Temporal coordination between initiation of HIV (+)-strand DNA synthesis and primer removal.
In this study, we have analyzed the interdependence between the polymerase and RNase H active sites of human immunodeficiency virus-1 reverse transcriptase (RT) using an in vitro system that closely mimics the initiation of (+)-strand DNA synthesis. Time course experiments show that RT pauses after addition of the 12th DNA residue, and at this stage the RNase H activity starts to cleave the RNA primer from newly synthesized DNA. Comparison of cleavage profiles obtained with 3'- and 5'-end-labeled primer strands indicates that RT now translocates in the opposite direction, i.e. in the 5' direction of the RNA strand. DNA synthesis resumes again in the 3' direction, after the RNA-DNA junction was efficiently cleaved. Moreover, we further characterized complexes generated before, during, and after position +12, by treating these with Fe2+ to localize the RNase H active site on the DNA template. Initially, when RT binds the RNA/DNA substrate, oxidative strand breaks were seen at a distance of 18 base pairs upstream from the primer terminus, whereas 17 base pairs were observed at later stages when the enzyme binds more and more DNA/DNA. These data show that the initiation of (+)-strand synthesis is accompanied by a conformational change of the polymerase-competent complex.


FGF2 specifies hESC-derived definitive endoderm into foregut/midgut cell lineages in a concentration-dependent manner.
Fibroblast growth factor (FGF) signaling controls axis formation during endoderm development. Studies in lower vertebrates have demonstrated that FGF2 primarily patterns the ventral foregut endoderm into liver and lung, whereas FGF4 exhibits broad anterior-posterior and left-right patterning activities. Furthermore, an inductive role of FGF2 during dorsal pancreas formation has been shown. However, whether FGF2 plays a similar role during human endoderm development remains unknown. Here, we show that FGF2 specifies hESC-derived definitive endoderm (DE) into different foregut lineages in a dosage-dependent manner. Specifically, increasing concentrations of FGF2 inhibits hepatocyte differentiation, whereas intermediate concentration of FGF2 promotes differentiation toward a pancreatic cell fate. At high FGF2 levels specification of midgut endoderm into small intestinal progenitors is increased at the expense of PDX1(+) pancreatic progenitors. High FGF2 concentrations also promote differentiation toward an anterior foregut pulmonary cell fate. Finally, by dissecting the FGF receptor intracellular pathway that regulates pancreas specification, we demonstrate for the first time to the best of our knowledge that induction of PDX1(+) pancreatic progenitors relies on FGF2-mediated activation of the MAPK signaling pathway. Altogether, these observations suggest a broader gut endodermal patterning activity of FGF2 that corresponds to what has previously been advocated for FGF4, implying a functional switch from FGF4 to FGF2 during evolution. Thus, our results provide new knowledge of how cell fate specification of human DE is controlled-facts that will be of great value for future regenerative cell therapies.


General and abdominal obesity and incident asthma in adults: the HUNT study.
Measures of body mass index (BMI) and waist circumference define general obesity and abdominal obesity respectively. While high BMI has been established as a risk factor for asthma in adults, waist circumference has seldom been investigated. To determine the association between BMI, waist circumference and incident asthma in adults, we conducted a prospective study (n=23,245) in a population living in Nord-Trøndelag, Norway in 1995-2008. Baseline BMI and waist circumference were measured and categorised as general obesity (BMI ≥30.0 kg·m(2)) and abdominal obesity (waist circumference ≥88 cm in females and ≥102 cm in males). Incident asthma was self-reported new-onset cases during an 11-yr follow-up period. Odds ratios for asthma associated with obesity were calculated using multivariable logistic regression. General obesity was a risk factor for asthma in females (OR 1.96, 95% CI 1.52-2.52) and males (OR 1.84, 95% CI 1.30-2.59). In females, after additional adjustment for BMI, abdominal obesity remained a risk factor for asthma development (OR 1.46, 95% CI 1.04-2.05). Abdominal obesity seems to increase the risk of incident asthma in females in addition to BMI, indicating that using both measures of BMI and waist circumference in females may be a superior clinical assessment for asthma risk than any measure alone.


Risk-taking behaviour of Cape Peninsula high-school students. Part III. Cigarette smoking.
The prevalence of a wide range of risk-taking behaviour among high-school students in the Cape Peninsula, South Africa, was investigated. In this article, the results for cigarette smoking are presented. Cluster sampling techniques produced a sample of 7,340 students from 16 schools in the three major education departments. A self-administered questionnaire was completed in a normal school period. Estimates for each education department were weighted to produce an overall estimate. Of the students 18.1% indicated that they smoked at least 1 cigarette per day. Of these, 66.9% had tried to stop. Of those who did not smoke at least 1 cigarette per day, 41.2% had smoked previously and 3.6% intended to start smoking. There were different trends according to gender, standard, and language(s) spoken at home. Of note was the small percentage of Xhosa-speaking females who smoked. There is an urgent need for smoking prevention programmes in schools.


Trophoblast-specific gene manipulation using lentivirus-based vectors.
The trophoblast layers of the mammalian placenta carry out many complex functions required to pattern the developing embryo and maintain its growth and survival in the uterine environment. Genetic disruption of many gene pathways can result in embryonic lethality because of placental failure, potentially confusing the interpretation of mouse knockout phenotypes. Development of tools to specifically and efficiently manipulate gene expression in the trophoblast lineage would greatly aid understanding of the relative roles of different genetic pathways in the trophoblast versus embryonic lineages. We show that short-term lentivirus-mediated infection of mouse blastocysts can lead to rapid expression of a green fluorescent protein (GFP) transgene specifically in the outer trophoblast progenitors and their later placental derivatives. Efficient trophoblast-specific gene knockdown can also be produced by lentivirus-mediated pol III-driven short hairpin RNA (shRNA) and efficient trophoblast-specific gene knockout by pol II-driven Cre recombinase lentiviral vectors. This lentivirus lineage-specific infection system thus facilitates both gain and loss of function studies during placental development in the mouse and potentially other mammalian species.


Disrupted in schizophrenia 1 regulates neuronal progenitor proliferation via modulation of GSK3beta/beta-catenin signaling.
The Disrupted in Schizophrenia 1 (DISC1) gene is disrupted by a balanced chromosomal translocation (1; 11) (q42; q14.3) in a Scottish family with a high incidence of major depression, schizophrenia, and bipolar disorder. Subsequent studies provided indications that DISC1 plays a role in brain development. Here, we demonstrate that suppression of DISC1 expression reduces neural progenitor proliferation, leading to premature cell cycle exit and differentiation. Several lines of evidence suggest that DISC1 mediates this function by regulating GSK3beta. First, DISC1 inhibits GSK3beta activity through direct physical interaction, which reduces beta-catenin phosphorylation and stabilizes beta-catenin. Importantly, expression of stabilized beta-catenin overrides the impairment of progenitor proliferation caused by DISC1 loss of function. Furthermore, GSK3 inhibitors normalize progenitor proliferation and behavioral defects caused by DISC1 loss of function. Together, these results implicate DISC1 in GSK3beta/beta-catenin signaling pathways and provide a framework for understanding how alterations in this pathway may contribute to the etiology of psychiatric disorders.


