points included access feasibility and complications; 254 of 300 patients completed follow-up at 30 days; of these, 128 were randomized to dRA and 126 to pRA with balanced demographic and procedural characteristics. Both groups had similar rates of access site bleeding (dRA 0% versus pRA 1.4%; P=0.25). Six patients with dRA failed access compared with 2 patients with pRA. Radial artery occlusion occurred in 2 pRA versus none in dRA. There were no significant differences in change in hand function, median hand-grip (dRA 0 [-3.2, 3.3] versus pRA 0.7 [-2.3, 3.3] kg; P=0.21), pinch-grip (dRA -0.3 [-1.2, 0.5] versus pRA 0 [-0.9, 0.9] kg; P=0.09), and QuickDASH (dRA 0 [-4.6, 2.3] versus pRA 0 [-4.6, 2.3] points, P=0.96). There was no significant difference in the composite of hand function between pRA and dRA. Conclusions dRA is a safe strategy for cardiac catheterization with a low complication rate. Compared with pRA, there is no increased risk of hand dysfunction at 30 days. Registration URL: https://www.ClinicalTrials.gov. Unique identifier: NCT04318990.


Failure of attenuated canine distemper virus (Rockborn strain) to suppress lymphocyte blastogenesis in dogs.
The attenuated Rockborn strain of canine distemper virus is commonly used in commercial vaccines. Since immunosuppression is a common feature of virulent (Snyder Hill) distemper virus infection of the dog, an evaluation of the cellular immune functions of dogs given inoculums of the less virulent Rockborn strain was done using lymphocyte blastogenesis responses to various mitogens. Unlike the viruslent Snyder Hill strain, the attenuated distemper virus did not alter lymphocyte blastogenesis responses to phytohemaglutinin (PHA) and pokeweed mitogen (PWM) which are considered in vitro correlates of T and B cell immunity.


Lifetime cigarette smoking is associated with striatal volume measures.
Nicotine, the primary addictive component of tobacco, affects the mammalian brain. Smokers' brains have smaller cortical grey matter volumes and/or lower densities compared with non-smokers'. Differences in subcortical structures like the striatum are however, less clear. A high concentration of nicotinic receptors makes the striatum a potential target for nicotine. In addition, striatal nuclei are essential components of the reward/reinforcement pathway involved in addiction. The aim of this study was to explore the relationship between striatal nuclei (caudate, putamen and nucleus accumbens area) volumes and lifetime smoking in a large community-based sample of 'young-old' individuals. Brain volumes were measured using a semi-automated method in 315 participants aged 64-70 years who were selected from a larger randomly sampled cohort and who consented to a magnetic resonance imaging scan. Multiple regression analysis was used to assess the relationship between striatal volumes and cigarette smoking measures while controlling for age, sex, intracranial and total brain volumes and general physical and mental health measures. Greater lifetime use of cigarettes (measured in pack-years) was associated with smaller left nucleus accumbens area volume (P = 0.018) and larger left putamen volume (P = 0.025). Greater putaminal volume was also associated with a lower age at smoking initiation (P = 0.004). In this generally healthy cohort, lifetime use of cigarettes is significantly associated with striatal volume measures. These changes could indicate predisposing factors for nicotine addiction, or an effect of chronic nicotine exposure or a combination of both.


Isolation of a lead tolerant novel bacterial species, Achromobacter sp. TL-3: assessment of bioflocculant activity.
Lead is one of the four heavy metals that has a profound damaging effects on human health. In the recent past there has been an increasing global concern for development of sustainable bioremediation technologies for detoxification of lead contaminant. Present investigation highlights for lead biosorption by a newly isolated novel bacterial species; Achromobacter sp. TL-3 strain, isolated from activated sludge samples contaminated with heavy metals (collected from oil refinery, Assam, North-East India). For isolation of lead tolerant bacteria, sludge samples were enriched into Luria Broth medium supplemented separately with a range of lead nitrate; 250, 500, 750, 1000, 1250 and 1500 ppm respectively. The bacterial consortium that could tolerate 1500 ppm of lead nitrate was selected further for purification of lead tolerant bacterial isolates. Purified lead tolerant bacterial isolates were then eventually inoculated into production medium supplemented with ethanol and glycerol as carbon and energy source to investigate for bioflocculant production. Bioflocculant production was estimated by monitoring the potential of lead tolerant bacterial isolate to flocculate Kaolin clay in presence of 1% CaCl2. Compared to other isolates, TL-3 isolate demonstrated for maximum bioflocculant activity of 95% and thus was identified based on 16S rRNA gene sequence analysis. TL3 isolate revealed maximum homology (98%) with Achromobacter sp. and thus designated as Achromobacter sp. TL-3. Bioflocculant activity of TL-3 isolate was correlated with the change in pH and growth. Achromobacter sp. TL-3 has significant potential for lead biosorption and can be effectively employed for detoxification of lead contaminated waste effluents/waste waters.


Antiherpes virus-specific treatment and cognition in schizophrenia: a test-of-concept randomized double-blind placebo-controlled trial.
To test our hypothesis that valacyclovir, an antiherpes virus-specific medication, added to antipsychotics (APs) would improve cognitive performance and psychopathology among schizophrenia subjects exposed to neurotropic herpes simplex virus, type 1 (HSV1). Using a double-blind placebo-controlled design, we randomized 24 HSV1-seropositive schizophrenia subjects to receive either valacyclovir (n = 12) or placebo (n = 12) for 18 weeks in addition to stable doses of APs. Valacyclovir dose was stabilized at 1.5 g twice daily orally. At each visit, subjects were evaluated for severity of psychopathology and side effects using standardized scales and a study-specific semistructured checklist. A computerized neurocognitive battery validated on both schizophrenia and healthy subjects was administered at baseline and follow-up. Intent-to-treat analysis, using linear regression models that included all randomized subjects, were used to examine differential changes in cognition and psychopathology scores over 18 weeks between valacyclovir and placebo, accounting for placebo response. Valacyclovir group improved in verbal memory, working memory, and visual object learning compared with placebo group. The effect sizes (Cohen's d) were 0.79 for working memory, 1.14 for immediate verbal memory, and 0.97 for the visual object learning. Psychotic symptom severity did not improve. Supplemental valacyclovir may alleviate impairments in cognitive domains that are often observed in schizophrenia but not psychotic symptoms in those exposed to HSV1. If replicated, this approach could provide a novel strategy to treat cognitive impairments in a subgroup of schizophrenia subjects who can be reliably identified using a blood test.


Atherosclerosis in smokers.
Cigarette smoking was found to be associated with an increased extent of raised lesions of the aorta and aortic calcification. The apparent association with aortic calcification was mainly due to co-existing alcohol consumption. The apparent association of smoking with the extent of coronary calcification was also related to co-existing alcohol consumption; smoking alone tended to be negatively associated with this lesion. There was no clear association between smoking and coronary stenosis, myocardial infarction, and heart weight.


