Notch signaling regulates formation of the three-dimensional architecture of intrahepatic bile ducts in mice.
Alagille syndrome, a chronic hepatobiliary disease, is characterized by paucity of intrahepatic bile ducts (IHBDs). To determine the impact of Notch signaling specifically on IHBD arborization, we studied the influence of both chronic gain and loss of Notch function on the intact three-dimensional IHBD structure using a series of mutant mouse models and a resin casting method. Impaired Notch signaling in bipotential hepatoblast progenitor cells (BHPCs) dose-dependently decreased the density of peripheral IHBDs, whereas activation of Notch1 results in an increased density of peripheral IHBDs. Although Notch2 has a dominant role in IHBD formation, there is also a redundant role for other Notch receptors in determining the density of peripheral IHBDs. Because changes in IHBD density do not appear to be due to changes in cellular proliferation of bile duct progenitors, we suggest that Notch plays a permissive role in cooperation with other factors to influence lineage decisions of BHPCs and sustain peripheral IHBDs. There is a threshold requirement for Notch signaling at multiple steps, including IHBD tubulogenesis and maintenance, during hepatic development that determines the density of three-dimensional peripheral IHBD architecture.


Betulinic acid induces a novel cell death pathway that depends on cardiolipin modification.
Cancer is associated with strong changes in lipid metabolism. For instance, normal cells take up fatty acids (FAs) from the circulation, while tumour cells generate their own and become dependent on de novo FA synthesis, which could provide a vulnerability to target tumour cells. Betulinic acid (BetA) is a natural compound that selectively kills tumour cells through an ill-defined mechanism that is independent of BAX and BAK, but depends on mitochondrial permeability transition-pore opening. Here we unravel this pathway and show that BetA inhibits the activity of steroyl-CoA-desaturase (SCD-1). This enzyme is overexpressed in tumour cells and critically important for cells that utilize de novo FA synthesis as it converts newly synthesized saturated FAs to unsaturated FAs. Intriguingly, we find that inhibition of SCD-1 by BetA or, alternatively, with a specific SCD-1 inhibitor directly and rapidly impacts on the saturation level of cardiolipin (CL), a mitochondrial lipid that has important structural and metabolic functions and at the same time regulates mitochondria-dependent cell death. As a result of the enhanced CL saturation mitochondria of cancer cells, but not normal cells that do not depend on de novo FA synthesis, undergo ultrastructural changes, release cytochrome c and quickly induce cell death. Importantly, addition of unsaturated FAs circumvented the need for SCD-1 activity and thereby prevented BetA-induced CL saturation and subsequent cytotoxicity, supporting the importance of this novel pathway in the cytotoxicity induced by BetA.


The impact of context on evidence utilization: a framework for expert groups developing health policy recommendations.
Should the same evidence lead to the same decision outcomes in different decision-making contexts? In order to improve comprehension of this issue, this study considers how context influences evidence utilization in the development of health policy recommendations. We used an embedded multiple case study design to study how four expert groups formulated policy recommendations for breast, cervical, colorectal, and prostate cancer screening in Ontario, Canada. We interviewed expert group members and analysed meeting agendas/minutes, interim/final reports and other case-related documents. Our analyses revealed varying policy objectives; the use, neglect, or overextended consideration of three key decision support tools; the varying skills/abilities of expert group members in using different decision support tools; the varying impact of effect modifiers, resource constraints and political interests; and the differing development/consideration of context-specific evidence to address uncertainty in the external decision-making context. While more work is needed to determine if these findings are generalizable beyond cancer screening policy, we believe the central challenge for evidence-based policy is not to develop international evidence, but rather to develop more systematic, rigorous, and global methods for identifying, interpreting, and applying evidence in different decision-making contexts. Our analyses suggest that identification of evidence must distinguish between different policy objectives in order to link a broad conceptualization of evidence to appropriate policy questions. Interpretation of evidence must acknowledge the varying nature of evidence for different policy objectives, balancing existing emphasis on evidentiary quality with more sophisticated methods for assessing the generalizability of evidence. The application of evidence must also acknowledge different policy objectives, appropriately employing rule-based grading schemes and agreement-based consensus methods that are sensitive to the nature of the evidence and contexts involved.


Influence of the pandemic dissemination of COVID-19 on radiotherapy practice: A flash survey in Germany, Austria and Switzerland.
The COVID-19 pandemic has already changed our globalised world and its long-term impact is not yet known. It is apparent that businesses and institutions are increasingly affected. COVID-19 discussions often focus on intensive care units in hospitals. However, COVID-19 also effects life-saving and -prolonging radiotherapy for patients suffering from cancer. We have conducted a structured online survey among medical physicists in Germany, Austria and Switzerland from March 23rd to 26th 2020. In total 154 responses (82 completed, 72 partially completed) were analysed in the context of the COVID-19 dissemination. 72.4% of the respondent's state that their processes are affected due to COVID-19, while the top three answers are longer processing times (54.2%), patient no-shows (42.5%) and staff reduction (36.7%). 75.8% expect further unavailability of their personnel in the upcoming weeks. All participants have already taken several measures, especially providing information for patients at the entrance (89.6%) or over the phone (73.6%), restricting access for accompanying persons (77.4%) and providing disinfectant at the entrance (72.6%). The results presented in this article aim to support business continuity and risk management for radiotherapy centres to prepare for future challenges. The results show that most radiotherapy centres has implemented initial contingency measures, applying them pragmatically. The main problem however remains, that is the high risk of infection both for patients and medical personnel along with the associated risk of temporarily loss of personnel and ordered closure of business.


Nourishing the Spirit: Exploratory Research on Ayahuasca Experiences along the Continuum of Recovery from Eating Disorders.
Eating disorders (EDs) are serious health conditions that are among the most difficult to treat. Innovative treatments are needed and modalities from across cultures must be considered. Ayahuasca is a psychoactive plant-based tea originally used by Amazonian indigenous groups. A growing body of research points to its promise in the healing of various mental health issues. This study explored the potential therapeutic value of ayahuasca in the context of EDs, including the perceived impact of the preparatory diet and the ayahuasca purge. Sixteen individuals previously diagnosed with an ED participated in a semi-structured interview relating to their experiences with ceremonial ayahuasca drinking. Interviews were analyzed using thematic analysis. Themes presented relate to the reduction or cessation of ED and mental health symptoms, shifts in body perception, and the importance of a ceremonial setting and after-care. For some, the preparatory diet resulted in familiar patterns of concern; however, none felt triggered by the purge in ayahuasca. Ceremonial ayahuasca drinking shows promise in the healing of EDs and warrants further research.


Word Processing in Children With Autism Spectrum Disorders: Evidence From Event-Related Potentials.
This investigation was conducted to determine whether young children with autism spectrum disorders exhibited a canonical neural response to word stimuli and whether putative event-related potential (ERP) measures of word processing were correlated with a concurrent measure of receptive language. Additional exploratory analyses were used to examine whether the magnitude of the association between ERP measures of word processing and receptive language varied as a function of the number of word stimuli the participants reportedly understood. Auditory ERPs were recorded in response to spoken words and nonwords presented with equal probability in 34 children aged 2-5 years with a diagnosis of autism spectrum disorder who were in the early stages of language acquisition. Average amplitudes and amplitude differences between word and nonword stimuli within 200-500 ms were examined at left temporal (T3) and parietal (P3) electrode clusters. Receptive vocabulary size and the number of experimental stimuli understood were concurrently measured using the MacArthur-Bates Communicative Development Inventories. Across the entire participant group, word-nonword amplitude differences were diminished. The average word-nonword amplitude difference at T3 was related to receptive vocabulary only if 5 or more word stimuli were understood. If ERPs are to ever have clinical utility, their construct validity must be established by investigations that confirm their associations with predictably related constructs. These results contribute to accruing evidence, suggesting that a valid measure of auditory word processing can be derived from the left temporal response to words and nonwords. In addition, this measure can be useful even for participants who do not reportedly understand all of the words presented as experimental stimuli, though it will be important for researchers to track familiarity with word stimuli in future investigations. https://doi.org/10.23641/asha.5614840.


Dominant expression of porcine Calbindin-D9k in the uterus during a luteal phase.
Calbindin-D9k (CaBP-9k) is a member of intracellular calcium binding proteins, which have a high affinity to calcium. CaBP-9k is mainly expressed in the mammalian intestine, uterus and placenta, and is regulated in tissue- and species-specific manners. Previous studies have shown that CaBP-9k expression is mainly controlled by steroid hormones and their receptors. Thus, we further investigated the expression and regulation of CaBP-9k during an estrus cycle in the pig uterus by Northern blot and immunoblot analysis in this study. In addition, serum levels of estrogen (E2) and progesterone (P4) were measured using ELISA. The CaBP-9k mRNA is highly expressed in the porcine uterus during a luteal phase compared to a follicular phase, and its mRNA level in a luteal phase is increased up to 10-fold compared to a follicular phase. In parallel to the level of CaBP-9k mRNA, the CaBP-9k protein is also dominantly expressed in the porcine uterus, and strongly expressed in the epithelium and glands of the porcine uterus during a luteal phase. Although, the localization of the CaBP-9k protein is scarcely detected at follicular phase, it is dominantly expressed in the porcine uterus during a luteal phase. In addition, the serum P4 level was significantly increased during a luteal phase compared to a follicular phase, whereas no difference was observed in E2 levels between follicular and luteal phases, indicating that the ratio of P4/E2 is remarkably increased in porcine uterus during a luteal phase compared to a follicular phase. In conclusion, these results suggest that P4 may play an important role in the up-regulation of CaBP-9k gene in the porcine uterus in a luteal phase, which is unlike the condition in the rat uterus. In addition, the porcine CaBP-9k may be dominantly expressed in the epithelium and glandular structure of pig uterus during a luteal phase. It may also be differentially regulated during this cycle presumably by steroid hormones, especially up-regulated P4 levels in this tissue.