Misreporting of energy: prevalence, characteristics of misreporters and influence on observed risk estimates in the Malmö Diet and Cancer cohort.
The present study investigates the prevalence of misreporting of energy in the Malmö Diet and Cancer cohort, and examines anthropometric, socio-economic and lifestyle characteristics of the misreporters. Further, the influence of excluding misreporters on risk estimates of post-menopausal breast cancer was examined. Information of reported energy intake (EI) was obtained from a modified diet history method. A questionnaire provided information on lifestyle and socio-economic characteristics. Individual physical activity level (PAL) was calculated from self-reported information on physical activity at work, leisure time physical activity and household work, and from estimates of hours of sleeping, self-care and passive time. Energy misreporting was defined as having a ratio of EI to BMR outside the 95 % CI limits of the calculated PAL. Logistic regression analysed the risk of being a low-energy reporter or a high-energy reporter. Almost 18 % of the women and 12 % of the men were classified as low-energy reporters, 2.8 % of the women and 3.5 % of the men were classified as high-energy reporters. In both genders high BMI, large waist circumference, short education and being a blue-collar worker were significantly associated with low-energy reporting. High-energy reporting was significantly associated with low BMI, living alone and current smoking. The results add support to the practice of energy adjustment as a means to reduce the influence of errors in risk assessment.


Intraluminal Flagellin Differentially Contributes to Gut Dysbiosis and Systemic Inflammation following Burn Injury.
Burn injury is associated with a loss of gut barrier function, resulting in systemic dissemination of gut-derived bacteria and their products. The bacterial protein and TLR5 agonist, flagellin, induces non-specific innate immune responses. Because we detected flagellin in the serum of burn patients, we investigated whether gut-derived flagellin was a primary or secondary contributor to intestinal dysfunction and systemic inflammation following burn injury. The apical surface of polarized human intestinal epithelial cells (IECs), Caco-2BBe, were exposed to 50 or 500 ng of purified flagellin and 1 x 105 of an intestinal E. coli (EC) isolate as follows: 1) flagellin added 30 min prior to EC, 2) flagellin and EC added simultaneously, or 3) EC added 30 min prior to flagellin. Our results showed that luminal flagellin and EC modulated each other's biological actions, which influenced their ability to induce basolateral secretion of inflammatory cytokines and subsequent translocation of bacteria and their products. A low dose of flagellin accompanied by an enteric EC in the lumen, tempered inflammation in a dose- and time-dependent manner. However, higher doses of flagellin acted synergistically with EC to induce both intestinal and systemic inflammation that compromised barrier integrity, increasing systemic inflammation following burn injury, a process we have termed flagellemia. In a murine model of burn injury we found that oral gavage of flagellin (1 μg/mouse) significantly affected the gut microbiome after burn injury. In these mice, flagellin disseminated out of the intestine into the serum and to distal organs (mesenteric lymph nodes and lungs) where it induced secretion of monocyte chemoattractant protein (MCP-1) and CXCL1/KC (mouse equivalent of human IL-8) at 24 and 48h post-burn. Our results illustrated that gut-derived flagellin alone or accompanied by a non-pathogenic enteric EC strain can function as an initiator of luminal and systemic inflammation following burn injury.


An fMRI-Neuronavigated Chronometric TMS Investigation of V5 and Intraparietal Cortex in Motion Driven Attention.
The timing of networked brain activity subserving motion driven attention in humans is currently unclear. Functional MRI (fMRI)-neuronavigated chronometric transcranial magnetic stimulation (TMS) was used to investigate critical times of parietal cortex involvement in motion driven attention. In particular, we were interested in the relative critical times for two intraparietal sulcus (IPS) sites in comparison to that previously identified for motion processing in area V5, and to explore potential earlier times of involvement. fMRI was used to individually localize V5 and middle and posterior intraparietal sulcus (mIPS; pIPS) areas active for a motion driven attention task, prior to TMS neuronavigation. Paired-pulse TMS was applied during performance of the same task at stimulus onset asynchronies (SOAs) ranging from 0 to 180 ms. There were no statistically significant decreases in performance accuracy for trials where TMS was applied to V5 at any SOA, though stimulation intensity was lower for this site than for the parietal sites. For TMS applied to mIPS, there was a trend toward a relative decrease in performance accuracy at the 150 ms SOA, as well as a relative increase at 180 ms. There was no statistically significant effect overall of TMS applied to pIPS, however, there appeared a potential trend toward a decrease in performance at the 0 ms SOA. Overall, these results provide some patterns of potential theoretical interest to follow up in future studies.


Atropa belladonna intoxication: a case report.
Atropa belladonna is a poisonous plant also called deadly nightshade. Its roots, leaves and fruits contain alkaloids: atropine, hyocyamine and scopolamine. The risk of poisoning in children is important because of possible confusion with other berries. Atropa belladonna acute intoxication is a severe condition, it's should be considered in the presence of anti-cholinergic toxidrome, the differential diagnosis include other plants or psychoactive drugs containing atropine. The treatment is mainly symptomatic including gastrointestinal decontamination with activated charcoal. In severe cases, physostigmine can be used as an antidote. We report the case of 11 year old girl with Atropa belladonna poisoning which was administrated in a therapeutic purpose as a remedy to jaundice. The child presented essentially a central anti-cholinergic syndrome. She was admitted in the intensive care unit, the progression was favorable with symptomatic treatment.


Expression and purification of recombinant Saccharomyces cerevisiae mitochondrial carrier protein YGR257Cp (Mtm1p).
The Saccharomyces cerevisiae mitochondrial carrier YGR257Cp (Mtm1p) is an integral membrane protein that plays an essential role in mitochondrial iron homeostasis and respiratory functions, but its carrier substrate has not previously been identified. Large amounts of pure protein are required for biochemical characterization, including substrate screening. Functional complementation of a Saccharomyces knockout by expression of TwinStrep tagged YGR257Cp demonstrates that an affinity tag does not interfere with protein function, but the expression level is very low. Heterologous expression in Pichia pastoris improves the yield but the product is heterogeneous. Expression has been screened in several Escherichia coli hosts, optimizing yield by modifying induction conditions and supplementing with rare tRNAs to overcome codon bias in the eukaryotic gene. Detection of an additional N-terminal truncation product in E. coli reveals the presence of a secondary intracistronic translation initiation site, which can be eliminated by silent mutagenesis of an alternative (Leu) initiation codon, resulting in production of a single, full-length polypeptide (∼30% of the total protein) as insoluble inclusion bodies. Purified inclusion bodies were successfully refolded and affinity purified, yielding approximately 40mg of pure, soluble product per liter of culture. Refolded YGR257Cp binds pyridoxal 5'-phosphate tightly (KD<1μM), supporting a new hypothesis that the mitochondrial carrier YGR237Cp and its homologs function as high affinity PLP transporters in mitochondria, providing the first evidence for this essential transport function in eukaryotes.


Lactate dehydrogenase, beta-glucuronidase and arylsulfatase activity in gingival crevicular fluid associated with experimental gingivitis in man.
Experimental gingivitis provides a useful model for studying the initiation of periodontal disease in man. This study evaluated over a 4-week period the Plaque Index (PLI), Gingival Bleeding Time Index (GBTI), and gingival crevicular fluid (GCF) for resting and flow volume as well as the concentration and total activity of three enzymes in the GCF (lactate dehydrogenase--LDH, beta-glucuronidase--BG and arylsulfatase--AS) from the maxillary right quadrant of eight subjects with healthy gingiva. After rising sharply during the 1st week, the PLI continued to increase during the 2nd week but remained constant during the 3rd and 4th weeks. The GBTI, and the resting and flow GCF volumes, increased steadily throughout the study. LDH concentration in GCF varied minimally during the experiment, while total LDH activity rose slightly over the 4-week period. BG concentration and total activity in GCF rose steadily from baseline to the 3rd week and then either fell or leveled off during the 4th week. AS concentration in GCF rose from baseline to the 2nd or 3rd week and then fell. AS total activity in GCF rose from baseline to the 2nd week and then remained constant. These data suggest that while clinical signs of inflammation increased over the 4 weeks of the experiment, a homeostatic mechanism in the crevicular environment may control ground substance-degrading enzyme activity during experimental gingivitis in man.