Quality of care and outcomes in women hospitalized for heart failure.
Although women account for a significant proportion of heart failure (HF) hospitalizations, data on the quality of care and in-hospital outcomes in women are limited. We examined The Joint Commission performance measures, other quality metrics, length of stay, and in-hospital mortality in women using 99 841 HF admissions (January 2005 to June 2009) at 248 hospitals participating in the American Heart Association Get With The Guidelines-Heart Failure registry. Women accounted for 50% of the HF admissions and were older (mean age, 74±14 versus 69±14 years), more likely to have hypertension (77% versus 72%), and less likely to have coronary disease (44% versus 53%) or renal insufficiency (18% versus 23%) than men (all P<0.001). The presenting symptoms were similar to men, but women had higher admission systolic blood pressure (mean, 144±31 versus 137±30 mm Hg; P<0.001) and ejection fraction (mean, 0.44±0.17% versus 0.34±0.16%; P<0.001). After adjustment for baseline differences, eligible women were less likely than men to have measurement of left ventricular function (adjusted odds ratio [OR], 0.81; 95% CI, 0.76 to 0.86) and to receive anticoagulation for atrial fibrillation (adjusted OR, 0.91; 95% CI, 0.86 to 0.96) or implantable cardioverter-defibrillators (adjusted OR, 0.70; 95% CI, 0.65 to 0.75) but were as likely to receive discharge instructions, angiotensin-converting enzyme inhibitors/angiotensin receptor blockers, β-blockers, and smoking cessation counseling at discharge. Although the median length of stay was 4 days, women were more likely than men to be hospitalized >4 days (adjusted OR, 1.13; 95% CI, 1.10 to 1.16) and >7 days (adjusted OR, 1.07; 95% CI, 1.04 to 1.11). Women had comparable in-hospital mortality to men (adjusted OR, 1.05; 95% CI, 0.96 to 1.14). Compared to men, women hospitalized for HF differ in many clinical characteristics and length of stay but have similar clinical presentations, receive similar quality of care for most but not all measures, and experience similar in-hospital mortality.


An outbreak of pertussis in rural Texas: an example of the resurgence of the disease in the United States.
During 2012, an increase in the number of pertussis cases or outbreaks was reported among most states within the United States. The majority of these cases included previously vaccinated children between the ages of 7-10 years. This underscores the growing concern regarding current immunization practices and vaccine efficacy, especially as it pertains to pertussis prevention within this age group. In the fall of 2012, an outbreak of pertussis occurred within a school district in a rural Texas county that was reflective of this national pattern. Our objective is to describe this outbreak, highlight the similarities with the national trend, and identify strategies for better disease prevention. The cases in this outbreak were interviewed and laboratory testing done. Information regarding exposure and immunization history among cases was obtained. Immunization audits of the affected institutions were also conducted. We performed a descriptive analysis of the collected data using EPI-INFO software v.3.5.3. A total of 34 cases were identified in this outbreak, of which 23 were PCR confirmed and 11 were epidemiologically linked. Ages ranged from 5 months to 12 years, and 62 % were among children aged 7-10 years. All cases were up-to-date on their pertussis vaccinations. Immunization coverage rate was over 90 % within each of the affected institutions. The characteristics of this outbreak bear striking similarities to the current national trend in terms of age groups and immunization status of the affected cases. Increased focus on this vulnerable target group, including heightened scrutiny of vaccine efficacy and delivery, is indicated.


Trend of Spine Surgeries in the Outpatient Hospital Setting Versus Ambulatory Surgical Center.
Retrospective database review. The aim of the present study was to examine how often spine surgery is being performed in an outpatient hospital setting versus a more "true" ambulatory setting, specifically ambulatory surgery centers (ASCs) in which admission and discharge are required on the same calendar day. Recent studies have assessed the safety, satisfactory clinical outcomes, and increasing utilization of both cervical and lumbar spinal surgeries performed in the outpatient setting. No studies have delineated between true ambulatory settings and outpatient hospitals when assessing the rates of these procedures. A retrospective review of the Truven Health Marketscan Research Databases was conducted for patients undergoing spine operations between 2003 and 2014. The frequency of each Common Procedural Terminology code was identified per year, and then categorized into each of "inpatient hospital," "outpatient hospital," or "ASC" in states that clearly define ASCs as facilities in which patients are discharged on the same calendar day of the operation, and do not stay overnight. During the period between 2003 and 2014, the procedures that had the most dramatic increase as an outpatient hospital procedure included lumbar decompression laminotomy first level (18.7%-68.5%) and posterior cervical decompression laminectomy without facetectomy discectomy first level (0%-46.7%). ASC procedures had more modest increases during this time period with the most significant increases in lumbar decompression laminotomy first level (0.7%-10.6%) and posterior cervical decompression laminotomy first level (0%-23.4%). "True" ambulatory surgeries are not increasing at the same rate as outpatient procedures with 23-hour observation capacity. Although prior studies have demonstrated the safety of outpatient spine surgery, one possible reason for this trend may be that surgeons feel that this safety may not be comparable to that of other outpatient procedures. 3.


Structure of the 116-kDa polypeptide of the clathrin-coated vesicle/synaptic vesicle proton pump.
A 116-kDa polypeptide has recently been found to be a common component of vacuolar proton pumps isolated from a variety of sources. The 116-kDa subunit of the proton pump was purified from clathrin-coated vesicles of bovine brain, and internal sequences were obtained from proteolytic peptides. Oligonucleotide probes designed from these peptide sequences were utilized in polymerase chain reactions to isolate partial bovine cDNA clones for the protein. Sequences from these were then utilized to isolate rat brain cDNA clones containing the full-length coding region. RNA blots indicate the presence of an abundant 3.9-kilobase message for the 116-kDa subunit in brain, and primer extension analysis demonstrates that the cloned sequence is full-length. The rat cDNA sequences predict synthesis of a protein of 96,267 Da. Analysis of the deduced amino acid sequence of the 116-kDa subunit suggests that it consists of two fundamental domains: a hydrophilic amino-terminal half that is composed of greater than 30% charged residues, and a hydrophobic carboxyl-terminal half that contains at least six transmembrane regions. The structural properties of the 116-kDa proton pump polypeptide agree well with its proposed function in coupling ATP hydrolysis by the cytoplasmic subunits to proton translocation by the intramembranous components of the pump.


Reversible dissociation of the poliovirus replication complex: functions and interactions of its components in viral RNA synthesis.
Membrane-bound replication complexes containing transcriptionally active replicative intermediates (RI) can be isolated from poliovirus-infected HEp-2 cells and consist of rosette-like structures of virus-induced vesicles surrounding the replicating viral RNA. At low ionic strength and low temperature, the rosettes reversibly dissociate into individual tubulated vesicles. As determined by immunoelectron microscopy and immunoprecipitation, the vesicles carry a set of viral structural and nonstructural proteins as well as RI RNA. At 30 degrees C, the vesicles reassociate into rosettes synthesizing plus-strand RNA in the RI. The in vitro transcriptional activities of rosettes and vesicles kept separated by high dilution were assessed by an RNase protection assay. The synthesis of the first 178 nucleotides at the 5' end of the plus strand was considered to reflect initiation, and the detection of a 530-nucleotide fragment in the P2 genomic region was considered to reflect elongation. It could be shown that the initiation and elongation of plus strands on individual vesicles are comparable to those in rosettes, with initiation proceeding in de novo-assembled initiation complexes. By use of detergent treatment it was found that initiation, but not elongation, is dependent on vesicular membranes.


Noninvasive Arterial Testing in Patients With Diabetes: A Guide for Foot and Ankle Surgeons.
This study was designed to compare the findings of noninvasive arterial testing in patients with and without diabetic foot pathology. The ABI (ankle brachial index), TBI (toe brachial index), and great toe pressures were measured in 207 patients. PAD (peripheral artery disease) was defined as an ABI < 0.91 on either extremity or a TBI < 0.7. PAD was identified in 103 of the 207 patients (49.8%), 80 patients with diabetic foot pathology and 23 patients with nondiabetic foot pathology. Although patients with diabetic foot pathology were 1.4 times more likely to have PAD compared to patients without diabetic pathology, this increased risk was not statistically significant (OR 1.41 [95% CI 0.75-2.64], P = .28). Patients with PAD and diabetic foot pathology were 4.9 times more likely to have ischemia (toe pressure < 60 mm Hg) than patients with PAD and nondiabetic foot pathology (OR 4.93 [95% CI 1.35-17.94], P < .05). Patients on dialysis had a 7.3 times increased likelihood of having PAD compared to patients not on dialysis (OR 7.3 [95% CI 1.6-33.6], P < .01). Patients with absent pedal pulses were 4.9 more likely to have PAD than patients with normal pulses (OR 4.9 [95% CI 2.6-9.4], P < .0001). PAD was identified in 97 of 188 patients (51.6%) with peripheral neuropathy compared to 6 of 19 patients (31.5%) without peripheral neuropathy (OR 2.31 [95% CI 0.84-6.33], P = .10). Combining the ABI with TBI improved the ability to diagnose PAD in diabetic patients because the ABI has high specificity (low false positives) and the TBI has high sensitivity (low false negatives). The TBI was more reliable in patients with noncompressible arteries, medial artery calcinosis and/or neuropathy. Due to the relative incompressibility of calcified distal arteries in patients with DM, the ABI may be within normal limits in patients with PAD. This false negative result may lead surgeons to assume that normal perfusion is present.


