Large outbreak of measles in a community with high vaccination coverage: implications for the vaccination schedule.
Attempts to eliminate measles from a country or region may be disrupted by an imported case that affects indigenous persons. The objective of this study was to analyze epidemiological and clinical characteristics of a measles outbreak in Catalonia, Spain, in 2006. Data on cases of measles reported to the Department of Health, Generalitat of Catalonia, during the period 28 August 2006 through 8 July 2007 were collected. Suspected cases were confirmed by determination of measles-specific immunoglobulin M antibodies and/or detection of virus genome. Incidences were calculated using the estimated population of Catalonia for 2006, and 95% confidence intervals were determined assuming a Poisson distribution. The association between proportions was determined using the chi(2) test and Fisher's exact test. The level of statistical significance was set at alpha = .05. A total of 381 cases were confirmed, for an incidence of 6.6 cases per 100,000 persons. A total of 89.5% of cases occurred in nonvaccinated persons, mainly those aged < or =15 months (incidence, 278.2 cases per 100,000 persons; mean age of patients, 12 months). Indigenous subjects accounted for 89.8% of cases, and laboratory confirmation of results was obtained for 87.1%. Measles genotype D4 was identified in all sequenced samples. The age distribution of cases of measles among children aged <15 months suggests that the first dose of vaccine should be routinely administered at the age of 12 months.


Silent coeliac disease is over-represented in children with type 1 diabetes and their siblings.
This study measured autoantibodies against tissue transglutaminase (anti-tTG) to detect untreated coeliac disease in children with type 1 diabetes and their siblings. Anti-tTG was measured in prospectively collected sera from 169 children at the onset of diabetes, 88 of their siblings and 96 matched control children. Coeliac disease was confirmed with a small intestinal biopsy. Coeliac disease was diagnosed in five children before diabetes onset. A further 12 children were diagnosed after diabetes onset, without any gastrointestinal symptoms, and 11 of these had anti-tTG at the onset of diabetes, with the remaining child showing seroconversion within 6 months. Hence, all the children with both diseases had anti-tTG at or before diabetes diagnosis, and the prevalence of coeliac disease was 10.1%. Moreover, 6.8% of the siblings and 3.1% of the control children had elevated levels of anti-tTG. None of the siblings reported any coeliac-related symptoms, despite being positive for anti-tTG, and coeliac disease has so far been biopsy confirmed in 4.5%. Silent coeliac disease is over-represented in children with type 1 diabetes and their siblings. All diabetes children and their siblings should be tested and followed for the presence of anti-tTG and coeliac disease.


Selection and characterization of T-cell variants lacking molecules involved in T-cell activation (T3 T-cell receptor, T44, and T11): analysis of the functional relationship among different pathways of activation.
A clone of the interleukin 2-producing Jurkat leukemia cell line termed JA3 (surface phenotype, T3+, Ti+, T44+, T11+, T40+) has been used to induce and select cell variants lacking surface molecules involved in T-cell activation. Following 200 rad of gamma-radiation (1 rad = 0.01 Gy), cells were treated with monoclonal antibodies (mAbs) directed to T3, Ti, T44, or T11 antigen and complement. After growth of the residual cells in culture, "negative" cells were cloned under limiting conditions. Depending on the specificity of the mAb used for the immunoselection, three groups of variants were obtained. (i) The use of mAbs directed to T3 or Ti resulted in cell variants that expressed the T3 Ti- T44+ Leu1+ T11+ T40+ 4F2+ HLA class I+ surface phenotype. (ii) Immunoselection with anti-T44 mAb resulted in 2 variants that shared the T3- Ti- T44- Leu1- T11+ T40+ 4F2+ HLA class I+ phenotype. (iii) Cell treatment with anti-T11 mAb resulted in 15 variants characterized by the lack of T11 antigen expression and of all the other T-cell-specific surface antigens. Therefore, it appears that the different sets of JA3 cell variants, like T cells at discrete stages of intrathymic differentiation, may follow a coordinated expression of surface differentiation antigens. Analysis of the functional responsiveness of the three distinct groups of JA3 cell variants to different stimuli showed that all produced interleukin 2 in response to A23187 calcium ionophore plus phorbol 12-myristate 13-acetate. The first group of variants (T3- Ti-) did not respond to stimulation with anti-T3, anti-Ti, or anti-T44 mAbs. Eight of 9 did not respond to phytohemagglutinin either; however, all responded to appropriate stimulatory combinations of anti-T11 mAbs (and to calcium ionophore). The second group of variants (T3-, Ti-, T44-, T11+), similar to the first group, did not respond to anti-T3, anti-Ti, anti-T44 mAbs, and phytohemagglutinin, but they were fully responsive to anti-T11 mAb. The last group of variants (lacking all the T-cell-specific surface antigens) only responded to calcium ionophore A23187.


Conformation of the C1 phorbol-ester-binding domain participates in the activating conformational change of protein kinase C.
The fluorescent phorbol ester 12-N-methylanthraniloylphorbol 13-acetate [sapintoxin D (SAPD)] was used as both the activator and the probe for the activating conformational change of the C1 domain of recombinant protein kinase C (PKC)alpha. Fluorescence emission spectra and steady-state anisotropy measurements of SAPD in fully active membrane-associated PKC show that there is a relatively hydrophobic environment and restricted motional freedom characterizing the phorbol-ester-binding site. SAPD also interacts with the membrane lipids so that it was necessary to resort to time-resolved anisotropy measurements to resolve the signals corresponding to PKC-bound SAPD from that associated with buffer and lipid. In the presence of membrane lipids (unilamellar vesicles of phosphatidylcholine and phosphatidylserine, 4:1 molar ratio) and Ca(2+), at a concentration sufficient to activate the enzyme fully, a long correlation time characteristic of highly restricted motion was observed for PKC-associated SAPD. The fraction of SAPD molecules displaying this restricted motion, in comparison with the total SAPD including that in lipids and in buffer, increased with increasing concentrations of Ca(2+) and paralleled the appearance of enzyme activity, whereas the rotational correlation time remained constant. This could be rationalized as an increase in the number of active PKC conformers in the total population of PKC molecules. It therefore seems that there is a distinct conformation of the C1 activator-binding domain associated with the active form of PKC. The addition of SAPD and dioleoyl-sn-glycerol together produced an activity higher than that achievable by either activator alone both at concentrations that alone induced maximal activity for the respective activator; this higher activity was associated with a further restriction in SAPD motion. Increasing the cholesterol concentration, the phosphatidylethanolamine concentration, the sn-2 unsaturation in phosphatidylcholine and the vesicle curvature each also elevated SAPD-induced PKC activity and again increased the PKC-associated SAPD rotational correlation time. In summary, the rotational correlation time of PKC-bound SAPD, extractable from a single time-resolved fluorescence anisotropy measurement, provides a novel probe for the involvement of interactions between the C1 domain and phorbol ester in the modulation of PKC activity.


Comparative analysis of the factors contributing to sustainability of a food and nutrition intervention programme: Two case studies from South Africa.
Sustainability of health interventions is a consistent and ongoing issue in Africa. Understanding key aspects of sustainable interventions provides the necessary methods for success. This research evaluates a nutrition intervention implemented in two peri-urban communities in the Free State and Gauteng province in South Africa (SA) respectively. A case study approach was employed using cross-case study analyses, including triangular data collection (empirical research through surveys of respondents; qualitative data collection through researcher journals, diaries, communications and photographs; and research team reflections). The information from these case studies is organized according to a framework from the United States (US) Office of Organizational Health (OAH) to evaluate the sustainability of intervention programmes. All eight of the OAH criteria were met in the Free State community and supported the importance of these key factors for sustainability compared to only three in the Gauteng community. The OAH framework provided a compelling rationale for the relative success of the intervention and clearly showed the Free State project was sustainable. The Gauteng project was not sustainable. Planned interventions should devote significant effort and time towards ensuring sustainability. The OAH guidelines provide key steps that the research shows is relevant to the South African context.


The roles of the shikimate pathway genes, aroA and aroB, in virulence, growth and UV tolerance of Burkholderia glumae strain 411gr-6.
Burkholderia glumae is the major causal agent of bacterial panicle blight of rice, which is a growing disease problem for rice growers worldwide. In our previous study, some B. glumae strains showed pigmentation phenotypes producing at least two (yellow-green and purple) pigment compounds in casein-peptone-glucose agar medium. The B. glumae strains LSUPB114 and LSUPB116 are pigment-deficient mutant derivatives of the virulent and pigment-proficient strain 411gr-6, having mini-Tn5gus insertions in aroA encoding 3-phosphoshikimate 1-carboxyvinyltransferase and aroB encoding 3-dehydroquinate synthase, respectively. Both enzymes are known to be involved in the shikimate pathway, which leads to the synthesis of aromatic amino acids. Here, we demonstrate that aroA and aroB are required for normal virulence in rice and onion, growth in M9 minimal medium and tolerance to UV light, but are dispensable for the production of the phytotoxin toxoflavin. These results suggest that the shikimate pathway is involved in bacterial pathogenesis by B. glumae without a significant role in the production of toxoflavin, a major virulence factor of this pathogen.


Spontaneous gastrosplenic fistula secondary to primary splenic lymphoma.
A gastrosplenic fistula is a rare complication of gastric and splenic lymphomas which can occur spontaneously or secondary to chemotherapy. We report a case of a spontaneous gastrosplenic fistula secondary to a diffuse splenic large B cell lymphoma in a previously well 43-year-old patient. CT imaging demonstrated the fistula, which was subsequently managed with chemotherapy. The clinical management of this rare condition is discussed with a review of the literature.


A randomized, controlled study of autologous therapy with bone marrow-derived aldehyde dehydrogenase bright cells in patients with critical limb ischemia.
The safety and efficacy of direct intramuscular injections of aldehyde dehydrogenase bright (ALDH(br)) cells isolated from autologous bone marrow mononuclear cells (ABMMNCs) and ABMMNCs were studied in patients with critical limb ischemia (CLI) who were not eligible for percutaneous or surgical revascularization. Many CLI patients are not candidates for current revascularization procedures, and amputation rates are high in these patients. Cell therapy may be a viable option for CLI patients. Safety was the primary objective and was evaluated by occurrence of adverse events. Efficacy, the secondary objective, was evaluated by assessment of Rutherford category, ankle-brachial index (ABI), transcutaneous partial pressure of oxygen (TcPO(2)), quality of life, and pain. ALDH(br) cells and ABMMNCs were successfully administered to all patients. No therapy-related serious adverse events occurred. Patients treated with ALDH(br) cells (n = 11) showed significant improvements in Rutherford category from baseline to 12 weeks (mean, 4.09 ± 0.30 to 3.46 ± 1.04; P = 0.05) and in ABI at 6 (mean, 0.22 ± 0.19 to 0.30 ± 0.24; P = 0.02), and 12 weeks (mean, 0.36 ± 0.18; P = 0.03) compared with baseline. Patients in the ABMMNC group (n = 10) showed no significant improvements at 6 or 12 weeks in Rutherford category but did show improvement in ABI from baseline to 12 weeks (0.38 ± 0.06 to 0.52 ± 0.16; P = 0.03). No significant changes from baseline were noted in ischemic ulcer grade or TcPO(2) in either group. Administration of autologous ALDH(br) cells appears to be safe and warrants further study in patients with CLI.