Shame predicts revictimization in victims of childhood violence: A prospective study of a general Norwegian population sample.
Victims of childhood violence often experience new victimization in adult life. However, risk factors for such revictimization are poorly understood. In this longitudinal study, we investigated whether violence-related shame and guilt were associated with revictimization. Young adults (age = 17-35) exposed to childhood violence (n = 505) were selected from a (Country) population study of 6,589 persons (Wave 1), and reinterviewed by telephone 12-18 months later (Wave 2). Wave 1 measures included shame, guilt, social support, posttraumatic stress, and binge drinking frequency, as well as childhood violence. Logistic regression was used to estimate associations between Wave 1 risk factors and Wave 2 revictimization (physical or sexual violence, or controlling partner behavior). In total, 31.5% (n = 159) had been revictimized during the period between Wave 1 and 2. Of these, 12.9% (n = 65) had experienced sexual assault, 22% (n = 111) had experienced physical assault and 7.1% (n = 36) had experienced controlling behavior from partner. Both shame and guilt were associated with revictimization, and withstood adjustment for other potentially important risk factors. In mutually adjusted models, guilt was no longer significant, leaving shame and binge drinking frequency as the only factors uniquely associated with revictimization. Violence-prevention aimed at victims of childhood violence should be a goal for practitioners and policymakers. This could be achieved by targeting shame, both on both on the individual level (clinical settings) and the societal level (changing the stigma of violence). (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Bronchoalveolar lavage cytology in children with chronic unexplained cough and severely neurologically impaired children.
We aimed to compare the cellular composition of bronchoalveolar lavage (BAL) fluids in children with chronic unexplained cough (group 1) and severely neurologically impaired children with chronic or recurrent respiratory problems (group 2) with the BAL cytology of children without pulmonary or systemic diseases (group 3). Bronchoscopy with BAL fluid analysis was performed in all subjects. Children with respiratory symptoms underwent 24-hour multichannel intraluminal impedance monitoring. A significant difference was found between the groups in the total number of cells in BAL fluid cytology (191 [range, 24-12,747], 747 [range, 53-13,000], and 105 [range, 41-233] cells/μL, P = .015), in the percentage of neutrophils (21.2 [SD = 32.4], 49.4 [SD = 36.6], and 3.6 [SD = 2.4], P < .001), and in the percentage of lipid-laden macrophages (10.3 [SD = 11.4], 13.7 [SD = 15.8] and 0.44 [SD = 1.0], P < .001). The BAL fluid cytology provides useful data for determining the cause of chronic unexplained cough and chronic or recurrent respiratory problems in severely neurologically impaired children.


Postoperative disc entrapment following cardiac valve replacement--a report of ten cases.
The sudden jamming of a prosthetic valve disc is one cause for postoperative hemodynamic deterioration. This complication occurred in 10 instances (2% of disc valves implanted), resulting in 4 fatalities. In 6 patients the disc entrapment was a complication following mitral valve replacement, and in 4 others the malfunction followed aortic valve surgery. The entrapment of the disc occurred on the second postoperative day in 3 patients. Two of these were due to an unresected chordal strand becoming wedged between the disc and valve rim of a Björk-Shiley mitral prosthesis and resulted in death. In the third patient, the aortic valve disc became attached to the Björk-Shiley composite aortic graft following the repair of an aortic dissection. A firm blood clot had formed between the graft and the oversewn aortic wall. This patient recovered after cardiopulmonary resuscitation and subsequent reoperation. The remaining cases developed while the patients were still either on the operating table or in the recovery room. The mechanisms of the disc entrapments are presented and the significance of an early correct diagnosis and urgent surgical correction is underlined.


Toward an understanding of the emotion-modulated startle eyeblink reflex: the case of anger.
Three studies investigated the effect of angering pictures on the startle eyeblink response, based on anger's unique identity as an approach-oriented negative affect. In Study 1, eyeblinks to startling noise probes during angering and neutral pictures did not differ, despite angering pictures being rated higher on arousal and anger and more negative in valence. Study 2 replicated Study 1; also, dysphoric participants exhibited potentiated eyeblinks to probes during angering pictures much like those to probes during fear/disgust stimuli. A follow-up study revealed that dysphoric participants rated angering pictures higher in fear. Study 3 again found that eyeblinks to probes during angering and neutral pictures did not differ. Taken together, these results suggest that probes during angering stimuli elicit eyeblinks much like those during neutral stimuli, perhaps due to the competing influences of arousal, valence, and motivation on the startle eyeblink reflex.


Tapping natural reservoirs of homing endonucleases for targeted gene modification.
Homing endonucleases mobilize their own genes by generating double-strand breaks at individual target sites within potential host DNA. Because of their high specificity, these proteins are used for "genome editing" in higher eukaryotes. However, alteration of homing endonuclease specificity is quite challenging. Here we describe the identification and phylogenetic analysis of over 200 naturally occurring LAGLIDADG homing endonucleases (LHEs). Biochemical and structural characterization of endonucleases from one clade within the phylogenetic tree demonstrates strong conservation of protein structure contrasted against highly diverged DNA target sites and indicates that a significant fraction of these proteins are sufficiently stable and active to serve as engineering scaffolds. This information was exploited to create a targeting enzyme to disrupt the endogenous monoamine oxidase B gene in human cells. The ubiquitous presence and diversity of LHEs described in this study may facilitate the creation of many tailored nucleases for genome editing.


Incidence and clinical relevance of TEL/AML1 fusion genes in children with acute lymphoblastic leukemia enrolled in the German and Italian multicenter therapy trials. Associazione Italiana Ematologia Oncologia Pediatrica and the Berlin-Frankfurt-Münster Study Group.
The molecular approach for the analysis of leukemia associated chromosomal translocations has led to the identification of prognostic relevant subgroups. In pediatric acute lymphoblastic leukemia (ALL), the most common translocations, t(9;22) and t(4;11), have been associated with a poorer clinical outcome. Recently the TEL gene at chromosome 12p13 and the AML1 gene at chromosome 21q22 were found to be involved in the translocation t(12;21)(p13;q22). By conventional cytogenetics, however, this chromosomal abnormality is barely detectable and occurs in less than 0.05% of childhood ALL. To investigate the frequency of the molecular equivalent of the t(12;21), the TEL/AML1 gene fusion, we have undertaken a prospective screening in the running German Berlin-Frankfurt-Münster (BFM) and Italian Associazione Italiana Ematologia Oncologia Pediatrica (AIEOP) multicenter ALL therapy trials. We have analyzed 334 unselected cases of pediatric ALL patients consecutively referred over a period of 5 and 9 months, respectively. The overall incidence of the t(12;21) in pediatric ALL is 18.9%. The 63 cases positive for the TEL/AML1 chimeric products ranged in age between 1 and 12 years, and all but one showed CD10 and pre-B immunophenotype. Interestingly, one case displayed a pre-pre-B immunophenotype. Among the B-lineage subgroup, the t(12;21) occurs in 22.0% of the cases. Fifteen of 61 (24.6%) cases coexpressed at least two myeloid antigens (CD13, CD33, or CDw65) in more than 20% of the gated blast cells. DNA index was available for 59 of the 63 TEL/AML1 positive cases; a hyperdiploid DNA content (> or = 1.16) was detected in only four patients, being nonhyperdiploid in the remaining 55. Based on this prospective analysis, we retrospectively evaluated the impact of TEL/AML1 in prognosis by identifying the subset of B-lineage ALL children enrolled in the closed German ALL-BFM-90 and Italian ALL-AIEOP-91 protocols who had sufficient material for analysis. A total of 342 children were investigated for the presence of TEL/AML1 fusion gene and 99 cases (28.9%) were positive. The patients expressing the TEL/AML1 fusion mRNA appeared to have a better event-free survival (EFS) than the patients who lacked this chimeric product. Whereas three of the TEL/AML1 positive cases (3.0%) have relapsed to date, 27 patients without TEL/AML1 rearrangement (11.1%) suffered from relapse. To date, the only subset of B-lineage ALL with a favorable prognosis has been the hyperdiploid group (DNA index > or = 1.16 < 1.6). Our findings reinforce the need to include the molecular screening of the t(12;21) translocation within ongoing prospective ALL trials to prove definitively its prognostic impact.


