Update on cardiothoracic surgery resident job opportunities.
Concerns regarding ample employment opportunities for graduating cardiothoracic surgery residents may affect perceptions of the field and recruitment into residency programs. We present the results of the Thoracic Surgery Residents Association/Thoracic Surgery Directors Association (TSRA/TSDA) 2008 Resident Survey, and compare them with the 2007 TSRA/TSDA survey and the 2006 interim report of the Society of Thoracic Surgeons Task Force on Job Opportunities. In April 2008, the TSRA/TSDA conducted an anonymous survey, linked to the cardiothoracic surgery resident online In-training Exam, with questions germane to resident job seeking and perceptions of the specialty. Results were compared with resident surveys from 2007 and 2006. Response rates for the 2008 and 2007 surveys were 100%, and 54.2% for 2006. Of graduating residents looking for employment, 61.6% had one or more job offers, compared with 64.6% and 83.5% from the 2007 and 2006 surveys, respectively. Of the respondents completing their job search, 24.5% entered private practice and 26.3% academia, compared with 12.1% and 30.1%, respectively, in the 2007 survey. Overall, 57.7% of all respondents had more than $50,000 education-related debt, compared with 54.2% of 2007 respondents. However, 71.5% of all 2008 respondents would recommend cardiothoracic surgery to a potential trainee, compared with 63.7% and 46.0% from 2007 and 2006 survey respondents, respectively. The 2008 survey suggests that although the majority of respondents found employment on completing residency, the percentage is less than 65%, reinforcing a need for formal networking programs or changes in residency training. Despite continued limited employment opportunities, resident impressions of cardiothoracic surgery have improved from 2006 to 2008.


Voices from Flint: Community Perceptions of the Flint Water Crisis.
We describe the self-reported socioeconomic and health impacts, as well as the coping mechanisms employed by a drug-using cohort of adults during the Flint water crisis (FWC) in Flint, Michigan. Participants from an ongoing longitudinal Emergency Department study were contacted between April 2016 and July 2016 and completed a survey focusing on exposure, consequences, and coping strategies. One hundred thirty-three participants (mean age = 26, 65% African-American, 61% public assistance) completed the survey (37.9% response rate). Of these, 75% reported exposure to water with elevated lead levels. Of these, 75% reported additional monthly expenses resulting from exposure. Almost 40% of parents reported changes in their children's health and 65% reported changes to their health since the FWC. Participants indicated the use of both positive (e.g., advice from trusted neighbors, 99.0%) and negative coping mechanisms (e.g., increased substance use, 20.0%) in response to this public health emergency. High-risk Flint residents reported multiple social, economic, and health-related consequences stemming from the FWC. Policymakers should consider additional resources for those affected, including increased access to mental health to aid recovery within the community.


Strategic planning in healthcare: the experience of the University of Wisconsin Hospital and Clinics.
In 1999, after 25 years of stable leadership from a single CEO, the University of Wisconsin Hospital and Clinics (UWHC) Authority Board named a new CEO. The 471-bed academic medical center had recently experienced significant change and challenges. In 1996, it had emerged as a public authority, a statutory designation by the state of Wisconsin that moved the hospital and clinics from the University of Wisconsin and the state of Wisconsin, and created it as a quasi-public entity with its own board. In 1999, when the new CEO was named, the hospital was experiencing a loss of revenue and market share, operating deficits, a 22 percent nurse vacancy rate, and patient satisfaction scores below the 40th percentile. The first task assigned to the new CEO by the board was the development of a new strategic plan that would reverse these trends and position UWHC as a premier academic hospital. The CEO began a strategic planning process that involved leaders, physicians, and staff from throughout the hospital and clinics, its affiliated medical school, and the physician practice plan. This article describes the collaborative, integrative, and communicative strategic planning process UWHC used; details the organization of the process; describes the results achieved by UWHC; and lists the lessons learned along the way.


Role of monocytes in mediating HIV-specific antibody-dependent cellular cytotoxicity.
Antibodies (Abs) that mediate antibody-dependent cellular cytotoxicity (ADCC) activity against HIV-1 are of major interest. A widely used method to measure ADCC Abs is the rapid and fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay. Antibody-dependent killing of a labelled target cell line by PBMC is assessed by loss of intracellular CFSE but retention of membrane dye PKH26 (CFSE-PKH26+). Cells of this phenotype are assumed to be derived from CFSE+PKH26+ target cells killed by NK cells. We assessed the effector cells that mediate ADCC in this assay. Backgating analysis and phenotyping of CFSE-PKH26+ revealed that the RFADCC assay's readout mainly represents CD3-CD14+ monocytes taking up the PKH26 dye. This was confirmed for 53 HIV+plasma-purified IgG samples when co-cultured with PBMC from three separate healthy donors. Emergence of the CFSE-PKH26+ monocyte population was observed upon co-culture of targets with purified monocytes but not with purified NK cells. Image flow cytometry and microscopy showed a monocyte-specific interaction with target cells without typical morphological changes associated with phagocytosis, suggesting a monocyte-mediated ADCC process. We conclude that the RFADCC assay primarily reflects Ab-mediated monocyte function. Further studies on the immunological importance of HIV-specific monocyte-mediated ADCC are warranted.


An outbreak of herpes simplex virus type 1 in an intensive care nursery.
Neonatal herpes simplex infection is not a common occurrence but one which warrants particular concern. An 1800-g premature infant who developed respiratory distress and died at 12 days unexpectedly yielded HSV from a culture of cerebrospinal fluid. There were no mucocutaneous lesions. Ten days later three other infants (ages 40, 69 and 11 days) developed vesicles which yielded herpes simplex. Health care staff cohorts were assigned to "clean" or "exposed" nursery areas. The three secondarily infected cases were treated with vidarabine and did not develop systemic symptoms. Typing of the isolates using immunofluorescence and monoclonal antibodies revealed all to be herpes simplex type 1. Restriction endonuclease cleavage of viral DNA determined that the isolates from the four infants were identical. The mothers of the infants denied any history of recent or recurrent herpes, and their cervical cultures were negative. The source of the outbreak has remained unknown. The possibility of manual transmission to the secondary cases remains likely despite standard infection control practices. Cohort isolation of all exposed patients prevented further spread.


Recent advances in glucose-lowering treatment to reduce diabetic kidney disease.
The epidemic of diabetes has now taken on epic proportions and therefore reducing the impact of diabetic complications represents one of the major global challenges in improving health and well-being worldwide. Preventing the development of diabetic kidney disease (DKD) is particularly important, as diabetes is one of the main risk factors for the development of chronic kidney disease, which in turn is strongly linked to development of cardiovascular disease, end-stage renal failure, hospitalization and premature death. Intensive glucose-lowering treatment has been shown to prevent and slow progression of DKD, yet to date, only certain populations have benefited from this intervention. We review the evidence for existing glucose-lowering treatments in the prevention of DKD, and research into techniques to better target individuals who will benefit from these therapies. Diabetic patients with established kidney disease may benefit from glucose-lowering treatment, particularly if a safer side-effect profile of these treatments is achieved. Better understanding of glucose homeostasis and evaluation of compounds inhibiting its downstream effects are required in order to improve the outlook for individuals with DKD. An additional approach to improve the success rate of glucose-lowering treatment is to improve the selection of individuals who may benefit from treatment. A potential means to identify such subjects could involve the use of biomarkers.


Axonal Transport Enables Neuron-to-Neuron Propagation of Human Coronavirus OC43.
Human coronaviruses (HCoVs) are recognized respiratory pathogens for which accumulating evidence indicates that in vulnerable patients the infection can cause more severe pathologies. HCoVs are not always confined to the upper respiratory tract and can invade the central nervous system (CNS) under still unclear circumstances. HCoV-induced neuropathologies in humans are difficult to diagnose early enough to allow therapeutic interventions. Making use of our already described animal model of HCoV neuropathogenesis, we describe the route of neuropropagation from the nasal cavity to the olfactory bulb and piriform cortex and then the brain stem. We identified neuron-to-neuron propagation as one underlying mode of virus spreading in cell culture. Our data demonstrate that both passive diffusion of released viral particles and axonal transport are valid propagation strategies used by the virus. We describe for the first time the presence along axons of viral platforms whose static dynamism is reminiscent of viral assembly sites. We further reveal that HCoV OC43 modes of propagation can be modulated by selected HCoV OC43 proteins and axonal transport. Our work, therefore, identifies processes that may govern the severity and nature of HCoV OC43 neuropathogenesis and will make possible the development of therapeutic strategies to prevent occurrences.IMPORTANCE Coronaviruses may invade the CNS, disseminate, and participate in the induction of neurological diseases. Their neuropathogenicity is being increasingly recognized in humans, and the presence and persistence of human coronaviruses (HCoV) in human brains have been proposed to cause long-term sequelae. Using our mouse model relying on natural susceptibility to HCoV OC43 and neuronal cell cultures, we have defined the most relevant path taken by HCoV OC43 to access and spread to and within the CNS toward the brain stem and spinal cord and studied in cell culture the underlying modes of intercellular propagation to better understand its neuropathogenesis. Our data suggest that axonal transport governs HCoV OC43 egress in the CNS, leading to the exacerbation of neuropathogenesis. Exploiting knowledge on neuroinvasion and dissemination will enhance our ability to control viral infection within the CNS, as it will shed light on underlying mechanisms of neuropathogenesis and uncover potential druggable molecular virus-host interfaces.


Narrative comprehension in 4-7-year-old children with autism: testing the Weak Central Coherence account.
Despite somewhat spared structural language development in high-functioning autism, communicative comprehension deficits persist. Comprehension involves the integration of meaning: global processing is required. The Weak Central Coherence theory suggests that individuals with autism are biased to process information locally. This cognitive style may impair comprehension, particularly if inferencing is required. However, task performance may be facilitated by this cognitive style if local processing is required. The current study was designed to examine the extent to which the 'weak central coherence' cognitive style affects comprehension and inferential processing of spoken narratives. The children with autism were expected to perform comparatively poorer on inferences relating to event scripts and comparatively better on inferences requiring deductive reasoning. Fourteen high-functioning children with autism were recruited from databases of various autism organizations (mean age = 6:7, 13 males, one female) and were matched on a receptive vocabulary and a picture-completion task with 14 typically developing children recruited from a local childcare centre (mean age = 4:10, seven males, seven females). The children were read short stories and asked questions about the stories. Results indicated that the children with autism were less able to make inferences based on event scripts, but the groups did not differ significantly on inferences requiring deductive logical reasoning. Despite similar group performance on questions relating to the main idea of the stories, only for the typically developing group was good performance on extracting the main idea of the narratives significantly correlated with performance on all other comprehension tasks. Findings provide some support for the Weak Central Coherence theory and demonstrate that young children with autism do not spontaneously integrate information in order to make script inferences, as do typically developing children. These findings may help to explain communicative problems of young children with autism and can be applied to intervention programme development. More research on the link between a 'weak central coherence' cognitive style and communicative comprehension in autism will be valuable in understanding the comprehension deficits associated with autism.