Reduction of baclofen-, but not sodium valproate-induced growth hormone release in type I diabetic men.
The effects of sodium valproate (a drug enhancing endogenous gamma aminobutyric acid (GABA)-ergic activity) and of the GABA analog baclofen (a GABA B receptor agonist) on serum GH levels was tested in 8 type I diabetic men and 8 normal controls. Sodium valproate (800 mg) or baclofen (10 mg) were given by mouth at 08.30 h on the experimental day. Control tests with a placebo were performed on different occasions. Basal GH levels were similar in controls and diabetic patients. Sodium valproate induced a 7 fold increase in serum GH concentrations in both groups. In contrast, baclofen-induced GH rise was significantly higher in normal controls (mean peak was 3.4 times higher than baseline) than in diabetic patients (mean peak was only 2.1 times higher than basal value). Serum GH levels did not change after placebo administration in any groups. These data suggest the presence of diabetes-induced alterations of a GABAergic pathway mediated by B receptors in the control of GH secretion. Alternatively, the data might indicate a change in diabetic men of other baclofen-sensitive neurotransmissions, different from GABA.


Is prolactin secreted ectopically?
The ectopic secretion of prolactin by neoplasms has been reported only rarely although in one preliminary report it has been found to be present commonly in tumor extracts. To ascertain whether this is due to a true rarity of prolactin-secreting tumors or to the fact that prolactin hypersecretion causes few notable clinical symptoms in patients with cancer, we measured prolactin levels by radioimmunoassay in the serums of 215 patients who had a variety of malignancies. Prolactin levels were elevated (greater than 25 ng/ml) in 15 patients. In 12 of these 15 patients, the elevations could be explained by the use of phenothiazine or opiates, or prior irradiation to the chest wall or head. Additional serum samples showed clearly normal prolactin levels in one of the patients. Of the entire series of 215 patients, therefore, only two patients (1 percent) had modestly elevated prolactin levels without having known causes for hyperprolactinemia: one patient with squamous cell carcinoma of the lung (prolactin 34.1 ng/ml) and one patient with breast cancer (prolactin 45.4 ng/ml). Even in these two patients the hyperprolactinemia could as likely be due to stimulation of afferent nerves in the chest wall as to ectopic secretion by the tumors. There is no clear evidence in our study or in the literature that prolactin is secreted ectopically; this is in sharp contrast to many studies showing the high frequencies of the ectopic secretion of a number of other hormones. Why this is so is unknown.


Short-Term Mortality Among Pediatric Patients With Heart Diseases Undergoing Veno-Arterial Extracorporeal Membrane Oxygenation: A Systematic Review and Meta-Analysis.
Veno-arterial extracorporeal membrane oxygenation serves as a crucial mechanical circulatory support for pediatric patients with severe heart diseases, but the mortality rate remains high. The objective of this study was to assess the short-term mortality in these patients. We systematically searched PubMed, Embase, and Cochrane Library for observational studies that evaluated the short-term mortality of pediatric patients undergoing veno-arterial extracorporeal membrane oxygenation. To estimate short-term mortality, we used random-effects meta-analysis. Furthermore, we conducted meta-regression and binomial regression analyses to investigate the risk factors associated with the outcome of interest. We systematically reviewed 28 eligible references encompassing a total of 1736 patients. The pooled analysis demonstrated a short-term mortality (defined as in-hospital or 30-day mortality) of 45.6% (95% CI, 38.7%-52.4%). We found a significant difference (P<0.001) in mortality rates between acute fulminant myocarditis and congenital heart disease, with acute fulminant myocarditis exhibiting a lower mortality rate. Our findings revealed a negative correlation between older age and weight and short-term mortality in patients undergoing veno-arterial extracorporeal membrane oxygenation. Male sex, bleeding, renal damage, and central cannulation were associated with an increased risk of short-term mortality. The short-term mortality among pediatric patients undergoing veno-arterial extracorporeal membrane oxygenation for severe heart diseases was 45.6%. Patients with acute fulminant myocarditis exhibited more favorable survival rates compared with those with congenital heart disease. Several risk factors, including male sex, bleeding, renal damage, and central cannulation contributed to an increased risk of short-term mortality. Conversely, older age and greater weight appeared to be protective factors.


Ellagic acid inhibits oxidized LDL-mediated LOX-1 expression, ROS generation, and inflammation in human endothelial cells.
LOX-1, a lectin-like receptor on endothelial cells, facilitates the uptake of oxidized low-density lipoprotein (oxLDL). Expression of LOX-1 is involved in the pathobiological effects of oxLDL in endothelial cells, including reactive oxygen species (ROS) generation, suppression of endothelial nitric oxide synthase (eNOS) activity, and leukocytic adhesion. Moderate consumption of phenolic-enriched food may have a protective effect against the development of atherosclerosis via the antioxidant capacity of phenolic compounds at the endothelial level. In this study, we determined whether ellagic acid, a polyphenolic compound widely distributed in fruits and nuts, protects against oxLDL-induced endothelial dysfunction by modulating the LOX-1-mediated signaling pathway. Human umbilical vein endothelial cells (HUVECs) were pretreated with ellagic acid at doses of 5, 10, 15, and 20 μM for 2 hours and then incubated with oxLDL (150 μg/mL) for an additional 24 hours. LOX-1 protein expression was markedly lower after exposure to oxLDL in HUVECs pretreated with ellagic acid or diphenyleneiodonium, a well-known inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, than in HUVECs exposed to oxLDL alone, suggesting that ellagic acid deactivates NADPH oxidase. We also found that oxLDL activated the membrane assembly of p47phox, Rac1, gp91 and p22phox, and the subsequent induction of ROS generation; however, ROS generation was markedly suppressed in cells pretreated with ellagic acid or anti-LOX-1 monoclonal antibody. In addition, oxLDL down-regulated eNOS and up-regulated inducible NO synthase (iNOS), thereby augmenting the formation of NO and protein nitrosylation. Furthermore, oxLDL induced the phosphorylation of p38 mitogen-activated protein kinase, activated the NF-κB-mediated inflammatory signaling molecules interleukin-(IL) 6 and IL-8 and the adhesion molecules intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin, and stimulated the adherence of THP-1 (a human acute monocytic leukemia cell line) to HUVECs. Pretreatment with ellagic acid, however, exerted significant cytoprotective effects in all events. Findings from this study may provide insight into a possible molecular mechanism by which ellagic acid inhibits LOX-1-induced endothelial dysfunction. Our data indicate that ellagic acid exerts its protective effects by inhibiting NADPH oxidase-induced overproduction of superoxide, suppressing the release of NO by down-regulating iNOS, enhancing cellular antioxidant defenses, and attenuating oxLDL-induced LOX-1 up-regulation and eNOS down-regulation.


Meis2 as a critical player in MN1-induced leukemia.
Meningioma 1 (MN1) is an independent prognostic marker for normal karyotype acute myeloid leukemia (AML), with high expression linked to all-trans retinoic acid resistance and poor survival. MN1 is also a potent and sufficient oncogene in murine leukemia models, strongly dependent on the MEIS1/AbdB-like HOX protein complex to transform common myeloid progenitors, block myeloid differentiation, and promote leukemic stem cell self-renewal. To identify key genes and pathways underlying leukemic activity, we functionally assessed MN1 cell phenotypic heterogeneity, revealing leukemic and non-leukemic subsets. Using gene expression profiling of these subsets combined with previously published comparisons of full-length MN1 and mutants with varying leukemogenic activity, we identified candidate genes critical to leukemia. Functional analysis identified Hlf and Hoxa9 as critical to MN1 in vitro proliferation, self-renewal and impaired myeloid differentiation. Although critical to transformation, Meis1 knockdown had little impact on these properties in vitro. However, we identified Meis2 as critical to MN1-induced leukemia, with essential roles in proliferation, self-renewal, impairment of differentiation and disease progression in vitro and in vivo. Here, we provide evidence of phenotypic and functional hierarchy in MN1-induced leukemic cells, characterise contributions of Hlf, Hoxa9 and Meis1 to in vitro leukemic properties, and reveal Meis2 as a novel player in MN1-induced leukemogenesis.


Patient and carer unmet needs: a survey of the British association of head and neck oncology nurses.
The aim of this survey was to ask members of the British Association of Head and Neck Oncology Nurses (BAHNON) about the identification of patients and carers unmet needs in the routine out-patient review clinic and the support services available during consultation. A national postal survey was sent out to the 210 current members of BAHNON in November 2009. Reminders were sent to non-responders in February 2010. The response rate was 61% (129/210). The vast majority (80%) were Clinical Nurse Specialists (CNS). The questionnaire data support the strong belief in attempts to identify unmet needs with over three-quarters feeling strongly about themselves being personally involved in attempting to identify unmet needs. Most of the responders used counselling and communication methods to elicit unmet concerns rather than specific tools such as questionnaires. The vast majority clearly felt that identifying unmet needs in clinic improves patients' perception of outcome post-treatment. Support services' readily' available at the time of consultation were as follows: H&N CNS (99%), Speech and Language (86%), Oncologist (84%), Dietician/Nutritionist (84%), Dentist (44%), Oral Rehabilitation consultant (27%), Dental Hygienist (26%), Physiotherapist (21%), Chaplain (20%), Emotional Support therapist (15%), Psychologist (15%), Occupational therapist (13%), Social worker (8%), other (11%). Although responders felt it very important to identify unmet needs in follow-up clinics, there is reliance on one to one discussion with the patient and carer. Hence in a busy clinic, needs might be easily missed and further research is required into ways to facilitate their identification.