[Regulation of fibrinolysis by lymphocytes in inflammatory diseases of the bronchopulmonary system].
Urokinase receptors on lymphocytes described in the paper link immune system of the body with fibrinolytic one, play a biological role in sano-++- and pathogenesis in aggravation of chronic purulent bronchitis, of focal and croupous pneumonia. A correlation between the trend in the expression of urokinase receptors on lymphocytes and changes in activity of plasminogen activator in the blood and urine was established: the more the expression, the more the activity. The method can serve a criterion of the treatment effectiveness+ in nonspecific inflammatory bronchial and pulmonary diseases and be helpful in designing methods of their treatment improvement.


Postsynthetic Selective Ligand Cleavage by Solid-Gas Phase Ozonolysis Fuses Micropores into Mesopores in Metal-Organic Frameworks.
Herein we report a novel, ozone-based method for postsynthetic generation of mesoporosity in metal-organic frameworks (MOFs). By carefully selecting mixed-ligand Zr-fcu-MOFs based on organic ligand pairs in which one ligand has ozone-cleavable olefin bonds and the other ligand is ozone-resistant, we were able to selectively break the cleavable ligand via ozonolysis to trigger fusion of micropores into mesopores within the MOF framework. This solid-gas phase method is performed at room-temperature, and, depending on the cleavable ligand used, the resultant ligand-fragments can be removed from the ozonated MOF by either washing or sublimation. Compared to the corresponding highly microporous starting MOFs, the highly mesoporous product MOFs exhibit radically distinct gas sorption properties.


Hybridization between crops and wild relatives: the contribution of cultivated lettuce to the vigour of crop-wild hybrids under drought, salinity and nutrient deficiency conditions.
With the development of transgenic crop varieties, crop-wild hybridization has received considerable consideration with regard to the potential of transgenes to be transferred to wild species. Although many studies have shown that crops can hybridize with their wild relatives and that the resulting hybrids may show improved fitness over the wild parents, little is still known on the genetic contribution of the crop parent to the performance of the hybrids. In this study, we investigated the vigour of lettuce hybrids using 98 F(2:3) families from a cross between cultivated lettuce and its wild relative Lactuca serriola under non-stress conditions and under drought, salinity and nutrient deficiency. Using single nucleotide polymorphism markers, we mapped quantitative trait loci associated with plant vigour in the F(2:3) families and determined the allelic contribution of the two parents. Seventeen QTLs (quantitative trait loci) associated with vigour and six QTLs associated with the accumulation of ions (Na(+), Cl(-) and K(+)) were mapped on the nine linkage groups of lettuce. Seven of the vigour QTLs had a positive effect from the crop allele and six had a positive effect from the wild allele across treatments, and four QTLs had a positive effect from the crop allele in one treatment and from the wild allele in another treatment. Based on the allelic effect of the QTLs and their location on the genetic map, we could suggest genomic locations where transgene integration should be avoided when aiming at the mitigation of its persistence once crop-wild hybridization takes place.


Increased error-prone non homologous DNA end-joining--a proposed mechanism of chromosomal instability in Bloom's syndrome.
BS is an inherited cancer predisposition disorder caused by inactivation of the RecQ family helicase, BLM. One of the defining features of cells from BS individuals is chromosomal instability, characterized by elevated sister chromatid exchanges (SCEs), as well as chromosomal breaks, deletions, and rearrangements. Although the basis for chromosomal instability is poorly understood, there is evidence that chromosomal abnormalities can arise through an alteration in the efficiency or fidelity of DNA double strand break (DSB) repair. Here, we show that BS cells demonstrate aberrant DSB repair mediated by the non-homologous end-joining (NHEJ) pathway for DNA repair, one of the two main pathways for the repair of DSBs in mammalian cells. Through a comparison of BS cell lines, and a derivative in which the BS phenotype has been reverted by expression of the BLM cDNA, we show that BS cells display aberrant end-joining of DSBs. Importantly, DNA end-joining in BS cells is highly error-prone and frequently results in DNA ligation at distant sites of microhomology, creating large DNA deletions. This aberrant repair is dependent upon the presence of the Ku70/86 heterodimer, a key component in the NHEJ pathway. We propose that aberrant NHEJ is a candidate mechanism for the generation of chromosomal instability in BS.


Acupuncture for nocturnal enuresis in children: a systematic review and exploration of rationale.
This review identified reports of acupuncture for childhood nocturnal enuresis, with the aim of ascertaining whether acupuncture is efficacious and or better than standard therapy for treating enuresis. Studies of children of either gender <18 years of age who received acupuncture treatment for nocturnal enuresis, were considered. The primary outcome measure was change in the mean number of wet episodes following treatment. Electronic searching was supplemented by hand searching of western medicine and traditional Chinese medicine (TCM) journals along with English language alternative medicine journals. Trials were assessed for quality and sources of bias. Meta-analysis was performed and the overall weighted odds ratio (OR) and associated 95% confidence interval (CI) were computed using the fixed effect model; the Forest plot was used to demonstrate results. Two hundred six abstracts were identified, of which 11 studies were eligible for data extraction. All the trials were of low methodological quality. There was some evidence that acupuncture is useful for nocturnal enuresis when used in conjunction with other treatment that may also include a different form of acupuncture (OR 3.98, CI: 2.2-7.2). When one form of acupuncture is compared with another there was marked heterogeneity, implying that some forms of acupuncture are effective. This review provides tentative evidence for the efficacy of acupuncture for the treatment of childhood nocturnal enuresis. Due to the low methodological quality of studies, evidence to identify, which parameters of acupuncture work best, is lacking. More rigorous trials are clearly warranted.


RAB25 confers resistance to chemotherapy by altering mitochondrial apoptosis signaling in ovarian cancer cells.
Ovarian cancer remains one of the most frequent causes of cancer-related death in women. Many patients with ovarian cancer suffer from de novo or acquired resistance to chemotherapy. Here, we report that RAB25 suppresses chemotherapy-induced mitochondrial apoptosis signaling in ovarian cancer cell lines and primary ovarian cancer cells. RAB25 blocks chemotherapy-induced apoptosis upstream of mitochondrial outer membrane permeabilization by either increasing antiapoptotic BCL-2 proteins or decreasing proapoptotic BCL-2 proteins. In particular, BAX expression negatively correlates with RAB25 expression in ovarian cancer cells. BH3 profiling assays corroborated that RAB25 decreases mitochondrial cell death priming. Suppressing RAB25 by means of RNAi or RFP14 inhibitory hydrocarbon-stapled peptide sensitizes ovarian cancer cells to chemotherapy as well as RAB25-mediated proliferation, invasion and migration. Our data suggest that RAB25 is a potential therapeutic target for ovarian cancer.