Participation in cancer clinical trials: why are patients not participating?
Participation in cancer clinical trials is low, particularly in racial and ethnic minorities in some cases, which has negative consequences for the generalizability for study findings. The objective of this study was to determine what factors are associated with patients' participation or willingness to participate and whether these factors vary by race/ethnicity. or . White, Hispanic, and black participants were obtained through the Florida cancer registry and who were diagnosed with breast, lung, colorectal, or prostate cancer (N = 1100). Participants were surveyed via telephone to obtain demographic information, past participation, and willingness to participate in clinical trials, as well as barriers and facilitators to participation. Logistic and Poisson regressions were performed. . Respondents were on average 67.4 years old, 42.7% were male, and 50.1% were married. In this population, 7.7% of respondents had participated in a clinical trial, and 36.5% stated that they would be willing to participate. In multivariate models, blacks and Hispanics were equally likely as whites to be willing to participate in cancer trials, but Hispanics were less likely to have participated, and this was especially more likely in non-English-speaking Hispanics compared with English-speaking Hispanics. Notable barriers across race/ethnicity were mistrust and lack of knowledge of clinical trials. Limitations. Cross-sectional design limits cause-and-effect conclusions. . There are racial differences in participation rates but not in willingness to participate. We hypothesize that willingness to participate is not very high because people are uninformed about participating, particularly in non-English-speaking Hispanics. Barriers and facilitators to participation vary by race. Improved understanding of cultural differences that can be addressed by physicians may restore faith, comprehension, and acceptability of clinical trials by all patients.


Evaluation of cool- and warm-season grasses for resistance to multiple chinch bug (Hemiptera: Blissidae) species.
Chinch bugs are common pests of many agronomic and horticulturally important crops and turfgrasses. The extensive overlap of plant hosts and geographic distribution of Blissus leucopterus leucopterus (Say), Blissus leucopterus hirtus Montandon, Blissus insularis Barber, and Blissus occiduus Barber underscores the importance of identifying resistant germplasm. Cool- and warm-season turfgrasses and sorghum, Sorghum bicolor (L.) Moench, were evaluated for resistance to chinch bugs in the Blissus complex, and the presence of multiple resistance was documented. Greenhouse studies established that B. occiduus-resistant ('Prestige', formerly NE91-118) and -susceptible ('378') buffalograsses,, Buchloë dactyloides (Nuttall) Engelmann, were susceptible to all other chinch bug species. KS94 sorghum exhibited resistance to both B. occiduus and B. l. leucopterus, whereas B. insularis-resistant St. Augustinegrass, Stenotaphrum secundatum (Walter) Kuntze ('Floratam'), was also resistant to B. occiduus. B. l. leucopterus-susceptible sorghum ('Wheatland') and B. insularis-susceptible St. Augustinegrasses ('Raleigh' and 'Amerishade') were highly resistant to B. occiduus. Endophyte-free and -enhanced fine fescues (Festuca spp.) were moderately to highly susceptible to B. l. hirtus but moderately to highly resistant to B. occiduus. The results of this research showed the buffalograsses evaluated, including B. occiduus-resistant Prestige, are moderately to highly susceptible to the three other chinch bug species. In contrast, B. occiduus did not cause considerable damage to any of the turfgrasses or sorghum cultivars evaluated, other than buffalograss, irrespective of whether or not they are resistant to another chinch bug species. This information is increasingly important as various grasses become adapted to regions that may possess chinch bug species other than those with which they are typically associated. These levels of Blissus resistance should be included when selecting resistant germplasm for managing Blissus species pests.


Altered expression of the glutamate transporter EAAT2b in neurological disease.
Functional studies suggest that up to 95% of all glutamate transport is handled by the glutamate transporter EAAT2. Amino and C-terminal antibodies demonstrate that under normal conditions EAAT2 is specific to astrocytes. A truncated splice variant of EAAT2, known as EAAT2b, also has been identified in astrocytes and some neurons. In vitro studies suggest EAAT2b transports glutamate similar to EAAT2, although the contribution of EAAT2b to normal clearance of extracellular glutamate is unknown. To investigate EAAT2b biology in pathological conditions, we examined the cellular and regional distribution of EAAT2b in amyotrophic lateral sclerosis. Using epitope-specific, affinity purified antibodies, we found that EAAT2b tissue levels were increased by more than twofold in amyotrophic lateral sclerosis motor cortex, whereas EAAT2 levels were decreased by up to 95%. EAAT2b distribution in normal human cortex was largely confined to the neuropil-like EAAT2, with occasional faint neuronal expression. In contrast, amyotrophic lateral sclerosis motor cortex had an obvious qualitative increase in neuropil EAAT2b staining and a drastic increase in neuronal soma and dendritic EAAT2b immunostaining. Despite these increases in EAAT2b immunostaining, functional transporter studies demonstrated a large loss of EAAT2 function. These studies clearly document altered regulation and splicing of the dominant glutamate transporter EAAT2 under conditions of neurological stress.


Functional prognosis of dizziness in older adults in primary care: a prospective cohort study.
To investigate the 6-month functional prognosis of dizziness in older adults in primary care, to identify important predictors of dizziness-related impairment, and to construct a score to assist risk prediction. Prospective cohort study with 6-month follow-up. Twenty-four primary care practices in the Netherlands. Four hundred seventeen older adults (mean age 78.5, range 65-95, 74% female) presenting consecutively to primary care with dizziness. Tests, including history and physical and additional examination, previously selected by an international expert panel and based on an earlier systematic review, were performed. The main outcome measure was 6-month dizziness-related impairment score measured using the Dizziness Handicap Inventory. Follow-up was complete for 92% of participants. Although 61% of participants experienced less impairment at 6 months, 130 participants (34%) showed persistent dizziness-related impairment. Factors most predictive of dizziness-related impairment at 6 months were onset of dizziness at least 6 months before inclusion, standing still as a dizziness-provoking circumstance, trouble with walking or (almost) falling (associated symptom), polypharmacy, absence of diabetes mellitus, presence of anxiety or depressive disorder, and impaired functional mobility. A score was constructed using these predictors to estimate the functional prognosis of dizziness at 6 months. A score based on the presence of easily obtainable clinical information facilitates identification of older adults in primary care with poor functional prognosis of their dizziness without exactly knowing the cause(s) of their dizziness. Clinical management might be most effective by treating factors that can be influenced, such as polypharmacy, anxiety and depression, and functional mobility.


The acute effects of static and ballistic stretching on vertical jump performance in trained women.
Traditionally stretching has been included as part of a warm-up that precedes athletic participation. However, there is mixed evidence as to whether stretching actually enhances or hinders athletic performance. Therefore, the purpose of this study was to examine the acute effects of static (SS) and ballistic stretching (BS) on vertical jump (VJ) performance and to investigate whether power was altered at 15 and 30 minutes after stretching. Sixteen actively trained women performed a series of vertical jumps (countermovement and drop jumps) after an initial nonstretching (NS) session and after participating in BS and SS sessions that were conducted in a balanced and randomized order. The results indicated that there was no significant difference (p < 0.05) in VJ scores as a result of static or ballistic stretching, elapsed time, or initial flexibility scores. This suggests that stretching prior to competition may not negatively affect the performance of trained women.


High spatiotemporal resolution in multifocal processing with femtosecond laser pulses.
We report spatial and temporal dispersion compensation for fan-out of femtosecond pulses with a low-frequency diffraction grating by means of a hybrid diffractive-refractive lens triplet. In this way, we achieve a multifocal light structure with nearly diffraction-limited light spots even for 20 fs pulse duration. The spatial chromatic compensation, which drastically reduces the lateral walk-off of the various spectral components, also allows us to improve the available bandwidth at the dispersion-compensated diffraction orders. In fact, the temporal width of the output pulse is essentially limited by the group-delay dispersion term, which is shown to be small. The high spatiotemporal resolution provided by our proposal permits parallel multifocal processing of materials with femtosecond pulses.


Gonadotropin-induced expression of receptors for growth hormone releasing factor in cultured granulosa cells.
The hypothalamic neuropeptide, GRF, is formed in the ovary and acts via specific receptors in granulosa cells to enhance cAMP production and steroidogenic responses to the pituitary gonadotropin, FSH. Granulosa cells cultured without hormonal treatment displayed low levels of binding sites for GRF and the related neuropeptide, vasoactive intestinal peptide. However, treatment with increasing concentrations (50-500 ng/ml) of FSH caused dose-dependent increases in cAMP production and expression of binding sites measured with radioiodinated [His1, Nle27]human GRF(1-32)NH2, with no change in binding affinity. The maximum increase in GRF binding sites (2.2-fold) was elicited by 250 ng/ml FSH after 72 h incubation. GRF binding sites were also increased by agents that elevate intracellular cAMP, including choleragen, vasoactive intestinal peptide, dibutyryl cAMP, and forskolin. Low doses of forskolin that did not alone increase [125I] [His1, Nle27] human GRF(1-32)NH2 binding potentiated the action of FSH on GRF binding sites, but the effects of maximal stimulatory doses of both agents were not additive. These findings demonstrate that FSH promotes the expression of GRF receptors in maturing granulosa cells through cAMP-dependent mechanisms. Since GRF enhances the actions of FSH on cAMP production and granulosa cell differentiation, and GRF receptors are increased by the cAMP-mediated actions of FSH, locally produced GRF could exert a positive autoregulatory action to accelerate follicular maturation by amplifying the granulosa cell response to FSH.


Parents' perceptions of staff competency in a neonatal intensive care unit.
* The aim of this study was to explore and describe parents' perceptions of staff competency in a neonatal intensive care unit (NICU). The study set out to use a grounded theory approach that was modified because of a number of constraints. Eight parents whose babies met a number of inclusion criteria were interviewed using focused conversational interviews. They were then transcribed and thematically analysed. The research approach was modified as the study developed because of practical and ethical access reasons: the sampling strategy and lack of opportunity to exploit fully the constant comparative method. * Four key themes which conceptualize competency as caring emerged from the data: parents are facilitated to integrate into the unit and do not feel a burden; parents feel in control whilst in the unit; parents have a choice to opt out from observing tasks and procedures on their baby; parents and the interprofessional team communicate well and provide appropriate information. These are discussed in the context of available literature. * In conclusion, the results of the study serve to highlight how parents' perceptions of competence in a professional are not based solely on skills and tasks but on many caring behaviours. The grounded theory approach has generated a number of areas for exploration, in particular, ideas about the conceptual basis of caring in an NICU context and its links to competence. The caring behaviours involve learning to share responsibility with families. The findings suggest that when there is 'a handing over' of control to parents and a greater emphasis on parent support, they feel less like 'guests'. These ideas are supported by the available literature. A number of methodological issues are raised.