Oligomycin-induced bioenergetic adaptation in cancer cells with heterogeneous bioenergetic organization.
Cancer cells constantly adapt to oxidative phosphorylation (OXPHOS) suppression resulting from hypoxia or mitochondria defects. Under the OXPHOS suppression, AMP-activated protein kinase (AMPK) regulates global metabolism adjustments, but its activation has been found to be transient. Whether cells can maintain cellular ATP homeostasis and survive beyond the transient AMPK activation is not known. Here, we study the bioenergetic adaptation to the OXPHOS inhibitor oligomycin in a group of cancer cells. We found that oligomycin at 100 ng/ml completely inhibits OXPHOS activity in 1 h and induces various levels of glycolysis gains by 6 h, from which we calculate the bioenergetic organizations of cancer cells. In glycolysis-dominant cells, oligomycin does not induce much energy stress as measured by glycolysis acceleration, ATP imbalance, AMPK activation, AMPK substrate acetyl-CoA carboxylase phosphorylation at Ser(79), and cell growth inhibition. In OXPHOS-dependent LKB1 wild type cells, oligomycin induces 5-8% ATP drops and transient AMPK activation during the initial 1-2 h. After AMPK activation is completed, oligomycin-induced increase of acetyl-CoA carboxylase phosphorylation at Ser(79) is still detected, and cellular ATP is back at preoligomycin treatment levels by sustained elevation of glycolysis. Cell growth, however, is inhibited without an increase in cell death and alteration in cell cycle distribution. In OXPHOS-dependent LKB1-null cells, no AMPK activation by oligomycin is detected, yet cells still show a similar adaptation. We also demonstrate that the adaptation to oligomycin does not invoke activation of hypoxia-induced factor. Our data suggest that cancer cells may grow and survive persistent OXPHOS suppression through an as yet unidentified regulatory mechanism.


Impact of Obesity on Clinical Outcomes in Urban Children Hospitalized for Status Asthmaticus.
The prevalence of both childhood asthma and obesity remain at historically high levels and disproportionately affect urban children. Asthma is a common and costly cause for pediatric hospitalization. Our objective was to determine the effect of obesity on outcomes among urban children hospitalized with status asthmaticus. A retrospective cohort study was performed by using billing system data and chart review to evaluate urban children admitted for asthma. Demographics, asthma severity, reported comorbidities, and outcomes were assessed. Obesity was defined by BMI percentile (lean<85%, overweight 85%-95%, obese≥95%). Outcomes were length of stay, hospitalization charges, ICU stay, repeat admissions, and subsequent emergency department (ED) visits. Bivariate analysis assessed for differences between overweight/obese and lean children. Multivariable regression assessed the relationship between overweight status and primary outcomes while controlling for other variables. Post hoc age-stratified analysis was also performed. The study included 333 subjects; 38% were overweight/obese. Overweight/obese children admitted with asthma were more likely than lean children to have subsequent ED visits (odds ratio 1.6, 95% confidence interval 1.0-2.6). When stratified by age, overweight/obese preschool-age children (<5 years) were >2 times as likely to have repeat ED visits than lean preschool-age children (odds ratio 2.3, 95% confidence interval 1.0-5.6). There were no differences in the other outcomes between overweight/obese and lean individuals within the entire cohort or within other age groups.


The preparation of activated factor X and its action on prothrombin.
The preparation of activated Factor X from reaction mixtures of bovine Factor X and Russell's-viper venom is described. The molecular weight of purified protein varies about a mean value of 40000; this variation is the result of at least two forms of Factor Xa. The action of activated Factor X, together with purified Factor V, was studied on purified prothrombin and the reaction products were isolated. In addition to thrombin, two other polypeptides with molecular weights of 16000 and 19500 were recovered.


[Working conditions and occupational pathology in electrolytic nickel refinery workers].
The working conditions during electrolytic nickel refinery correspond to Class 3, Grade 3-4, which may give rise to health problems in workers of all occupations employed in this production. Their risk and prevalence are significantly higher in the most exposed occupational group, such as electrolysis workers (40.1% of all cases). The structure of occupational diseases is dominated by respiratory diseases (68.8%), mainly due to exposure to aerosols of nickel compounds. It is concluded that working conditions and medical prevention interventions should be improved for this group of workers.


Identification of a novel nuclear factor-binding site in the Pisum sativum sad gene promoters.
DNA fragments containing the 5' promoter regions of the Pisum sativum sadA and sadC genes were amplified from genomic DNA, cloned and sequenced. These sequences contain a number of conserved cis-acting elements, which are potentially involved in stress-induced transcription of the sad genes. To determine whether any of the identified elements are active in binding nuclear factors in vitro, 11 60-bp overlapping (by 30 bp) DNA probe fragments covering the proximal sadC promoter sequence (360 bp) were used in electrophoretic mobility shift assays with competition. Binding activities were compared in nuclear extracts from control, UV-B-stressed and wounded pea leaves. The pattern of DNA binding was almost identical with all three extracts, with one 30-bp region being the predominant site for factor binding. Using overlapping sub-fragments of this region, the majority of the specific binding could be attributed to the novel 11-bp GC-rich sequence GTGGCGCCCAC. An almost identical sequence is conserved in the sadA promoter. This motif has features in common with a number of recognised cis-elements, which suggests a possible binding site for factors which play a role in regulating sad gene transcription.


From Systemic Inflammation to Myocardial Fibrosis: The Heart Failure With Preserved Ejection Fraction Paradigm Revisited.
In accordance with the comorbidity-inflammation paradigm, comorbidities and especially metabolic comorbidities are presumed to drive development and severity of heart failure with preserved ejection fraction through a cascade of events ranging from systemic inflammation to myocardial fibrosis. Recently, novel experimental and clinical evidence emerged, which strengthens the validity of the inflammatory/profibrotic paradigm. This evidence consists among others of (1) myocardial infiltration by immunocompetent cells not only because of an obesity-induced metabolic load but also because of an arterial hypertension-induced hemodynamic load. The latter is sensed by components of the extracellular matrix like basal laminin, which also interact with cardiomyocyte titin; (2) expression in cardiomyocytes of inducible nitric oxide synthase because of circulating proinflammatory cytokines. This results in myocardial accumulation of degraded proteins because of a failing unfolded protein response; (3) definition by machine learning algorithms of phenogroups of patients with heart failure with preserved ejection fraction with a distinct inflammatory/profibrotic signature; (4) direct coupling in mediation analysis between comorbidities, inflammatory biomarkers, and deranged myocardial structure/function with endothelial expression of adhesion molecules already apparent in early preclinical heart failure with preserved ejection fraction (HF stage A, B). This new evidence paves the road for future heart failure with preserved ejection fraction treatments such as biologicals directed against inflammatory cytokines, stimulation of protein ubiquitylation with phosphodiesterase 1 inhibitors, correction of titin stiffness through natriuretic peptide-particulate guanylyl cyclase-PDE9 (phosphodiesterase 9) signaling and molecular/cellular regulatory mechanisms that control myocardial fibrosis.


Cuisine and health: a new initiative for science and technology 'The Zhejiang Report' from Hangzhou.
There has been a rapid and remarkable recovery in Hangzhou, Zhejiang Province, China, during the 1990s, of a distinctive cuisine whose origins can be traced back to, at least, the building of the Grand Canal between Beijing and Hangzhou in the 600s AD and the stimulus to trade and migration, that was provided. This, along with exceptional health indices in the region, provides a basis for a new integrative study of the food chain, food culture and food science in regard to the human condition, which would be manifest by the degree of environmental sustainability, economic progress, social cohesion, health status, well-being and happiness that these dimensions of "food for humans " should support. Collectively, these have rarely been subject to systematic scholarly pursuit. Through the endeavours of the Zhejiang Association for Science and Technology, representing some 150,000 professionals and the international community, represented principally by the International Union of Nutritional Sciences (IUNS) and its partners in WHO, the FAO Network of Excellence and the Hangzhou University of Commerce, this field has been brought together through the papers in this issue of Asia Pacific Journal of Clinical Nutrition. The report builds on related endeavours in recent times, notably 'The Okinawan Roundtable on Nutrition and Cardiovascular Disease' which recognizes the exceptional and distinguishing features of the Okinawan diet in health protection, and the assessment of the place of fatty fruits, like red palm fruit amongst 'North and West African Foods'.


Characterization of Cardiac Amyloidosis by Atrial Late Gadolinium Enhancement Using Contrast-Enhanced Cardiac Magnetic Resonance Imaging and Correlation With Left Atrial Conduit and Contractile Function.
The diagnosis of cardiac amyloidosis (CA) often necessitates invasive myocardial biopsy. We sought to evaluate whether late gadolinium enhancement (LGE) of the atrial myocardium by cardiac magnetic resonance imaging was associated with impaired left atrial (LA) function and whether the extent of LA LGE may enhance diagnostic differentiation of CA from other cardiomyopathies. Twenty-two patients with biopsy-proven CA, 37 with systemic hypertension (SH), and 22 with nonischemic dilated cardiomyopathy (NIDC) underwent cardiac magnetic resonance imaging and echocardiographic evaluation. Patients with CA had greater minimal LA volume (57 ± 53 vs 24 ± 18 in SH and 19 ± 25% in NIDC, p = 0.003), and significantly lower total LA emptying function (19 ± 14 vs 40 ± 14 in SH and 33 ± 20% in NIDC, p = 0.0006). The mean proportion of atrial enhancement (LGELA%) was significantly greater in patients with CA than with SH and NIDC (59 ± 36% vs 7.4 ± 2.1 and 2.9 ± 9.0%, p <0.0001, respectively). There was also a strong inverse correlation between both active and total atrial emptying (r = -0.69, p = 0.001; r = -0.67, p = 0.01, respectively) with LGELA% for patients with CA. In multivariate regression analysis, LGELA% was the strongest adjusted predictor for CA diagnosis. Using receiver operating characteristic analysis, LGELA% ≥33% produced the greatest diagnostic utility for CA (sensitivity 76%, specificity 94%). Patients with CA may have extensive LGE of the LA myocardium, which is associated with marked reduction in LA emptying function. The extent of LA LGE was highly predictive for the diagnosis of CA.