[Long-term postoperative course and surgical indications in patients with hypertrophic obstructive cardiomyopathy].
Operative results and late follow-up of 63 patients undergoing surgery for hypertrophic obstructive cardiomyopathy (HOCM) were analyzed in relation to the surgical indication. There were 64 postoperative follow-ups because one patient had two myectomy operations. 27 patients without limiting symptoms (group A) were operated on because of a pressure gradient of more than 50 mm Hg. 31 patients (group B) underwent myectomy operation because of limiting symptoms, and 5 patients (group C) required additional surgery because of mitral incompetence or left ventricular aneurysm. Comparison of preoperative clinical and hemodynamic data indicate that patients of group B were in a more advanced stage of disease than patients of group A. Operative mortality was low, with one perioperative death. Operative complications were rare, and after adequate treatment caused no significant morbidity. The left ventricular systolic pressure gradient was relieved in almost every case, with simultaneous reduction of left ventricular enddiastolic pressure. With a few exceptions the patients experienced longstanding improvement of symptoms. Durability of symptomatic improvement and survival were significantly better in patients of group A. Late results in patient group C were unfavorable, mainly because of the additional lesions requiring surgery. Atrial fibrillation and congestive heart failure were relieved by surgery, but recurrence was frequent in late follow-up. Ventricular arrhythmia during postoperative follow-up was common in both group A and group B, without obvious prognostic significance. In the light of these results the operative indications were reconsidered. Surgery is indicated in patients with limiting symptoms not responding to medical therapy, if a left ventricular pressure gradient of more than 50 mm Hg is present. Simultaneous mitral incompetence of severe degree requires primary mitral valve replacement by low profile prostheses. Risk factors, such as reanimation for ventricular fibrillation or a family history of sudden death, may militate in favour of surgical treatment. The operative indication is doubtful in patients with a high left ventricular pressure gradient but without consistent symptoms and without additional risk factors.


Clinical efficacy of hydroxychloroquine in patients with covid-19 pneumonia who require oxygen: observational comparative study using routine care data.
To assess the effectiveness of hydroxychloroquine in patients admitted to hospital with coronavirus disease 2019 (covid-19) pneumonia who require oxygen. Comparative observational study using data collected from routine care. Four French tertiary care centres providing care to patients with covid-19 pneumonia between 12 March and 31 March 2020. 181 patients aged 18-80 years with documented severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pneumonia who required oxygen but not intensive care. Hydroxychloroquine at a dose of 600 mg/day within 48 hours of admission to hospital (treatment group) versus standard care without hydroxychloroquine (control group). The primary outcome was survival without transfer to the intensive care unit at day 21. Secondary outcomes were overall survival, survival without acute respiratory distress syndrome, weaning from oxygen, and discharge from hospital to home or rehabilitation (all at day 21). Analyses were adjusted for confounding factors by inverse probability of treatment weighting. In the main analysis, 84 patients who received hydroxychloroquine within 48 hours of admission to hospital (treatment group) were compared with 89 patients who did not receive hydroxychloroquine (control group). Eight additional patients received hydroxychloroquine more than 48 hours after admission. In the weighted analyses, the survival rate without transfer to the intensive care unit at day 21 was 76% in the treatment group and 75% in the control group (weighted hazard ratio 0.9, 95% confidence interval 0.4 to 2.1). Overall survival at day 21 was 89% in the treatment group and 91% in the control group (1.2, 0.4 to 3.3). Survival without acute respiratory distress syndrome at day 21 was 69% in the treatment group compared with 74% in the control group (1.3, 0.7 to 2.6). At day 21, 82% of patients in the treatment group had been weaned from oxygen compared with 76% in the control group (weighted risk ratio 1.1, 95% confidence interval 0.9 to 1.3). Eight patients in the treatment group (10%) experienced electrocardiographic modifications that required discontinuation of treatment. Hydroxychloroquine has received worldwide attention as a potential treatment for covid-19 because of positive results from small studies. However, the results of this study do not support its use in patients admitted to hospital with covid-19 who require oxygen.


Estradiol enhances neurogenesis following ischemic stroke through estrogen receptors alpha and beta.
Neurogenesis persists throughout life under normal and degenerative conditions. The adult subventricular zone (SVZ) generates neural stem cells capable of differentiating to neuroblasts and migrating to the site of injury in response to brain insults. In the present study, we investigated whether estradiol increases neurogenesis in the SVZ in an animal model of stroke to potentially promote the ability of the brain to undergo repair. Ovariectomized C57BL/6J mice were implanted with capsules containing either vehicle or 17beta-estradiol, and 1 week later they underwent experimental ischemia. We utilized double-label immunocytochemistry to identify the phenotype of newborn cells (5-bromo-2'-deoxyuridine-labeled) with various cellular markers; doublecortin and PSA-NCAM as the early neuronal marker, NeuN to identify mature neurons, and glial fibrillary acidic protein to identify astrocytes. We report that low physiological levels of estradiol treatment, which exert no effect in the uninjured state, significantly increase the number of newborn neurons in the SVZ following stroke injury. This effect of estradiol is limited to the dorsal region of the SVZ and is absent from the ventral SVZ. The proliferative actions of estradiol are confined to neuronal precursors and do not influence gliosis. Furthermore, we show that both estrogen receptors alpha and beta play pivotal functional roles, insofar as knocking out either of these receptors blocks the ability of estradiol to increase neurogenesis. These findings clearly demonstrate that estradiol stimulates neurogenesis in the adult SVZ, thus potentially facilitating the brain to remodel and repair after injury.


The rise and fall of the autochthonous self: from Italian Renaissance art and Shakespeare to Heidegger, Lacan, and intersubjectivism.
This article addresses the unresolved question of the existence of a private core autochthonous self, as it has been described by Winnicott, Modell, and others. The postmodern version of the self has eliminated this concept entirely, relegating the self to a changing and unstable display, or regarding it as totally chaotic, or even an illusion. The question is raised whether by returning to the origins of this notion of a private self and then tracing its apparent dissolution it might be possible to discover some evidence that it still exists. The methodology used is that of obtaining knowledge directly through the arts and the claim is made that because empirical science has clamored to be the only source of knowledge, we have lost what could be obtained by direct intuitive seeing and experiencing the works of creative geniuses. To explore the rise of the autochthonous self this article provides an examination of the shift from Gothic art to Italian Renaissance art, a time which engendered the origin of "man" with his or her elusive private individual self that then became expressed in changing works of art. As this spread north, Shakespeare appeared and similarly invented and illustrated in his characters the private individual self, a concept not appreciated or recognized before the renaissance. But as science arose and Western civilization began to decline, a corresponding disillusionment with "man" took place. The self began to be viewed as solely a social construction with no core except perhaps a genetic endowment. This was accompanied by a reduction in the concept of the human as a valuable and precious living being and was replaced by regarding the human as an object of control and exploitation. After the Second World War a movement in contemporary United States psychoanalysis gradually replaced the ideas of Freud and his emphasis on the "I" in the psychoanalytic process, with forms of relational therapy, assuming that the self was ab initio intersubjectively formed and could be altered fundamentally by focus on intersubjective processes. The author contends that this attitude makes it less likely for the psychoanalyst to focus on the regressive transferences from which derivatives of the private self arise and to grasp the phenomenological whole of the patient.