The Impact of Presentation Mode and Technology on Reading Comprehension among Blind and Sighted Individuals.
Technological advancements have made distributing reading materials in audio formats more common. Investigating how presentation mode impacts comprehension among sighted and blind individuals will inform the distribution of information to enhance comprehension. The aims were (1) to investigate the hypothesis that reading comprehension is enhanced by increased physical engagement and cognitive effort through text or braille and (2) to explore how assistive technology impacts comprehension for blind individuals. In a within-subjects design, 31 sighted and 34 blind participants read and listened to scientific passages and verbally answered free-response questions about what they read and heard. For sighted participants, passages were presented in text and human voice actor recordings. For blind participants, passages were presented with hard-copy braille, a refreshable braille display, voice actor recordings, and a screen reader. Comprehension scores were analyzed using mixed-effects regression and pairwise comparisons on the estimated marginal means. In study 1, the comprehension difference between text or hard-copy braille and the voice actor formats was assessed to address the first aim. Sighted participants had better comprehension with text (mean, 74.8%; 95% confidence interval [CI], 70.5 to 79.1%) than with a voice actor (mean, 69.7%; 95% CI, 65.4 to 74.0%; P = .02), and blind participants had superior comprehension with hard-copy braille (mean, 70.4%; 95% CI, 63.3 to 77.5%) than with a voice actor (mean, 61.9%; 95% CI, 54.7 to 69.0%; P = .03). In study 2, the comprehension differences among blind participants between the four formats were investigated to address the second aim. Comprehension was better with hard-copy braille (mean, 70.6%; 95% CI, 63.4 to 77.7%) than with a screen reader (mean, 60.7%; 95% CI, 53.5 to 67.9%; P = .02) and better with a braille display (mean, 69.7%; 95% CI, 62.5 to 76.9%) than with a screen reader (P = .04). Study 1 supports the hypothesis that more physically engaging tasks enhance comprehension, and study 2 suggests that listening to scientific materials using a synthesized voice may reduce comprehension ability compared with hard-copy braille and braille displays.


Evaluation of Bayesian models of substitution rate evolution--parental guidance versus mutual independence.
We have evaluated the performance of two classes of probabilistic models for substitution rate variation over phylogenetic trees. In the first class, branch rates are considered to be independent and identically distributed (i.i.d.) stochastic variables. Three versions with respect to the underlying distribution (Gamma, Inverse Gaussian, and LogNormal) are considered. The i.i.d. models are compared with the autocorrelated (AC) model, where rates of adjacent nodes in the tree are AC, so that a node rate is LogNormal distributed around the rate of the parent node. The performance of different models is evaluated using three empirical data sets. For all data sets, it was clear that all tested models extracted substantial knowledge from data when posterior divergence time distributions were compared with the prior distributions and, furthermore, that they clearly outperformed a molecular clock. Moreover, the descriptive power of the i.i.d. models, as evaluated by Bayes factors, was either equal to or clearly better than that of the AC model. The latter effect increased with extended taxon sampling. Likewise, under none of the models could we find compelling evidence, in any of the data sets, for rate correlation between adjacent branches/nodes. These findings challenge previous suggestions of universality of autocorrelation in sequence evolution. We also performed an additional comparison with a divergence time prior including calibration information from fossil evidence. Adding fossil information to the prior had negligible effect on Bayes factors and mainly affected the width of the posterior distribution of the divergence times, whereas the relative position of the mean divergence times were largely unaffected.


A novel C2H2 transcription factor that regulates gliA expression interdependently with GliZ in Aspergillus fumigatus.
Secondary metabolites are produced by numerous organisms and can either be beneficial, benign, or harmful to humans. Genes involved in the synthesis and transport of these secondary metabolites are frequently found in gene clusters, which are often coordinately regulated, being almost exclusively dependent on transcription factors that are located within the clusters themselves. Gliotoxin, which is produced by a variety of Aspergillus species, Trichoderma species, and Penicillium species, exhibits immunosuppressive properties and has therefore been the subject of research for many laboratories. There have been a few proteins shown to regulate the gliotoxin cluster, most notably GliZ, a Zn2Cys6 binuclear finger transcription factor that lies within the cluster, and LaeA, a putative methyltransferase that globally regulates secondary metabolism clusters within numerous fungal species. Using a high-copy inducer screen in A. fumigatus, our lab has identified a novel C2H2 transcription factor, which plays an important role in regulating the gliotoxin biosynthetic cluster. This transcription factor, named GipA, induces gliotoxin production when present in extra copies. Furthermore, loss of gipA reduces gliotoxin production significantly. Through protein binding microarray and mutagenesis, we have identified a DNA binding site recognized by GipA that is in extremely close proximity to a potential GliZ DNA binding site in the 5' untranslated region of gliA, which encodes an efflux pump within the gliotoxin cluster. Not surprisingly, GliZ and GipA appear to work in an interdependent fashion to positively control gliA expression.


Distinct gastric phenotype in patients with pathogenic variants in SMAD4: A nationwide cross-sectional study.
Background and study aims In most patients with juvenile polyposis Syndrome, it is possible to detect a pathogenic germline variant in SMAD4 or BMPR1A . It is well known that patients with a pathogenic variant in SMAD4 have a higher risk of gastric polyposis and gastric cancer compared to BMPR1A carriers, but the natural history of gastric involvement is poorly described. We aimed to systematically review endoscopic and histopathological gastric findings in Danish patients with pathogenic variants in SMAD4. Patients and methods This was a retrospective, cross-sectional study including endoscopic and histological gastric findings in all known Danish patients with pathogenic variants in SMAD4 . The patients were identified by data from various registries as well as from clinical genetic departments and laboratories. Results We identified 41 patients (2-72 years) with a pathogenic SMAD4 variant . In 31 patients, we were able to retrieve information on upper gastrointestinal endoscopy. Eighty-seven percent had at least one gastric abnormality including erythema (72 %) and edema (72 %). Half of the patients also had vulnerability of the mucosa and 68 % had gastric polyposis. An increasing frequency of abnormalities were observed with increasing age. Gastric cancer was diagnosed in 5 % of the cases and 22 % had a gastrectomy mainly because of massive polyposis. Conclusions This study showed that most patients with pathogenic SMAD4 variants have a distinct phenotype of the gastric mucosa, and with an increasing severity in the elderly patients. These findings provide new insights into the natural history of gastric manifestations in patients with pathogenic SMAD4 variants.


[Resistance, exchange and capacitance vessels of the skeletal musculature in acute hypoxic hypoxia].
In cats under conditions of autoperfusion of hemodynamically isolated m. gastrocnemius, acute hypoxia (10% O2 in nitrogen) increased the precapillary and decreased the postcapillary resistance, reduced the capillary hydrostatic pressure and induced shifts of the capillary filtration coefficient (CFC) by 30% on the average. In cross-circulation performed for analysis of the neurogenic component in the vascular responses, analogous changes of the above parameters were revealed, the CFC decreasing, however. In denervated vascular bed of the m. gastrocnemius, the acute hypoxia decreased the precapillary and increased the postcapillary resistance, the hydrostatic pressure and the CFC increasing. In decentralized vessels of skeletal muscle, hypoxia induced prevalence of filtration while absorption of fluid prevailed in perfusion of the preparation with donor's arterial blood and in recipient's hypoxia.