Identification and characterization of two sigma-like binding sites in the mouse neuroblastoma x rat glioma hybrid cell line NG108-15.
NG108-15 cells were shown to possess high affinity binding sites for 1,3-di(2-[5-3H]tolyl)guanidine ([3H]DTG), a selective sigma ligand (Kd = 23 nM; maximal number of binding sites = 15.6 pmol/mg). The rank order of potency of drugs at this site was DTG greater than haloperidol greater than pentazocine greater than 3-(3-hydroxyphenyl)-N-(1-propyl)piperidine [(+)-3-PPP] greater than phencyclidine (PCP) greater than metaphit greater than (-)-3-PPP greater than (-)-N-allylnormetazocine [(-)-SKF 10,047] greater than (-)-butaclamol greater than (+)-butaclamol greater than (+)-SKF 10,047 greater than dizolcipine (MK 801 [5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine- maleate]) greater than ketamine. Both, Kd value and pattern of ligand selectivity suggest a close relationship to sigma sites in rodent brain. However, in comparison to sigma sites in brain stereoselectivity for the benzomorphan, SKF 10,047 was reversed and the affinities for benzomorphans were only moderate to low. Thus, sigma binding sites in NG108-15 cells seem to correspond to recently detected sigma sites in a pheochromocytoma cell line (PC12). [3H]-1-(2-thienyl)-cyclohexyl]piperidine ([3H]TCP), a PCP receptor-selective ligand binds to NG108-15 cells with moderate affinity (Kd = 139 nM; maximal number of binding sites = 4.7 pmol/mg of protein).(ABSTRACT TRUNCATED AT 250 WORDS)


A survey of putative secreted and transmembrane proteins encoded in the C. elegans genome.
Almost half of the Caenorhabditis elegans genome encodes proteins with either a signal peptide or a transmembrane domain. Therefore a substantial fraction of the proteins are localized to membranes, reside in the secretory pathway or are secreted. While these proteins are of interest to a variety of different researchers ranging from developmental biologists to immunologists, most of secreted proteins have not been functionally characterized so far. We grouped proteins containing a signal peptide or a transmembrane domain using various criteria including evolutionary origin, common domain organization and functional categories. We found that putative secreted proteins are enriched for small proteins and nematode-specific proteins. Many secreted proteins are predominantly expressed in specific life stages or in one of the two sexes suggesting stage- or sex-specific functions. More than a third of the putative secreted proteins are upregulated upon exposure to pathogens, indicating that a substantial fraction may have a role in immune response. Slightly more than half of the transmembrane proteins can be grouped into broad functional categories based on sequence similarity to proteins with known function. By far the largest groups are channels and transporters, various classes of enzymes and putative receptors with signaling function. Our analysis provides an overview of all putative secreted and transmembrane proteins in C. elegans. This can serve as a basis for selecting groups of proteins for large-scale functional analysis using reverse genetic approaches.


The Use of Alcohol, Tobacco, Illegal Drugs and Medicines: An Estimate of Consumption and Substance-Related Disorders in Germany.
Prevalence estimates of the use of tobacco, alcohol, illegal drugs, and psychoactive medications and of substance-related disorders enable an assessment of the effects of substance use on health and society. The data used for this study were derived from the 2018 Epidemiological Survey of Substance Abuse (Epidemiologischer Suchtsurvey, ESA). The sample of the German adult population comprised 9267 persons aged 18 to 64 (response rate, 42%). Population estimates were obtained by extrapolation to a total resident population of 51 544 494 people. In the 30 days prior to the survey, 71.6% of the respondents (correspond- ing to 36.9 million persons in the population) had consumed alcohol, and 28.0% (14.4 million) had consumed tobacco. 4.0% reported having used e-cigarettes, and 0.8% reported having used heat-not-burn products. Among illegal drugs, cannabis was the most commonly used, with a 12-month prevalence of 7.1% (3.7 million), followed by amphetamines (1.2%; 619 000). The prevalence of the use of anal- gesics without a prescription (31.4%) was markedly higher than that of the use of prescribed analgesics (17.5%, 26.0 million); however, analgesics were taken daily less commonly than other types of medication. 13.5% of the sample (7.0 million) had at least one dependence diagnosis (12-month prevalence). Substance use and the consumption of psychoactive medications are widespread in the German population. Substance-related disorders are a major burden to society, with legal substances causing greater burden than illegal substances.


Premature adrenarche leads to polycystic ovary syndrome? Long-term consequences.
Premature adrenarche is characterized by an early increase in adrenal androgen production that results in the development of pubic hair before the age of 8 years in girls and 9 years in boys, with or without axillary hair, and with no other signs of sexual development. Premature adrenarche has no adverse effects on the onset and progression of gonadarche and final height. However, it can no longer be considered a benign condition as it has been associated with hyperinsulinemia, dyslipidemia, and obesity already in the prepubertal period and polycystic ovary syndrome (PCOS) at adolescence. Furthermore, a possible association between premature adrenarche and metabolic and endocrine abnormalities with low birth weight has been postulated. PCOS, as recently redefined, is the most common endocrine disorder to affect women of reproductive age and has been associated with increased risk for type 2 diabetes and increased prevalence of cardiovascular risk factors at an earlier age than expected. Premature adrenarche and PCOS share similar metabolic disturbances. It may be that metabolic abnormalities start very early in life during the prenatal or prepubertal period and premature adrenarche may be a forerunner of PCOS and the metabolic syndrome in some girls. Large long-term epidemiological studies are needed to allow clear association of the two conditions and assessment of the risk of disease in later life.


Effect of sublingual nitroglycerin on cardiac performance in patients with coronary artery disease and non-dyskinetic left ventricular contraction.
In 8 patients with coronary artery disease and symmetrical left ventricular contraction, an echocardiographic study of left ventricular function was performed before and 3 minutes after the administration of 0-6 mg nitroglycerin sublingually. The left ventricular end-diastolic diameter decreased from 5-2 +/- 0-2 to 4-9 +/- 0-2 cm (P less than 0-05) and the end-systolic diameter from 4-2 +/- 0-2 to 3-7 +/- 0-2 cm (P less than 0-001). The estimated stroke volume did not change significantly, while the cardiac output increased, 5-8 +/- 0-6 to 7-7 +/- 0-6 l min-1 (P less than 0-001) and the heart rate increased from 72 +/- 5 to 90 +/- 6 (P less than 0-001). The mean arterial blood pressure decreased from 105 +/- 4 to 88 +/- 3 mmHg (P less than 0-001). The ejection fraction increased from 53 +/- 3 per cent to 65 +/- 6 per cent (P less than 0-001) and the mean velocity of circumferential fibre shortening (VCF) from 0-81 +/- 0-05 to 1-15 +/- 0-10 circumferences per second (P less than 0-001). The estimated midsystolic midwall stress decreased from 155 +/- 14 g cm-2 to 102 +/- 12 g cm-2 after mitroglycerin (P less than 0-001). The administration of nitroglycerin was associated with a significant decrease in left ventricular preload and afterload. A vasodilating effect is suggested by the fall in peripheral resistance. The overall improvement in ejection fraction and VCF may not reflect a true increase contractility, because of the concomitant fall in wall stress.


Global and regional lung function assessment using portable electrical impedance tomography (EIT) system: clinical study.
Recent development of affordable, portable and self-administrable electrical impedance tomography (EIT) system demonstrated the feasibility of using standalone EIT and subject's anthropometrics to predict the gold standard spirometry indicators for lung-function assessment. Compared to spirometry, the system showed the advantage of providing spatial mapping of the spirometry indicators. Nevertheless, the previous study was limited to healthy subjects. Here, we recruited (N=88): 47 lung disease patients and 41 healthy controls to perform simultaneous EIT and spirometry measurements to validate the capabilities of the system. Lung disease patients include 13 interstitial lung disease (ILD), 10 asthma, 8 chronic obstructive pulmonary disease (COPD), 8 bronchiectasis, and 8 with other diseases including left pneumonectomy, lung cancer, lung tumor, lymphangioleiomyomatosis, motor neuron disease, heart failure and bronchiolitis obliterans syndrome. The results showed significant correlation of the predicted global spirometry indicators (p<0.0001) and significant distinguishability between most disease groups and healthy subjects demonstrating the capability of the EIT system in diagnostic screening. Furthermore, the regional mapping of the spirometry indicators is evaluated and shown to be distinct for each disease group, providing an additional dimension for medical professionals to diagnose and monitor lung disease patients.Clinical Relevance- This establishes the significance of EIT-based global and regional indicators for assessing lung function on lung disease patients.


Mouse polyclonal and monoclonal antibody to scrapie-associated fibril proteins.
Antibody response in mice to scrapie-associated fibril proteins (protease-resistant proteins [PrPs]) was generated to different epitopes depending on the source of antigen. Mice responded differently to PrPs isolated from scrapie-infected animals of homologous (mouse) versus heterologous (hamster) species. An enzyme-linked immunosorbent assay established to monitor this antibody response in mice immunized with PrPs was unable to detect such a response in scrapie-infected mice. A monoclonal antibody (MAb), 263K 3F4, derived from a mouse immunized with hamster 263K PrPs reacted with hamster but not mouse PrPs. MAb 263K 3F4 also recognized normal host protein of 33 to 35 kilodaltons in brain tissue from hamsters and humans but not from bovine, mouse, rat, sheep, or rabbit brains. This is the first demonstration of epitope differences on this host protein in different species. The defining of various epitopes on PrP through the use of MAbs will lead to a better understanding of the relationship of PrPs to their host precursor protein and to the infectious scrapie agent.


[The ways of harmonization of clinical laboratory measurement techniques].
The results of implementation of different clinical laboratory techniques are to be equal in clinically significant limits to be optimally applied in diagnostics of diseases and treatment of patients. When the results of laboratory tests are not standardized and harmonized for the very same clinical assay the results can be expressed by unmatched numbers. Unfortunately, in some handbooks the values are presented based on the results of application of specific laboratory techniques without considering possibility or likelihood of differences between various techniques. When this is a case, accumulation of data of diferent clinical research studies and working out of clinical handbooks on this basis will be inconsistent. Inadequate understanding of issue that the results of laboratory tests are not standardized and harmonized can lead to incorrect clinical, financial, managerial or technical decisions. The standardization of clinical laboratory techniques was applied to many measurands related to primary referent techniques (standard specimen of pure substance) or/and developed referent measurement techniques. However, harmonization of clinical laboratory techniques for those measurands which are not related any developed measurement techniques is quite problematic due to inadequate determination of measurand, its inadequate analytical specificity, insufficient attention to commutability of referent materials and poor systematic approach to harmonization. To overcome these issues an infrastructure is to be developed to support systematic approach to identification and prioritization of measurands which are to be harmonized on the basis of clinical importance and technical applicability. The management of technical implementation harmonization process for specific measurands.