Tumor Tissue-Specific Biomarkers of Colorectal Cancer by Anatomic Location and Stage.
The progress in the discovery and validation of metabolite biomarkers for the detection of colorectal cancer (CRC) has been hampered by the lack of reproducibility between study cohorts. The majority of discovery-phase biomarker studies have used patient blood samples to identify disease-related metabolites, but this pre-validation phase is confounded by non-specific disease influences on the metabolome. We therefore propose that metabolite biomarker discovery would have greater success and higher reproducibility for CRC if the discovery phase was conducted in tumor tissues, to find metabolites that have higher specificity to the metabolic consequences of the disease, that are then validated in blood samples. This would thereby eliminate any non-tumor and/or body response effects to the disease. In this study, we performed comprehensive untargeted metabolomics analyses on normal (adjacent) colon and tumor tissues from CRC patients, revealing tumor tissue-specific biomarkers (n = 39/group). We identified 28 highly discriminatory tumor tissue metabolite biomarkers of CRC by orthogonal partial least-squares discriminant analysis (OPLS-DA) and univariate analyses (VIP > 1.5, p < 0.05). A stepwise selection procedure was used to identify nine metabolites that were the most predictive of CRC with areas under the curve (AUCs) of >0.96, using various models. We further identified five biomarkers that were specific to the anatomic location of tumors in the colon (n = 236). The combination of these five metabolites (S-adenosyl-L-homocysteine, formylmethionine, fucose 1-phosphate, lactate, and phenylalanine) demonstrated high differentiative capability for left- and right-sided colon cancers at stage I by internal cross-validation (AUC = 0.804, 95% confidence interval, CI 0.670-0.940). This study thus revealed nine discriminatory biomarkers of CRC that are now poised for external validation in a future independent cohort of samples. We also discovered a discrete metabolic signature to determine the anatomic location of the tumor at the earliest stage, thus potentially providing clinicians a means to identify individuals that could be triaged for additional screening regimens.


Temporarily increased TGFβ following radon spa correlates with reduced pain while serum IL-18 is a general predictive marker for pain sensitivity.
Sustained pain relief following radon spa therapy in patients suffering from chronic painful diseases has been well described. But still, the underlying mechanisms are not fully understood. We conducted the prospective and explorative RAD-ON01 study which included 103 patients who suffered from chronic painful musculoskeletal disorders of the spine and/or joints and present here the data of the examination of pro- and anti-inflammatory cytokines in the serum of the patients before and at weeks 6, 12 and 30 after therapy. While TNFα, IL-1β, IFNγ, IL-1Ra and IL-10 were not altered, TGFβ was temporarily significantly (p = 0.013) elevated 6 weeks after therapy. Importantly, this elevation positively correlated with lowered pain sensitivity (r = 0.41). Further, the amount of IL-18 in the serum positively correlated with lowered pain sensitivity. Therefore, IL-18 can be considered as predictive marker for pain sensitivity of radon spa patients. We conclude that alterations in TGFβ and general IL-18 levels in serum have prognostic and predictive value in situations of lowered pain by exposure of patients to very low-doses of radiation as it is the case in radon spa.


Direct costs of diabetes care: a survey in Ottawa, Ontario 1986.
205 unselected Ottawa area patients with diabetes completed questionnaires indicating their annual utilization of equipment, supplies, professional and institutional services. Respondents' average age was 47.3 and average duration of diabetes 18.3 years. About 70% used self-testing of blood glucose. Calculations of costs were based on prevailing local prices. Annual treatment-related costs for the 9.3% on anti-diabetic tablets averaged $236.40, for the 79% on injected insulin $362.34, and for the 7.3% on insulin-by-pump $1603.20. Averaged over all respondents, routine care costs were $962.01 annually, of which 23.4% was for at-home testing, 45.3% for treatment supplies, 20.9% for physicians' services and 10.4% for miscellaneous items. Non-routine care costs averaged $1982.24 with all except $45.84 for emergency room charges being in-hospital care costs. Cost of routine plus non-routine care averaged $2944.25 annually of which 64.4% was for hospital services, 13.9% for treatment needs, 9.6% for physicians' services and 7.7% for testing supplies. This type of survey, particularly if carried out with help from diabetes association volunteers, offers an affordable means to monitor utilization trends and costs of supplies and services for patient care.


Attenuation of highly pathogenic avian influenza A(H5N1) viruses in Indonesia following the reassortment and acquisition of genes from low pathogenicity avian influenza A virus progenitors.
The highly pathogenic avian influenza (HPAI) A(H5N1) virus is endemic in Indonesian poultry and has caused sporadic human infection in Indonesia since 2005. Surveillance of H5N1 viruses in live bird markets (LBMs) during 2012 and 2013 was carried out to provide epidemiologic and virologic information regarding viral circulation and the risk of human exposure. Real-time RT-PCR of avian cloacal swabs and environmental samples revealed influenza A-positive specimens, which were then subjected to virus isolation and genomic sequencing. Genetic analysis of specimens collected at multiple LBMs in Indonesia identified both low pathogenicity avian influenza (LPAI) A(H3N8) and HPAI A(H5N1) viruses belonging to clade 18.104.22.168a. Comparison of internal gene segments among the LPAI and HPAI viruses revealed that the latter had acquired the PB2, PB1, and NS genes from LPAI progenitors and other viruses containing a wild type (wt) genomic constellation. Comparison of murine infectivity of the LPAI A(H3N8), wt HPAI A(H5N1) and reassortant HPAI A(H5N1) viruses showed that the acquisition of LPAI internal genes attenuated the reassortant HPAI virus, producing a mouse infectivity/virulence phenotype comparable to that of the LPAI virus. Comparison of molecular markers in each viral gene segment suggested that mutations in PB2 and NS1 may facilitate attenuation. The discovery of an attenuated HPAI A(H5N1) virus in mice that resulted from reassortment may have implications for the capability of these viruses to transmit and cause disease. In addition, surveillance suggests that LBMs in Indonesia may play a role in the generation of reassortant A(H5) viruses and should be monitored.


Cytotoxic effect of Eucalyptus citriodora resin on human hepatoma HepG2 cells.
The aim of this study was to evaluate the antiproliferative effect of Eucalyptus citriodora resin (ECR) on human hepatoma HepG2 cells. The results from MTT assay and LDH leakage analysis showed that water extracts of ECR (WEECR) in the dose range of 0-500 μg/ml displayed stronger cytotoxic effects on HepG2 cells than other organic solvent extracts of ECR. By flow cytometry analysis, WEECR slowed down the cell cycle at the G0/G1 phase after 24 h of incubation. Moreover, WEECR treatment induced an apoptotic response in HepG2 cells. WEECR-induced apoptosis was in association with the attenuation of mitochondrial transmembrane potentials (ΔΨ(m)), increased Bax/Bcl-2 ratio and activation of caspase-3. In addition, WEECR contained high concentration of phenolics and flavonoids, which may be responsible for the potent cytotoxicity of WEECR on HepG2 cells. Taken together, WEECR may be a potent antihepatoma agent due to apoptosis in HepG2 cells.


[Diagnostic importance of the ergometrine test in ischemic heart disease].
The ergometrine test was performed in 102 patients with ischemic heart disease (IHD) and cardialgias of various genesis. The diagnosis was verified in all patients by the bicycle exercise test and coronary arteriography. The variant of angina pectoris was diagnosed when there were precordial pains and the ST-segment elevation after intravenous ergometrine injection. The appearance of angina pectoris and ischemic ST-segment depression was observed in patients with angina of effort and at rest. This type of reaction in the ergometrine test indicates obstructive coronary heart disease. The appearance of precordial pain without ECG changes is probably the sign of latent coronary insufficiency. The ergometrine testing is proposed for the differential diagnosis of different forms of angina pectoris.


Role of the endocannabinoid system in the mechanisms involved in the LPS-induced preterm labor.
Prematurity is the leading cause of perinatal morbidity and mortality worldwide. There is a strong causal relationship between infection and preterm births. Intrauterine infection elicits an immune response involving the release of inflammatory mediators like cytokines and prostaglandins (PG) that trigger uterine contractions and parturition events. Anandamide (AEA) is an endogenous ligand for the cannabinoid receptors CB1 and CB2. Similarly to PG, endocannabinoids are implicated in different aspects of reproduction, such as maintenance of pregnancy and parturition. Little is known about the involvement of endocannabinoids on the onset of labor in an infectious milieu. Here, using a mouse model of preterm labor induced by lipopolysaccharide (LPS), we explored changes on the expression of components of endocannabinoid system (ECS). We have also determined whether AEA and CB antagonists alter PG production that induces labor. We observed an increase in uterine N-acylphosphatidylethanolamine-specific phospholipase D expression (NAPE-PLD, the enzyme that synthesizes AEA) upon LPS treatment. Activity of catabolic enzyme fatty acid amide hydrolase (FAAH) did not change significantly. In addition, we also found that LPS modulated uterine cannabinoid receptors expression by downregulating Cb2 mRNA levels and upregulating CB1 protein expression. Furthermore, LPS and AEA induced PGF2a augmentation, and this was reversed by antagonizing CB1 receptor. Collectively, our results suggest that ECS may be involved in the mechanism by which infection causes preterm birth.


Glioblastoma and Increased Survival with Longer Chemotherapy Duration.
The five-year survival rate for patients with glioblastoma (GBM) is low at approximately 4.7%. Radiotherapy plus concomitant and adjuvant temozolomide (TMZ) remains the standard of care. The optimal duration of therapy with TMZ is unknown. This study sought to evaluate the survival benefit of two years of treatment. This was a retrospective chart review of all patients diagnosed with GBM and treated with TMZ for up to two years between January 1, 2002 and December 31, 2011. The Kaplan-Meier method with log-rank test was used to estimate the progression-free survival (PFS) and the overall survival (OS). The results were compared to historical controls and data from previous clinical trials of patients treated up to one year. Data from 56 patients with confirmed GBM were evaluated. The OS probability was 54% (SE = 0.068) at one year, 28.3% (SE = 0.064) at two years, 17.8% (SE = 0.059) at three years, and 4% (SE = 0.041) at five years. Seven patients (12.5%) were treated with TMZ for two years. Their median time-to-progression was 28 months (95% CI = 5.0 - 28.0), and they had an increased survival probability at three years compared to other patients (log-rank test χ2 (1, N = 56) = 19.2, p < 0.0001). There may be an advantage for a longer duration of TMZ therapy among patients with GBM, but the sample size was too small for generalization. A multicenter prospective study is needed to identify optimal duration of TMZ therapy.


ROLE OF PRIMARY HEALTH CARE IN RE-HOSPITALIZATION OF PATIENTS WITH HEART FAILURE.
Re-hospitalization of cardiac patients is a great financial burden not only for healthcare system, but also for patients. Main aim of the research is to identify features and reasons of re-hospitalization and to determine ways of reducing. Within the Quantitative research, we have analyzed data of re-hospitalized patients during 30 days. Within the qualitative study, in-depth survey of medical staff and patient conducted. Main reason of re-hospitalization is that patients do not fulfill doctor's prescriptions after discharging from hospital. This is because of financial difficulties and lack of developing family doctor institution in the country. Usually, after discharging patient from hospital, for medical supervision he/she addresses to family doctor very seldom. There is no coordination between family and hospital doctors. Primary health care plays a significant role in the reduction of re-hospitalization rate. In this regard, it is important to increase the role of the family doctor, to improve procedure of discharging patients, continuous medical supervision on patients, to expand outpatient services package in state healthcare programs, to improve day care centers.