Establishment of epilepsy surgery in Peru.
Epilepsy surgery is a well-established treatment for certain types of intractable epilepsy. While there is a relatively high number of epilepsy surgery centers in Canada and the United States, the same cannot be said about many other parts of the world, such as South America. Although there are notable exceptions, such as Brazil and Colombia, formal epilepsy surgery centers have been lacking in many South American countries, including Peru. Although sporadic cases have been performed in Peru, there was no formal epilepsy surgery center in the country as of 2011. Beginning in 2008, with the support of the Partnering Epilepsy Centers in America program of the North American Commission of the International League Against Epilepsy, the Epilepsy Program at Western University in London, Canada, was partnered with the Department of Epilepsy at the Instituto Nacional de Ciencias Neurologicas in Lima. This was the beginning of a long-term relationship that culminated in the establishment of the first 2 formal epilepsy surgery centers in Peru. The purpose of this communication is to briefly summarize the establishment of 2 independent epilepsy surgery programs in Peru and to document the methods with which this accomplishment was achieved.


Optimal and safe upper limits of iodine intake for early pregnancy in iodine-sufficient regions: a cross-sectional study of 7190 pregnant women in China.
The WHO Technical Consultation recommends urinary iodine concentrations (UIC) from 250 to 499 μg/L as more-than-adequate iodine intake and UIC ≥ 500 μg/L as excessive iodine for pregnant and lactating women, but scientific evidence for this is weak. We investigated optimal and safe ranges of iodine intake during early pregnancy in an iodine-sufficient region of China. Seven thousand one hundred ninety pregnant women at 4-8 weeks gestation were investigated and their UIC, serum thyroid stimulating hormone (TSH), free thyroxine (FT4), thyroid-peroxidase antibody (TPOAb), thyroglobulin antibody (TgAb), and thyroglobulin (Tg) were measured. The prevalence of overt hypothyroidism was lowest in the group with UIC 150-249 μg/L, which corresponded to the lowest serum Tg concentration (10.18 μg/L). Prevalences of subclinical hypothyroidism (2.4%) and isolated hypothyroxinemia (1.7%) were lower in the group with UIC 150-249 μg/L. Multivariate logistic regression indicated that more-than-adequate iodine intake (UIC 250-499 μg/L) and excessive iodine intake (UIC ≥ 500 μg/L) were associated with a 1.72-fold and a 2.17-fold increased risk of subclinical hypothyroidism, respectively. Meanwhile, excessive iodine intake was associated with a 2.85-fold increased risk of isolated hypothyroxinemia. Moreover, the prevalence of TPOAb positivity and TgAb positivity presented a U-shaped curve, ranging from mild iodine deficiency to iodine excess. The upper limit of iodine intake during early pregnancy in an iodine-sufficient region should not exceed UIC 250 μg/L, because this is associated with a significantly high risk of subclinical hypothyroidism, and a UIC of 500 μg/L should not be exceeded, as it is associated with a significantly high risk of isolated hypothyroxinemia.


Telephone directory listings of presumptive Chinese surnames: an appropriate sampling frame for a dispersed population with characteristic surnames.
Choosing an appropriate sampling frame for an epidemiologic study depends on the characteristics of the population under investigation and the sources available for identification of potential subjects. Not all countries maintain (or permit sampling from) population registers. In studies of dispersed populations, electoral rolls may lead to sampling bias, and random-digit dialing is inefficient. A dispersed population characterized by distinctive surnames might be appropriately identified and sampled through telephone directory listings, however. The method proved feasible and efficient for a prevalence study of health-related characteristics of Chinese residents in Melbourne, Australia, yielding a sample demographically comparable to the corresponding census population.


Temporal dependence of local motion induced shifts in perceived position.
It has been shown that a moving visual pattern can influence the perceived position of outlying, briefly flashed objects. Using a rotating bar as an inducing stimulus we observed a shift, in the direction of motion, of the perceived position of small bars flashed together on either side of the moving bar. The greatest shift occurred when the 13 ms flashes were presented 60 ms before the rotating bar came closest to their locations. By varying rotation speed we showed that the peak effect was determined by the temporal rather than the spatial interval. The motion induced shift could be attenuated by introducing background flickering dots. The perceived shift decreased with distance from motion when the eccentricity of the flashes was kept constant. We conclude that the shift reflects feedback to primary visual cortex from motion selective cells in extrastriate cortex with receptive fields that overlap the retinal location of the flash.


Expression of alternate growth hormone receptor messenger RNA in ovary and uterus of cattle.
Growth hormone (GH) receptor mRNA is found within the corpus luteum and endometrium of cattle. However, binding sites for placental lactogen (PL) but not GH are found within these tissues. The objectives were to isolate cDNA for the GH receptor within the reproductive tissues of cattle and to examine these cDNA as potential variants of the GH receptor that bind PL. Ten cDNA clones were isolated from a bovine endometrial cDNA library with a 32P-labeled cDNA of the GH receptor extracellular domain. On the basis of restriction enzyme digestion, 2 of the 10 cDNA clones contained exon 1. The DNA sequence of these clones was determined by dideoxy nucleotide sequencing. The exon 1 DNA sequence of each clone (exon 1B) was different from the previously reported exon 1 for the bovine GH receptor cDNA isolated from liver (exon 1A). Analyses of these cDNA sequences showed that exon 1B contained significant homology with placental forms of the GH receptor found in mouse and human. Unlike the human cDNA, the bovine cDNA isolated from endometrium contained an intact third exon. Amplification of GH receptor mRNA by reverse transcriptase polymerase chain reaction, with exon 1A- and 1B-specific forward primers, showed that exon 1B was expressed in liver, corpus luteum, ovary, endometrium, and myometrium. Exon 1A was found almost exclusively in liver, and little was found in reproductive tissues. The predicted initiation of protein coding for the GH receptor was within the second exon and was not changed by the splicing of the alternate first exon. This suggests that the alternate mRNA results in the expression of intact GH receptor protein that is similar to that found within liver. Alternative promoters (1B) may control the expression of the receptor outside the liver. Furthermore, mechanisms other than the differential splicing of GH receptor protein may dictate the specificity of PL binding within the endometrium and corpus luteum.


Mammary carcinogenesis in the rat by topical application of fluorenylhydroxamic acids and their acetates.
This work confirms the previous observation that a single application of N-hydroxy-2-fluorenylacetamide or N-hydroxy-3-fluorenylacetamide to the mammary gland of the rat induced a high incidence of tumors, whereas the corresponding arylamides, N-2-fluorenylacetamide (2-FAA) and N-3-fluorenylacetamide, were only weakly active. The results suggested N-hydroxylation of the arylamides as a prerequisite for mammary carcinogenesis. Since N-hydroxylation of 2-FAA by hepatic microsomes is catalyzed by the mixed-function oxidase containing cytochrome P-450 or the 2-methylcholanthrene-inducible cytochrome P1-450, we examined whether these cytochromes are present in mammary microsomes. In contrast to liver, neither cytochrome nor N-hydroxylation of 2-FAA was detected in the mammary gland of normal and 3-methylcholanthrene-treated rats. These experiments indicated that the N-hydroxylation of 2-FAA, although obligatory for induction of mammary neoplasia, is not performed in the mammary gland but may take place in the liver. We also examined the carcinogenicity of N-acetoxy-2-fluorenylacetamide and N-acetoxy-3-fluorenylacetamide for the mammary gland upon topical application. Since both acetates were carcinogenic and since the acetyl group of acetyl coenzyme A is transferred to fluorenylhydroxamic acids at pH 7.4, these esters may be ultimate carciogens in mammary carcinogenesis. Ovariectomized rats did not develop mammary tumors after a single application of the fluorenylhydroxamic acids, and administration of estradiol and fluorenylhydroxamic acids to the ovariectomized rats did not improve the tumor yield. These results indicate that induction of mammary tumors by fluorenylhydroxamic acids is under hormonal control.