Caregiver burden in Alzheimer disease: cross-sectional and longitudinal patient correlates.
Alzheimer disease (AD) imposes a severe burden on patients and their caregivers. Although there is substantial evidence of the adverse impact of burden, considerably less is known about its specific correlates and potential causes. The authors use data from the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE)-AD study to examine the relationship of burden and depression among AD caregivers to patient and caregiver sociodemographic characteristics, patients' cognitive status, psychiatric and behavioral symptoms, functional abilities, quality of life, and intensity of care provided by caregivers. CATIE-AD included outpatients in usual care settings and assessed treatment outcomes during 9 months. Data were examined from 421 ambulatory outpatients with a diagnosis of dementia of the Alzheimer type or probable AD with agitation or psychosis. The Burden Interview, the Beck Depression Inventory, and the Caregiver Distress Scale were used to evaluate caregiver burden. More severe psychiatric and behavioral problems and decreased patient quality of life, as well as lower functional capability were significantly associated with higher levels of burden and depression among caregivers at baseline. Six-month changes showed that decreased symptoms and improved quality of life were associated with decreased burden and accounted for most of the explained variance in change in burden measures. Severity of psychiatric symptoms, behavioral disturbances, and patients' quality of life are the main correlates of caregivers' experience burden. Psychosocial and pharmacologic interventions targeting these two aspects of the disorder are likely to not only alleviate patient suffering but also promote caregiver well-being.


The efficacy of somatosensory evoked potentials in evaluating new neurological deficits after spinal thoracic fusion and decompression.
Posterior thoracic fusion (PTF) is used as a surgical treatment for a wide range of pathologies. The monitoring of somatosensory evoked potentials (SSEPs) is used to detect and prevent injury during many neurological surgeries. The authors conducted a study to evaluate the efficacy of SSEPs in predicting perioperative lower-extremity (LE) neurological deficits during spinal thoracic fusion surgery. The authors included patients who underwent PTF with SSEP monitoring performed throughout the entire surgery from 2010 to 2015 at the University of Pittsburgh Medical Center (UPMC). The sensitivity, specificity, odds ratio, and receiver operating characteristic curve were calculated to evaluate the diagnostic accuracy of SSEP changes in predicting postoperative deficits. Univariate analysis was completed to determine the impact of age exceeding 65 years, sex, obesity, abnormal baseline testing, surgery type, and neurological deficits on the development of intraoperative changes. From 2010 to 2015, 771 eligible patients underwent SSEP monitoring during PTF at UPMC. Univariate and linear regression analyses showed that LE SSEP changes significantly predicted LE neurological deficits. Significant changes in LE SSEPs had a sensitivity and specificity of 19% and 96%, respectively, in predicting LE neurological deficits. The diagnostic odds ratio for patients with new LE neurological deficits who had significant changes in LE SSEPs was 5.86 (95% CI 2.74-12.5). However, the results showed that a loss of LE waveforms had a poor predictive value for perioperative LE deficits (diagnostic OR 1.58 [95% CI 0.19-12.83]). Patients with new postoperative LE neurological deficits are 5.9 times more likely to have significant changes in LE SSEPs during PTF. Surgeon awareness of an LE SSEP loss may alter surgical strategy and positively impact rates of postoperative LE neurological deficit status. The relatively poor sensitivity of LE SSEP monitoring may indicate a need for multimodal neurophysiological monitoring, including motor evoked potentials, in thoracic fusion surgery.


Prime-boost vaccination with HIV-1 Gag protein and cytosine phosphate guanosine oligodeoxynucleotide, followed by adenovirus, induces sustained and robust humoral and cellular immune responses.
A prophylactic vaccine for HIV-1 will probably require the induction and maintenance of both humoral and cellular immunity. One current strategy to achieve such long term immune responses is a prime-boost vaccination approach using a DNA priming inoculation, followed by recombinant viral boost. In this report we use a novel prime-boost approach in which the priming injections consist of recombinant HIV-1 Gag protein mixed with cytosine phosphate guanosine oligodeoxynucleotide (CpG ODN), followed by recombinant adenoviral boost expressing HIV-1 Gag. Analysis of the immune responses indicates that HIV-1 Gag protein plus CpG ODN immunization alone induces potent humoral as well as Th1 and CD8+ T cell responses. Boosting with recombinant adenovirus strikingly enhances CD8+, but not Th1, T cell responses, resulting in CD8+ T cell responses far greater in magnitude than Th1 responses. Furthermore, the Th1 and CD8+ T cell responses following prime-boost immunization were seen in both lymphoid and peripheral mucosal organs and were sustained over several months. Together, these data suggest a new immunization approach for elicitation of long term humoral and cellular immune responses.


Nutrient flows in agriculture in The Netherlands with special emphasis on pig production.
Annual nitrogen (N), phosphorus (P), and potassium (K) flows in agriculture in The Netherlands were identified and quantified in 1990, with special emphasis on pig production. Also, the effects that various management strategies in pig production have on NPK emission in 1990 were compared using a static deterministic simulation model. Ammonia emission from pig production in 1990 (60.9 Gg N) exceeded the defined target for the year 2000 (12.7 Gg N). Measures that affect volatilization of ammonia directly (i.e., introduction of low-emission stables, manure storage facilities, or manure application techniques) reduced ammonia emission most effectively. These measures, however, should be combined with a reduction in application of artificial N fertilizer to avoid an increase in N losses through leaching, run-off, or denitrification. Targets for ammonia emission in the year 2010 require a reduction in the pig population of 24 to 62%, in addition to implications of measures described in this article. National NPK losses in 1990 through leaching, run-off, or denitrification, predicted at 223.5 kg/ha for N, 32.7 kg/ha for P, and 67 kg/ha for K, exceeded government targets for the year 2010 (185 kg N/ha; 8.7 kg P/ha; norm not set for K). Reducing application of artificial NPK fertilizer reduced national NPK losses most effectively. For P, use of phytase and feeding pigs in accordance with their P requirements is required, in addition to limited use of artificial P fertilizer to meet targets for the year 2010. Hence, from an environmental point of view, pig production in The Netherlands is limited primarily by ammonia emission targets for the year 2010.


Circadian rest-activity rhythm disturbances in Alzheimer's disease.
Previous studies showed circadian rhythm disturbances in patients with Alzheimer's disease. Rest-activity rhythm disturbances manifest themselves through a fragmentation of the rhythm, a weak coupling with Zeitgebers, and high levels of activity during the night. The aim of the present study was to investigate which factors contribute to the presence of these disturbances. Therefore, several rest-activity rhythm, constitutional, and environmental variables were assessed in a heterogeneous group of 34 patients with Alzheimer's disease, including presenile and senile patients living at home or in a nursing home, as well as in 11 healthy controls. Circadian rest-activity rhythm disturbances were most prominent in institutionalized patients. Regression analyses showed the involvement of the following variables. First stability of the rest-activity rhythm is associated with high levels of daytime activity and high levels of environmental light resulting from seasonal effects as well as from indoor illumination. Presenile onset contributed to instability of the rhythm. Second, fragmentation of periods of activity and rest is associated with low levels of daytime activity, and is most prominent in moderately severe dementia. Third, night-time activity level is higher during the times of the year when the days are getting shorter and lower when the days are growing longer. These findings indicate that rest-activity rhythm disturbances may improve by increasing environmental light and daytime activity, an assumption for which empirical evidence has recently been published.


Evaluation of 5-fluorouracil metabolic enzymes as predictors of response to adjuvant chemotherapy outcomes in patients with stage II/III colorectal cancer: a decision-curve analysis.
The effectiveness of 5-fluorouracil (5-FU)-based adjuvant chemotherapy is reported in patients with colorectal cancer (CRC), but the usefulness of 5-FU metabolic enzymes as predictive biomarkers of the efficacy of this chemotherapy remains unclear. This study aims to verify whether 5-FU metabolic enzymes are predictive biomarkers in the clinical setting of adjuvant chemotherapy for stage II/III CRC. In total, 179 patients with stage II/III CRC who were treated at our institute between 2000 and 2010 were enrolled. Messenger RNA (mRNA) expression of major 5-FU metabolic enzymes, namely thymidylate synthase, dihydropyrimidine dehydrogenase, thymidine phosphorylase (TP), orotate phosphoribosyl transferase, and β-actin (control) was evaluated using the Danenberg Tumor Profile method. mRNA expression and other clinicopathological data were investigated with regard to CRC relapse. A total of 78 patients underwent surgery alone, while 101 underwent adjuvant chemotherapy (5-FU plus leucovorin [LV] or tegafur plus uracil /LV) following surgery. Relapse-free survival was longer and risk of recurrence was lower in association with high TP mRNA expression than in association with low TP mRNA expression in the adjuvant chemotherapy group (hazard ratio 0.66; 95 % confidence interval 0.47-0.92; p = 0.016), but not in the surgery alone group. mRNA expression of no other enzymes was associated with relapse in both groups. In decision-curve analyses, the predictive efficiency of TP mRNA expression plus clinicopathological factors was slightly better than that of clinicopathological factors only. TP mRNA expression in tumors predicted the effects of adjuvant chemotherapy for stage II/III CRC, although the beneficial effects were marginal.


[Age-adaptable to any age].
A definition separating the "elderly" from younger people should be avoided as well as any manifestation of any type of segregation. The environment must provide equal chances and opportunities for every person, regardless of age or abilities. Self-determined independence throughout life and the chance to remain in one's established home, community, and familiar environment require barrier-free design in all areas of the person's life-space. Dependencies and disabling factors can be diminished, and abilities can be restored by intensive rehabilitation and support of personal dignity, assisted by ambulant care and by products designed as barrier-free. They, lower the need for special aids and save costs. Such solutions are available. They must be applied with all consequences.