Body fat distribution and metabolic variables in patients with neonatal progeroid syndrome.
Neonatal progeroid syndrome (NPS), also known as Wiedemann-Rautenstrauch Syndrome, is a rare autosomal recessive disorder characterized by accelerated aging and lipodystrophy from birth. Affected children have extreme intrauterine growth retardation, poor postnatal weight gain, and characteristic facial dysmorphic features such as a triangular shape, pinched nose, pseudohydrocephalus with wide fontanelles and prominent subcutaneous (sc) veins. Generalized loss of sc fat has been reported as a cardinal feature; however, the pattern of fat loss and its association with insulin resistance and its metabolic complications have not been systematically studied. The aim of the current study was to examine body fat distribution and body composition in two girls with NPS using anthropometric measures, whole-body magnetic resonance imaging (MRI) and dual energy X-ray absorptiometry (DEXA), and to assess metabolic complications such as hyperinsulinemia and dyslipidemia. Both the girls (aged 17 years and 10 years, respectively) had generalized paucity of sc fat on physical examination. However, measurements of skin-fold thickness revealed that sc fat was decreased over the extremities, but preserved over the chest and abdomen. MRI studies confirmed the presence of normal amounts of sc truncal fat, and marked loss of fat from the face and distal extremities. Striking fat loss was also noted in the paravertebral and lateral gluteal regions. Interestingly, body composition analysis with DEXA scan revealed a marked reduction in both the fat and lean tissue mass. Fasting glucose, lipids and insulin levels were not elevated. We conclude that patients with NPS do not have generalized lipodystrophy as previously reported, but fat loss is confined to the face, distal extremities, and possibly the paravertebral and lateral gluteal regions. Metabolic abnormalities related to insulin resistance are also uncommon in this condition.


The self-reported health condition of women after their participation in a stress management programme: a pilot study.
From a public health perspective, it is important to develop effective measures to deal with stress which are based on the individual's participation, such as stress management provided in group sessions. Therefore, the aim of the present study was to compare and evaluate the self-reported health condition of women in terms of their general symptoms, stress and sense of coherence (SOC) after participation in a stress management programme. The intervention, which had a modified crossover design and involved 40 women divided into two groups (G1 and G2), comprised eight meetings, the content of which was both theoretical and practical, and included information about stress, stress management, massage and mental training. A questionnaire was filled in on three occasions: before and after the intervention (8 weeks later), and after another 8 weeks (16 weeks later). The questionnaire contained 60 items comprising background factors, general symptoms, stress and SOC. No significant differences existed between the groups at baseline. In favour of the intervention, significant differences were found between the groups with regard to fewer general symptoms (P = 0.035) as well as a tendency to stress reduction (P = 0.060). A comparison within groups showed that both groups had a significant reduction in symptoms after the intervention (G1, P = 0.002; and G2, P = 0.003) and in reduced stress (both P = 0.001). After a further 8 weeks, both groups still showed significantly fewer general symptoms and reduced stress, as well as significant improvements with regard to SOC (G1, P = 0.012; and G2, P = 0.026). These findings indicate that the combination of mental training and massage in this stress management programme had a positive influence on the women's health condition. The pilot study design could be used in a full-scale study with randomised groups.


The Arabidopsis thaliana cysteine-rich receptor-like kinase CRK20 modulates host responses to Pseudomonas syringae pv. tomato DC3000 infection.
In plants, the cysteine-rich repeat kinases (CRKs) are a sub-family of receptor-like protein kinases that contain the DUF26 motif in their extracellular domains. It has been shown that in Arabidopsis thaliana, CRK20 is transcriptionally induced by pathogens, salicylic acid and ozone (O(3)). However, its role in responses to biotic and abiotic stress remains to be elucidated. To determine the function of CRK20 in such responses, two CRK20 loss-of-function mutants, crk20-1 and crk20-2, were isolated from public collections of Arabidopsis T-DNA tagged lines and examined for responses to O(3) and Pseudomonas syringae pv. tomato (Pst) DC3000. crk20-1 and crk20-2 showed similar O(3) sensitivities and no differences in the expression of defense genes when compared with the wild-type. However, pathogen growth was significantly reduced, while there were no differences in the induction of salicylic acid related defense genes or salicylic acid accumulation. Furthermore, correlation analysis of CRK20 gene expression suggests that it has a role in the control of H(2)O and/or nutrient transport. We therefore propose that CRK20 promotes conditions that are favorable for Pst DC3000 growth in Arabidopsis, possibly through the regulation of apoplastic homeostasis, and consequently, of the environment of this biotrophic pathogen.


Drosophila ptip is essential for anterior/posterior patterning in development and interacts with the PcG and trxG pathways.
Development of the fruit fly Drosophila depends in part on epigenetic regulation carried out by the concerted actions of the Polycomb and Trithorax group of proteins, many of which are associated with histone methyltransferase activity. Mouse PTIP is part of a histone H3K4 methyltransferase complex and contains six BRCT domains and a glutamine-rich region. In this article, we describe an essential role for the Drosophila ortholog of the mammalian Ptip (Paxip1) gene in early development and imaginal disc patterning. Both maternal and zygotic ptip are required for segmentation and axis patterning during larval development. Loss of ptip results in a decrease in global levels of H3K4 methylation and an increase in the levels of H3K27 methylation. In cell culture, Drosophila ptip is required to activate homeotic gene expression in response to the derepression of Polycomb group genes. Activation of developmental genes is coincident with PTIP protein binding to promoter sequences and increased H3K4 trimethylation. These data suggest a highly conserved function for ptip in epigenetic control of development and differentiation.


Corneal thickness changes in myopic children during and after short-term orthokeratology lens wear.
To evaluate thickness changes in the central and mid-peripheral cornea (CCT and mPCT), corneal epithelium (CET and mPET) and stroma (CST and mPST) of myopic children during and after short-term orthokeratology (ortho-k) lens wear, with conventional (CCF, 0.75 D) and increased compression factors (ICF, 1.75 D). This was a self-controlled case series study. Subjects wore a CCF lens in one eye and an ICF lens in the other. Anterior segment optical coherence tomography images were captured weekly for 1 month during lens wear and for 2 weeks after discontinuing lens wear. CCT and CET (central 3-mm cornea) and mPCT and mPET (within a 4-6 mm diameter annulus) were measured. Stromal thickness (ST) was determined by subtracting epithelium thickness (ET) from corneal thickness (CT). The repeatability of the analytical software was also investigated on age-matched spectacle-wearing subjects (n = 98). Excluding three outliers (>3 S.D.s), the coefficient of repeatability and intraclass correlation coefficients of 98 spectacle-wearing subjects ranged from 2.63 to 4.64 μm and from 0.90 to 0.99, respectively. For the weekly-change study, CCT and CET in both eyes were significantly thinner after lens wear (p < 0.001) and CET thinning in the ICF eyes were significantly higher (p < 0.02). CCT changes were mainly contributed by CET. CST, mPCT, mPET and mPST changes were not significant (p > 0.20) in either eye. CT (all sublayers) rebounded to baseline values 2 weeks after discontinuing lens wear (0.99 > p > 0.12). Significant reductions in CT and ET, but not ST, were observed within 1 month of ortho-k lens wear. Wearing ICF lenses resulted in a higher reduction in CET. Corneal thickness changes were reversible after discontinuing lens wear.


[Experiences and health needs of men in the period post-birth of a child].
This research addresses men's perceptions about their experiences and health needs in the post-birth period. The goal is to distinguish analytically and from a gender perspective the health needs related to the event, experienced, expressed and/or denied by men. This is an exploratory, descriptive and qualitative study carried out in two municipal health areas of Cuiabá, state of Mato Grosso, using a semi structured interview and thematic analysis of the data. The participants were eight men who were experiencing the post-birth period of a child. Above all, they expressed the need to provide financial security to the family, emphasizing also the need of their children's well-being, with the affective return that this experience brings. They did not perceive themselves with health needs during the post-birth period. The support provided by local health services is important so that men can understand the experience of parenthood and its relationship with social and cultural aspects in order to distinguish their own needs, appreciate self-care, and adopt a gender equity perspective.


Ischemic colitis secondary to inferior mesenteric arteriovenous fistula and portal vein stenosis in a liver transplant recipient.
Arteriovenous fistula (AVF) involving the inferior mesenteric vessels is rare, and the affected patients usually present with abdominal pain, mass, or features of established portal hypertension. Colonic ischemia is a less common and more serious manifestation of AVF. We report a case of ischemic colitis secondary to inferior mesenteric AVF in a patient who underwent a previous liver transplantation, subsequently developed portal vein stenosis, and then presented with acute lower gastrointestinal bleeding. He underwent percutaneous transhepatic placement of a portal vein stent and left colectomy.


Intra-operative 3D imaging system for robot-assisted fracture manipulation.
Reduction is a crucial step in the treatment of broken bones. Achieving precise anatomical alignment of bone fragments is essential for a good fast healing process. Percutaneous techniques are associated with faster recovery time and lower infection risk. However, deducing intra-operatively the desired reduction position is quite challenging due to the currently available technology. The 2D nature of this technology (i.e. the image intensifier) doesn't provide enough information to the surgeon regarding the fracture alignment and rotation, which is actually a three-dimensional problem. This paper describes the design and development of a 3D imaging system for the intra-operative virtual reduction of joint fractures. The proposed imaging system is able to receive and segment CT scan data of the fracture, to generate the 3D models of the bone fragments, and display them on a GUI. A commercial optical tracker was included into the system to track the actual pose of the bone fragments in the physical space, and generate the corresponding pose relations in the virtual environment of the imaging system. The surgeon virtually reduces the fracture in the 3D virtual environment, and a robotic manipulator connected to the fracture through an orthopedic pin executes the physical reductions accordingly. The system is here evaluated through fracture reduction experiments, demonstrating a reduction accuracy of 1.04 ± 0.69 mm (translational RMSE) and 0.89 ± 0.71 ° (rotational RMSE).