Computational biophysical characterization of the SARS-CoV-2 spike protein binding with the ACE2 receptor and implications for infectivity.
SARS-CoV-2 is a novel highly virulent pathogen which gains entry to human cells by binding with the cell surface receptor - angiotensin converting enzyme (ACE2). We computationally contrasted the binding interactions between human ACE2 and coronavirus spike protein receptor binding domain (RBD) of the 2002 epidemic-causing SARS-CoV-1, SARS-CoV-2, and bat coronavirus RaTG13 using the Rosetta energy function. We find that the RBD of the spike protein of SARS-CoV-2 is highly optimized to achieve very strong binding with human ACE2 (hACE2) which is consistent with its enhanced infectivity. SARS-CoV-2 forms the most stable complex with hACE2 compared to SARS-CoV-1 (23% less stable) or RaTG13 (11% less stable). Notably, we calculate that the SARS-CoV-2 RBD lowers the binding strength of angiotensin 2 receptor type I (ATR1) which is the native binding partner of ACE2 by 44.2%. Strong binding is mediated through strong electrostatic attachments with every fourth residue on the N-terminus alpha-helix (starting from Ser19 to Asn53) as the turn of the helix makes these residues solvent accessible. By contrasting the spike protein SARS-CoV-2 Rosetta binding energy with ACE2 of different livestock and pet species we find strongest binding with bat ACE2 followed by human, feline, equine, canine and finally chicken. This is consistent with the hypothesis that bats are the viral origin and reservoir species. These results offer a computational explanation for the increased infection susceptibility by SARS-CoV-2 and allude to therapeutic modalities by identifying and rank-ordering the ACE2 residues involved in binding with the virus.


Using biological traits to assess diet selection: the case of the Pyrenean Desman.
Traditionally, researchers have assessed diet selection by comparing consumed versus available taxa. However, taxonomic assignment is probably irrelevant for predators, who likely base their selection on characteristics including prey size, habitat, or behavior. Here, we use an aquatic insectivore, the threatened Pyrenean Desman (Galemys pyrenaicus), as a model species to assess whether biological traits help unravel the criteria driving food and habitat preferences. We reanalyzed data from a previous taxonomy-based study of prey selection in two contrasting streams, one with excellent conservation status and the other affected by diversion for hydropower and forestry. Available and consumed prey were characterized according to nine biological traits, and diet selection was estimated by comparing availability-measured from Surber net samples, and consumption-analyzed by metabarcoding desman feces. Traits offered a biologically coherent image of diet and almost identical selection patterns in both streams, depicting a highly specialized rheophilic predator. Desmans positively selected prey with a preference for fast flow and boulder substrate, indicating their preferred riffle habitat. On the other hand, they positively selected prey with larger but not the largest potential size, living in the water column or the litter, and not inside sediments. They also chose agile prey, swimmers or prey attached to the substrate, prey with high body flexibility, and prey living exposed and clustered in groups. Overall, our results offer a picture of desman diet preference and point to biological traits as being better than taxonomic identity to describe the diet preference of consumers.


Decreased myeloperoxidase expressing cells in the aged rat brain after excitotoxic damage.
Brain aging is associated to several morphological and functional alterations that influence the evolution and outcome of CNS damage. Acute brain injury such as an excitotoxic insult induces initial tissue damage followed by associated inflammation and oxidative stress, partly attributed to neutrophil recruitment and the expression of oxidative enzymes such as myeloperoxidase (MPO), among others. However, to date, very few studies have focused on how age can influence neutrophil infiltration after acute brain damage. Therefore, to evaluate the age-dependent pattern of neutrophil cell infiltration following an excitotoxic injury, intrastriatal injection of N-methyl-d-aspartate was performed in young and aged male Wistar rats. Animals were sacrificed at different times between 12h post-lesion (hpl) to 14 days post-lesion (dpl). Cryostat sections were processed for myeloperoxidase (MPO) immunohistochemistry, and double labeling for either neuronal cells (NeuN), astrocytes (GFAP), perivascular macrophages (ED-2), or microglia/macrophages (tomato lectin histochemistry). Our observations showed that MPO + cells were observed in the injured striatum from 12 hpl (when maximum values were found) until 7 dpl, when cell density was strongly diminished. However, at all survival times analyzed, the overall density of MPO + cells was lower in the aged versus the adult injured striatum. MPO + cells were mainly identified as neutrophils (especially at 12 hpl and 1 dpl), but it should be noted that MPO + neurons and microglia/macrophages were also found. MPO + neurons were most commonly observed at 12 hpl and reduced in the aged. MPO + microglia/macrophages were the main population expressing MPO from 3 dpl, when density was also reduced in aged subjects. These results point to neutrophil infiltration as another important factor contributing to the different responses of the adult and aged brain to damage, highlighting the need of using aged animals for the study of acute age-related brain insults.


Plakophilin-2 Haploinsufficiency Causes Calcium Handling Deficits and Modulates the Cardiac Response Towards Stress.
Human variants in plakophilin-2 (PKP2) associate with most cases of familial arrhythmogenic cardiomyopathy (ACM). Recent studies show that PKP2 not only maintains intercellular coupling, but also regulates transcription of genes involved in Ca2+ cycling and cardiac rhythm. ACM penetrance is low and it remains uncertain, which genetic and environmental modifiers are crucial for developing the cardiomyopathy. In this study, heterozygous PKP2 knock-out mice (PKP2-Hz) were used to investigate the influence of exercise, pressure overload, and inflammation on a PKP2-related disease progression. In PKP2-Hz mice, protein levels of Ca2+-handling proteins were reduced compared to wildtype (WT). PKP2-Hz hearts exposed to voluntary exercise training showed right ventricular lateral connexin43 expression, right ventricular conduction slowing, and a higher susceptibility towards arrhythmias. Pressure overload increased levels of fibrosis in PKP2-Hz hearts, without affecting the susceptibility towards arrhythmias. Experimental autoimmune myocarditis caused more severe subepicardial fibrosis, cell death, and inflammatory infiltrates in PKP2-Hz hearts than in WT. To conclude, PKP2 haploinsufficiency in the murine heart modulates the cardiac response to environmental modifiers via different mechanisms. Exercise upon PKP2 deficiency induces a pro-arrhythmic cardiac remodeling, likely based on impaired Ca2+ cycling and electrical conduction, versus structural remodeling. Pathophysiological stimuli mainly exaggerate the fibrotic and inflammatory response.


Bone marrow involvement by non-Hodgkin's lymphoma: the clinical significance of morphologic discordance between the lymph node and bone marrow. Nebraska Lymphoma Study Group.
Bone marrow specimens from 317 patients with non-Hodgkin's lymphoma (NHL) obtained at initial staging were evaluated for the presence of lymphoma or benign lymphoid aggregates. Thirty-two percent (102 patients) had lymphoma in their bone marrow, and 9% had benign lymphoid aggregates. Bone marrow lymphoma was present in 39% of low-grade, 36% of intermediate-grade, and 18% of high-grade lymphomas. The bone marrow was involved in 25% of patients with diffuse large-cell or immunoblastic NHL (ie, diffuse histiocytic lymphoma of Rappaport). Bone marrow involvement did not affect survival of patients with low-grade NHL, but survival was significantly shorter (P = .03) for patients with intermediate- and high-grade NHL with bone marrow involvement. Bone marrow involvement was equally common in B-cell and T-cell NHL (31% v 32%). However, patients with T-cell NHL and bone marrow involvement had shorter survival than B-cell NHL with marrow involvement (P = .02) or T-cell NHL without marrow involvement (P = .05). A high incidence of morphologic discordance between lymph node and bone marrow was observed (ie, 40%), always with a more aggressive subtype in the lymph node than in the bone marrow. Presence of large-cell lymphoma in the bone marrow predicted for short survival. Survival for patients with small-cell lymphoma in their bone marrow did not differ significantly from patients with negative bone marrows. We conclude that bone marrow involvement in large-cell NHL, especially in those of T-cell origin, portends a poor prognosis. However, the subgroup of patients with an aggressive histologic subtype of NHL in a lymph node biopsy and small-cell NHL in the bone marrow do not have a poorer outlook than those without bone marrow involvement.


Sensorineural hearing loss in Kawasaki disease.
Kawasaki disease is a common nonspecific vasculitis seen in childhood. The most significant long-term sequela is coronary artery aneurysm. However, the spectrum of complications involves not only the heart, but also other organs such as the eyes, skin, kidneys, gallbladder, liver, and central nervous system. Sensorineural hearing loss (SNHL) is a relatively unrecognized complication of the disease. Although most of the complications (except coronary artery aneurysm) are self-limiting, SNHL can be persistent. It is, especially important in infants and young children who might not be able to report the hearing deficits and are most likely to have cognitive and speech delays if this hearing loss is not addressed in a timely manner. We report a child with Kawasaki disease who had SNHL during the 2(nd) week of the illness. The aim of this article is to briefly review the pathophysiology behind this hearing loss and strongly emphasize the importance of universal hearing evaluation in all children diagnosed with Kawasaki disease. This screening in children with Kawasaki disease may provide some timely intervention if needed. Since most Kawasaki disease patients will be seen by cardiologists, we hope to create more awareness about this complication to the cardiology community as well.


Colonoscopic lesions in veterans with spinal cord injury.
The overall goal of this observational study was to determine the type and prevalence of colonoscopic lesions encountered in veterans with traumatic spinal cord injury (SCI) and to examine their relationship to lesion level, completeness, and duration postinjury. We retrospectively reviewed the electronic charts of veterans with SCI who are regularly followed in our SCI clinic. Colonoscopy in veterans with SCI was undertaken for their gastrointestinal (GI) complaints. Of the 87 veterans with SCI, 71 who were 50 years of age or older were included in this study. Of these 71, 28 underwent colonoscopies (39.4%). Demographic variables were matched between patients who underwent colonoscopies and those who did not for age, sex, race/ethnicity, age of onset of SCI, and duration of SCI (p > 0.05). The colonoscopic lesions seen in 26 (93%) of the 28 veterans with SCI who underwent colonoscopies included diverticulae, internal hemorrhoids, and polyps. No relationship was found between colonoscopic lesion type and SCI lesion location or severity. A relationship was found between total colonoscopic lesions and duration of SCI (p < 0.001). Age-, sex-, and race/ethnicity-matched veterans without SCI (controls) who underwent colonoscopy for their GI complaints had more colonoscopic lesions than the veterans with SCI who underwent colonoscopies (p < 0.001).


Discrimination learning.
Eight rats were run through discrimination training sessions in which responses in the dark were not reinforced whereas the first response after the onset of a light was reinforced. The procedure generated orderly learning and latency data for the individual animal. The latency distributions are adequately described by a simple mathematical formulation.