The Impact of Modifiable Preoral Factors on Swallowing and Nutritional Outcomes in Healthy Adults: A Scoping Review.
Swallowing has previously been characterized as consisting of four phases; however, it has become apparent that these four phases are not truly discrete and may be influenced by factors occurring prior to bolus entrance into the oral cavity (i.e., preoral factors). Still, the relationship between these factors and swallowing remains poorly understood. The aim of this review was to synthesize and characterize the literature pertaining to the influence of preoral factors on swallowing and nutritional outcomes in healthy individuals. We performed a scoping review, searching the databases of PubMed, CINAHL, Cochrane, and Scopus. Search terms included those related to swallowing, experience of preoral factors, and exclusionary terminology to reduce animal and pediatric literature. Our initial search revealed 5,560 unique articles, of which 153 met our inclusionary criteria and were accepted into the review. Of the accepted articles, 78% were focused exclusively on nutritional outcomes, 17% were focused on both swallowing and nutritional outcomes, and 5% were focused on solely swallowing outcomes. Of the preoral factors examined, 99% were exteroceptive in nature (17% olfactory, 44% visual, 21% auditory, 7% tactile, 11% other), while 1% were proprioceptive in nature. This review supports the influence of preoral factors on swallowing and nutritional outcomes. However, there is a large emphasis on the visual modality and on nutritional outcomes. Nearly none of the literature found in this review directly measured swallowing safety, efficiency, or physiology. Future work will benefit from a larger focus on proprioceptive preoral factors as they relate to swallowing outcomes.


Chronically Elevating Circulating Ketones Can Reduce Cardiac Inflammation and Blunt the Development of Heart Failure.
Previous studies have shown beneficial effects of acute infusion of the primary ketone body, β-hydroxybutyrate, in heart failure (HF). However, whether chronic elevations in circulating ketones are beneficial remains unknown. To chronically elevate circulating ketones in mice, we deleted the expression of the ketolytic, rate-limiting-enzyme, SCOT (succinyl-CoA:3-ketoacid-CoA transferase 1; encoded by Oxct1), in skeletal muscle. Tamoxifen-inducible skeletal muscle-specific Oxct1Muscle-/- knockout (n=32) mice and littermate controls (wild type; WT; n=35) were subjected to transverse aortic constriction (TAC) surgery to induce HF. Deletion of SCOT in skeletal, but not cardiac muscle resulted in elevated concentrations of fasted circulating β-hydroxybutyrate in knockout mice compared with WT mice (P=0.030). Five weeks following TAC, WT mice progressed to HF, whereas knockout mice with elevated fasting circulating ketones were largely protected from the TAC-induced effects observed in WT mice (ejection fraction, P=0.011; mitral E/A, P=0.012). Furthermore, knockout mice with TAC had attenuated expression of markers of sterile inflammation and macrophage infiltration, which were otherwise elevated in WT mice subjected to TAC. Lastly, addition of β-hydroxybutyrate to isolated hearts was associated with reduced NLRP3 (nucleotide-binding domain-like receptor protein 3)-inflammasome activation, which has been previously shown to play a role in contributing to HF-induced cardiac inflammation. These data show that chronic elevation of circulating ketones protects against the development of HF that is associated with the ability of β-hydroxybutyrate to directly reduce inflammation. These beneficial effects of ketones were associated with reduced cardiac NLRP3 inflammasome activation, suggesting that ketones may modulate cardiac inflammation via this mechanism.


7-Hydroxylated epiandrosterone (7-OH-EPIA) reduces ischaemia-induced neuronal damage both in vivo and in vitro.
Recent evidence suggests that steroids such as oestradiol reduce ischaemia-induced neurodegeneration in both in vitro and in vivo models. A cytochrome P450 enzyme termed cyp7b that 7-hydroxylates many steroids is expressed at high levels in brain, although the role of 7-hydroxylated steroids is unknown. We have tested the hypothesis that the steroid-mediated neuroprotection is dependent on the formation of 7-hydroxy metabolites. Organotypic hippocampal slice cultures were prepared from Wistar rat pups and maintained in vitro for 14 days. Cultures were then exposed to 3 h hypoxia and neuronal damage assessed 24 h later using propidium iodide fluorescence as a marker of cell damage. Neurodegeneration occurred primarily in the CA1 pyramidal cell layer. The steroids oestradiol, dehydroepiandrosterone and epiandrosterone (EPIA) were devoid of neuroprotective efficacy when present at 100 nM pre-, during and post-hypoxia. The 7-hydroxy metabolites of EPIA, 7alpha-OH-EPIA and 7beta-OH-EPIA significantly reduced neurotoxicity at 100 nM and 10 nM. 7beta-OH-EPIA was also neuroprotective in two in vivo rat models of cerebral ischaemia: 0.1 mg/kg 7beta-OH-EPIA significantly reduced hippocampal cell loss in a model of global forebrain ischaemia, whereas 0.03 mg/kg was neuroprotective in a model of focal ischaemia even when administration was delayed until 6 h after the onset of ischaemia. Taken together, these data demonstrate that 7-hydroxylation of steroids confers neuroprotective efficacy, and that 7beta-OH-epiandrosterone represents a novel class of neuroprotective compounds with potential for use in acute neurodegenerative diseases.


Epitope Mapping of the Anti-Diacylglycerol Kinase Monoclonal Antibody DhMab-4 for Immunohistochemical Analysis.
Diacylglycerol kinase (DGK) plays a pivotal role in intracellular signaling pathways in mammals. Activated G protein-coupled receptor activates phospholipase C (PLC) through heterotrimeric G protein, following which PLC hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) into diacylglycerol (DG) and inositol 1,4,5-trisphosphate (IP3). DGK catalyzes DG phosphorylation to produce phosphatidic acid. DG and phosphatidic acid function as second messengers and their intracellular concentrations are regulated by DGK; therefore, DGK plays an important role in regulating many biological processes. There are ten DGK isozymes, of which DGKη is classified as a type II DGK. Reports have shown that DGKη is associated with several diseases; for example, it is highly expressed in the hippocampus and cerebellum and is a key element in bipolar disorder. Although a DGKη-specific monoclonal antibody (mAb) is necessary to reveal the association between the expression of DGKη and diseases, an anti-DGKη mAb for immunohistochemistry has not yet been established. In this study, we established a specific anti-human DGKη (hDGKη) mAb, DhMab-4 (mouse IgG2b, kappa). DhMab-4 strongly stained Purkinje cells of human cerebellum in immunohistochemistry analysis. For epitope mapping of DhMab-4, we produced deletion or point mutants of hDGKη and performed western blotting to determine the binding epitope of DhMab-4. DhMab-4 reacted with dN745 mutant but not with dN750 mutant, indicating that the N-terminus of the DhMab-4 epitope is located between amino acids 745 and 750. More detailed analysis using point mutants demonstrated that five mutants, that is, D747A, P748A, F749A, G750A, and T752A, were not detected by DhMab-4. These results indicate that Asp747, Pro748, Phe749, Gly750, and Thr752 are important for DhMab-4 binding to hDGKη.


Lead-Free Mixed Tin and Germanium Perovskites for Photovoltaic Application.
The power-conversion efficiency (PCE) of lead halide perovskite photovoltaics has reached 22.1% with significantly improved structural stability, thanks to a mixed cation and anion strategy. However, the mixing element strategy has not been widely seen in the design of lead-free perovskites for photovoltaic application. Herein, we report a comprehensive study of a series of lead-free and mixed tin and germanium halide perovskite materials. Most importantly, we predict that RbSn0.5Ge0.5I3 possesses not only a direct bandgap within the optimal range of 0.9-1.6 eV but also a desirable optical absorption spectrum that is comparable to those of the state-of-the-art methylammonium lead iodide perovskites, favorable effective masses for high carrier mobility, as well as a greater resistance to water penetration than the prototypical inorganic-organic lead-containing halide perovskite. If confirmed in the laboratory, this new lead-free inorganic perovskite may offer great promise as an alternative, highly efficient solar absorber material for photovoltaic application.


Evidence for frequency-dependent selection maintaining polymorphism in the Batesian mimic Papilio polytes in multiple islands in the Ryukyus, Japan.
Batesian mimicry is a well-studied adaptation for predation avoidance, in which a mimetic species resembles an unpalatable model species. Batesian mimicry can be under positive selection because of the protection gained against predators, due to resemblance to unpalatable model species. However, in some mimetic species, nonmimetic individuals are present in populations, despite the benefits of mimicry. The mechanism for evolution of such mimetic polymorphism remains an open question. Here, we address the hypothesis that the abundance of mimics is limited by that of the models, leading to mimetic polymorphism. In addition, other forces such as the effects of common ancestry and/or isolation by distance may explain this phenomenon. To investigate this question, we focused on the butterfly, Papilio polytes, that exhibits mimetic polymorphism on multiple islands of the Ryukyus, Japan, and performed field surveys and genetic analysis. We found that the mimic ratio of P. polytes was strongly correlated with the model abundance observed on each of the five islands, suggesting negative frequency-dependent selection is driving the evolution of polymorphism in P. polytes populations. Molecular phylogenetic analysis indicated that the southern island populations are the major source of genetic diversity, and the middle and northern island populations arose by relatively recent migration. This view was also supported by mismatch distribution and Tajima's D analyses, suggesting a recent population expansion on the middle and northern islands, and stable population persistence on the southern islands. The frequency of the mimetic forms within P. polytes populations is thus explained by variations in the model abundance rather than by population structure. Thus, we propose that predation pressure, rather than neutral forces, have shaped the Batesian mimicry polymorphism in P. polytes observed in the Ryukyus.


Recombinant flagellins with deletions in domains D1, D2, and D3: Characterization as novel immunoadjuvants.
Bacterial flagellin activates the innate immune system and ultimately the adaptive immune system through a Toll-like receptor 5 (TLR5)-dependent signaling mechanism. Given that TLR5 is widely distributed in epithelia, flagellin is currently being developed as a mucosal adjuvant. Flagellin FliC from Salmonella enterica has four domains: the conserved D0 and D1 domains and the hypervariable D2 and D3 domains. The deletion of D3 and partial deletion of D2 in the recombinant FliCΔ174-400 strongly impairs flagellin's intrinsic antigenicity but does not affect the TLR5-dependent immunostimulation activity, i.e., the capacity to promote innate responses and adaptive responses to co-administered antigens. Here, we describe the development of novel recombinant flagellins with various deletions encompassing all of D2 and D3, and part of D1. Most of the recombinant molecules conserved an α-helical secondary structure that was as resistant to heat denaturation as the native protein. Whereas the recombinant flagellins' ability to trigger TLR5 varied markedly in vitro, most gave equivalent in vivo TLR5-dependent innate immune responses following intranasal administration of 2 μg of flagellin to mice. Concordantly, the recombinant flagellins were also valuable respiratory adjuvants for eliciting antibody responses to the foreign antigen ovalbumin, although their intrinsic antigenicity was decreased compared to the native flagellin and not increased compared to FliCΔ174-400. Our results show that the additional deletions of D2 and the distal part of D1 of FliCΔ174-400 does not impact on antigenicity and does not significantly modify the immunostimulatory adjuvant activity. Altogether, this study generated a novel set of recombinant flagellin that constitutes a portfolio of TLR5-dependent candidate adjuvants for vaccination.