The corticotropin-releasing factor-like diuretic hormone 44 (DH44) and kinin neuropeptides modulate desiccation and starvation tolerance in Drosophila melanogaster.
Malpighian tubules are critical organs for epithelial fluid transport and stress tolerance in insects, and are under neuroendocrine control by multiple neuropeptides secreted by identified neurons. Here, we demonstrate roles for CRF-like diuretic hormone 44 (DH44) and Drosophila melanogaster kinin (Drome-kinin, DK) in desiccation and starvation tolerance. Gene expression and labelled DH44 ligand binding data, as well as highly selective knockdowns and/or neuronal ablations of DH44 in neurons of the pars intercerebralis and DH44 receptor (DH44-R2) in Malpighian tubule principal cells, indicate that suppression of DH44 signalling improves desiccation tolerance of the intact fly. Drome-kinin receptor, encoded by the leucokinin receptor gene, LKR, is expressed in DH44 neurons as well as in stellate cells of the Malpighian tubules. LKR knockdown in DH44-expressing neurons reduces Malpighian tubule-specific LKR, suggesting interactions between DH44 and LK signalling pathways. Finally, although a role for DK in desiccation tolerance was not defined, we demonstrate a novel role for Malpighian tubule cell-specific LKR in starvation tolerance. Starvation increases gene expression of epithelial LKR. Also, Malpighian tubule stellate cell-specific knockdown of LKR significantly reduced starvation tolerance, demonstrating a role for neuropeptide signalling during starvation stress.


Monitoring of cerebral oximetry during head-up tilt test in adults with history of syncope and orthostatic intolerance.
We applied near-infrared-spectroscopy (NIRS) to measure absolute frontal cerebral tissue oxygen saturation (SctO2) during head-up tilt test (HUT) in patients investigated for unexplained syncope. Synchronized non-invasive beat-to-beat haemodynamic monitoring, ECG, SctO2 (NIRS; normal range: 60-80%), and peripheral oxygen saturation (left hand, SpO2) were applied during HUT in a random sample of patients with unexplained syncope. Tracings of 54 patients (mean-age: 55 ± 19 years, 39% male) with negative HUT, vasovagal syncope (VVS), or orthostatic hypotension (OH) were analysed. In 44 patients HUT was diagnostic, in 10 HUT was negative. Thirty-one experienced VVS. Of these, 6 had spontaneous and 25 nitroglycerin-induced syncope. Thirteen patients had orthostatic hypotension (OH). Although there was no significant change in mean-arterial pressure from baseline to 1 min before syncope or end of passive HUT phase (-1.4 ± 13.9 mmHg; P = 0.45), there was a significant fall in SctO2 during the same period (-3.2 ± 3.2%; P ≤ 0.001). Among patients who experienced syncope, a decrease in SctO2 from 71 ± 5% at baseline to 53 ± 9% (P < 0.001) at syncope was observed. During HUT, there was a significant difference in delta SctO2 between spontaneous VVS (-4.5 ± 3.0%) and negative HUT (-1.3 ± 1.9%; P = 0.021), but not between spontaneous VVS and OH (-5.4 ± 4.2%; P = 0.65). In spontaneous VVS, progressive decrease of SctO2 was independent of mean arterial pressure decrease (P = 0.22). Progressive decrease in cerebral tissue oxygenation independent of mean-arterial pressure may precede spontaneous vasovagal reflex during tilt. Patients experience syncope when SctO2 falls below 60%. These data confirm clinical utility of absolute cerebral oximetry monitoring for syncope investigation. We applied NIRS to measure frontal cerebral tissue oxygen saturation (SctO2) during head-up tilt test (HUT) in patients with unexplained syncope. In 44 of 54 patients, HUT was diagnostic. In patients with syncope, a significant SctO2-decrease was observed. Different patterns of SctO2 can be detected.


Comparison of SPET brain perfusion and 18F-FDG brain metabolism in patients with chronic fatigue syndrome.
Chronic fatigue syndrome is a clinically defined condition of uncertain aetiology. We compared 99Tcm-HMPAO single photon emission tomography (SPET) brain perfusion with dual-head 18F-FDG brain metabolism in patients with chronic fatigue syndrome. Eighteen patients (14 females, 4 males), who fulfilled the diagnostic criteria of the Centers for Disease Control for chronic fatigue syndrome, were investigated. Thirteen patients had abnormal SPET brain perfusion scans and five had normal scans. Fifteen patients had normal glucose brain metabolism scans and three had abnormal scans. We conclude that, in chronic fatigue syndrome patients, there is discordance between SPET brain perfusion and 18F-FDG brain uptake. It is possible to have brain perfusion abnormalities without corresponding changes in glucose uptake.


Increased vagal tone accounts for the observed immune paralysis in patients with traumatic brain injury.
Traumatic brain injury (TBI) is a leading cause of death and disability, especially in the younger population. In the acute phase after TBI, patients are more vulnerable to infection, associated with a decreased immune response in vitro. The cause of this immune paralysis is poorly understood. Apart from other neurologic dysfunction, TBI also results in an increase in vagal activity. Recently, the vagus nerve has been demonstrated to exert an anti-inflammatory effect, termed the cholinergic anti-inflammatory pathway. The anti-inflammatory effects of the vagus nerve are mediated by the alpha 7 nicotinic acetylcholine receptor present on macrophages and other cytokine-producing cells. From these observations, we hypothesize that the immune paralysis observed in patients with TBI may, at least in part, result from augmented vagal activity and subsequent sustained effects of the cholinergic anti-inflammatory pathway. This pathway may counteract systemic proinflammation caused by the release of endogenous compounds termed alarmins as a result of tissue trauma. However, sustained activity of this pathway may severely impair the body's ability to combat infection. Since the cholinergic anti-inflammatory pathway can be pharmacologically modulated in humans, it could represent a novel approach to prevent infections in patients with TBI.


Molecular architecture of the ErbB2 extracellular domain homodimer.
Human epidermal growth factor receptors (HERs or ErbBs) play crucial roles in numerous cellular processes. ErbB2 is a key member of ErbB family, and its overexpression is recognized as a frequent molecular abnormality. In cancer, this overexpression correlates with aggressive disease and poor patient outcomes. Dimer-dependent phosphorylation is a key event for the signal transduction of ErbBs. However, the molecular mechanism of the dimerization of ErbB2 remains elusive. In the present work, we report the homodimer architecture of the ErbB2 extracellular domain (ECD) which is unique compared with other dimer-models of ErbBs. The structure of the ErbB2 ECD homodimer represents a "back to head" interaction, in which a protruding β-hairpin arm in domain II of one ErbB2 protomer is inserted into a C-shaped pocket created by domains I-III of the adjacent ErbB2 protomer. This dimerized architecture and its impact on the phosphorylation of ErbB2 intracellular domain were further verified by a mutagenesis study. We also elucidated the different impacts of two clinically administered therapeutic antibodies, trastuzumab and pertuzumab, on ErbB2 dimerization. This information not only provides an understanding of the molecular mechanism of ErbBs dimerization but also elucidates ErbB2-targeted therapy at the molecular level.