Glutamine (Q)-peptide screening for transglutaminase reaction using mRNA display.
Information on subsite specificity of the transglutaminase (TG) is important to design any specific peptides for TG's applications and inhibitor studies. Here, mRNA display was introduced for identifying the subsite specificity of TG from Streptomyces mobaraensis (STG). Functionally active peptides expressed from mRNA display library were differentially conjugated to hexa lysine (K₆-beads according to their relative activities for STG. The active peptide substrates for STG were enriched through six rounds of screening, and its corresponding cDNA/mRNA sequences were identified by DNA sequencing. The results showed that tripeptides such as LQQ and TQP do not show any activity for STG, but the minimum size of the peptide displaying STG activity is pentapeptide. One such predicted peptide sequence, that is, RLQQP (TQ1), showed higher reactivity (ca. 182% conjugation yield) to STG than that of the highly active sequence, that is, control-Q (PQPQLPYPQPQLPY), well-known previously for mammalian TG2. Furthermore, when recombinant DsRed was tagged with TQ1 sequence at its C-terminal, DsRed-TQ1 underwent efficient covalent-immobilization onto alginate-gelatin bead by STG reaction, showing a Q-peptide application as a useful tagging molecule.


Attachment-Based Family Therapy to Improve Family Functioning in Adolescent Binge-Spectrum Eating Disorders: An Initial Evaluation Via Case Series Design.
Binge-spectrum eating disorders (EDs; bulimia nervosa, binge eating disorder) often develop during adolescence and are associated with serious psychological and physical consequences. Current treatments for adolescents are highly behavioral in nature and while efficacious, many patients do not reach remission indicating that current treatments fail to target a key maintenance factor for EDs. One potential maintenance factor is poor family functioning (FF). In particular, high family conflict (e.g., arguing, critical comments) and low family cohesion (e.g., warmth, support) are known to maintain ED behaviors. Poor FF can (1) cause or exacerbate an adolescent's use of ED behaviors to cope with life stress and/or (2) inhibit parents from being a resource to adolescents during ED treatment. Attachment-Based Family Therapy (ABFT) is specifically designed to improve FF, and thus may be a promising adjunct to behavioral ED intervention strategies. ABFT, however, has not been tested in adolescents with binge-spectrum EDs. Thus, the current study is the first to evaluate a 16-week adapted ABFT treatment for adolescents with EDs (N = 8, Mage = 16.00, 71.43% female, 71.43% White) fusing together behavioral treatment for EDs with ABFT for highest possible impact. Eight families were treated in an open pilot trial to examine treatment feasibility, acceptability, and preliminary efficacy on FF and eating pathology. Overall, findings were promising. ABFT + B treatment was feasible and acceptable and showed preliminary evidence that it could improve FF and ED behaviors. Future research will test this intervention in a larger sample and further examine the role of FF in maintaining ED symptoms.


Bilateral cochlear implantation: current concepts, indications, and results.
The optimal treatment for bilateral hearing loss continues to evolve as cochlear implant (CI) and hearing aid technologies advance, as does our understanding of the central auditory system. Ongoing discussions continue on the validity and feasibility of bilateral CI in terms of performance, justification of need, medical/surgical safety concerns, and economics. The purpose of this review article is to provide an update on the advantages and disadvantages of bilateral CI and to provide a discussion on timing (simultaneous vs. sequential), technology (bimodal vs. binaural) and feasibility. Binaural advantages are found in both adult and pediatric bilateral CI recipients, the greatest being the head shadow effect and improvements in localization and loudness summation. This theoretically offers an advantage over their unilateral implanted counterparts in terms of improved sound localization and enhanced speech perception under noisy conditions. Most investigators agree that bilateral stimulation during critical periods of development is paramount for optimizing auditory functioning in children. Currently, bilateral CI is widely accepted as a safe and effective means of bilateral auditory stimulation.


Cytokine-in-adjuvant steering of the immune response phenotype to HIV-1 vaccine constructs: granulocyte-macrophage colony-stimulating factor and TNF-alpha synergize with IL-12 to enhance induction of cytotoxic T lymphocytes.
To enhance and steer immune responses to synthetic peptide vaccines toward selected functional types and to understand the cooperative action of cytokines in fine-tuning the immune response, we attempted to influence the in vivo cytokine environment by delivering cytokines directly to the microenvironment in which the immune response is initiated. Here we study the effects of IL-2, IL-4, IL-7, IL-1beta, IL-12, IFN-gamma, TNF-alpha, and granulocyte-macrophage CSF (GM-CSF) incorporated with peptide in adjuvant on a variety of responses elicited: CTL, T cell proliferation, cytokine production and message, and Ab isotype. We show GM-CSF to be the single most effective cytokine for enhancing both cellular and humoral immunity to two previously characterized HIV-1 MN vaccine constructs. Novel synergies were also detected. GM-CSF synergized with IL-12 for CTL induction in BALB/c mice concomitant with suppression of Th2 cytokines IL-4 and IL-10. TNF-alpha also synergized with IL-12, but by a different mechanism, inducing IFN-gamma production in BALB/c mice and thus shifting the response to a Th1 phenotype. The results presented here suggest that in addition to IL-2, optimum induction of CD8+ CTL in vivo requires a combination of cytokines, including GM-CSF (probably acting to enhance Ag presentation and CD4+ cell help) and IL-12 (steering the Th response toward Th1 cytokines).


Activation mechanism of endothelin ETB receptor by endothelin-1.
Endothelin, a 21-amino-acid peptide, participates in various physiological processes, such as regulation of vascular tone, humoral homeostasis, neural crest cell development and neurotransmission. Endothelin and its G-protein-coupled receptor are involved in the development of various diseases, such as pulmonary arterial hypertension, and thus are important therapeutic targets. Here we report crystal structures of human endothelin type B receptor in the ligand-free form and in complex with the endogenous agonist endothelin-1. The structures and mutation analysis reveal the mechanism for the isopeptide selectivity between endothelin-1 and -3. Transmembrane helices 1, 2, 6 and 7 move and envelop the entire endothelin peptide, in a virtually irreversible manner. The agonist-induced conformational changes are propagated to the receptor core and the cytoplasmic G-protein coupling interface, and probably induce conformational flexibility in TM6. A comparison with the M2 muscarinic receptor suggests a shared mechanism for signal transduction in class A G-protein-coupled receptors.


Creative forms: booklets by the hospital senses collective.
This article details the creation of a series of booklets designed to explore sensory encounters with hospitals and healthcare environments. The booklets were devised as a series of prompts or provocations, created to attend to and examine embodied, sensory encounters with health/care settings rather than to present research findings. Bringing together an expansive range of backgrounds and skill sets the booklets were created to sit within and beyond language through their design, form and content. Within this article we share the ways in which the works are deliberately unfinished and exploratory as this necessitates that those interacting with them create their own meanings and explore how they think and feel about health/care environments. The form and design promote a certain attentiveness and embodied engagement. For example, users must engage with the works carefully, gently turning and unfurling the fragile pages. This is further illustrated through qualitative insights collected from users of the booklets. Throughout this paper we argue for multiplicity in the ways in which we explore and present sensory-focused research. Our attention to multiplicity is supported not only through the design, form and content of the physical booklets but through the creative audio description, text and images created to complement and support these works. These are available online to ensure that our provocations are widely accessible. Within this paper we critique how a reliance on narrative form can limit the ways in which we engage with spatial, sensory and emotional concepts. Such concepts are by their very nature challenging to articulate and arguably require more-than-text-based approaches. We propose that embracing creative, exploratory and seemingly risky routes to examining and presenting such concepts is critical in expanding research.


A health impact assessment framework for assessing vulnerability and adaptation planning for climate change.
This paper presents a detailed description of an approach designed to investigate the application of the Health Impact Assessment (HIA) framework to assess the potential health impacts of climate change. A HIA framework has been combined with key climate change terminology and concepts. The fundamental premise of this framework is an understanding of the interactions between people, the environment and climate. The diversity and complexity of these interactions can hinder much needed action on the critical health issue of climate change. The objectives of the framework are to improve the methodology for understanding and assessing the risks associated with potential health impacts of climate change, and to provide decision-makers with information that can facilitate the development of effective adaptation plans. While the process presented here provides guidance with respect to this task it is not intended to be prescriptive. As such, aspects of the process can be amended to suit the scope and available resources of each project. A series of working tables has been developed to assist in the collation of evidence throughout the process. The framework has been tested in a number of locations including Western Australia, Solomon Islands, Vanuatu and Nauru.


Calcium-activated potassium channels in native endothelial cells from rabbit aorta: conductance, Ca2+ sensitivity and block.
1. Isolated native endothelial cells, obtained by treatment of rabbit aortic endothelium with papain and dithiothreitol, were voltage clamped, and single channel (unitary) and spontaneous transient outward currents (STOCs) were recorded from both whole cells and excised membrane patches. 2. In inside-out patches, the reversal potential of unitary currents was dependent on the extracellular K+ concentration and had a single-channel slope conductance of 220 pS in symmetrical 140 mM-K+ solutions. The open-state probability (Po) of the unitary K+ currents was sensitive to the intracellular Ca2+ concentration with half-maximal activation at approximately 1 microM at +20 mV. The ionic selectivity and Ca2+ sensitivity indicate that a large conductance, Ca(2+)-activated K+ channel is present in freshly dissociated rabbit aortic endothelial cells. 3. The frequency and amplitude of whole-cell unitary currents and amplitude of spontaneous transient outward currents were voltage-dependent. Whole-cell outward K+ currents evoked by depolarizing voltage ramps had amplitudes often corresponding to the simultaneous opening of more than five single Ca(2+)-activated K+ channels. Lowering the intracellular EGTA concentration tenfold, and hence the Ca2+ buffering capacity of the cell, increased unitary K+ current activity and shifted the relationship between Po and membrane potential by approximately -20 mV. 4. Bradykinin (1 microM), adenosine 5'-triphosphate (3 microM) and acetylcholine (3 microM) applied extracellularly evoked a biphasic increase in N Po (where N is number of channels activated) of the Ca(2+)-activated K+ channel studied in the whole-cell recording configuration. The development of a biphasic response to agonist stimulation requires a source of extracellular Ca2+. The sustained increase in N Po of the Ca(2+)-activated K+ channel was attenuated upon the removal of external Ca2+ (Mg2+ replacement) or in the presence of the Ca2+ entry blocker, Ni2+, and the potassium channel blockers tetrabutylammonium (TBA) or tetraethylammonium (TEA). 5. Unitary and spontaneous transient outward currents were inhibited by extracellularly applied TEA (0.5 mM), TBA (0.5-5 mM) and charybdotoxin (100 nM). Ca(2+)-activated K+ currents were blocked completely by 5 mM-TEA, whereas 3,4-diaminopyridine (1 mM), Ba2+ (10 mM) and apamin (0.1-1 microM) did not abolish these K+ currents. 6. The K+ channel opener cromakalim (10 microM) evoked a sustained increase in N Po of the Ca(2+)-activated K+ channels which was not potentiated by the addition of bradykinin. Glibenclamide (10 microM) alone increased N Po and partially inhibited the cromakalim-induced increase in N Po with respect to control.(ABSTRACT TRUNCATED AT 400 WORDS)