UV radiation-sensitive norin 1 rice contains defective cyclobutane pyrimidine dimer photolyase.
Norin 1, a progenitor of many economically important Japanese rice strains, is highly sensitive to the damaging effects of UVB radiation (wavelengths 290 to 320 nm). Norin 1 seedlings are deficient in photorepair of cyclobutane pyrimidine dimers. However, the molecular origin of this deficiency was not known and, because rice photolyase genes have not been cloned and sequenced, could not be determined by examining photolyase structural genes or upstream regulatory elements for mutations. We therefore used a photoflash approach, which showed that the deficiency in photorepair in vivo resulted from a functionally altered photolyase. These results were confirmed by studies with extracts, which showed that the Norin 1 photolyase-dimer complex was highly thermolabile relative to the wild-type Sasanishiki photolyase. This deficiency results from a structure/function alteration of photolyase rather than of nonspecific repair, photolytic, or regulatory elements. Thus, the molecular origin of this plant DNA repair deficiency, resulting from a spontaneously occurring mutation to UV radiation sensitivity, is defective photolyase.


[Effect of ethanol on central mechanisms of appetite induced by electric stimulation of the lateral hypothalamus].
The effect of ethanol on food motivation evoked by electric stimulation of the "food center" in the hypothalamus was studied in experiments on rabbits. Intravenous injections of ethanol (0.5 g/kg) were accompanied by a short-term (up to 30 min) increase in the threshold of the hypothalamic "food center". Analysis of food motivation latency demonstrated that ethanol eliminated both inhibitory effects of the dorsal hippocamp and facilitating effects of the mesencephalic reticular formation on the excitability of the lateral hypothalamus. It is inferred that ethanol induces profound changes in monoaminergic and cholinergic mechanisms of the brain that provide for food motivation.


Interaction of rhodamine 123 with living cells studied by flow cytometry.
The cationic fluorochrome rhodamine 123 (R123), reported to bind specifically to mitochondria of living cells, was presently investigated with respect to its uptake by a variety of cell types in various functional states and the subsequent effect of the dye on cell growth. The emission spectrum of R123 taken up by cells undergoes a 12-nm red shift, suggesting formation of a complex. Cells accumulate R123 rapidly; near maximum binding is reached after 5 to 10 min, regardless of the temperature (0-37 degrees) of incubation. There is a dose-dependent relationship between R123 concentration in the medium and the dye accumulation in the cell that covers the range of 0.1 to 10.0 and 0.1 to 5.0 microgram of R123 per ml under equilibrium and nonequilibrium conditions, respectively. Some leakage of the dye from cells occurs, following their transfer into dye-free medium. Despite the leakage, the intracellular dye can be detected after at least two cell divisions, thus indicating that: (a) the R123-labeled cells divide; (b) during division, labeled mitochondria are distributed into the daughter cells; and (c) R123 may be used as a cell tracer. Cell death often is accompanied by a transient increase in R123 fluorescence. Dead cells exhibit either uniform, strong fluorescence or show a patchy labeling pattern suggesting swollen mitochondria. With time (4 to 8 hr), dead cells lose ability to retain R123 and lyse. Uptake of R123 by living cells is increased during the transition from quiescence into the cycle, and a decrease is seen when Friend leukemia cells undergo erythroid differentiation; in all cases, changes in R123 uptake are correlated with changes in cellular RNA content. Simultaneous cell staining with R123 and ethidium or propidium provides a rapid assay of the viability of the cells and their metabolic state, i.e., as related to proliferation or motility. Pulse-labeling of cells with up to 10 microgram of R123 per ml has no significant effect on their immediate growth and cloning efficiency. In the continuous presence of R123, however, cells become specifically arrested in the G1A compartment, i.e., in early G1 phase. Detailed analysis of the cell cycle kinetics reveals that cell progression through all phases is slowed 4 hr after addition of R123. Cell exit from G1A, however, is affected as early as 2 hr following addition of R123, and with time the cells are unable to leave this compartment at all. Uncharged rhodamine dyes (rhodamine 110 and rhodamine B) do not accumulate in mitochondria and are without effect on the cell cycle. The cytostatic effect of R123 is discussed in light of the dye specificity for mitochondrial membranes and the disruption of cell energy metabolism, resulting in the inability of the cells to attain a critical content of essential components (i.e., ribosomal RNA), necessary for cell entrance into the prereplicative (G1B) compartment of G1 phase.


High-resolution NMR studies of structure and dynamics of human ERp27 indicate extensive interdomain flexibility.
ERp27 (endoplasmic reticulum protein 27.7 kDa) is a homologue of PDI (protein disulfide-isomerase) localized to the endoplasmic reticulum. ERp27 is predicted to consist of two thioredoxin-fold domains homologous with the non-catalytic b and b' domains of PDI. The structure in solution of the N-terminal b-like domain of ERp27 was solved using high-resolution NMR data. The structure confirms that it has the thioredoxin fold and that ERp27 is a member of the PDI family. (15)N-NMR relaxation data were obtained and ModelFree analysis highlighted limited exchange contributions and slow internal motions, and indicated that the domain has an average order parameter S(2) of 0.79. Comparison of the single-domain structure determined in the present study with the equivalent domain within full-length ERp27, determined independently by X-ray diffraction, indicated very close agreement. The domain interface inferred from NMR data in solution was much more extensive than that observed in the X-ray structure, suggesting that the domains flex independently and that crystallization selects one specific interdomain orientation. This led us to apply a new rapid method to simulate the flexibility of the full-length protein, establishing that the domains show considerable freedom to flex (tilt and twist) about the interdomain linker, consistent with the NMR data.


Elevated intracellular calcium levels in cerebellar granule neurons of weaver mice.
Weaver mice carry a mutation in the pore domain of the Girk2 (Kcnj6) gene. The mutation causes GIRK2 containing channels to lose ion selectivity and to become constitutively active. It is not known how this alteration in ion channel activity causes in cerebellar granule cells the defects in neurite extension, cell migration and induction of cell death that are characteristic of weaver mice. One possibility is that the mutation causes an inability to regulate intracellular calcium levels properly. We tested this hypothesis by measuring intracellular calcium levels in granule cells and Purkinje cells in slices from the cerebellum of weaver mice. We report here that weaver mice have increases in resting calcium levels in their granule cells, which may account for the multiple effects of the weaver mutation upon these cells.


Genetic variation in CACNA1C affects brain circuitries related to mental illness.
The CACNA1C gene (alpha-1C subunit of the L-type voltage-gated calcium channel) has been identified as a risk gene for bipolar disorder and schizophrenia, but the mechanism of association has not been explored. To identify the neural system mechanism that explains the genetic association between the CACNA1C gene and psychiatric illness using neuroimaging and human brain expression. We used blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging (fMRI) to measure brain activation in circuitries related to bipolar disorder and schizophrenia by comparing CACNA1C genotype groups among healthy subjects. We tested the effect of genotype on messenger RNA (mRNA) levels of CACNA1C in postmortem human brain. A case-control analysis was used to determine the association of CACNA1C genotype with schizophrenia. National Institutes of Health Clinical Center. Healthy men and women of white race/ethnicity participated in the fMRI study. Postmortem samples from normal human brains were used for the brain expression study. Patients with schizophrenia and healthy subjects were used in the case-control analysis. BOLD fMRI, mRNA levels in postmortem brain samples, and genetic association with schizophrenia. The risk-associated single-nucleotide polymorphism (SNP rs1006737) in CACNA1C predicted increased hippocampal activity during emotional processing (P = .001 uncorrected, P((false recovery rate [FDR])) = .05, z = 3.20) and increased prefrontal activity during executive cognition (P = 2.8e-05 uncorrected, P(FDR) = .01, z = 4.03). The risk-associated SNP also predicted increased expression of CACNA1C mRNA in human brain (P = .002). CACNA1C was associated with schizophrenia in our case-control sample (odds ratio, 1.77; P = .03). The risk-associated SNP in CACNA1C maps to circuitries implicated in genetic risk for bipolar disorder and schizophrenia. Its effects in human brain expression implicate a molecular and neural system mechanism for the clinical genetic association.


Replacing Animal Protein with Soy-Pea Protein in an "American Diet" Controls Murine Crohn Disease-Like Ileitis Regardless of Firmicutes: Bacteroidetes Ratio.
The current nutritional composition of the "American diet" (AD; also known as Western diet) has been linked to the increasing incidence of chronic diseases, including inflammatory bowel disease (IBD), namely Crohn disease (CD). This study investigated which of the 3 major macronutrients (protein, fat, carbohydrates) in the AD has the greatest impact on preventing chronic inflammation in experimental IBD mouse models. We compared 5 rodent diets designed to mirror the 2011-2012 "What We Eat in America" NHANES. Each diet had 1 macronutrient dietary source replaced. The formulated diets were AD, AD-soy-pea (animal protein replaced by soy + pea protein), AD-CHO ("refined carbohydrate" by polysaccharides), AD-fat [redistribution of the ω-6:ω-3 (n-6:n-3) PUFA ratio; ∼10:1 to 1:1], and AD-mix (all 3 "healthier" macronutrients combined). In 3 separate experiments, 8-wk-old germ-free SAMP1/YitFC mice (SAMP) colonized with human gut microbiota ("hGF-SAMP") from CD or healthy donors were fed an AD, an AD-"modified," or laboratory rodent diet for 24 wk. Two subsequent dextran sodium sulfate-colitis experiments in hGF-SAMP (12-wk-old) and specific-pathogen-free (SPF) C57BL/6 (20-wk-old) mice, and a 6-wk feeding trial in 24-wk-old SPF SAMP were performed. Intestinal inflammation, gut metagenomics, and MS profiles were assessed. The AD-soy-pea diet resulted in lower histology scores [mean ± SD (56.1% ± 20.7% reduction)] in all feeding trials and IBD mouse models than did other diets (P < 0.05). Compared with the AD, the AD-soy-pea correlated with increased abundance in Lactobacillaceae and Leuconostraceae (1.5-4.7 log2 and 3.0-5.1 log2 difference, respectively), glutamine (6.5 ± 0.8 compared with 3.9 ± 0.3 ng/μg stool, P = 0.0005) and butyric acid (4:0; 3.3 ± 0.5 compared with 2.54 ± 0.4 ng/μg stool, P = 0.006) concentrations, and decreased linoleic acid (18:2n-6; 5.4 ± 0.4 compared with 8.6 ± 0.3 ng/μL plasma, P = 0.01). Replacement of animal protein in an AD by plant-based sources reduced the severity of experimental IBD in all mouse models studied, suggesting that similar, feasible adjustments to the daily human diet could help control/prevent IBD in humans.