[Characteristics of lipid composition of erythroid and myeloid cells of the bone marrow of rabbits].
Lipid composition of erythroid and myeloid cells of rabbit bone marrow was studied by thin layer chromatography. The myelocaryocytes contained phospholipids, free and esterified cholesterol, free fatty acids and triglycerides. The main components of the total lipid fraction were phospholipids and triglycerides. The ratio of phospholipids was lower and content of triglycerides and cholesterol esters was higher in myeloid cells, as compared with erythroid cells. Also myeloid components of bone marrow contained 1.8-fold more lipids per one cell than the erythroid cells.


Addressing innovative off-label medication use at an academic medical center.
A large hospital's systematic and evidence-based approach to adjudicating, monitoring, and ensuring the safety of off-label medication use is described. In 2003 the University of Pittsburgh Medical Center (UPMC)-Presbyterian implemented a policy that created a formal process for the systematic evaluation of formulary requests and drug-utilization patterns indicating or suggesting off-label use. Explicit criteria were developed for differentiating "innovative off-label use" (i.e., use based on a reasonable rationale yet lacking definitive scientific support in the form of fully published randomized controlled trials) from medication use more appropriately classified as clinical research. The UPMC-Presbyterian policy also outlined a process for the development, implementation, and evaluation of guidelines on innovative off-label use, including the collection of efficacy and safety outcomes. As of October 2012, 31 proposals for off-label medication use had been evaluated by the medical center's pharmacy and therapeutics committee and formulary subcommittee. Thirteen requests resulted in a determination of innovative off-label use and the development of prescribing guidelines, and 10 prompted the extension of an agent's current formulary status; in 6 cases, proposed off-label uses were determined to constitute clinical research. In some instances, innovative off-label medication use generated safety and outcomes data that led to changes in local standards of care. An algorithm to guide decision-making with regard to requests and proposals for off-label medication use is provided. The UPMC-Presbyterian experience indicates that off-label medication use can be effectively managed using evidence-based principles and peer review mechanisms.


Impact of socioeconomic status on disease phenotype, genomic landscape and outcomes in myelodysplastic syndromes.
Genetic and epigenetic alterations contribute to the biological and clinical characteristics of myelodysplastic syndromes (MDS), but a role for socioeconomic environment remains unclear. Here, socioeconomic status (SES) for 283 MDS patients was estimated using the Scottish Index of Multiple Deprivation tool. Indices were assigned to quintile categorical indicators ranked from SES1 (lowest) to SES5 (highest). Clinicopathological features and outcomes between SES quintiles containing 15%, 20%, 19%, 30% and 16% of patients were compared. Prognostic scores identified lower-risk MDS in 82% of patients, with higher-risk disease in 18%. SES quintiles did not associate with age, gender, cytogenetics, International Prognostic scores or, in sub-analysis (n = 95), driver mutations. The odds ratio of a diagnosis of refractory anaemia was greater than other MDS sub-types in SES5 (OR 1·9, P = 0·024). Most patients (91%) exclusively received supportive care. SES did not associate with leukaemic transformation or cause of death. Cox regression models confirmed male gender (P < 0·05), disease-risk (P < 0·0001) and age (P < 0·01) as independent predictors of leukaemia-free survival, with leukaemic transformation an additional determinant of overall survival (P = 0·07). Thus, if access to healthcare is equitable, SES does not determine disease biology or survival in MDS patients receiving supportive treatment; additional studies are required to determine whether outcomes following disease-modifying therapies are influenced by SES.


Provenance does matter: links between winter trophic segregation and the migratory origins of European robins.
Amongst migratory species, it is common to find individuals from different populations or geographical origins sharing staging or wintering areas. Given their differing life histories, ecological theory would predict that the different groups of individuals should exhibit some level of niche segregation. This has rarely been investigated because of the difficulty in assigning migrating individuals to breeding areas. Here, we start by documenting a broad geographical gradient of hydrogen isotopes (δ 2H) in robin Erithacus rubecula feathers across Europe. We then use δ 2H, as well as wing-tip shape, as surrogates for broad migratory origin of birds wintering in Iberia, to investigate the ecological segregation of populations. Wintering robins of different sexes, ages and body sizes are known to segregate between habitats in Iberia. This has been attributed to the despotic exclusion of inferior competitors from the best patches by dominant individuals. We find no segregation between habitats in relation to δ 2H in feathers, or to wing-tip shape, which suggests that no major asymmetries in competitive ability exist between migrant robins of different origins. Trophic level (inferred from nitrogen isotopes in blood) correlated both with δ 2H in feathers and with wing-tip shape, showing that individuals from different geographic origins display a degree of ecological segregation in shared winter quarters. Isotopic mixing models indicate that wintering birds originating from more northerly populations consume more invertebrates. Our multi-scale study suggests that trophic-niche segregation may result from specializations (arising in the population-specific breeding areas) that are transported by the migrants into the shared wintering grounds.


GABA(A) receptor downregulation in brains of subjects with autism.
Gamma-aminobutyric acid A (GABA(A)) receptors are ligand-gated ion channels responsible for mediation of fast inhibitory action of GABA in the brain. Preliminary reports have demonstrated altered expression of GABA receptors in the brains of subjects with autism suggesting GABA/glutamate system dysregulation. We investigated the expression of four GABA(A) receptor subunits and observed significant reductions in GABRA1, GABRA2, GABRA3, and GABRB3 in parietal cortex (Brodmann's Area 40 (BA40)), while GABRA1 and GABRB3 were significantly altered in cerebellum, and GABRA1 was significantly altered in superior frontal cortex (BA9). The presence of seizure disorder did not have a significant impact on GABA(A) receptor subunit expression in the three brain areas. Our results demonstrate that GABA(A) receptors are reduced in three brain regions that have previously been implicated in the pathogenesis of autism, suggesting widespread GABAergic dysfunction in the brains of subjects with autism.


Diverse effects of P-glycoprotein inhibitory agents on human leukemia cells expressing the multidrug resistance protein (MRP).
Multidrug resistance (MDR) to cancer chemotherapy is frequently associated with decreased drug accumulation in cancer cells due to drug expulsion by multidrug transporters such as P-glycoprotein (Pgp) and multidrug resistance protein (MRP). The novel resistance modifying agents PSC 833, 280-446, and LY 335979 are primarily targeted at inhibition of Pgp, and their MRP inhibitory potential is largely unknown. In the present study we addressed the effect of these agents on MRP-derived drug resistance. Drug-resistant human leukemia cells with Pgp+/MRP- (KG1a/200, K562/150) and Pgp-/MRP+ (HL60/130) phenotypes were maintained in suspension cultures for experimental studies of drug accumulation and drug sensitization by Pgp inhibitors. Intracellular accumulation of the fluorescent anthracycline daunorubicin was measured by flow cytometry and fluorescence detection. Daunorubicin dose-response curves were generated by non-linear regression of electronically measured cell counts of 72- - 96-h cultures. The half-maximal growth inhibitory dose (GI50) was used as measure of growth inhibition. All MDR phenotypes studied exercised significant resistance to daunorubicin. PSC 833, 280-446 and LY335979 were equal in sensitizing Pgp+/MRP- cells to daunorubicin-induced growth inhibition (p < 0.0001). The Pgp-/MRP+ cells responded to PSC 833 and 280-446 by increased accumulation of daunorubicin (p = 0.0022 and p = 0.0005, respectively) and sensitization to the drug (p = 0.0009 and p = 0.0007, respectively). Conversely, LY335979 did not affect accumulation of daunorubicin in Pgp-/MRP+ cells nor sensitize these cells to daunorubicin. Pgp inhibitory agents have differential effects on MRP-derived drug resistance which could be exploited in treatment of multidrug resistance in cancer patients.


[Influence of prodigiozan-dependent comuton on the resistance of liver mitochondria against damage by protonofor].
An effector of tissue stress of hepatocytes, prodigiozan-dependent comuton (PDC), provokes deenergiezation of liver mitochondria, preloaded by Ca2+ ions. In this case a decrease of membrane potential (MP) and Ca2+ efflux by cyclosporine A sensitive mechanism of megapore is observed. If megapore is blocked by cyclosporin A, protonofor FCCP provoked decrease of MP and Ca2+ efflux by cyclosporin A-insensitive mechanism. It is shown that PDC increases resistance of mitochondria to mentioned protonofor action by inhibition of both these effects. An inhibitory action of PDC is realized by K+ and NADH-dependent mechanism. The effector of hepatocyte tissue stress, prodigiozan-dependent comuton (PDC), evokes deenergizing liver mitochondria preloaded with Ca2+, both membrane potential (MP) decrease and Ca2+ release in according to cyclosporine A-sensitive mechanism of megapore being observed. If megapore is blocked by cyclosporin A, protonophore FCCP reduces of MP and Ca2+ release in according to cyclosporin A-insensitive mechanism. PDC is shown to increase the resistance of mitochondria against protonophore action mentioned above by means of inhibition of both these effects. Inhibitory action of PDC is realized due to both K+ and NADH-dependent mechanism. Protective effect takes place only in intact mitochondria of these cells providig (on condition that) its megapore mechanism is not activated. Moreover, the results obtained are evidence of PDC can function as protector due to intensification of energy generation in damaged.


Thermocapillary phenomena and performance limitations of a wickless heat pipe in microgravity.
A counterintuitive, thermocapillary-induced limit to heat- pipe performance was observed that is not predicted by current thermal-fluid models. Heat pipes operate under a number of physical constraints including the capillary, boiling, sonic, and entrainment limits that fundamentally affect their performance. Temperature gradients near the heated end may be high enough to generate significant Marangoni forces that oppose the return flow of liquid from the cold end. These forces are believed to exacerbate dry out conditions and force the capillary limit to be reached prematurely. Using a combination of image and thermal data from experiments conducted on the International Space Station with a transparent heat pipe, we show that in the presence of significant Marangoni forces, dry out is not the initial mechanism limiting performance, but that the physical cause is exactly the opposite behavior: flooding of the hot end with liquid. The observed effect is a consequence of the competition between capillary and Marangoni-induced forces. The temperature signature of flooding is virtually identical to dry out, making diagnosis difficult without direct visual observation of the vapor-liquid interface.