[Prion diseases and a new variant of Creutzfeldt-Jakob disease].
The causal link of a new variant of CJD (v-CJD) with bovine spongiform encephalopathy (BSE) has led to world-wide panic. BSE emerged in 1986 through dietary products contaminated with scrapie pathogen, BSE case reports increased in number up to 37,000/year in 1993, then declined in 1994 when the first case of v-CJD emerged. There is a 3-year gap between the emergence of BSE and the introduction of a ban on the use of specified bovine offal in human food. People might have consumed dietary products contaminated with BSE pathogen for the period. Species barrier which has protected human from being transmitted with sheep scrapie pathogen might not work so well in cow-to-human transmission as in sheep-to-human. There are familial forms and sporadic forms of human prion diseases. The familial prion diseases are always related to prion protein (PrP) gene mutations. Sporadic forms of prion diseases are composed of sporadic CJD, iatrogenic CJD and kuru. Sporadic CJD occurs in elder people and shows cortical signs and periodic synchronous discharges (PSD) in short clinical course and pathologically spongiform degeneration and abnormal PrP deposition in synapse structures, but no amyloid plaque deposition in the brain is observed. Since the v-CJD patients had no mutations in PrP gene, v-CJD belongs to sporadic forms of prion diseases. v-CJD shows quite different clinicopathological features from those of sporadic CJD, rather shows similar features to iatrogenic CJD of pituitary hormone and kuru. These three diseases share several features, which are younger patient age, main clinical manifestation of cerebellar signs, negative or rare record of PSD, and PrP amyloid plaque deposition in the brain. The patients of v-CJD, however, had no history of exposure to iatrogenic factors and of cannibalism. Also they did not have a valine polymorphism at PrP codon 129 which is frequently observed in the patients with iatrogenic CJD and kuru. Thus, v-CJD is a quite novel entity in human prion diseases.


Residue 82 of the Chikungunya virus E2 attachment protein modulates viral dissemination and arthritis in mice.
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that has reemerged to cause profound epidemics of fever, rash, and arthralgia throughout sub-Saharan Africa, Southeast Asia, and the Caribbean. Like other arthritogenic alphaviruses, mechanisms of CHIKV pathogenesis are not well defined. Using the attenuated CHIKV strain 181/25 and virulent strain AF15561, we identified a residue in the E2 viral attachment protein that is a critical determinant of viral replication in cultured cells and pathogenesis in vivo. Viruses containing an arginine at E2 residue 82 displayed enhanced infectivity in mammalian cells but reduced infectivity in mosquito cells and diminished virulence in a mouse model of CHIKV disease. Mice inoculated with virus containing an arginine at this position exhibited reduced swelling at the site of inoculation with a concomitant decrease in the severity of necrosis in joint-associated tissues. Viruses containing a glycine at E2 residue 82 produced higher titers in the spleen and serum at early times postinfection. Using wild-type and glycosaminoglycan (GAG)-deficient Chinese hamster ovary (CHO) cell lines and soluble GAGs, we found that an arginine at residue 82 conferred greater dependence on GAGs for infection of mammalian cells. These data suggest that CHIKV E2 interactions with GAGs diminish dissemination to lymphoid tissue, establishment of viremia, and activation of inflammatory responses early in infection. Collectively, these results suggest a function for GAG utilization in regulating CHIKV tropism and host responses that contribute to arthritis. CHIKV is a reemerging alphavirus of global significance with high potential to spread into new, immunologically naive populations. The severity of CHIKV disease, particularly its propensity for chronic musculoskeletal manifestations, emphasizes the need for identification of genetic determinants that dictate CHIKV virulence in the host. To better understand mechanisms of CHIKV pathogenesis, we probed the function of an amino acid polymorphism in the E2 viral attachment protein using a mouse model of CHIKV musculoskeletal disease. In addition to influencing glycosaminoglycan utilization, we identified roles for this polymorphism in differential infection of mammalian and mosquito cells and targeting of CHIKV to specific tissues within infected mice. These studies demonstrate a correlation between CHIKV tissue tropism and virus-induced pathology modulated by a single polymorphism in E2, which in turn illuminates potential targets for vaccine and antiviral drug development.


Mutation analysis of the short cytoplasmic domain of the cell-cell adhesion molecule CEACAM1 identifies residues that orchestrate actin binding and lumen formation.
CEACAM1-4S (carcinoembryonic antigen cell adhesion molecule 1, with 4 ectodomains and a short, 12-14 amino acid cytoplasmic domain) mediates lumen formation via an apoptotic and cytoskeletal reorganization mechanism when mammary epithelial cells are grown in a three-dimensional model of mammary morphogenesis. We show by quantitative yeast two-hybrid, BIAcore, NMR HSQC and STD, and confocal analyses that amino acids phenylalanine (Phe(454)) and lysine (Lys(456)) are key residues that interact with actin orchestrating the cytoskeletal reorganization. A CEACAM1 membrane model based on vitamin D-binding protein that predicts an interaction of Phe(454) at subdomain 3 of actin was supported by inhibition of binding of actin to vitamin D-binding protein by the cytoplasmic domain peptide. We also show that residues Thr(457) and/or Ser(459) are phosphorylated in CEACAM1-transfected cells grown in three-dimensional culture and that mutation analysis of these residues (T457A/S459A) or F454A blocks lumen formation. These studies demonstrate that a short cytoplasmic domain membrane receptor can directly mediate substantial intracellular signaling.


Time-independent indices of circadian blood pressure and heart rate regulation from ambulatory blood pressure monitoring.
We hypothesized that addressing the 24-h relationship between RR interval and systolic arterial pressure (SAP) during ambulatory blood pressure monitoring could detect alterations in circadian arterial pressure and heart rate profile and could also be linked to the ambulatory arterial stiffness index. We analyzed retrospectively 676 ambulatory blood pressure monitoring 24-h recordings. Individuals were median split into low (n = 338) and high (n = 338) SAP groups (mean 24 h of 120 +/- 7, 140 +/- 10 mmHg, respectively). We estimated for each individual ambulatory blood pressure monitoring, the slope (ms/mmHg) of the regression of RR interval on SAP over 24-h (defined as theta index). Theta index was significantly (P < 0.001) steeper in individuals with lower pressure than those with higher pressure, whereas day-night difference in SAP did not differ significantly. Theta index was also less negative in nondipper than in dipper individuals and displayed a significant interaction (P < 0.001) with hypertension. The theta index correlated significantly with age and ambulatory arterial stiffness index. Moreover, approximately 14-month reproducibility, as assessed by intraclass correlation coefficient, for theta index was similar to SAP (0.703 and 0.757, respectively). These results indicate that theta can detect differences in circadian arterial pressure profile that could assess the efficiency of arterial pressure (and heart rate) regulatory mechanisms over the whole day in individuals with different arterial pressure levels. Theta can be easily obtained under ambulatory conditions without any assumption on underlying mathematical models or operator-dependent attribution of arterial pressure and heart rate values to daytime or night-time subperiods. Cross-sectional and prospective studies relating theta to target organ damage and cardiovascular outcomes will inform us about its clinical relevance.


Arabidopsis FHY3/FAR1 gene family and distinct roles of its members in light control of Arabidopsis development.
FHY3 (far-red elongated hypocotyls 3) and FAR1 (far-red-impaired response) are two homologous proteins essential for phytochrome A controlled far-red responses in Arabidopsis (Arabidopsis thaliana). There are 12 additional FHY3/FAR1-related genes in the Arabidopsis genome. The predicted sizes of this family of proteins range from 531 amino acids to 851 amino acids, and they share 12.0% to 82.4% amino acid identities over their entire lengths. In addition, most FRS proteins contain one to three coiled-coil domains and one or two putative nuclear localization signals. Semiquantitative reverse transcription-polymerase chain reaction analyses revealed that all FRS genes except FRS10 are expressed in all tissues examined, including rosette leaves, cauline leaves, inflorescence stems, flowers, and siliques. Analyses of gene specific promoterGUS fusion reporter gene expression revealed that all FRS genes except FRS1 are expressed in hypocotyls, and their expression in hypocotyl is induced by far-red light treatment. Transient expression of green fluorescent protein tagged FRS fusion proteins in onion (Allium cepa) epidermal cells revealed that all FRS proteins are targeted into the nucleus. T-DNA knockout frs6 and frs8 mutants flowered early under both long-day and short-day conditions (with much more drastic effects under short-day conditions), suggesting that FRS6 and FRS8 regulate flowering time. In addition, FRS9 RNAi transgenic plants showed a specific hypersensitivity to red light inhibition of hypocotyl elongation and light-regulated gene expression, indicating that FRS9 is a specific negative regulator of phyB signaling mediating seedling deetiolation. In summary, our results support the notion that FRS family members play distinct roles in light control of Arabidopsis development, most likely by regulating nuclear gene expression.


Phenotypic and genotypic within-host diversity of Pseudomonas aeruginosa urinary isolates.
This study aimed to assess phenotypic and molecular inter-patient and within-host diversity of Pseudomonas aeruginosa isolates responsible for urinary tract infection (UTI) or asymptomatic bacteriuria (AB). Clinical data of 120 consecutive P. aeruginosa UTI (n = 40) and AB (n = 80) were prospectively analyzed. Up to five P. aeruginosa isolates per sample were collected. Antimicrobial susceptibility testing (AST) was determined for all isolates (n = 591); a subset of 358 was characterized by multilocus sequence typing. 444 isolates (75%) were non-multidrug resistant (MDR), 113 (19%) were MDR, and 34 (6%) were extensively drug resistant. A genetically highly diverse population was observed (64 sequence types [STs]), without strict correlation between genotypes and clinical settings. 35 patients (28%; 12 UTIs and 23 ABs) presented distinct antimicrobial resistance (AMR) profiles within a given urine sample, significantly associated with previous carbapenem and fluroquinolones exposure; five of them also exhibited polyclonal UTI or AB (with isolates belonging to two STs). P. aeruginosa urinary isolates of these 120 patients were highly diverse, in terms of AMR as well as genetic background. Both within-host AMR and molecular diversity can complicate AST, treatment and control of P. aeruginosa UTI.