2-difluoromethyloestrone 3-O-sulphamate, a highly potent steroid sulphatase inhibitor.
Steroid sulphatase is a target enzyme of growing therapeutic importance. The synthesis and in vitro biological evaluation of three novel 2-substituted analogues of oestrone 3-O-sulphamate (EMATE), an established steroid sulphatase inhibitor, are described. One inhibitor, 2-difluoromethyloestrone 3-O-sulphamate (6), was found to have an IC50 of 100 pM and be some 90-fold more potent than EMATE in inhibiting steroid sulphatase activity in a placental microsomal preparation, rendering this agent the most potent steroidal STS inhibitor in vitro reported to date. Lowering of the pKa value of the leaving parent steroid phenol by the 2-difluoromethyl group during irreversible enzyme sulphamoylation most likely facilitates the potent inactivation of steroid sulphatase by (6). However, our preliminary molecular docking studies using the X-ray crystal structure of steroid sulphatase suggest that F.......H interactions between the 2-difluoromethyl group of (6) and hydrogen bond donor residues lining the catalytic site of STS might also contribute to the high potency observed for (6).


Genetic organization of the mecA region in methicillin-susceptible and methicillin-resistant strains of Staphylococcus sciuri.
A homolog of the Staphylococcus aureus methicillin resistance gene mecA was recently shown to be ubiquitous in independent isolates of the animal species Staphylococcus sciuri. The mecA gene homolog and regions flanking it were cloned and sequenced from four strains of S. sciuri: strain K1 (ATCC 29062), a representative of S. sciuri subsp. sciuri; two strains (K3 and K8) representing S. sciuri subsp. rodentius; and strain K11, a representative of S. sciuri subsp. carnaticum. Strains K1 and K11 were susceptible to methicillin, while strains K3 and K8 showed heterogeneous resistance. The mecA genes of strains K1 and K11 and one of the two copies of mecA (mecA1) present in strain K3 had virtually identical DNA sequences in the mecA gene and were similar in genetic organization in the flanking regions. In contrast, the single copy of mecA in strain K8 and the second copy of mecA (mecA2) in strain K3 had mecA DNA sequences identical to that of S. aureus mecA, and the mecA region in these two strains was also similar to that of the same region in the S. aureus strain used for comparison. Interestingly, an open reading frame defining an N-terminal truncated polypeptide, NTORF101, with a high degree of homology to a DNA segment in the hypervariable region of methicillin-resistant S. aureus (and also similar to the Escherichia coli gene ugpQ) was also identified downstream of the mecA homolog of strain K11, representing S. sciuri subsp. carnaticum. The ugpQ-like gene is not present in methicillin-susceptible strains of S. aureus. The presence of such a ugpQ-like gene together with the homolog of mecA in strain K11 supports the speculation that these genetic elements may be evolutionary relatives and/or precursors of the genetic determinant of methicillin resistance in S. aureus.


Transcription factor 4 (TCF4) and schizophrenia: integrating the animal and the human perspective.
Schizophrenia is a genetically complex disease considered to have a neurodevelopmental pathogenesis and defined by a broad spectrum of positive and negative symptoms as well as cognitive deficits. Recently, large genome-wide association studies have identified common alleles slightly increasing the risk for schizophrenia. Among the few schizophrenia-risk genes that have been consistently replicated is the basic Helix-Loop-Helix (bHLH) transcription factor 4 (TCF4). Haploinsufficiency of the TCF4 (formatting follows IUPAC nomenclature: TCF4 protein/protein function, Tcf4 rodent gene cDNA mRNA, TCF4 human gene cDNA mRNA) gene causes the Pitt-Hopkins syndrome-a neurodevelopmental disease characterized by severe mental retardation. Accordingly, Tcf4 null-mutant mice display developmental brain defects. TCF4-associated risk alleles are located in putative coding and non-coding regions of the gene. Hence, subtle changes at the level of gene expression might be relevant for the etiopathology of schizophrenia. Behavioural phenotypes obtained with a mouse model of slightly increased gene dosage and electrophysiological investigations with human risk-allele carriers revealed an overlapping spectrum of schizophrenia-relevant endophenotypes. Most prominently, early information processing and higher cognitive functions appear to be associated with TCF4 risk genotypes. Moreover, a recent human study unravelled gene × environment interactions between TCF4 risk alleles and smoking behaviour that were specifically associated with disrupted early information processing. Taken together, TCF4 is considered as an integrator ('hub') of several bHLH networks controlling critical steps of various developmental, and, possibly, plasticity-related transcriptional programs in the CNS and changes of TCF4 expression also appear to affect brain networks important for information processing. Consequently, these findings support the neurodevelopmental hypothesis of schizophrenia and provide a basis for identifying the underlying molecular mechanisms.


Transforming service employees and climate: a multilevel, multisource examination of transformational leadership in building long-term service relationships.
This longitudinal field study integrates the theories of transformational leadership (TFL) and relationship marketing to examine how TFL influences employee service performance and customer relationship outcomes by transforming both (at the micro level) the service employees' attitudes and (at the macro level) the work unit's service climate. Results revealed that, at the individual level, managers' TFL was positively related to employee service performance, which, in turn, positively predicted customers' expressed intention to maintain a long-term service relationship with the service employee and manager-reported number of the employee's long-term customers measured 9 months later. In addition, the relationship between TFL and employee service performance was partially mediated by employee self-efficacy. Furthermore, store-level TFL was positively associated with store-level service climate, and service climate further enhanced the relationship between individual-level TFL and employee service performance.


Perpetration of intimate partner violence and mental health outcomes: sex- and gender-disaggregated associations among adolescents and young adults in Nigeria.
The association between intimate partner violence (IPV) victimisation and poor mental health outcomes is well established. Less is known about the correlation between IPV perpetration and mental health, particularly among adolescents and young adults. Using data from the nationally representative Violence Against Children Survey, this analysis examines the association between IPV perpetration and mental health for male and female adolescents and young adults in Nigeria. Multivariate logistic regression models were used to examine associations between ever-perpetration of IPV and four self-reported mental health variables: severe sadness, feelings of worthlessness, suicide ideation, and alcohol use. Models were sex-disaggregated, controlled for age, marital status, and schooling, and tested with and without past exposure to violence. Standard errors were adjusted for sampling stratification and clustering. Observations were weighted to be representative of 13-24 year-olds in Nigeria. Males were nearly twice as likely as females to perpetrate IPV (9% v. 5%, respectively; P < 0.001), while odds of perpetration for both sexes were higher for those ever experiencing IPV (adjusted odds ratio (aOR) = 4.60 for males; aOR = 2.71 for females). Female perpetrators had 2.73 higher odds of reporting severe sadness (95% confidence interval CI = 1.44, 5.17; P = 0.002) and 2.72 times greater odds of reporting suicide ideation (1.28, 5.79; P = 0.010) than non-perpetrating females, even when controlling for past-year violence victimisation. In contrast, male perpetrators had 2.65 times greater odds of feeling worthless (1.09, 6.43; P = 0.031), and 2.36 times greater odds of reporting alcohol use in the last 30 days (1.50, 3.73; P < 0.001), as compared to non-perpetrating males. Among adolescents and young adults in Nigeria, IPV perpetration and negative mental health outcomes are associated but differ for males and females. Mindful of the cross-sectional nature of the data, it is possible that socially determined gender norms may shape the ways in which distress from IPV perpetration is understood and expressed. Additional research is needed to clarify these associations and inform violence prevention efforts.


Genetic landscape and novel disease mechanisms from a large LGMD cohort of 4656 patients.
Limb-girdle muscular dystrophies (LGMDs), one of the most heterogeneous neuromuscular disorders (NMDs), involves predominantly proximal-muscle weakness with >30 genes associated with different subtypes. The clinical-genetic overlap among subtypes and with other NMDs complicate disease-subtype identification lengthening diagnostic process, increases overall costs hindering treatment/clinical-trial recruitment. Currently seven LGMD clinical trials are active but still no gene-therapy-related treatment is available. Till-date no nation-wide large-scale LGMD sequencing program was performed. Our objectives were to understand LGMD genetic basis, different subtypes' relative prevalence across US and investigate underlying disease mechanisms. A total of 4656 patients with clinically suspected-LGMD across US were recruited to conduct next-generation sequencing (NGS)-based gene-panel testing during June-2015 to June-2017 in CLIA-CAP-certified Emory-Genetics-Laboratory. Thirty-five LGMD-subtypes-associated or LGMD-like other NMD-associated genes were investigated. Main outcomes were diagnostic yield, gene-variant spectrum, and LGMD subtypes' prevalence in a large US LGMD-suspected population. Molecular diagnosis was established in 27% (1259 cases; 95% CI, 26-29%) of the patients with major contributing genes to LGMD phenotypes being: CAPN3(17%), DYSF(16%), FKRP(9%) and ANO5(7%). We observed an increased prevalence of genetically confirmed late-onset Pompe disease, DNAJB6-associated LGMD subtype1E and CAPN3-associated autosomal-dominant LGMDs. Interestingly, we identified a high prevalence of patients with pathogenic variants in more than one LGMD gene suggesting possible synergistic heterozygosity/digenic/multigenic contribution to disease presentation/progression that needs consideration as a part of diagnostic modality. Overall, this study has improved our understanding of the relative prevalence of different LGMD subtypes, their respective genetic etiology, and the changing paradigm of their inheritance modes and novel mechanisms that will allow for improved timely treatment, management, and enrolment of molecularly diagnosed individuals in clinical trials.