Proteolytic cleavage of [3H]nitrobenzylthioinosine-labelled nucleoside transporter in human erythrocytes.
The transmembrane topology of the nucleoside transporter of human erythrocytes, which had been covalently photolabelled with [3H]nitrobenzylthioinosine, was investigated by monitoring the effect of proteinases applied to intact erythrocytes and unsealed membrane preparations. Treatment of unsealed membranes with low concentrations of trypsin and chymotrypsin at 1 degree C cleaved the nucleoside transporter, a band 4.5 polypeptide, apparent Mr 66 000-45 000, to yield two radioactive fragments with apparent Mr 38 000 and 23 000. The fragment of Mr 38 000, in contrast to the Mr 23 000 fragment, migrated as a broad peak (apparent Mr 45 000-31 000) suggesting that carbohydrate was probably attached to this fragment. Similar treatment of intact cells under iso-osmotic saline conditions at 1 degree C had no effect on the apparent Mr of the [3H]nitrobenzylthioinosine-labelled band 4.5, suggesting that at least one of the trypsin cleavage sites resulting in the apparent Mr fragments of 38 000 and 23 000 is located at the cytoplasmic surface. However, at low ionic strengths the extracellular region of the nucleoside transporter is susceptible to trypsin proteolysis, indicating that the transporter is a transmembrane protein. In contrast, the extracellular region of the [3H]cytochalasin B-labelled glucose carrier, another band 4.5 polypeptide, was resistant to trypsin digestion. Proteolysis of the glucose transporter at the cytoplasmic surface generated a radiolabelled fragment of Mr 19 000 which was distinct from the Mr 23 000 fragment radiolabelled with [3H]nitrobenzylthioinosine. The affinity for the reversible binding of [3H]cytochalasin B and [3H]nitrobenzylthioinosine to the glucose and nucleoside transporters, respectively, was lowered 2-3-fold following trypsin treatment of unsealed membranes, but the maximum number of inhibitor binding sites was unaffected despite the cleavage of band 4.5 to lower-Mr fragments.


Relation between fragmented QRS and collateral circulation in patients with chronic total occlusion without prior myocardial infarction.
It has been shown that the fragmented QRS (fQRS) on electrocardiogram (ECG) signifies regional myocardial scar in patients with non-Q-wave myocardial infarction (MI). We hypothesized that presence of fQRS on ECG may be related with poorly-grown collateral coronary circulation (CCC) in patients with chronic total coronary occlusion (CTO) without prior MI. This retrospective observational study is included 56 patients (mean age 61.73±7.96 years; 67.9% men) with CTO in one of the major coronary arteries. Collateral circulation was graded according to Rentrop's classification. The fQRS was defined as the presence of an additional R wave or notching of R or S wave or the presence of fragmentation in two contiguous ECG leads corresponding to a major coronary artery territory. Patients with pathological Q-wave or history of MI, typical bundle brunch blocks (BBB) and incomplete right BBB were excluded from study. Statistical analysis was performed using Chi-square test, Student's t-test and logistic regression analysis. Fifteen patients had Rentrop grade 1, 15 patients had grade 2 and 26 patients had grade 3 CCC. Five (19%) of the patients who have grade 3 CCC, seven (47%) of the patients who had grade 2 CCC, ten (67%) of the patients who had grade 1 CCC had fQRS (p=0.002). Logistic regression analysis showed high predictive value of the presence of fQRS for Rentrop 1 CCC (OR=8.4, 95% CI 1.97-35.7; p=0.004). Results of our study may implicate the presence of fQRS on electrocardiogram as a predictor of a poorly grown CCC in patients with chronic total occlusion without prior MI.


Responsiveness of the Child Perceptions Questionnaire11-14 for Cambodian children undergoing basic dental care.
Oral-Health-Related Quality of Life (OHRQoL) instruments are useful outcome measures in dental health services research; however, there are no reports of the use OHRQoL measures in evaluating the outcome of basic dental care in children anywhere. The aims of this study are to evaluate the responsiveness of the Khmer version of the Child Perceptions Questionnaire11-14 (CPQ11-14) and to describe changes in OHRQoL associated with the provision of basic dental care. A pre-test/post-test design, with a clinical convenience sample of children aged from 8 to 14 years of age who received basic dental care from a local nongovernmental organisation (NGO). Of the 140 children in the baseline sample, 116 (82.9%) were followed up. The mean change in CPQ11-14 overall score for those with caries was 1.7, representing a small improvement in OHRQoL. There was a moderate (one in three) statistically significant reduction in the prevalence of impacts across the whole sample (P < 0.001; McNemar test), and one in five improved by the minimally important difference (MID) of five-scale points. The Khmer version of the CPQ11-14 appears to be a valid and responsive measure for assessing treatment-associated changes in OHRQoL in children with dental caries in Cambodia.


Diabetic retinopathy: Role of inflammation and potential therapies for anti-inflammation.
Diabetic retinopathy is a leading cause of blindness among working-age adults. Despite many years of research, treatment options for diabetic retinopathy remain limited and with adverse effects. Discovery of new molecular entities with adequate clinical activity for diabetic retinopathy remains one of the key research priorities in ophthalmology. This review is focused on the therapeutic effects of cannabidiol (CBD), a non-psychoactive native cannabinoid, as an emerging and novel therapeutic modality in ophthalmology based on systematic studies in animal models of inflammatory retinal diseases including diabetic retinopathy - a retinal disease associated with vascular-neuroinflammation. Special emphasis is placed on novel mechanisms which may shed light on the pharmacological activity associated with CBD preclinically. These include a self-defence system against inflammation and neurodegeneration mediated by inhibition of equilibrative nucleoside transporter and activation of adenosine receptor by treatment with CBD.


Relationship between objective and subjective sleep measures in depressed patients and healthy controls.
The purpose of this study was to correlate subjective sleep characteristics based on questionnaire response, and objective sleep EEG features based on polysomnography, in 52 patients with major depressive disorders (MDD) and 49 healthy controls. With the exception of the number of awakenings, subjective and objective sleep measures were strongly correlated in both groups. Patients and controls were able to accurately judge time in bed, total sleep time and sleep latency. However, sleep quality, depth, and how rested participants felt upon awakening were not strongly correlated with objective sleep characteristics, particularly in those with MDD. The findings suggest that estimates such as total sleep time and sleep latency, obtained from questionnaire data, bear a strong resemblance to objective polysomnographic characteristics in both those with MDD and healthy controls. Patients with MDD do not show sleep-state misperceptions although depressed women are more accurate in estimating sleep characteristics than depressed men.


Acute hemorrhagic pulmonary infarction and necrotizing pneumonia in horses: 21 cases (1967-1993).
To characterize history, clinical signs, and pathologic findings in horses with histologically confirmed acute hemorrhagic pulmonary infarction and necrotizing pneumonia. Retrospective study. 21 horses. 19 of the 21 horses were Thoroughbred racehorses in training. Eighteen horses had had strenuous exercise immediately prior to onset of illness. Fifteen horses had a serosanguineous nasal discharge during hospitalization. Seventeen horses had radiographic evidence of pulmonary consolidation and pleural effusion. Nine of 14 horses had ultrasonographic evidence of large pulmonary parenchymal defects consistent with consolidation. Pleurocentesis yielded a suppurative, serosanguineous effusion in the 14 horses in which it was performed. Bacteria were isolated from all transtracheal aspirates (14) and from 6 of 12 pleural fluid samples. Actinobacillus suis-like organisms and Streptococcus equi subsp zooepidemicus were most commonly isolated. Nineteen horses were hospitalized and treated. Mean duration of treatment was 5 days, and most horses were euthanatized because of secondary complications, continued costs of medical treatment, or poor prognosis for future performance. Pathologic lesions included well-demarcated regions of hemorrhagic pulmonary infarction with necrosis and a serosanguineous pleural effusion. Thrombosis of pulmonary vessels was found in 11 horses. An acute or peracute onset of severe respiratory distress, with serosanguineous nasal discharge, ultrasonographic and radiographic evidence of severe pulmonary consolidation, and serosanguineous suppurative pleural effusion, is strongly suggestive of pulmonary infarction in horses. Horses with pulmonary infarction responded poorly to conventional treatment for pleuropneumonia and had a poor prognosis for recovery.