Human Platelet Lysate Supports Efficient Expansion and Stability of Wharton's Jelly Mesenchymal Stromal Cells via Active Uptake and Release of Soluble Regenerative Factors.
Manufacturing of mesenchymal stromal cell (MSC)-based therapies for regenerative medicine requires the use of suitable supply of growth factors that enhance proliferation, cell stability and potency during cell expansion. Human blood derivatives such as human platelet lysate (hPL) have emerged as a feasible alternative for cell growth supplement. Nevertheless, composition and functional characterization of hPL in the context of cell manufacturing is still under investigation, particularly regarding the content and function of pro-survival and pro-regenerative factors. We performed comparative analyses of hPL, human serum (hS) and fetal bovine serum (FBS) stability and potency to support Wharton's jelly (WJ) MSC production. We demonstrated that hPL displayed low inter-batch variation and unique secretome profile that was not present in hS and FBS. Importantly, hPL-derived factors including PDGF family, EGF, TGF-alpha, angiogenin and RANTES were actively taken up by WJ-MSC to support efficient expansion. Moreover, hPL but not hS or FBS induced secretion of osteoprotegerin, HGF, IL-6 and GRO-alpha by WJ-MSC during the expansion phase. Thus, hPL is a suitable source of factors supporting viability, stability and potency of WJ-MSC and therefore constitutes an essential raw material that in combination with WJ-MSC introduces a great opportunity for the generation of potent regenerative medicine products.


The Expanding Frontier of Outpatient Spine Surgery.
In 2014, inpatient spinal fusion surgery had the highest aggregate cost of any inpatient surgery performed in the United States, costing 12 billion dollars. As the national health care system seeks to improve value-based care, there is increased motivation to perform surgery on an outpatient basis. To ensure improved patient outcomes with this transition, patient selection has become increasingly important to identify who would most benefit from outpatient spine fusion, for example. This demands an improved understanding of the demographics of patients who have been receiving outpatient spine fusion on which the spine surgery community can build to improve cost-effective care delivered. The Healthcare Cost and Utilization Project, State Ambulatory Surgery Databases, and Agency for Healthcare Research and Quality databases were queried for demographic data regarding all-cause outpatient spine surgery between 2012 and 2014. Outpatient surgery volume was compared with inpatient surgery volume-which was provided by the State Inpatient Databases. A total of 1,164,040 spine fusion procedures were identified between 2012 and 2014, of which 132,900 procedures were performed as outpatient surgery (11.4%). Of all fusion procedures amongst 18- to 44-year-old patients, 18.4% were outpatient. A larger proportion of white patients, rather than black or Hispanic patients, underwent ambulatory procedures (12.14% vs 9.53% vs 7.46%, respectively); 16.54% of spinal fusion procedures for patients with private insurance was performed on an outpatient basis. Based on patient income, 76% of all outpatient fusions were performed on patients who live in "not low" income ZIP codes. There has been a gradual trend toward performing more outpatient spinal fusion procedures over the studied period. This study has also revealed unique trends in the demographics of patients who have received outpatient spine fusion during this time. 3.


Effect of Mg2+ on stress, myosin phosphorylation, and ATPase activity in detergent-skinned swine carotid media.
Smooth muscle contraction has a relatively high requirement for free magnesium (Mg2+). In this study we examined the effect of Mg2+ concentration ([Mg2+]) on Ca2+-dependent stress development and stress maintenance, myosin ATPase activity, and myosin light chain (MLC) phosphorylation levels in Triton X-100 detergent-skinned fibers of the swine carotid media. Increasing [Mg2+] in a stepwise fashion from 0.1 to 6 mM 1) decreased the magnitude and Ca2+ sensitivity of stress development but augmented the amount of stress maintained without proportional MLC phosphorylation, 2) produced a greater decrease in the Ca2+ sensitivity of MLC phosphorylation than that of stress development, and 3) decreased myosin ATPase activity. These findings demonstrate that Mg2+ differentially modulates the MLC phosphorylation-dependent development of stress and the MLC phosphorylation-independent maintenance of stress. We suggest that increases in [Mg2+] enhance stress maintenance by increasing [MgADP], thus increasing the number of cross bridges in a force-generating state, and by a direct effect on the pathway responsible for Ca2+-dependent, MLC phosphorylation-independent contractions.


STO-609, a specific inhibitor of the Ca(2+)/calmodulin-dependent protein kinase kinase.
STO-609, a selective inhibitor of Ca(2+)/calmodulin-dependent protein kinase kinase (CaM-KK) was synthesized, and its inhibitory properties were investigated both in vitro and in vivo. STO-609 inhibits the activities of recombinant CaM-KK alpha and CaM-KK beta isoforms, with K(i) values of 80 and 15 ng/ml, respectively, and also inhibits their autophosphorylation activities. Comparison of the inhibitory potency of the compound against various protein kinases revealed that STO-609 is highly selective for CaM-KK without any significant effect on the downstream CaM kinases (CaM-KI and -IV), and the IC(50) value of the compound against CaM-KII is approximately 10 microg/ml. STO-609 inhibits constitutively active CaM-KK alpha (glutathione S-transferase (GST)-CaM-KK-(84-434)) as well as the wild-type enzyme. Kinetic analysis indicates that the compound is a competitive inhibitor of ATP. In transfected HeLa cells, STO-609 suppresses the Ca(2+)-induced activation of CaM-KIV in a dose-dependent manner. In agreement with this observation, the inhibitor significantly reduces the endogenous activity of CaM-KK in SH-SY5Y neuroblastoma cells at a concentration of 1 microg/ml (approximately 80% inhibitory rate). Taken together, these results indicate that STO-609 is a selective and cell-permeable inhibitor of CaM-KK and that it may be a useful tool for evaluating the physiological significance of the CaM-KK-mediated pathway in vivo as well as in vitro.


Effect of a molasses-based liquid supplement on gastrointestinal tract barrier function, inflammation, and performance of newly received feedlot cattle before and after a transport stress.
The objective of this study was to determine the effect of a dry versus a molasses-based liquid supplement on ruminal butyrate concentration, gastrointestinal tract (GIT) barrier function, inflammatory status, and performance of newly received feedlot cattle. In experiment 1, 60 mixed breed steers (234 ± 2.1 kg) were weaned, held overnight at a sale barn, then transported 14 h to Purdue University. After arrival, steers were weighed, blocked by body weight, and allotted within block to treatments (six pens per treatment and five steers per pen). Diets consisted of 45% roughage and 55% concentrate (dry matter basis). Treatments differed in the supplement source as follows: DRY: 10% dry supplement or LIQUID: 10% liquid molasses-based supplement. Feed intake, average daily gain (ADG), and gain:feed were determined for the three 21-d periods and overall. In experiment 2, 16 crossbred heifers (246 ± 7.5 kg) were used (8 heifers per treatment). Diets were the same as in experiment 1 and were fed for 60 d. On d 56 ruminal fluid samples were collected at 0, 3, 6, and 9 h after feeding. To mimic a stress event, heifers were transported for 4 h on d 61, rested overnight, and transported 12 h on d 62. Blood was collected from heifers immediately prior to transport and immediately upon their return. Gut barrier function using a Cr-EDTA marker was determined after transportation. Data were analyzed using the GLIMMIX procedure of SAS. Steers fed the liquid supplement had greater (P ≤ 0.03) ADG through d 42 and overall compared to steers fed the dry supplement. Feed intake did not differ (P = 0.25) between treatments from d 0 to d 21. However, steers fed the liquid supplement showed greater (P < 0.001) dry matter intake after d 21 and overall compared to those fed the dry supplement. Steers fed the liquid supplement tended (P < 0.09) to have reduced serum haptoglobin and lipopolysaccharide-binding protein (LBP) compared to those fed the dry supplement. Heifers fed the liquid supplement had greater (P = 0.02) Cr in urine and tended (P = 0.07) to have lower serum LBP after transport compared to those fed the dry supplement. Heifers fed the liquid supplement had 72% lower serum haptoglobin before, but only a 19% lower serum haptoglobin after transport compared to animals fed the dry supplement (treatment × time; P = 0.07). Therefore, the liquid supplement altered GIT barrier function, and improved inflammatory status, resulting in increased growth of receiving cattle.