Betula etnensis Raf. (Betulaceae) Extract Induced HO-1 Expression and Ferroptosis Cell Death in Human Colon Cancer Cells.
Betula etnensis Raf. (Birch Etna) belonging to the Betulaceae family grows on the eastern slope of Etna. Many bioactive compounds present in Betula species are considered promising anticancer agents. In this study, we evaluated the effects of B. etnensis Raf. bark methanolic extract on a human colon cancer cell line (CaCo2). In order to elucidate the mechanisms of action of the extract, cellular redox status, cell cycle, and heme oxygenase-1 (HO-1) expression in ferroptosis induction were evaluated. Cell viability and proliferation were tested by tetrazolium (MTT) assayand cell cycle analysis, while cell death was evaluated by annexin V test and lactic dehydrogenase (LDH) release. Cellular redox status was assessed by measuring thiol groups (RSH) content, reactive oxygen species (ROS) production, lipid hydroperoxide (LOOH) levels and (γ-glutamylcysteine synthetase) γ-GCS and HO-1 expressions. The extract significantly reduced cell viability of CaCo2, inducing necrotic cell death in a concentration-depending manner. In addition, an increase in ROS levels and a decrease of RSH content without modulation in γ-GCS expression were detected, with an augmentation in LOOH levels and drastic increase in HO-1 expression. These results suggest that the B. etnensis Raf. extract promotes an oxidative cellular microenvironment resulting in CaCo2 cell death by ferroptosis mediated by HO-1 hyper-expression.


Variable stromal periductular expression of CD34 and smooth muscle actin (SMA) in intraductal carcinoma of the breast.
In breast carcinoma, the stromal loss of CD34 expression and acquisition of SMA myofibroblastic features may constitute a prerequisite for tumor invasiveness. However, this hypothesis remains controversial, with some authors describing the loss of CD34 fibrocytes in the absence of SMA myofibroblastic-like cells in the stroma of invasive carcinoma. Others have also described the disappearance of CD34 fibrocytes from in situ carcinoma. To clarify this issue, we compared the distribution of CD34 fibrocytes and SMA reactive myofibroblasts between stromal areas of tumor-free mammary tissue, ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC). In addition to 28 IDC, 300 normal duct-lobular units and 600 ducts with DCIS (158 low-grade, 266 intermediate, and 176 high-grade) were scored. The relationships between staining patterns and different histological features (grade of DCIS and presence or absence of necrosis) were compared. Loss of CD34 expression and acquisition of SMA expression were more frequent in high-grade in situ lesions than in intermediate and low-grade lesions (p<0.001). When necrosis was found in association with grade 2 or 3 DCIS, the decrease in CD34 expression was higher than in lesions without necrosis and that independently of the grade of DCIS (p<0.05). Necrosis did not appear to play a significant role in the expression of SMA (p = 0.35). In all cases, the stroma of invasive carcinomas showed a complete loss of CD34 fibrocytes. Future research on both CD34 fibrocytes and mechanisms stromal changes are essential in the future and may potentially lead to new treatment approaches.


In vitro antimicrobial and antimycobacterial activity and HPLC-DAD screening of phenolics from Chenopodium ambrosioides L.
The main objective of this study was to demonstrate the antimicrobial potential of the crude extract and fractions of Chenopodium ambrosioides L., popularly known as Santa-Maria herb, against microorganisms of clinical interest by the microdilution technique, and also to show the chromatographic profile of the phenolic compounds in the species. The Phytochemical screening revealed the presence of cardiotonic, anthraquinone, alkaloids, tannins and flavonoids. The analysis by HPLC-DAD revealed the presence of rutin in the crude extract (12.5±0.20mg/g), ethyl acetate (16.5±0.37mg/g) and n-butanol (8.85±0.11mg/g), whereas quercetin and chrysin were quantified in chloroform fraction (1.95±0.04 and 1.04±0.01mg/g), respectively. The most promising results were obtained with the ethyl acetate fraction, which inhibited a greater number of microorganisms and presented the lowest values of MIC against Staphylococcus aureus and Enterococcus faecalis (MIC=0.42mg/mL), Pseudomonas aeruginosa (MIC=34.37mg/mL), Paenibacillus apiarus (MIC=4.29mg/mL) and Paenibacillus thiaminolyticus (MIC=4.29mg/mL). Considering mycobacterial inhibition, the best results were obtained by chloroform fraction against M. tuberculosis, M. smegmatis, and M. avium (MIC ranging from 156.25 to 625μg/mL). This study proves, in part, that the popular use of C. ambrosioides L. can be an effective and sustainable alternative for the prevention and treatment of diseases caused by various infectious agents.


Neuroleptic malignant syndrome in a patient with corticobasal degeneration.
Parkinson's disease is a principal underlying disease of neuroleptic malignant syndrome (NMS) occurring in parkinsonian disorders, but NMS may occur in patients with progressive supranuclear palsy and multiple system atrophy. We report first patient with corticobasal degeneration (CBD) who developed NMS after abrupt reduction of antiparkinsonian medication and concurrent infection. It should be kept in mind that the prevention of infectious illness, which is common complication in parkinson-plus syndrome, is important, and dose reduction or withdrawal of anti-parkinsonian medications should be carefully performed even in the patients with CBD who are expected to be unresponsive to levodopa treatment.


Positive inotropic action of NMDA receptor antagonist (+)-MK801 in rat heart.
(+)-MK801, a noncompetitive NMDA receptor antagonist, was reported to exhibit anticonvulsive and neuroprotective activities during the postischemic period. Intravenous administration of (+)-MK801 produced tachycardia in rats, but bradycardia in pigs. We examined the mechanical and electrophysiological effects of (+)-MK801 on rat cardiac tissues. (+)-MK801 dose-dependently increased (3-100 microM) twitch tension in rat atria and ventricular strips. The spontaneous beating rate in rat right atria, however, was dose-dependently decreased by (+)-MK801. The inotropic effect of (+)-MK801 was affected neither by alpha(1)-antagonist (1 microM prazosin) nor by beta(1)-adrenoceptor antagonist (3 microM atenolol), but significantly by a transient outward K(+) channel blocker (3 mM 4-aminopyridine). (+)-MK801 did not cause any significant change of intracellular cAMP content. Electrophysiological study in rat ventricular cells revealed that (+)-MK801 concentration-dependently prolonged the action potential duration with a concomitant decrease in the maximum rate of the action potential upstroke (V(max)) and an increase in the recovery time constant of V(max). Voltage clamp study showed that (+)-MK801 (3 microM) reduced inward Na(+) current (I(Na)), along with a slowing of its recovery from inactivation and a slight negative shift of its voltage-dependent steady-state inactivation curves. At a much higher concentration (30 microM), (+)-MK801 slightly reduced the amplitude of L-type calcium inward current (I(Ca)), although the voltage dependence of its steady-state inactivation was unaffected. For the potassium currents in rat ventricular cells, 3 microM of (+)-MK801 reduced the peak transient outward current (I(to)), steady-state outward current (I(ss)) and inward current through K(1) channels. The inhibition of I(to) was associated with a prominent negative shift in the voltage dependence of its steady-state inactivation curve. The outward current through K(1) channels was unaffected. These results indicate that (+)-MK801 may be a strong I(Na) and I(to) blocker with some I(Ca) blocking activity. The inhibition of I(to) and other K(+) efflux would prolong action potential duration, produce positive inotropic action and contribute to the negative chronotropic effect of (+)-MK801.


Tissue responses to molecularly reinforced polylactide-co-glycolide implants.
Plates for internal fixation fabricated from biodegradable polymers degrade via an autocatalytic route. When they are used in bone implants of significant size and thickness, hollowing of the implant may occur while the overall dimensions appear unchanged. We hypothesized that incorporation of a cross-linked polypropylene fumarate matrix into polylactide-co-glycolide bone plates may provide an internal molecular network which prevents implant collapse. Cross-linking reagents of varying hydrophilicity including N-vinylpyrrolidone (VP), hydroxyethylmethacrylate (HEMA), and ethyleneglycol dimethacrylate (EGDMA) were employed. With the objective of determining the most biocompatible and structurally sound composition for molecular reinforcement, we investigated tissue responses in both subcutaneous and orthotopic rodent implantation models in relation to maintenance of implant integrity by histologic, histomorphometric, and stereomicroscopic analysis. Results showed that tissue responses were correlated with dimensional stability of the implants. The most favorable results were seen with the hydrophobic cross-linker EGDMA; this may have been related to the initial reduction of the water uptake by the implant. Cross-linking of polypropylene fumarate with EGDMA within a polylactide-co-glycolide bone plate may offer a means to maintain excellent biocompatibility while improving dimensional stability of biodegradable bone plates.


Bronchial thermoplasty.
Asthma, by definition is a variable disease. When there is more than normal natural variation in airflow, asthma can be provoked by a wide range of stimuli that include infectious, allergic, and environmental agents. Bronchoconstriction determines much of the short-term variability in airflow that characterizes asthma. Current treatments do not redress the excess smooth muscle mass that is present in the remodeled airway in chronic asthma. Thus, it is intriguing to consider the potential contribution of bronchial thermoplasty (a procedure that involves controlled heat treatment to reduce the mass of the airway smooth muscle) as an effective therapy for poorly controlled asthma.


Constitutive internalization of cystic fibrosis transmembrane conductance regulator occurs via clathrin-dependent endocytosis and is regulated by protein phosphorylation.
Although the cystic fibrosis transmembrane conductance regulator (CFTR) is primarily implicated in the regulation of plasma-membrane chloride permeability, immunolocalization and functional studies indicate the presence of CFTR in the endosomal compartment. The mechanism of CFTR delivery from the cell surface to endosomes is not understood. To delineate the internalization pathway, both the rate and extent of CFTR accumulation in endosomes were monitored in stably transfected Chinese hamster ovary (CHO) cells. The role of clathrin-dependent endocytosis was assessed in cells exposed to hypertonic medium, potassium depletion or intracellular acid-load. These treatments inhibited clathrin-dependent endocytosis by >90%, as verified by measurements of 125I-transferrin uptake. Functional association of CFTR with newly formed endosomes was determined by an endosomal pH dissipation protocol [Lukacs, Chang, Kartner, Rotstein, Riordan and Grinstein (1992) J. Biol. Chem. 267, 14568-14572]. As a second approach, endocytosis of CFTR was determined after cell-surface biotinylation with the cleavable sulphosuccinimidyl-2-(biotinamido)ethyl-1,3-dithio- propionate. Both the biochemical and the functional assays indicated that arresting the formation of clathrin-coated vesicles inhibited the retrieval of the CFTR from the plasma membrane to endosomes. An overall arrest of membrane traffic cannot account for the inhibition of CFTR internalization, since the fluid-phase endocytosis was not effected by the treatments used. Thus the efficient, constitutive internalization of surface CFTR (5% per min) occurs, predominantly by clathrin-dependent endocytosis. Stimulation of protein phosphorylation by cAMP-dependent protein kinase A and by protein kinase C decreased the rate of internalization of cell-surface biotinylated CFTR, and contributed to a substantial diminution of the internal CFTR pool compared with that of unstimulated cells. These results suggest that the rate of CFTR internalization may participate in the determination of the CFTR channel density, and consequently, of the cAMP-stimulated chloride conductance of the plasma membrane.