The effect of reproductive events and alterations of sex hormone levels on the symptoms of fibromyalgia.
The fibromyalgia syndrome (FS) is a chronic pain disorder frequently affecting women of fertile age. However, the relationship of FS and pregnancy has been given little attention. In the present retrospective analysis, based on personal interviews, the influence on FS symptomatology by pregnancy, abortion, menstruation, use of oral contraceptives, and breast feeding was investigated. Twenty-six women with an established diagnosis of FS and a total of 40 pregnancies during disease were included in the study. With the exception of one patient, all women described worsening fibromyalgia symptoms during pregnancy with the last trimester experienced as the worst period. A new change of fibromyalgia symptoms within 6 months after delivery was reported for 37 of the 40 pregnancies, to the better in four and to the worse in 33 cases, resulting in a prolonged sick leave for 14 patients. An increase in depression and anxiety was a prominent problem in the post partum period. FS had no adverse effect on the outcome of pregnancy or the health of the neonate. In the majority of patients with FS, hormonal changes connected with abortion, use of hormonal contraceptives, and breast feeding did not modulate symptom severity. A pre-menstrual worsening of symptoms was recorded by 72% of the patients. Comparing the 26 patients who had borne children during disease with 18 patients who had all their children before the onset of FS revealed a negative effect of pregnancy and the post partum period of FS and increased functional impairment and disability in the 26 patients.


[Expression of DNA methyltransferase in myeloma U266 cells and its significance].
In order to study the activity and gene expression of DNA methyltransferase (DNMT) on U266 myeloma cells and to analyze their significance, the activity of DNMT was detected by ELISA, and the expressions of DNMT1, DNMT3a and 3b were analyzed by RT-PCR. U266 cells were treated by phenylhexyl isothiocyanate (PHI), and the change of activity and gene expression of DNMT were determined. The results indicated that the activity and expression of DNMT in U266 myeloma cells were higher, compared with normal control. After being treated by different concentration of PHI, U266 cells were driven into apoptosis and the activity of DNMT decreased obviously and the mRNA level of DNMT declined. It is concluded that the activity and gene expression of DNMT on U266 myeloma cells are higher, and DNMT may be a new therapeutic target of multiple myeloma.


Gender Parity Remains To Be Achieved for the Range of Editorial Roles Associated with Current Australasian Medical Journals.
With gender parity of medical school graduates having been achieved for well over two decades, it is timely to assess whether this has translated into gender parity for all of the editorial type roles of Australasian medical journals, reflecting a move toward gender equity. Data analysis was undertaken of the gender ratios of the current editorial roles of Australasian medical journals as compared to available Australian Health Workforce data. This reveals some variation in the gender ratios for all of the current range of editorial type positions and, hence, an absence of parity. There are no women holding formal editorial positions at all for 27.7% of these journals, whilst 77.7% of the chief editors' roles are occupied by men. For five out of 18 (27.7%) of the journals, gender parity has been or is close to having been achieved for these particular roles. These gender ratios do not mirror the gender ratios of the wider community of practice for at least 50% of the journals. Hence, it can be seen that gender parity is yet to be achieved for the range of editorial roles of Australasian medical journals, which carries implications for gender equity initiatives.


The effect of hypophysectomy and subsequent replacement therapy with sheep prolactin or bovine growth hormone on the lactose synthetase activity of rabbit mammary gland.
1. The effects of hypophysectomy and replacement therapy with sheep prolactin and bovine growth hormone on the lactose synthetase activity of the mammary glands of lactating rabbits were studied. 2. There was an approximately fourfold decline in the lactose synthetase activity of homogenates calculated on a DNA basis within 6-7 days of hypophysectomy. Prolactin reversed this decline but growth hormone had no effect. 3. Changes in the properties of a particulate fraction isolated from the glands indicated that a decline in the effective concentration of alpha-lactalbumin was one factor contributing to the decreased lactose synthetase activity after hypophysectomy. 4. As the changes in lactose output produced by hypophysectomy and prolactin therapy are much greater than the changes in total lactose synthetase activity it is concluded that the activity of this enzyme is not the main factor controlling lactose output under these conditions.


Ubiquitination by SAG regulates macrophage survival/death and immune response during infection.
The checkpoint between the life and death of macrophages is crucial for the host's frontline immune defense during acute phase infection. However, the mechanism as to how the immune cell equilibrates between apoptosis and immune response is unclear. Using in vitro and ex vivo approaches, we showed that macrophage survival is synchronized by SAG (sensitive to apoptosis gene), which is a key member of the ubiquitin-proteasome system (UPS). When challenged by pathogen-associated molecular patterns (PAMPs), we observed a reciprocal expression profile of pro- and antiapoptotic factors in macrophages. However, SAG knockdown disrupted this balance. Further analysis revealed that ubiquitination of Bax and SARM (sterile α- and HEAT/armadillo-motif-containing protein) by SAG-UPS confers survival advantage to infected macrophages. SAG knockdown caused the accumulation of proapoptotic Bax and SARM, imbalance of Bcl-2/Bax in the mitochondria, induction of cytosolic cytochrome c and activation of caspase-9 and -3, all of which led to disequilibrium between life and death of macrophages. In contrast, SAG-overexpressing macrophages challenged with PAMPs exhibited upregulation of protumorigenic cytokines (IL-1β, IL-6 and TNF-α), and downregulation of antitumorigenic cytokine (IL-12p40) and anti-inflammatory cytokine (IL-10). This suggests that SAG-dependent UPS is a key switch between immune defense and apoptosis or immune overactivation and tumorigenesis. Altogether, our results indicate that SAG-UPS facilitates a timely and appropriate level of immune response, prompting future development of potential immunomodulators of SAG-UPS.


Differential hemodynamic effects of beta-adrenoceptor blockers, Ca antagonists and combined alpha-beta-receptor blockade in ischemic heart disease.
The hemodynamic responses to 3 different therapeutical regimens: beta-adrenoceptor blockade, calcium inflow inhibition and combined alpha-beta-blockade were evaluated in 3 matched randomized groups of patients with ischemic heart disease and typical exercise-induced angina. The groups consisted of 22, 16 and 15 men, mean age 55-59 years. They were studied at rest and during ischemia-inducing exercise, before and after single oral doses of 100 mg metoprolol, 10 mg nifedipine and 200 mg labetalol. Pressures in the brachial artery and the pulmonary circulation were recorded by means of percutaneously introduced catheters. Cardiac output was determined according to the Fick principle. Metoprolol reduced mean arterial pressures, heart rate and cardiac output. Systemic vascular resistance and left ventricular filling pressure increased. Nifedipine resulted under all conditions in a distinct reduction of systemic vascular resistance and arterial pressures and a slight increase in heart rate and cardiac output. Left ventricular filling pressure was significantly lowered, the more the higher the initial level. The effect of labetalol was similar to that of nifedipine; however, cardiac output was unchanged and heart rate was slightly reduced. Left ventricular filling pressure was significantly lower. It is apparent that suppression of adrenergic stimulation by beta-receptor blockade alone may have adverse hemodynamic effects in ischemic heart disease and prompt further functional deterioration. Conversely, both calcium and combined alpha-beta-receptor blockade tend to improve left ventricular function by lowering both left ventricular preload and total systemic vascular resistance. The results strongly suggest that in patients in whom beta-receptor blockers appear indicated, their adverse hemodynamic effects can be offset by concomitant alpha1-receptor blockade or vasodilation without losing symptomatic efficacy. Combined alpha-beta-receptor blockade has the advantage over calcium antagonists alone to prevent any increase in adrenergic activity and related hyperkinetic response.


PKC activation increases Ca²⁺ sensitivity of permeabilized lymphatic muscle via myosin light chain 20 phosphorylation-dependent and -independent mechanisms.
The contractile activity of muscle cells lining the walls of collecting lymphatics is responsible for generating and regulating flow within the lymphatic system. Activation of PKC signaling contributes to the regulation of smooth muscle contraction by enhancing sensitivity of the contractile apparatus to Ca(2+). It is currently unknown whether PKC signaling contributes to the regulation of lymphatic muscle contraction. We hypothesized that the activation of PKC signaling would increase the sensitivity of the lymphatic myofilament to Ca(2+). To test this hypothesis, we determined the effects of PKC activation with phorbol esters [PMA or phorbol dibutyrate (PDBu)] on the contractile behavior of α-toxin-permeabilized rat mesenteric and cervical lymphatics or the thoracic duct. The addition of PMA or PDBu induced a significant increase in the contractile force of submaximally activated α-toxin-permeabilized lymphatic muscle independent of a change in intracellular Ca(2+) concentration, and the Ca(2+)-force relationship of lymphatic muscle was significantly left shifted, indicating greater myofilament Ca(2+) sensitivity. Phorbol esters increased the maximal rate of force development, whereas the rate of relaxation was reduced. Western blot and immunohistochemistry data indicated that the initial rapid increase in tension development after stimulation by PDBu was associated with myosin light chain (MLC)20 phosphorylation; however, the later, steady-state Ca(2+) sensitization of permeabilized lymphatic muscle was not associated with increased phosphorylation of MLC20 at Ser(19), 17-kDa C-kinase-potentiated protein phosphatase-1 inhibitor at Thr(38), or caldesmon at Ser(789). Thus, these data indicate that PKC-dependent Ca(2+) sensitization of lymphatic muscle may involve MLC20 phosphorylation-dependent and -independent mechanism(s).


Disability education and implications for genetic counselor training.
The genetic counseling (GC) community has faced criticism about the duality of promoting patient autonomy while also advocating for individuals with disabilities. This study assessed the attitudes of the disability community and GCs to identify content that should be included in GC disability education and evaluate the landscape of GC disability education. Members of the disability community and GCs completed an electronic survey distributed through electronic listservs and partnering organizations. A total of 672 responses were analyzed from both the disability community (n = 596) and the GC community (n = 76). Members of the disability community noted differences in GC comfort level discussing different aspects of disability with GCs being perceived as being very knowledgeable about medical aspects 71% of the time versus 49% of the time when discussing social/lifestyle aspects of disability. This discordance was reflected in GCs reported comfort level in discussing medical aspects (89%) and social aspects of disability (65%) during a session. Most GC respondents (71%) felt they received adequate knowledge during their disability education and variation was reported in the execution of disability education by training programs. Disability education content recommendations from the disability community and GCs included emphasizing four key aspects of disability: medical, social/lifestyle, lived experience, and the disability rights movement. Respondents of both cohorts stressed the inclusion of and exposure to persons with disabilities in disability education to understand the lived experience of persons with disabilities. The disability community identified additional disability education content to be included such as empathy training, family hardships, and mental health. The results of this study have practice implications and provide a foundation for training expectations to ensure future GCs possess the necessary skills to improve the quality of services provided to families and persons with disabilities.