Age-related effects of executive function on takeover performance in automated driving.
The development of highly automated vehicles can meet elderly drivers' mobility needs; however, worse driving performance after a takeover request (TOR) is frequently found, especially regarding non-driving related tasks (NDRTs). This study aims to detect the correlation between takeover performance and underlying cognitive factors comprising a set of higher order cognitive processes including executive functions. Thirty-five young and 35 elderly participants were tested by computerized cognitive tasks and simulated driving tasks to evaluate their executive functions and takeover performance. Performance of n-back tasks, Simon tasks, and task switching were used to evaluate updating, inhibition, and shifting components of executive functions by principal component analysis. The performance of lane changing after TOR was measured using the standard deviation of the steering wheel angle and minimum time-to-collision (TTC). Differences between age groups and NDRT engagement were assessed by two-way mixed analysis of variance. Older participants had significantly lower executive function ability and were less stable and more conservative when engaged in NDRT. Furthermore, a significant correlation between executive function and lateral driving stability was found. These findings highlight the interaction between age-related differences in executive functions and takeover performance; thus, provide implications for designing driver screening tests or human-machine interfaces.


Performance and Profitability of Fungicides for Managing Soybean White Mold: A 10-Year Summary of Cooperative Trials.
White mold, caused by Sclerotinia sclerotiorum, is a yield-limiting disease of soybean in Brazil. Uniform fungicide trials have been conducted annually since 2009. Data from 74 cooperative field trials conducted over a 10-year period were assembled. We selected five fungicides applied two times around flowering: dimoxystrobin plus boscalid (DIMO+BOSC), carbendazim plus procymidone (CARB+PROC), fluazinam (FLUZ), fluopyram (FLUO), and procymidone (PROC). For comparison, thiophanate-methyl (TMET) applied four times was also included as a low-cost treatment. Network models were fitted to the log of white mold incidence (percentages) and log of sclerotia mass data (grams/hectare) and to the nontransformed yield data (kilograms/hectare) for each treatment, including the untreated check. Back-transformation of the meta-analytic estimates indicated that the lowest and highest mean (95% confidence interval [CI]) percent reductions in incidence and sclerotia mass were 54.2 (49.3 to 58.7) and 51.6% (43.7 to 58.3) for TMET and 83.8 (79.1 to 87.5) and 87% (81.9 to 91.6) for CARB+PROC, respectively. The overall mean (95% CI) yield responses ranged from 323 kg/ha (247.4 to 400.3) for TMET to 626 kg/ha (521.7 to 731.7) for DIMO+BOSC, but the variance was significantly reduced by a binary variable (30% threshold) describing disease incidence in the untreated check. On average, an increment of 352 kg/ha was estimated for trials where the incidence was >30% compared with the low-disease scenario. Hence, the probability of breaking even on fungicide costs for the high-disease scenario was >65% for the more effective, but more expensive fungicide (FLUZ) than TMET. For the low-disease scenario, profitability was less likely and depended more on variations in fungicide cost and soybean price.


CD8 T-cell recognition of human 5T4 oncofetal antigen.
The 5T4 oncofetal antigen is expressed by a wide variety of human carcinomas, including colorectal, ovarian and gastric carcinomas. The restricted expression of 5T4 on tumor tissues as well as its implication in tumor progression and bad prognosis makes 5T4 a promising new candidate for immunotherapy. An MVA vaccine encoding 5T4 antigen has been successfully evaluated in preclinical studies in a murine tumor model. Here, we report the generation of human CD8 T cells specific for the 5T4 antigen by stimulation with autologous monocyte derived DC infected with a replication defective adenovirus encoding the 5T4 cDNA (Ad5T4). Analysis of several donors confirms a repertoire of such CD8 responses. In a parallel approach, incorporating the results of proteasome-mediated digestion of 5T4 derived 35-mer peptides and the potential high affinity epitopes predicted by a computer-based algorithm, we identified 8 putative HLA-A*0201-presented CD8 MHC class I epitopes of 5T4 antigen. Two of these generated specific CD8 T cells after restimulation with peptide loaded autologous DC and assay by cytotoxicity and IFN gamma ELISPOT. Moreover these particular peptide generated T cells recognized naturally 5T4 positive tumor cells only if they expressed HLA-A*0201 as judged by IFN gamma ELISPOT or ELISA. Also, HLA-A*0201 CD8 T cells recognized these peptides in a DC-Ad5T4 polyclonal response. In conclusion, there is a repertoire of CD8 T cell recognition of 5T4 in normal human donors and some candidate HLA-A*0201 epitopes have been identified.


COVID-19 attributed mortality and ambient temperature: a global ecological study using a two-stage regression model.
A negative correlation between ambient temperature and COVID-19 mortality has been observed. However, the World Meteorological Organization (WMO) has reinforced the importance of government interventions and warned countries against relaxing control measures due to warmer temperatures. Further understanding of this relationship is needed to help plan vaccination campaigns opportunely. Using a two-stage regression model, we conducted cross-sectional and longitudinal analyses to evaluate the association between monthly ambient temperature lagged by one month with the COVID-19 number of deaths and the probability of high-level of COVID-19 mortality in 150 countries during time t = 60, 90, and 120 days since the onset. First, we computed a log-linear regression to predict the pre-COVID-19 respiratory disease mortality to homogenize the baseline disease burden within countries. Second, we employed negative binomial and logistic regressions to analyze the linkage between the ambient temperature and our outcomes, adjusting by pre-COVID-19 respiratory disease mortality rate, among other factors. The increase of one Celsius degree in ambient temperature decreases the incidence of COVID-19 deaths (IRR = 0.93; SE: 0.026, p-value<0.001) and the probability of high-level COVID-19 mortality (OR = 0.96; SE: 0.019; p-value<0.001) over time. High-income countries from the northern hemisphere had lower temperatures and were most affected by pre-COVID respiratory disease mortality and COVID-19 mortality. This study provides a global perspective corroborating the negative association between COVID-19 mortality and ambient temperature. Our longitudinal findings support the statement made by the WMO. Effective, opportune, and sustained reaction from countries can help capitalize on higher temperatures' protective role including the timely rollout of vaccination campaigns.


[Study of 2.5S RNA of 1,4-alpha-glucan branching enzyme by fluorescent methods using ethidium bromide].
The fluorescence yield and lifetime of ethidium bromide complexes with 1,4-alpha-glucan branching enzyme and its free nucleic acid component 2.5S RNA were measured. Both fluorescence parameters showed a 10-fold increase in comparison with those characteristics for the free dye. This increase allows to suggest the existence of double-stranded regions in 2.5S RNA both in the free as well as in the protein bound state. The coefficients of fluorescence polarization were also determined for ethidium bromide complexed with free and protein bound 2.5S RNA. They proved to be 13 and 18% respectively. No concentration depolarization was observed in both types of ethidium bromide and ethidium bromide--enzyme--RNA complexes. This proves that the double-stranded regions are rather short and that two ethidium bromide molecules can't be bound to each of them. The binding isotherms were measured for ethidium bromide absorbed on 2.5S RNA and on the holoenzyme. Their parameters napp and rmax are identical in the cases of free and protein bound 2,5S RNA (rmax = 0.046 +/- 0.001). However the binding constants of ethidium bromide complexes with free and protein bound 2.5S RNA differ significantly (Kapp = 2.2 X 10(6) M-1 for free 2.5S RNA and Kapp = 1.6 X 10(6) M-1 for the holoenzyme). The quantity of nucleotides involved in the two double-stranded regions accessible for ethidium binding is estimated to be about 28%. Increasing of Mg2+ ion concentration up to 10(-3) results in a decrease of ethidium bromide binding with double stranded regions. It may be due to a more compact tertiary structure of 2.5S RNA in the presence of Mg2+ in the free as well as in protein bound state.