Translocation of a hybrid YopE-adenylate cyclase from Yersinia enterocolitica into HeLa cells.
Pathogenic bacteria of the genus Yersinia release in vitro a set of antihost proteins called Yops. Upon infection of cultured epithelial cells, extracellular Yersinia pseudotuberculosis transfers YopE across the host cell plasma membrane. To facilitate the study of this translocation process, we constructed a recombinant Yersinia enterocolitica strain producing YopE fused to a reporter enzyme. As a reporter, we selected the calmodulin-dependent adenylate cyclase of Bordetella pertussis and we monitored the accumulation of cyclic AMP (cAMP). Since bacteria do not produce calmodulin, cyclase activity marks the presence of hybrid enzyme in the cytoplasmic compartment of the eukaryotic cell. Infection of a monolayer of HeLa cells by the recombinant Y. enterocolitica strain led to a significant increase of cAMP. This phenomenon was dependent not only on the integrity of the Yop secretion pathway but also on the presence of YopB and/or YopD. It also required the presence of the adhesin YadA at the bacterial surface. In contrast, the phenomenon was not affected by cytochalasin D, indicating that internalization of the bacteria themselves was not required for the translocation process. Our results demonstrate that Y. enterocolitica is able to transfer hybrid proteins into eukaryotic cells. This system can be used not only to study the mechanism of YopE translocation but also the fate of the other Yops or even of proteins secreted by other bacterial pathogens.


Oscillations in cerebral blood flow detected with a transcranial Doppler index.
Although transcranial Doppler ultrasound (TCD) has been used to detect oscillations in CBF, interpretation is severely limited, since only blood velocity and not flow is measured. Oscillations in vessel diameter could, therefore, mask or alter the detection of those in flow by TCD velocities. In this report, the authors use a TCD-derived index of flow to detect and quantify oscillations of CBF in humans at rest. A flow index (FI) was calculated from TCD spectra by averaging the intensity weighted mean in a beat-by-beat manner over 10 seconds. Both FI and TCD velocity were measured in 16 studies of eight normal subjects at rest every 10 seconds for 20 minutes. End tidal CO2 and blood pressure were obtained simultaneously in six of these studies. The TCD probe position was meticulously held constant. An index of vessel area was calculated by dividing FI by velocity. Spectral estimations were obtained using the Welch method. Spectral peaks were defined as peaks greater than 2 dB above background. The frequencies and magnitudes of spectral peaks of FI, velocity, blood pressure, and CO2 were compared with t tests. The Kolmogorov-Smirnov test was used to further confirm that the data were not white noise. In most cases, three spectral peaks (a, b, c) could be identified, corresponding to periods of 208+/-93, 59+/-31, and 28+/-4 (SD) seconds for FI, and 196+/-83, 57+/-20, and 28+/-6, (SD) seconds for velocity. The magnitudes of the spectral peaks for FI were significantly greater (P<0.02) than those for velocity. These magnitudes corresponded to variations of at least 15.6%, 9.8%, and 6.8% for FI, and 4.8%, 4.2%, and 2.8% for velocity. The frequencies of the spectral peaks of CO2 were similar to those of FI with periods of 213+/-100, 60+/-46, and 28+/-3.6 (SD) seconds. However, the CO2 spectral peak magnitudes were small, with an estimated maximal effect on CBF of (+/-) 2.5+/-0.98, 1.5+/-0.54, and 1.1+/-0.31 (SD) percent. The frequencies of the blood pressure spectral peaks also were similar, with periods of 173+/-81, 44+/-8, and 26+/-2.5 (SD) seconds. Their magnitudes were small, corresponding to variations in blood pressure of (+/-) 2.1+/-0.55, 0.97+/-0.25, and 0.72+/-0.19 (SD) percent. Furthermore, coherence analysis showed no correlation between CO2 and FI, and only weak correlations at isolated frequencies between CO2 and velocity, blood pressure and velocity, or blood pressure and FI. The Kolmogorov-Smirnov test distinguished our data from white noise in most cases. Oscillations in vessel flow occur with significant magnitude at three distinct frequencies in normal subjects at rest and can be detected with a TCD-derived index. The presence of oscillations in blood velocity at similar frequencies but at lower magnitudes suggests that the vessel diameters oscillate in synchrony with flow. Observed variations in CO2 and blood pressure do not explain the flow oscillations. Ordinary TCD velocities severely underestimate these oscillations and so are not appropriate when small changes in flow are to be measured.


Evaluation of a topical herbal drug for its in-vivo immunomodulatory effect on cytokines production and antibacterial activity in bovine subclinical mastitis.
Antibiotics have been in use in the treatment of bovine mastitis since decades; however, their use is associated with cost issues and human health concern. Use of herbal drugs does not generally carry these disadvantages. Many plants/herbs have been evaluated in the treatment of bovine mastitis with additional property of immunomodulation in affected mammary gland. To evaluate a topical herbal drug in two breeds of cattle for its in-vivo immunomodulatory effect on cytokines production and antibacterial activity in bovine subclinical mastitis. The response to treatment was evaluated by enumerating somatic cell count (SCC), determining total bacterial load, and studying the expression of different cytokines (interleukin [IL]-6, IL-8, IL-12, granulocyte macrophage-colony stimulating factor, interferon (IFN)-γ and tumor necrosis factor [TNF]-α). The pre- and post-treatment SCC in mastitic quarters statistically did not differ significantly, however, total bacterial load declined significantly from day 0 onwards in both the breeds. Highly significant differences (P < 0.01) were observed in all the cytokines on day 0, 5, and 21 postlast treatment in both the breeds. The expression level of all the cytokines showed a significant increase on day 5, while a decrease was noticed on day 21 in both the breeds of cattle. The comparison of cytokine expression profiles between crossbred and Gir cattle revealed a significant difference in expression of IL-6 and TNF-α. However, other cytokines exhibited a similar pattern of expression in both breeds, which was non-significant. The topical herbal drug exhibited antibacterial and immunomodulatory activities in subclinical mastitis and thus the work supports its use as alternative herbal therapy against subclinical udder infection in bovines.


[CME Sonography 96/Answers: Spleen Changes].
CME Sonography 96/Answers: Spleen Changes Abstract. The spleen is a somewhat neglected organ but there are some changes that must not be neglected. Splenomegaly, caused worldwide most commonly by malaria, can occur in other infections, in portal vein thrombosis or in portal hypertension in the context of liver cirrhosis. An accessory spleen is also often found. Sometimes after splenectomy, small remains are found which may later hypertrophy. Where focal changes are concerned, we differentiate between various forms of spleen cysts, lymphomas, metastases, and benign tumors.


The conserved Mec1/Rad53 nuclear checkpoint pathway regulates mitochondrial DNA copy number in Saccharomyces cerevisiae.
How mitochondrial DNA (mtDNA) copy number is determined and modulated according to cellular demands is largely unknown. Our previous investigations of the related DNA helicases Pif1p and Rrm3p uncovered a role for these factors and the conserved Mec1/Rad53 nuclear checkpoint pathway in mtDNA mutagenesis and stability in Saccharomyces cerevisiae. Here, we demonstrate another novel function of this pathway in the regulation of mtDNA copy number. Deletion of RRM3 or SML1, or overexpression of RNR1, which recapitulates Mec1/Rad53 pathway activation, resulted in an approximately twofold increase in mtDNA content relative to the corresponding wild-type yeast strains. In addition, deletion of RRM3 or SML1 fully rescued the approximately 50% depletion of mtDNA observed in a pif1 null strain. Furthermore, deletion of SML1 was shown to be epistatic to both a rad53 and an rrm3 null mutation, placing these three genes in the same genetic pathway of mtDNA copy number regulation. Finally, increased mtDNA copy number via the Mec1/Rad53 pathway could occur independently of Abf2p, an mtDNA-binding protein that, like its metazoan homologues, is implicated in mtDNA copy number control. Together, these results indicate that signaling through the Mec1/Rad53 pathway increases mtDNA copy number by altering deoxyribonucleoside triphosphate pools through the activity of ribonucleotide reductase. This comprises the first linkage of a conserved signaling pathway to the regulation of mitochondrial genome copy number and suggests that homologous pathways in humans may likewise regulate mtDNA content under physiological conditions.


The effect of starvation on the incorporation of palmitate into glycerides and phospholipids of rat liver homogenates.
1. Glyceride biosynthesis from glycerol phosphate and [1-(14)C]palmitate was studied in liver homogenates of rats that were fed ad libitum or starved for 36-40hr. The changes in enzyme activity were related to total DNA content or total liver homogenate as these were found to be equivalent and to be the most meaningful parameters. 2. In liver homogenates from fed rats, labelled palmitate was incorporated mainly into phosphatidate (58% of the total incorporation into lipids), diglycerides (25%) and triglycerides (16%), whereas monoglycerides, cholesterol esters and phospholipids other than phosphatidate were labelled only to a small extent. Addition of particle-free supernatant to full homogenates increased the total incorporation of palmitate by 45% and the pattern of incorporation altered to 53% incorporated into triglycerides, 24% into diglycerides and 17% into phosphatidate. This result suggested that, in liver homogenates, phosphatidate phosphohydrolase (EC 18.104.22.168) may be rate-limiting in the biosynthesis of glycerides via the glycerol phosphate pathway. 3. Upon starvation, the amount of palmitate incorporated per liver into total phospholipids plus glycerides was decreased to between 68% and 75% of that observed with fed animals. In homogenates from fed animals 41-44% of the labelled phospholipids plus glycerides was in glycerides; this value increased to between 63% and 75% with starved rats. Of the palmitate incorporated into total phospholipids, between 85% and 86% was found in phosphatidate, independent of the nutritional state of the animal. The ratio of palmitate incorporated into triglycerides/diglycerides rose from 0.7, obtained with fed rats, to 1.0 with starved animals. 4. These results indicate that starvation caused a decrease in the activity (per total liver) of acyl-CoA-glycerol phosphate acyltransferase(s) (EC 22.214.171.124) and an increase in the activity of acyl-CoA-diglyceride acyltransferase (EC 188.8.131.52). The largest change, however, seemed to be related to the increased activity of the phosphatidate phosphohydrolase in the particle-free supernatant. 5. The latter enzyme was assayed in the particle-free supernatant with membrane-bound phosphatidate as substrate. In starvation, the activity per total liver was increased to between 130% and 190% and the specific activity to between 180% and 320% of the values for fed rats.