[Need for improvement in education on appropriate prescribing in elderly patients].
Prescribing errors can cause great harm to patients. In the Netherlands, it is estimated that 7000 preventable medication-related hospitals admissions occur annually, caused in many cases by prescribing errors. Elderly patients are at greatest risk, since this patient demographic is most likely to be prescribed multiple medications. Robust education on appropriate prescribing is essential for all clinicians with the authority to prescribe. Currently, some issues still require improvement in the Netherlands: a) education continues to focus heavily on basic pharmacology knowledge instead of patient related pharmacotherapy skills, b) an appropriate assessment procedure on pharmacotherapy knowledge and skills is often lacking, c) there is no mandatory requirement for physicians to maintain their knowledge and skills in the field of pharmacotherapy during their working career. In this article we discuss means to improve this situation, with the overall aim to ensure that all vulnerable elderly patients are in safe hands with each physician.


Kinetic selection of HPV 16 E6/E7-directed antisense nucleic acids: anti-proliferative effects on HPV 16-transformed cells.
The E6/E7-coding sequences of the human papillomavirus type 16 (HPV 16) were probed for kinetic accessibility in vitro by pools of catalytic antisense RNA. Only long-chain complementary RNA and very few antisense sequences with a 3' portion complementary to a 10 nt window within unspliced and spliced E6-coding target sequences showed fast annealing with k(ass) values of up to 10(4) M-1s-1 indicating that the majority of E6/E7 RNA sequences are unfavourable targets for antisense inhibitors and ribozymes. Fast-annealing antisense oligodeoxyribonucleotides directed against the window of 10 nt inhibited cell proliferation of HPV 16-transformed SiHa cells but not slow-annealing antisense species. Antisense RNA of several hundred nucleotides in length also showed significant anti-proliferative activity. Biological effects of antisense oligodeoxyribonucleotides were specific for the antisense sequence, could only be found in HPV-positive but not in HPV-negative cell lines, and were related to decreased levels of E7 protein and E6/E7-specific transcripts. This work suggests that HPV 16 E7/E6 sequences exhibit a low accessibility for antisense oligonucleotides. This can be overcome, however, by exploiting the relationship between fast annealing of antisense species and their increased efficacy in human cells.


Adhesion force between cyclopentane hydrate and mineral surfaces.
Clathrate hydrate adhesion forces play a critical role in describing aggregation and deposition behavior in conventional energy production and transportation. This manuscript uses a unique micromechanical force apparatus to measure the adhesion force between cyclopentane hydrate and heterogeneous quartz and calcite substrates. The latter substrates represent models for coproduced sand and scale often present during conventional energy production and transportation. Micromechanical adhesion force data indicate that clathrate hydrate adhesive forces are 5-10× larger for calcite and quartz minerals than stainless steel. Adhesive forces further increased by 3-15× when increasing surface contact time from 10 to 30 s. In some cases, liquid water from within the hydrate shell contacted the mineral surface and rapidly converted to clathrate hydrate. Further measurements on mineral surfaces with physical control of surface roughness showed a nonlinear dependence of water wetting angle on surface roughness. Existing adhesive force theory correctly predicted the dependence of clathrate hydrate adhesive force on calcite wettability, but did not accurately capture the dependence on quartz wettability. This comparison suggests that the substrate surface may not be inert, and may contribute positively to the strength of the capillary bridge formed between hydrate particles and solid surfaces.


Atrial natriuretic factor and body water distribution.
In the rat, the effects of an atrial natriuretic factor (ANF) (Rat, 8-33 Peninsula Lab) on body water distribution have been evaluated. The ANF administration to nephrectomized animals produced a decrease in plasma volume and a slight increase in haematocrit and in plasma albumin concentration. No modifications were observed in total and intracellular water. The fluid efflux from the capillaries appeared to be located in the interstitial space. These results suggest that ANF could regulate plasma volume and systemic blood pressure, concurrently with its other known effects.


A steroid receptor-microRNA switch regulates life span in response to signals from the gonad.
Although the gonad primarily functions in procreation, it also affects animal life span. Here, we show that removal of the Caenorhabditis elegans germ line triggers a switch in the regulatory state of the organism to promote longevity, co-opting components involved in larval developmental timing circuits. These components include the DAF-12 steroid receptor, which is involved in the larval stage two-to-stage three (L2-L3) transition and up-regulates members of the let-7 microRNA (miRNA) family. The miRNAs target an early larval nuclear factor lin-14 and akt-1/kinase, thereby stimulating DAF-16/FOXO signaling to extend life. Our studies suggest that metazoan life span is coupled to the gonad through elements of a developmental timer.


Use of problem-oriented medical record in psychiatry: a survey of university-based residency training programs.
A survey of university-based psychiatric residency training programs revealed that nearly one-half of them use the problem-oriented medical record (POMR) and that conversion to this system appears to be growing rapidly. Experiences with the method have in general been positive. The author identifies factors associated with successful operation of the POMR and suggests that an integrated, hospital-wide adoption of the system may be crucial.


The transcriptomic basis of oviposition behaviour in the parasitoid wasp Nasonia vitripennis.
Linking behavioural phenotypes to their underlying genotypes is crucial for uncovering the mechanisms that underpin behaviour and for understanding the origins and maintenance of genetic variation in behaviour. Recently, interest has begun to focus on the transcriptome as a route for identifying genes and gene pathways associated with behaviour. For many behavioural traits studied at the phenotypic level, we have little or no idea of where to start searching for "candidate" genes: the transcriptome provides such a starting point. Here we consider transcriptomic changes associated with oviposition in the parasitoid wasp Nasonia vitripennis. Oviposition is a key behaviour for parasitoids, as females are faced with a variety of decisions that will impact offspring fitness. These include choosing between hosts of differing quality, as well as making decisions regarding clutch size and offspring sex ratio. We compared the whole-body transcriptomes of resting or ovipositing female Nasonia using a "DeepSAGE" gene expression approach on the Illumina sequencing platform. We identified 332 tags that were significantly differentially expressed between the two treatments, with 77% of the changes associated with greater expression in resting females. Oviposition therefore appears to focus gene expression away from a number of physiological processes, with gene ontologies suggesting that aspects of metabolism may be down-regulated during egg-laying. Nine of the most abundant differentially expressed tags showed greater expression in ovipositing females though, including the genes purity-of-essence (associated with behavioural phenotypes in Drosophila) and glucose dehydrogenase (GLD). The GLD protein has been implicated in sperm storage and release in Drosophila and so provides a possible candidate for the control of sex allocation by female Nasonia during oviposition. Oviposition in Nasonia therefore clearly modifies the transcriptome, providing a starting point for the genetic dissection of oviposition.


The Impact of Drug Interactions in Patients with Community-Acquired Pneumonia on Hospital Length of Stay.
(1) Background: An aging society is frequently affected by multimorbidity and polypharmacy, which, in turn, leads to an increased risk for drug interaction. The aim of this study was to evaluate the influence of drug interactions on the length of stay (LOS) in hospitals. (2) Methods: This retrospective, single-centre study is based on patients treated for community-acquired pneumonia in the hospital. Negative binomial regression was used to analyse the association between drug interactions and the LOS in the hospital. (3) Results: The total cohort contained 503 patients, yet 46 inpatients (9%) that died were not included in the analyses. The mean age was 74 (±15.3) years, 35% of patients older than 65 years were found to have more than two drug interactions, and 55% had a moderate, severe, or contraindicated adverse drug reaction. The regression model revealed a significant association between the number of drug interactions (rate ratio (RR) 1.02; 95%-CI 1.01-1.04) and the severity of drug interactions (RR 1.22; 95%-CI 1.09-1.37) on the LOS for the overall cohort as well as for the subgroup of patients aged 80 years and older. (4) Conclusion: Drug interactions are an independent risk factor for prolonged hospitalisation. Standardised assessment tools to avoid drug interactions should be implemented in clinical routines.


Protective effects of propolis on cryopreservation of common carp (Cyprinus carpio) sperm.
Cryopreservation of sperm is common procedures in aquaculture, particularly used for routine in artificial insemination. However, these application cause damages and adversely affected sperm motility, viability and consequently lower hatching rates. The objective of this study is to determine whether propolis has an effect on cryopreservation and fertilization ability and to investigate the potential protective effect of propolis on spermatozoa of Cyprinus carpio. Many studies have been done in cryopreservation offish spermatozoa, but none of them contain propolis in extender composition. The extenders were prepared by using modified Kurokura Solution to which 10% Me(2)SO added with different levels of propolis (0.2, 0.4, 0.6, 0.8 and 1 mgml(-1)) and 10% egg yolk (as a control without propolis). The pooled semen samples diluted at the ratio of 1:9 by the extenders were subjected to cryopreservation. The percentage and duration of motility and fertilization tests of cryopreserved sperm samples have been done immediately after thawing and compared with control and fresh semen. The extenders containing propolis exhibited higher percentage motility and motility duration than control group (P < 0.05). Especially the group IV (0.8 mgml(-1) propolis) and the group V (1 mgml(-1) propolis) showed significant positive effects on both post thaw motility and hatching ability. The propolis maintained the integrity of the spermatozoa during the cryopreservation process. Evaluating with its contents, it has been shown that propolis is an appropriate cryoprotective agent in fish semen.


Nitrile-containing pharmaceuticals: target, mechanism of action, and their SAR studies.
The nitrile group is an important functional group widely found in both pharmaceutical agents and natural products. More than 30 nitrile-containing pharmaceuticals have been approved by the FDA for the management of a broad range of clinical conditions in the last few decades. Incorporation of a nitrile group into lead compounds has gradually become a promising strategy in rational drug design as it can bring additional benefits including enhanced binding affinity to the target, improved pharmacokinetic profile of parent drugs, and reduced drug resistance. This paper reviews the existing drugs with a nitrile moiety that have been approved or in clinical trials, involving their targets, molecular mechanism of pharmacology and SAR studies, and classifies them into different categories based on their clinical usages.


Comparative analysis of functional diversity of rumen microbiome in bison and beef heifers.
Ruminants play a key role in the conversion of cellulolytic plant material into high-quality meat and milk protein for humans. The rumen microbiome is the driver of this conversion, yet there is little information on how gene expression within the microbiome impacts the efficiency of this conversion process. The current study investigates gene expression in the rumen microbiome of beef heifers and bison and how transplantation of ruminal contents from bison to heifers alters gene expression. Understanding interactions between the host and the rumen microbiome is the key to developing informed approaches to rumen programming that will enhance production efficiency in ruminants.