Danlou Recipe promotes cholesterol efflux in macrophages RAW264.7 and reverses cholesterol transport in mice with hyperlipidemia induced by P407.
Liver X Receptor (LXR) agonists could attenuate the development of atherosclerosis but bring excess lipid accumulation in the liver. Danlou Recipe was believed to be a benefit for improving the lipid profile. Thus, it is unclear whether Danlou Recipe could attenuate hyperlipidemia without excess lipid accumulated in the liver of mice. This study aimed to clarify if Danlou Recipe could alleviate the progression of hyperlipidemia in mice without extra lipids accumulated in the liver. Male murine macrophage RAW264.7 cells and murine peritoneal macrophages were used for the in vitro experiments. Cellular cholesterol efflux was determined using the fluorescent cholesterol labeling method. Those genes involved in lipid metabolism were evaluated by qRT-PCR and western blotting respectively. In vivo, a mouse model of hyperlipidemia induced by P407 was used to figure out the effect of Danlou Recipe on reverse cholesterol transport (RCT) and hyperlipidemia. Ethanol extract of Danlou tablet (EEDL) was prepared by extracting the whole powder of Danlou Prescription from ethanol, and the chemical composition was analyzed by ultra-performance liquid chromatography (UPLC). EEDL inhibits the formation of RAW264.7 macrophage-derived foam cells, and promotes ABCA1/apoA1 conducted cholesterol efflux in RAW264.7 macrophages and mouse peritoneal macrophages. In the P407-induced hyperlipidemia mouse model, oral administration of EEDL can promote RCT in vivo and improve fatty liver induced by a high-fat diet. Consistent with the findings in vitro, EEDL promotes RCT by upregulating the LXR activities. Our results demonstrate that EEDL has the potential for targeting RCT/LXR in the treatment of lipid metabolism disorders to be developed as a safe and effective therapy.


Pathophysiology and treatment of nonfamilial hyperparathyroidism.
Primary hyperparathyroidism (HPT) is the main cause of hypercalcemia and the most common parathyroid glands disease. The diagnosis is easy in patients with hypercalcemia and elevated PTH serum level. Minimally invasive parathyroidectomy (PTx) represents the treatment of choice for symptomatic patients, leading to several advantages, including immediate normalization of hypercalcemia and significant improvement of bone mineral density, cardiovascular dysfunctions, neuropsychological symptoms and quality of life. Secondary and tertiary HPT are relatively common complications in patients with chronic kidney disease (CKD) or advanced kidney failure, and in kidney transplant recipients who did not achieve complete calcium/phosphate metabolism normalization, respectively. The drugs available for patients with secondary HPT, and to treat hyperphosphatemia include non-calcium-containing phosphate binder, calcitriol analogues, calcimimetic agents, or a combination of two or more drugs. Although recent studies report that PTx significantly improves survival also in patients with CKD and severe secondary HPT, the indications for surgery are not yet well established. Subtotal or total PTx with or without autotransplantation are the surgical options for treating all patients with secondary HPT. Total PTx leads to a faster reduction in serum calcium level and normalization of PTH, but the risk of hypoparathyroidism is higher than after subtotal PTx. Further studies are needed to confirm the usefulness of the drugs currently recommended, and others will have to be tested in the near future.


Methylphenidate ameliorates depressive comorbidity in ADHD children without any modification on differences in serum melatonin concentration between ADHD subtypes.
The vast majority of Attention-deficit/hyperactivity disorder (ADHD) patients have other associated pathologies, with depressive symptoms as one of the most prevalent. Among the mediators that may participate in ADHD, melatonin is thought to regulate circadian rhythms, neurological function and stress response. To determine (1) the serum baseline daily variations and nocturnal excretion of melatonin in ADHD subtypes and (2) the effect of chronic administration of methylphenidate, as well as the effects on symptomatology, 136 children with ADHD (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision: DSM-IV-TR criteria) were divided into subgroups using the "Children's Depression Inventory" (CDI). Blood samples were drawn at 20:00 and 09:00 h, and urine was collected between 21:00 and 09:00 h, at inclusion and after 4.61 ± 2.29 months of treatment. Melatonin and its urine metabolite were measured by radioimmunoassay RIA. Factorial analysis was performed using STATA 12.0. Melatonin was higher predominantly in hyperactive-impulsive/conduct disordered children (PHI/CD) of the ADHD subtype, without the influence of comorbid depressive symptoms. Methylphenidate ameliorated this comorbidity without induction of any changes in the serum melatonin profile, but treatment with it was associated with a decrease in 6-s-melatonin excretion in both ADHD subtypes. In untreated children, partial homeostatic restoration of disrupted neuroendocrine equilibrium most likely led to an increased serum melatonin in PHI/CD children. A differential cerebral melatonin metabolization after methylphenidate may underlie some of the clinical benefit.


Thoughts and consideration regarding immigrant clinicians: is cultural preservation influencing providers' practice in HPV vaccination.
Immigrant clinicians are vital to population healthcare delivery and therefore population health. One in four physicians in the USA is foreign-born and notably represented in family and pediatric medicine-specialties charged with administering childhood/adolescent vaccines, such as human papillomavirus vaccine (HPVV). Our examination suggests there may be unique cultural and socialization factors that influence clinician HPVV recommendation practice; however, immigrant clinicians have not been adequately engaged within the national HPVV agenda. Given the volume and significance of immigrant clinicians, engagement of these clinicians, in both community and nation-wide efforts to increase HPVV, is a necessary step for improving and achieving the national health goal of optimizing HPVV for cancer prevention.


Immunoelectronmicroscopy of soluble and membrane proteins with a sensitive postembedding method.
The application of immunoelectronmicroscopy to soluble proteins is limited because soluble proteins can redistribute during fixation. Fixation may also adversely affect the recognition of proteins associated with membranes. We show here how displacements of soluble proteins can be prevented and antigen sensitivity improved by freeze-substitution immunocytochemistry. The usefulness of this method for soluble cytoplasmic proteins is demonstrated for the twitchin protein in Aplysia muscle and the kinesin motor proteins in squid giant axons, in which the sizes of various cytoplasmic pools of kinesins are estimated. The utility for membrane proteins present in small numbers of copies is demonstrated by labeling a glutamate receptor subunit in mouse cerebellar cortex and the ZO-1 protein in tight junctions between MDCK cells. Thus, freeze-substitution immunocytochemistry can show the native distribution of both soluble and membrane proteins labeled with polyclonal antibodies and, at the same time, can reveal structural features comparable to those in chemically fixed or osmium freeze-substituted samples.