Quantitative immunoproteomics analysis reveals novel MHC class I presented peptides in cisplatin-resistant ovarian cancer cells.
Platinum-based chemotherapy is widely used to treat various cancers including ovarian cancer. However, the mortality rate for patients with ovarian cancer is extremely high, largely due to chemo-resistant progression in patients who respond initially to platinum based chemotherapy. Immunotherapy strategies, including antigen specific vaccines, are being tested to treat drug resistant ovarian cancer with variable results. The identification of drug resistant specific tumor antigens would potentially provide significant improvement in effectiveness when combined with current and emerging therapies. In this study, using an immunoproteomics method based on iTRAQ technology and an LC-MS platform, we identified 952 MHC class I presented peptides. Quantitative analysis of the iTRAQ labeled MHC peptides revealed that cisplatin-resistant ovarian cancer cells display increased levels of MHC peptides derived from proteins that are implicated in many important cancer pathways. In addition, selected differentially presented epitope specific CTL recognize cisplatin-resistant ovarian cancer cells significantly better than the sensitive cells. These over-presented, drug resistance specific MHC class I associated peptide antigens could be potential targets for the development of immunotherapeutic strategies for the treatment of ovarian cancer including the drug resistant phenotype.


Genetic predictors of response to treatment with citalopram in depression secondary to traumatic brain injury.
To determine which serotonergic system-related single nucleotide polymorphisms (SNPs) predicted variation in treatment response to citalopram in depression following a traumatic brain injury (TBI). Ninety (50 M/40 F, aged 39.9, SD = 18.0 years) post-TBI patients with a major depressive episode (MDE) were recruited into a 6-week open-label study of citalopram (20 mg/day). Six functional SNPs in genes related to the serotonergic system were examined: serotonin transporter (5HTTLPR including rs25531), 5HT1A C-(1019)G and 5HT2A T-(102)C, methylene tetrahydrofolate reductase (MTHFR) C-(677)T, brain-derived neurotrophic factor (BDNF) val66met and tryptophan hydroxylase-2 (TPH2) G-(703)T. Regression analyses were performed using the six SNPs as independent variables: Model 1 with response (percentage Hamilton Depression (HAMD) change from baseline to endpoint) as the dependent variable and Model 2 with adverse event index as the dependent variable (Bonferroni corrected p-value < 0.025). MTHFR and BDNF SNPs predicted greater treatment response (R(2)= 0.098, F = 4.65, p = 0.013). The 5HTTLPR predicted greater occurrence of adverse events (R(2)= 0.069, F = 5.72, p = 0.020). Results suggest that polymorphisms in genes related to the serotonergic system may help predict short-term response to citalopram and tolerability to the medication in patients with MDE following a TBI.


Genetic relationships and variation in reproductive strategies in four closely related bromeliads adapted to neotropical 'inselbergs': Alcantarea glaziouana, A. regina, A. geniculata and A. imperialis (Bromeliaceae).
Bromeliads (Bromeliaceae) adapted to rock outcrops or 'inselbergs' in neotropical rain forests have been identified as suitable plant models for studying population divergence and speciation during continental plant radiations. Little is known about genetic relationships and variation in reproductive strategies within and among inselberg-adapted species, yet knowledge of these parameters is important for understanding divergence processes and for conservation planning. Nuclear microsatellites were used to assess the role of clonal reproduction, estimate genetic diversity and explore genetic relationships and variation in reproductive strategies for a total of 15 populations of four closely related Alcantarea inselberg species in south-eastern Brazil: A. glaziouana, A. regina, A. geniculata and A. imperialis. Clonal propagation is frequent in coastal populations of A. glaziouana and A. regina, but absent in the high-altitude species A. geniculata and A. imperialis. Considerable variation in clonal diversity, gene diversity (H(e)), allelic richness, and Wright's inbreeding coefficient (F(IS)) exists within and between species of Alcantarea. A Bayesian analysis of coastal inselberg species indicated pronounced genetic structure. A neighbor-joining analysis grouped populations of each species together with moderate bootstrap support, except for the high altitude species A. imperialis. The coastal inselberg species A. glaziouana and A. regina tend to propagate asexually via vegetative clonal growth, and both reproductive strategies and breeding systems vary greatly between populations and species of Alcantarea. The microsatellite data indicate a history of hybridization and reticulation involving the high-altitude species A. geniculata and A. imperialis in areas of co-occurrence. The results highlight the need to understand similarities and differences in reproductive strategies both within and between related species for conservation planning and as a basis for understanding evolutionary processes in tropical radiations.


Latent factor structure of a behavioral economic cigarette demand curve in adolescent smokers.
Behavioral economic demand curves, or quantitative representations of drug consumption across a range of prices, have been used to assess motivation for a variety of drugs. Such curves generate multiple measures of drug demand that are associated with cigarette consumption and nicotine dependence. However, little is known about the relationships among these facets of demand. The aim of the study was to quantify these relationships in adolescent smokers by using exploratory factor analysis to examine the underlying structure of the facets of nicotine incentive value generated from a demand curve measure. Participants were 138 adolescent smokers who completed a hypothetical cigarette purchase task, which assessed estimated cigarette consumption at escalating levels of price/cigarette. Demand curves and five facets of demand were generated from the measure: Elasticity (i.e., 1/α or proportionate price sensitivity); Intensity (i.e., consumption at zero price); O(max) (i.e., maximum financial expenditure on cigarettes); P(max) (i.e., price at which expenditure is maximized); and Breakpoint (i.e., the price that suppresses consumption to zero). Principal components analysis was used to examine the latent structure among the variables. The results revealed a two-factor solution, which were interpreted as "Persistence," reflecting insensitivity to escalating price, and "Amplitude," reflecting the absolute levels of consumption and price. These findings suggest a two factor structure of nicotine incentive value as measured via a demand curve. If supported, these findings have implications for understanding the relationships among individual demand indices in future behavioral economic studies and may further contribute to understanding of the nature of cigarette reinforcement.


The impact of unfractionated heparin or bivalirudin on patients with stable coronary artery disease undergoing percutaneous coronary intervention.
To compare bleeding and clinical events of patients with stable angina or silent ischemia undergoing percutaneous coronary intervention (PCI) treated with unfractionated heparin (UFH) or bivalirudin. Few direct comparisons between UFH monotherapy versus bivalirudin exist for patients with stable ischemic heart disease undergoing PCI. A prospective, investigator-initiated, single-center, single-blinded, randomized trial of UFH versus bivalirudin was conducted. The primary endpoint was all bleeding (major and minor) from index-hospitalization to 30 days post discharge. Secondary endpoints included major adverse cerebral and cardiovascular events (MACCE) and net adverse clinical events (NACE). Two-hundred-sixty patients were randomized for treatment with either UFH (n = 123) (47%) or bivalirudin (n = 137) (53%) There were no significant differences in baseline clinical and angiographic characteristics between the two groups. Primary endpoint was similar in both groups (10.9% with bivalirudin vs 7.3% with UFH [P = 0.31]). Major bleeding rates were 5.8% and 2.4%, respectively (P = 0.17). There was a higher MACCE (3.5% vs 0%, P = 0.03) and NACE (8.8% vs 2.4%, P = 0.03) rate with bivalirudin compared to UFH, respectively. Bivalirudin had increased odds of NACE (OR = 3.65, 95% CI: 1.00-13.3.6). Death and stent thrombosis rates were low and similar in both groups. Radial access was associated with fewer bleeding events compared to femoral access but not statistically significant (P = 0.29). Among patients with stable angina or silent ischemia, there was no difference between UFH and bivalirudin in bleeding rates up to 30-days post-PCI. MACCE and NACE were higher among the bivalirudin group. Radial access was associated with a numerically lower rate of bleeding compared with femoral access.


Skeletogenesis in the Persian sturgeon Acipenser persicus and its correlation with gene expression of vitamin K-dependent proteins during larval development.
The present study describes morphological development of the skeleton in the Persian sturgeon Acipenser persicus and discusses the hypothesis that expression of genes encoding vitamin K-dependent proteins (VKDP) might be correlated with the mineralization of skeletal tissues during early development in sturgeons. Results showed that development of cartilage started just after hatching (mean ± S.D., 10·9 ± 0·7 mm in total length, LT ) in the head and notochord, whereas the first signs of mineralization occurred in the dentary and in the dermopalatine and palatopterygoid elements of the upper jaw, coinciding with the onset of exogenous feeding (20·1 ± 1·5 mm LT ). All branchial arch elements developed between 19·3 and 22·3 mm LT , whereas mineralization was only observed in tooth plates associated with the hypobranchial 1 and gill rakers at 20·8 ± 1·5 mm LT and 48·4 ± 6·4 mm LT , respectively. Quantitative real-time PCR showed that transcripts of VKDP genes including bone Gla protein (bgp), matrix Gla protein (mgp) and Gla rich protein (grp) genes were significantly up-regulated during the transition to exogenous feeding, supporting hypotheses about relevance of the above-mentioned genes in chondrogenesis at early developmental stages. The strong mineralization of skeletal elements from 21·5 to 27·3 mm LT (20 days post hatch) was in accordance with the maximal levels of bgp, mgp and grp expression indicating a correlation between development of the skeleton and the expression of VKDP genes. These data are important for evaluating A. persicus larval quality, understanding the influence of rearing biotic and abiotic factors on skeletogenesis and recognizing the occurrence of skeletal deformities in this species.


Role of angiotensinogen gene polymorphism on Helicobacter pylori infection-related gastric cancer risk in Japanese.
The renin-angiotensin (RA) system including angiotensinogen (AGT), angiotensin I and angiotensin II influences the regulation of cell proliferation, angiogenesis and inflammation. AGT-20 A/C polymorphism is associated with the plasma AGT and angiotensin II levels. The aim of this study was to clarify the association of AGT-20 A/C polymorphism with susceptibility to gastric cancer and peptic ulcer in Japanese. We assessed the AGT-20 A/C polymorphism in Helicobacter pylori-positive patients with gastric cancer (n = 135), gastric ulcer (n = 148) and duodenal ulcer (n = 113) and controls (n = 292) consisting of H.pylori-positive gastritis alone (n = 160) and H.pylori-negative subjects (n = 132). The age- and sex-adjusted odds ratios (ORs) of AGT-20 A/C and C/C genotypes relative to A/A genotype for gastric cancer risk were 1.695 [95% confidence interval (CI): 1.035-2.777] and 2.259 (95% CI: 0.351-14.533), respectively. The AGT-20 C allele increased the gastric cancer risk (OR: 1.685, 95% CI: 1.037-2.736), especially the intestinal type of gastric cancer (OR: 1.792, 95% CI: 1.040-3.089). However, there was no association between the AGT-20 polymorphism and susceptibility to peptic ulcer. The carriage of AGT-20 C allele was associated with an increased risk for H.pylori-related gastric cancer development in Japanese, indicating that the RA system plays an important role in the pathogenesis of gastric cancer.