Antibody responses in humans infected with newly emerging strains of West Nile Virus in Europe.
Infection with West Nile Virus (WNV) affects an increasing number of countries worldwide. Although most human infections result in no or mild flu-like symptoms, the elderly and those with a weakened immune system are at higher risk for developing severe neurological disease. Since its introduction into North America in 1999, WNV has spread across the continental United States and caused annual outbreaks with a total of 36,000 documented clinical cases and ∼1,500 deaths. In recent years, outbreaks of neuroinvasive disease also have been reported in Europe. The WNV strains isolated during these outbreaks differ from those in North America, as sequencing has revealed that distinct phylogenetic lineages of WNV concurrently circulate in Europe, which has potential implications for the development of vaccines, therapeutics, and diagnostic tests. Here, we studied the human antibody response to European WNV strains responsible for outbreaks in Italy and Greece in 2010, caused by lineage 1 and 2 strains, respectively. The WNV structural proteins were expressed as a series of overlapping fragments fused to a carrier-protein, and binding of IgG in sera from infected persons was analyzed. The results demonstrate that, although the humoral immune response to WNV in humans is heterogeneous, several dominant peptides are recognized.


[Distribution of the kinin-breakdown activity in areas of the rat brain in the dynamics of spontaneous hypertension].
Kinin-damaging activity (KDA) has been studied in 8 brain areas of normotensive rats and rats with spontaneous hypertension, aged 3 to 12 months. A significant depression in KDA was revealed in midbrain, striatum, thalamus, and pituitary body of normotensive rats 6 months of age, as compared to 3-month-old animals. A tendency towards KDA increase was noted in the hypothalamus. In 12-month-old normotensive rats KDA level returned to baseline (3 months of age). Comparison of KDA in rats with spontaneous hypertension aged 3 to 12 months has revealed no age-dependent differences and it is, therefore, believed that rats with spontaneous hypertension lack certain mechanisms inducing a considerable decrease of KDA in 6-month-old normotensive rats.


Improving the timeliness of care for patients with acute ST-elevation myocardial infarction: implications of "self-transport" versus use of EMS.
Early reperfusion of the myocardium is crucial in treating patients with acute ST segment elevation myocardial infarction (STEMI). Emergency medical services (EMS) plays an important role in minimizing delays to reperfusion. This study demonstrates that a large proportion of patients presenting with STEMI do not use EMS. Thus, such patients cannot benefit from many mechanisms designed to minimize time delays. Healthcare providers are challenged with both implementing strategies for improving care (minimizing delays to reperfusion) for patients not using EMS, and encouraging patients to access care via EMS in the event of acute chest pain.


Evolution of the Sterol Biosynthetic Pathway of Pythium insidiosum and Related Oomycetes Contributes to Antifungal Drug Resistance.
Pythiosis is a life-threatening infectious disease caused by the oomycete Pythium insidiosum Direct exposure to Py. insidiosum zoospores can initiate infections of the eye, limb, gastrointestinal tract, or skin/subcutaneous tissue. Treatments for pythiosis have mostly relied on surgery. Antifungal drugs are generally ineffective against Py. insidiosum However, one patient with an invasive Py. insidiosum infection recovered completely following treatment with terbinafine and itraconazole. Additionally, the drug target sterol biosynthetic enzymes have been identified in the oomycete Aphanomyces euteiches It remains an open question whether Py. insidiosum is susceptible to the antifungal drugs and harbors any of the known drug target enzymes. Here, we determined the in vitro susceptibilities of terbinafine and itraconazole against 30 isolates of Py. insidiosum We also analyzed endogenous sterols and searched for genes encoding the sterol biosynthetic enzymes in the genomes of Py. insidiosum and related oomycetes. The susceptibility assay showed that the growth of each of the Py. insidiosum isolates was inhibited by the antifungal agents, but only at difficult-to-achieve concentrations, which explains the clinical resistance of the drugs in the treatment of pythiosis patients. Genome searches of Py. insidiosum and related oomycetes demonstrated that these organisms contained an incomplete set of sterol biosynthetic enzymes. Gas chromatographic mass spectrometry did not detect any sterol end products in Py. insidiosum In conclusion, Py. insidiosum possesses an incomplete sterol biosynthetic pathway. Resistance to antifungal drugs targeting enzymes in the ergosterol biosynthetic pathway in Py. insidiosum was due to modifications or losses of some of the genes encoding the drug target enzymes.


Presentation and management of community-onset vs hospital-onset first seizures.
New-onset seizures are frequently encountered in community and hospital settings. It is likely that seizures presenting in these distinct settings have different etiologies and prognoses, requiring different investigation and treatment approaches. We directly compare the presentation and management of patients with community- and hospital-onset first seizures attending the same hospital. We reviewed the medical records of patients aged 18 years or older with discharge International Classification of Diseases, Australian Classification (ICD-10-AM) codes of G40 (epilepsy), G41 (status epilepticus), and R56.8 (unspecified convulsions), who attended a general hospital in Melbourne, Australia, from January 1, 2008, through November 30, 2016. Patients with new-onset seizures were included for analysis. A total of 367 patients were discharged with a relevant ICD-10-AM code. Among them, 151 patients met the inclusion criteria: 97 presented to the emergency department with community-onset seizure (median age 70 years), and 54 experienced seizures during hospitalization for other indications (median age 80.5 years). Provoked seizures were more common in the latter group (26.8% vs 63.0%, p < 0.001), with exposure to proconvulsant drugs a major risk factor. Despite not fulfilling the International League Against Epilepsy (ILAE) diagnostic criteria, 72.5% (58/80) who survived to discharge were prescribed antiepileptic drug (AED) therapy, whereas 19.0% (12/63) of those who met the ILAE criteria were not. Hospitalized elderly patients are at an increased risk of provoked seizures, and caution should be exercised when prescribing potential proconvulsant medications and procedures. A more standardized approach to AED prescribing is needed. Further studies should consider morbidity, mortality, and health economic effects of first seizures and assess optimal management to improve outcomes in this cohort.


Test-retest reliability measures for curve data: an overview with recommendations and supplementary code.
The purpose of this paper is to provide an overview of available methods for reliability investigations when the outcome of interest is a curve. Curve data, or functional data, is commonly collected in biomechanical research in order to better understand different aspects of human movement. Using recent statistical developments, curve data can be analysed in its most detailed form, as functions. However, an overview of appropriate statistical methods for assessing reliability of curve data is lacking. A review of contemporary literature of reliability measures for curve data within the fields of biomechanics and statistics identified the following methods: coefficient of multiple correlation, functional limits of agreement, measures of distance and similarity, and integrated pointwise indices (an extension of univariate reliability measures to curve data, inclusive of Pearson correlation, intraclass correlation, and standard error of measurement). These methods are briefly presented, implemented (R-code available as supplementary material) and evaluated on simulated data to highlight advantages and disadvantages of the methods. Among the identified methods, the integrated intraclass correlation and standard error of measurement are recommended. These methods are straightforward to implement, enable results over the domain, and consider variation between individuals, which the other methods partly neglect.


Multiphoton multifocal microscopy exploiting a diffractive optical element.
Multiphoton multifocal microscopy (MMM) usually has been achieved through a combination of galvo scanners with microlens arrays, with rotating disks of microlens arrays, and cascaded beam splitters with asynchronous rastering of scanning mirrors. Here we describe the achievement of a neat and compact MMM by use of a high-diffraction-efficiency diffractive-optic element that generates a multiple-spot grid of uniform intensity to achieve higher fidelity in imaging of live cells at adequate speeds.


[Metabolic memory enhances hormesis effect to the copper ions in age-depended manner].
The ability of young and old rats to manifest the hormesis effect to lethal doses of copper sulphate and the ability to save the induced "adaptive" pattern of redistribution of copper ions after the transfer of animals in the standard conditions is the mechanism of metabolic memory. It was found that pretreatment of animals with low-dose (1 mg per 100 g body mass, i.e. 33% of the lethal dose) of copper sulfate induced the formation of their resistance to lethal doses (3 mg per 100 g), so the hormesis effect was manifested. Hormesis effect depended on the number of pre injections of small doses of copper sulphate in an S-shaped manner. The protective effect increased after 1 to 3 of preliminary injections of copper sulfate, and after four or more injections the hormesis effect decreased. It is shown that the cardinal role in intracellular pattern of copper ion redistribution play heat-stable copper binding proteins 12 kDa cytosolic proteins. The formed "adaptive" pattern of intracellular distribution of the copper ions may be reproduced, after at least, one month. The prolonged hormesis effect can be attributed to the forming metabolic memory. The intracellular distribution pattern of the copper ions was age-dependent. Age-related differences were found in hormesis effect induced by copper ions, which results in increased binding capacity of copper binding proteins in old animals, with a higher content of copper ions in the mitochondria and microsomes as compared to young animals.


Polyneuritis cranialis: oculopharyngeal subtype of Guillain-Barré syndrome.
Guillain-Barré syndrome (GBS) constitutes a spectrum of related post-infectious neuropathies, which are characterized by their anatomical patterns of weakness and neurological involvement. Historically, the term polyneuritis cranialis has been used to describe some patients with GBS presenting with multiple cranial neuropathies in the absence of limb weakness. We examine previous reports of polyneuritis cranialis to determine disease characteristics and define new diagnostic criteria. Disease characteristics were determined from 15 historical case reports of patients presenting with isolated 'polyneuritis cranialis', 'cranial polyneuropathy', 'multiple cranial neuropathy' and 'multiple cranial neuropathies' due to GBS. Median age was 40 years. 80% displayed antecedent infectious symptoms. In all cases, disease course was monophasic with clinical improvement within weeks or months. Initial symptoms were ocular (73%) or bulbar (33%). Mean number of cranial nerves involved was 6 (range, 3-10). 93% displayed ocular signs, 73% facial weakness or numbness and 87% bulbar signs. In 3 patients (20%), there was significant asymmetry. Cerebrospinal fluid albuminocytological dissociation was present in 67% of cases. Serum anti-ganglioside antibodies were tested for in 8 of 15 patients and anti-GQ1b antibodies were found in 3 patients, whilst anti-GT1a antibodies were found in 1 patient. Polycranial neuritis (the oculopharyngeal subtype of GBS) can be defined in patients with disease characteristics of GBS who display ocular and pharyngeal weakness in the absence of limb weakness or ataxia. In half of cases tested for, anti-ganglioside antibodies were present and most frequently against GQ1b.