Breast Cancer Cells Metastasize to the Tissue-Engineered Premetastatic Niche by Using an Osteoid-Formed Polycaprolactone/Nanohydroxyapatite Scaffold.
It has been deemed that the premetastatic niche (PMN) plays a critical role in facilitating bone metastasis of breast cancer cells. Tissue engineering scaffolds provide an advantageous environment to promote osteogenesis that may mimic the bony premetastatic niches (BPMNs). In this study, human mesenchymal stem cells (hMSCs) were seeded onto designed polycaprolactone/nanohydroxyapatite (PCL-nHA) scaffolds for osteogenic differentiation. Subsequently, a coculture system was used to establish the tissue-engineered BPMNs by culturing breast cancer cells, hMSCs, and osteoid-formed PCL-nHA scaffolds. Afterwards, a migration assay was used to investigate the recruitment of MDA-MB-231, MCF-7, and MDA-MB-453 cells to the BPMNs' supernatants. The cancer stem cell (CSC) properties of these migrated cells were investigated by flow cytometry. Our results showed that the mRNA expression levels of alkaline phosphatase (ALP), Osterix, runt-related transcription factor 2 (Runx2), and collagen type I alpha 1 (COL1A1) on the PCL-nHA scaffolds were dramatically increased compared to the PCL scaffolds on days 11, 18, and 32. The expression of CXCL12 in these BPMNs was increased gradually over coculturing time, and it may be a feasible marker for BPMNs. Furthermore, migration analysis results showed that the higher maturation of BPMNs collectively contributed to the creation of a more favorable niched site for the cancerous invasion. The subpopulation of breast cancer stem cells (BCSCs) was more likely to migrate to fertile BPMNs. The proportion of BCSCs in metastatic MDA-MB-231, MCF-7, and MDA-MB-453 cells were increased by approximately 63.47%, 149.48%, and 127.60%. The current study demonstrated that a designed tissue engineering scaffold can provide a novel method to create a bone-mimicking environment that serves as a useable platform to recapitulate the BPMNs and help interrogate the scheme of bone metastasis by breast cancer.


Toxicity data for modeling impacts of oil components in an Arctic ecosystem.
Ecological impact assessment modeling systems are valuable support tools for managing impacts from commercial activities on marine habitats and species. The inclusion of toxic effects modeling in these systems is predicated on the availability and quality of ecotoxicology data. Here we report on a data gathering exercise to obtain toxic effects data on oil compounds for a selection of cold-water marine species of fish and plankton associated with the Barents Sea ecosystem. Effects data were collated from historical and contemporary literature resources for the endpoints mortality, development, growth, bioaccumulation and reproduction. Evaluating the utility and applicability of these data for modeling, we find that data coverage is limited to a sub-set of the required endpoints. There is a need for new experimental studies for zooplankton focused on the endpoints development and bioaccumulation and for larvae and juvenile fish focused on growth and development.


Secondary hyperparathyroidism: benign bystander or culpable contributor to adverse health outcomes?
Elevation in serum parathyroid hormone (PTH) often accompanies vitamin D deficiency and renal impairment. PTH elevation in renal failure is viewed as an unfavorable development. Evidence is increasing that PTH elevation may be associated with increased morbidity and mortality. In many instances these PTH effects appear to be independent of vitamin D status. PTH mediates its effects through the ubiquitous type 1 PTH/PTH-related peptide receptor, which is notably present in the cardiovascular system. Increased PTH may promote cardiovascular disease through diminished cardiac contractility, enhanced coronary risk, and cardiac valvular and vascular calcification. High PTH levels appear to be linked to the metabolic syndrome and are aligned with hyperlipidemia, decreased insulin sensitivity, and, perhaps, decreased insulin secretion. Increased PTH also is associated with neuroendocrine activation, increased sympathetic activity, and endothelial stress. The relation between PTH and vitamin D is complex and may show significant threshold variations, especially when calcium intake, age, and race are considered. Moreover, evidence is increasing that fragments of PTH may not only be hormonally active but also may have opposing effects to PTH. Despite these caveats, PTH values provide useful clinical diagnostic and prognostic information in monitoring many chronic ailments such as heart and renal failure and multiple sclerosis.


Genes, Environments, and Time: The Biology of Adversity and Resilience.
Exposures to adverse environments, both psychosocial and physicochemical, are prevalent and consequential across a broad range of childhood populations. Such adversity, especially early in life, conveys measurable risk to learning and behavior and to the foundations of both mental and physical health. Using an interactive gene-environment-time (GET) framework, we survey the independent and interactive roles of genetic variation, environmental context, and developmental timing in light of advances in the biology of adversity and resilience, as well as new discoveries in biomedical research. Drawing on this rich evidence base, we identify 4 core concepts that provide a powerful catalyst for fresh thinking about primary health care for young children: (1) all biological systems are inextricably integrated, continuously "reading" and adapting to the environment and "talking back" to the brain and each other through highly regulated channels of cross-system communication; (2) adverse environmental exposures induce alterations in developmental trajectories that can lead to persistent disruptions of organ function and structure; (3) children vary in their sensitivity to context, and this variation is influenced by interactions among genetic factors, family and community environments, and developmental timing; and (4) critical or sensitive periods provide unmatched windows of opportunity for both positive and negative influences on multiple biological systems. These rapidly moving frontiers of investigation provide a powerful framework for new, science-informed thinking about health promotion and disease prevention in the early childhood period.


APLP1 Is a Synaptic Cell Adhesion Molecule, Supporting Maintenance of Dendritic Spines and Basal Synaptic Transmission.
The amyloid precursor protein (APP), a key player in Alzheimer's disease, belongs to the family of synaptic adhesion molecules (SAMs) due to its impact on synapse formation and synaptic plasticity. These functions are mediated by both the secreted APP ectodomain that acts as a neurotrophic factor and full-length APP forming trans-cellular dimers. Two homologs of APP exist in mammals: the APP like proteins APLP1 and APLP2, exhibiting functions that partly overlap with those of APP. Here we tested whether APLP1 and APLP2 also show features of SAMs. We found that all three family members were upregulated during postnatal development coinciding with synaptogenesis. We observed presynaptic and postsynaptic localization of all APP family members and could show that heterologous expression of APLP1 or APLP2 in non-neuronal cells induces presynaptic differentiation in contacting axons of cocultured neurons, similar to APP and other SAMs. Moreover, APP/APLPs all bind to synaptic-signaling molecules, such as MINT/X11. Furthermore, we report that aged APLP1 knock-out mice show impaired basal transmission and a reduced mEPSC frequency, likely resulting from reduced spine density. This demonstrates an essential nonredundant function of APLP1 at the synapse. Compared to APP, APLP1 exhibits increased trans-cellular binding and elevated cell-surface levels due to reduced endocytosis. In conclusion, our results establish that APLPs show typical features of SAMs and indicate that increased surface expression, as observed for APLP1, is essential for proper synapse formation in vitro and synapse maintenance in vivoSIGNIFICANCE STATEMENT According to the amyloid-cascade hypothesis, Alzheimer's disease is caused by the accumulation of Aβ peptides derived from sequential cleavage of the amyloid precursor protein (APP) by β-site APP cleaving enzyme 1 (BACE1) and γ-secretase. Here we show that all mammalian APP family members (APP, APLP1, and APLP2) exhibit synaptogenic activity, involving trans-synaptic dimerization, similar to other synaptic cell adhesion molecules, such as Neuroligin/Neurexin. Importantly, our study revealed that the loss of APLP1, which is one of the major substrates of BACE1, causes reduced spine density in aged mice. Because some therapeutic interventions target APP processing (e.g., BACE inhibitors), those strategies may alter APP/APLP physiological function. This should be taken into account for the development of pharmaceutical treatments of Alzheimer's disease.


Informing evidence: claimsmaking in midwives' and clients' talk about interventions.
Communication for informed choice is particularly challenging in clinical settings such as direct-entry midwifery, where the care model embraces diverse therapies and forms of knowledge. We identified three discursive moves (explanation, invocation, and evaluation) that Ontario midwives and clients used in making claims about proposed interventions. The analysis was informed by an understanding of communication as an interactionally situated and socially constructed interpretive practice. Both midwives and women called on the authority of biomedical discourse, but they also turned to sources such as women's wisdom to support their cases. The flexible use of these moves afforded participants considerable latitude in accepting or rejecting forms of evidence as authoritative. However, strategies designed to empower clients in making choices could unintentionally serve to enhance the authority of the care provider. Talk about interventions brings into view both the knowledge systems and the broader relations within which regulated midwifery practice operates.