Studies on the relationship between pulsed UV light irradiation and the simultaneous occurrence of molecular and cellular damage in clinically-relevant Candida albicans.
This constitutes the first study to report on the relationship between pulsed UV light (PL) irradiation and the simultaneous occurrence of molecular and cellular damage in clinical strains of Candida albicans. Microbial protein leakage and propidium iodide (PI) uptake assays demonstrated significant increases in cell membrane permeability in PL-treated yeast that depended on the amount of UV pulses applied. This finding correlated well with the measurement of increased levels of lipid hydroperoxidation in the cell membrane of PL-treated yeast. PL-treated yeast cells also displayed a specific pattern of intracellular reactive oxygen species (ROS) generation, where ROS were initially localised in the mitochondria after low levels of pulsing (UV dose 0.82 μJ/cm(2)) before more wide-spread cytosolic ROS production occurred with enhanced pulsing. Intracellular ROS levels were measured using the specific mitochondrial peroxide stain dihydrorhodamine 123 and the cytosolic oxidation stain dichloroflurescin diacetate. Use of the dihydroethidium stain also revealed increased levels of intracellular superoxide as a consequence of augmented pulsing. The ROS bursts observed during the initial phases of PL treatment was consistent with the occurrence of apoptotic cells as confirmed by detection of specific apoptotic markers, abnormal chromatin condensation and externalisation of cell membrane lipid phosphatidylserine. Increased amount of PL-irradiation (ca. UV does 1.24-1.65 μJ/cm(2)) also resulted in the occurrence of late apoptotic and necrotic yeast phenotypes, which coincided with the transition from mitochondrial to cytosolic localisation of ROS and with irreversible cell membrane leakage. Use of the comet assay also revealed significant nuclear damage in similarly treated PL samples. Although some level of cellular repair was observed in all test strains during sub-lethal exposure to PL-treatments (≤20 pulses or UV dose 0.55 μJ/cm(2)), this was absent in similar samples exposed to increased amounts of pulsing. This study showed that PL-irradiation inactivates C. albicans test strains through a multi-targeted process with no evidence of microbial ability to support cell growth after ≤20 pulses. Implications of our findings in terms of application of PL for contact-surface disinfection are discussed.


Fluoxetine-induced inhibition of male rat copulatory behavior: modification by lesions of the nucleus paragigantocellularis.
In Experiment 1, the 5-HT uptake blocker fluoxetine (FLX; 20 mg/kg) reduced the proportion of sexually experienced male rats displaying ejaculations. Among those animals that did ejaculate there was an increase in intromission frequency (IF), ejaculation latency (EL), and postejaculatory interval (PEI) and a reduction in copulatory efficiency (CE) during the final copulatory sequence prior to sexual exhaustion. In Experiment 2, we found similar inhibitory effects of FLX as well as facilitating effects of lesions of the nucleus paragigantocellularis (PGi) on male rat copulatory behavior. Males with PGi lesions displayed more ejaculations and a longer latency to sexual exhaustion compared to intact animals. When FLX was given to rats with PGi lesions, it did not influence the proportion of rats ejaculating nor did it alter IF, EL, or PEI during the final copulatory series prior to exhaustion. These findings suggest that the inhibitory influences of FLX on male rat copulatory behavior are mediated in part by the interaction of FLX with neurons originating in the PGi.


[Tuberculosis in the prisons of the Sverdlovsk Region].
In 1997 to 2002, the Sverdlovsk Region showed a reduction in the proportion of the accused and prisoners among adults and adolescents from 37.2 to 17.3% in the structure of tuberculosis morbidity and from 18.2 to 3.2% in that of its mortality. In 2003, prophylactic examinations revealed tuberculosis in 95% of the accused and prisoners. As compared with the general population, in adult and adolescent prisoners, morbidity due to tuberculosis was 8 times higher, its relapses were 9 times greater (early and late relapses being 28 and 3 times greater, respectively), its incidence was 9-fold and mortality, 1.3-fold.


How do you know you've done any better?
Three points are made in this paper. It is found that concurrent controlled randomized trials are most useful (especially Phase II trials) in speeding up the introduction of better treatments for cancer patients. A scheme of continuing comparative trials, proposed by Theodore Colton, which require no statistical significance tests, is strongly recommended. The "historical-controls" approach of Gehan and Freireich is examined and is largely rejected because it appears to be unworkable. It is urged that patient characteristics that are associated with good response be determined, for the process of giving each patient the optimal treatment for him. This is referred to as "the epidemiology of treatment." Two examples of how early uncontrolled results are misleading in evaluating potential cancer treatments are given.


Duration of dual antiplatelet therapy after implantation of drug-eluting stents.
The potential benefits and risks of the use of dual antiplatelet therapy beyond a 12-month period in patients receiving drug-eluting stents have not been clearly established. In two trials, we randomly assigned a total of 2701 patients who had received drug-eluting stents and had been free of major adverse cardiac or cerebrovascular events and major bleeding for a period of at least 12 months to receive clopidogrel plus aspirin or aspirin alone. The primary end point was a composite of myocardial infarction or death from cardiac causes. Data from the two trials were merged for analysis. The median duration of follow-up was 19.2 months. The cumulative risk of the primary outcome at 2 years was 1.8% with dual antiplatelet therapy, as compared with 1.2% with aspirin monotherapy (hazard ratio, 1.65; 95% confidence interval [CI], 0.80 to 3.36; P=0.17). The individual risks of myocardial infarction, stroke, stent thrombosis, need for repeat revascularization, major bleeding, and death from any cause did not differ significantly between the two groups. However, in the dual-therapy group as compared with the aspirin-alone group, there was a nonsignificant increase in the composite risk of myocardial infarction, stroke, or death from any cause (hazard ratio, 1.73; 95% CI, 0.99 to 3.00; P=0.051) and in the composite risk of myocardial infarction, stroke, or death from cardiac causes (hazard ratio, 1.84; 95% CI, 0.99 to 3.45; P=0.06). The use of dual antiplatelet therapy for a period longer than 12 months in patients who had received drug-eluting stents was not significantly more effective than aspirin monotherapy in reducing the rate of myocardial infarction or death from cardiac causes. These findings should be confirmed or refuted through larger, randomized clinical trials with longer-term follow-up. (ClinicalTrials.gov numbers, NCT00484926 and NCT00590174.)


Contribution of Proteins and Peptides to the Impact of a Soy Protein Isolate on Oxidative Stress and Inflammation-Associated Biomarkers in an Innate Immune Cell Model.
The innate and adaptative immune systems are involved in the regulation of inflammatory and oxidative processes and mediators such as reactive oxygen species (ROS) and nitric oxide (NO). The exacerbated action of these players results in an oxidative stress status and chronic inflammation, which is responsible for the development of non-communicable diseases (NCDs). By modulating these mediators, bioactive compounds in food can exert a key role in the prevention of several NCDs. Among these compounds, soybean proteins and peptides such as lunasin have been considered to be among the most promising. The aim of this study was to obtain and characterize a soluble protein-enriched extract from a commercial soybean protein isolate and fractionate it into different fractions through ultrafiltration. Their antioxidant and immunomodulatory properties were then evaluated using biochemical and cell models. A total of 535 proteins (from 282 protein groups) were identified in the extract, in which the presence of the peptide lunasin was confirmed. The enrichment of this peptide was achieved in the 3-10 kDa fraction. The protective effects against the oxidative stress induced by LPS in the macrophage model could have been mediated by the radical scavenging capacity of the peptides present in the soybean samples. Under basal conditions, the extract and its ultrafiltered fractions activated macrophages and induced the release of NO. However, under challenged conditions, the whole extract potentiated the NO-stimulating effects of LPS, whereas the fraction containing 3-10 kDa peptides, including lunasin, counteracted the LPS-induced NO increase. Our findings suggest a promising role of soybean protein as an ingredient for functional foods and nutraceuticals aimed at promoting health and preventing oxidative stress and/or immune-alteration-associated diseases.


Decay of unstable equilibrium and nonequilibrium states with inverse probability current taken into account.
We study the causes of noise delayed decay of unstable states in nonlinear dynamic systems within the framework of the overdamped Brownian motion model. For the analysis, we use the exact expressions for the decay times of unstable states, which take into account the inverse probability current in contrast to the well-known mean first passage time method. These expressions are valid for any intensity of fluctuations and for arbitrary potential profiles. The effect of delay is shown to arise under the decay of unstable nonequilibrium states due to the action of two different mechanisms. These mechanisms are caused by the inverse probability current and by the nonlinearity of potential describing an unstable state.


Cell-Type Transcriptomes of the Multicellular Green Alga Volvox carteri Yield Insights into the Evolutionary Origins of Germ and Somatic Differentiation Programs.
Germ-soma differentiation is a hallmark of complex multicellular organisms, yet its origins are not well understood. Volvox carteri is a simple multicellular green alga that has recently evolved a simple germ-soma dichotomy with only two cell-types: large germ cells called gonidia and small terminally differentiated somatic cells. Here, we provide a comprehensive characterization of the gonidial and somatic transcriptomes of V. carteri to uncover fundamental differences between the molecular and metabolic programming of these cell-types. We found extensive transcriptome differentiation between cell-types, with somatic cells expressing a more specialized program overrepresented in younger, lineage-specific genes, and gonidial cells expressing a more generalist program overrepresented in more ancient genes that shared striking overlap with stem cell-specific genes from animals and land plants. Directed analyses of different pathways revealed a strong dichotomy between cell-types with gonidial cells expressing growth-related genes and somatic cells expressing an altruistic metabolic program geared toward the assembly of flagella, which support organismal motility, and the conversion of storage carbon to sugars, which act as donors for production of extracellular matrix (ECM) glycoproteins whose secretion enables massive organismal expansion. V. carteri orthologs of diurnally controlled genes from C. reinhardtii, a single-celled relative, were analyzed for cell-type distribution and found to be strongly partitioned, with expression of dark-phase genes overrepresented in somatic cells and light-phase genes overrepresented in gonidial cells- a result that is consistent with cell-type programs in V. carteri arising by cooption of temporal regulons in a unicellular ancestor. Together, our findings reveal fundamental molecular, metabolic, and evolutionary mechanisms that underlie the origins of germ-soma differentiation in V. carteri and provide a template for understanding the acquisition of germ-soma differentiation in other multicellular lineages.