A study on the heterogeneity of histone H1 from dry maize embryos.
Maize dry embryo cells have been chosen as model for the study of H1 histone complement of metabolically inactive plant chromatin. H1 has been fractionated into two distinct bands in acetic acid/urea gels. Analysis of each of these bands in SDS-containing gels has shown that maize embryo H1 is heterogeneous, consisting of at least six proteins. Cross-reactivity of maize H1 species with antibodies against mouse liver total H1 and against its individual variants indicates that they share common immunological determinants but differ substantially from each other when compared by peptide mapping. It is concluded that a couple of the plant H1 subfractions are related to animal H1A histone and that another one is related to H1B histone. The other three maize H1 variants are closely related to each other and they also share common immunological determinants with mouse H1A, and possibly with H1B. All maize H1 species slightly cross-react with antibodies against H1 degree, suggesting that no one of the plant subfractions could be characterized as H1 degree in particular. One of the proteins co-extracted and comigrating with H1 is most probably an embryo storage protein.


A possible mechanism of detoxification of copper, in the fresh water mollusc, Lymnaea luteola.
The cidal effect of copper sulphate on pulmonate snail host Lymnaea luteola was studied in relation to lipid metabolism. Alterations in the levels of glycerol, phospholipids, glycerides, sterols, sterol esters and free fatty acids due to copper sulphate treatment are recorded in foot, mantle and digestive gland of this mollusc. These results have been interpreted as part of a mechanism of detoxification, prevalent in this fresh water mollusc.


Effectiveness of a community-based intervention to prevent childhood TB in Lesotho.
BACKGROUND: Child contact management (CCM) is a recognized strategy to prevent TB; however, implementation is suboptimal. PREVENT was a cluster-randomized trial that evaluated the effectiveness and acceptability of a community-based intervention (CBI) to improve CCM in Lesotho.METHODS: Ten health facilities (HFs) were randomized to CBI or standard-of-care (SOC). CBI included nurse training/mentorship, health education by village health workers (VHW), adherence support, and multidisciplinary team meetings. Information on TB cases registered from February 2016 to June 2018 and their child contacts was abstracted. Outcomes were TB preventive treatment (TPT) initiation, TPT completion, and CBI acceptability. Generalized linear mixed models were used to test for differences between study arms and qualitative interview thematic analysis for acceptability.RESULTS: Among 547 registered children (CBI: n = 399; SOC: n = 148) of 426 adult TB patients, 46% were <2 years, 48% female, and 3% HIV-exposed/positive, with no significant differences between study arms. A total of 501 children initiated TPT-98% at CBI and 88% at SOC HFs (P < 0.0001). TPT completion was 82% in CBI vs. 59% in SOC sites (P = 0.048). Caregivers and providers reported that CBI was acceptable.CONCLUSION: The CBI was acceptable and significantly improved TPT initiation and completion in Lesotho, offering the opportunity to mitigate the threat of TB among children.


Tetramethylpyrazine reverses intracerebroventricular streptozotocin-induced memory deficits by inhibiting GSK-3β.
Brain dysfunction, especially cognitive impairment, is one of the main complications in Alzheimer's disease (AD), which threatens the health of 46.8 million people worldwide. At present, the pathogenesis of cognitive dysfunction is only partially understood, and effective therapies for memory loss in AD remain elusive. Tetramethylpyrazine (TMP) is one of the major bioactive compounds purified from Chuanxiong, a Chinese herb used for the treatment of neurovascular and cardiovascular diseases. The neuroprotective properties of TMP are evident in some neurodegenerative diseases, including Parkinson's disease. However, whether TMP plays a neuroprotective role in AD is still unknown. Here, we report that 2-week treatment with TMP rescued both short-term and long-term fear memory impairment induced by intracerebroventricular injection of streptozotocin in a well-known AD rat model. Administration of TMP also restored spatial learning and memory retention abilities in streptozotocin-injected rats. Furthermore, TMP inhibited the activity of GSK-3β, an important kinase that mediates hippocampal synaptic and memory disorders in diabetes mellitus. Finally, we found that TMP treatment restored the function of cholinergic neurons. Our data suggest that dietary uptake of TMP can provide protection against memory loss in AD, and the inhibition of GSK-3β may play an important role in this protective effect.


Identifying cis-regulatory elements by statistical analysis and phylogenetic footprinting and analyzing their coexistence and related gene ontology.
Discovery of cis-regulatory elements in gene promoters is a highly challenging research issue in computational molecular biology. This paper presents a novel approach to searching putative cis-regulatory elements in human promoters by first finding 8-mer sequences of high statistical significance from gene promoters of humans, mice, and Drosophila melanogaster, respectively, and then identifying the most conserved ones across the three species (phylogenetic footprinting). In this study, a conservation analysis on both closely related species (humans and mice) and distantly related species (humans/mice and Drosophila) is conducted not only to examine more candidates but also to improve the prediction accuracy. We have found 124 putative cis-regulatory elements and grouped these into 20 clusters. The investigation on the coexistence of these clusters in human gene promoters reveals that SP1, EGR, and NRF-1 are the dominant clusters appearing in the combinatorial combination of up to five clusters. Gene Ontology (GO) analysis also shows that many GO categories of transcription factors binding to these cis-regulatory elements match the GO categories of genes whose promoters contain these elements. Compared with previous research, the contribution of this study lies not only in the finding of new cis-regulatory elements, but also in its pioneering exploration on the coexistence of discovered elements and the GO relationship between transcription factors and regulated genes. This exploration verifies the putative cis-regulatory elements that have been found from this study and also gives new insight on the regulation mechanisms of gene expression.


Generation and characterization of a monoclonal antibody that cross-reacts with the envelope protein from the four dengue virus serotypes.
Dengue viruses (DENVs; serotypes 1-4) are members of the flavivirus family. The envelope protein (E) of DENV has been defined as the principal antigenic target in terms of protection and diagnosis. Antibodies that can reliably detect the E surface glycoprotein are necessary for describing and mapping new DENV epitopes as well as for developing more reliable and inexpensive diagnostic assays. In this study, we describe the production and characterization of a monoclonal antibody (mAb) against a recombinant DENV-2 E protein that recognizes a sequential antigen in both native and recombinant form located in domain II of the E protein of all four DENV serotypes. We confirmed that this mAb, C21, recognizes a sequence located in the fusion peptide. In addition, C21 does not have neutralizing activity against DENV-2 in an in vitro system. Furthermore, the C21 mAb is an ideal candidate for the development of research reagents for studying DENV biology because it cross-reacts with the four dengue serotypes.


Mouse Gastric Epithelial Cells Resist CagA Delivery by the Helicobacter pylori Type IV Secretion System.
The initial step in bacterial infection is adherence of the bacterium to the target cell surface. Helicobacter pylori exploits the interaction of bacterial adhesin protein HopQ with human epithelial CEACAMs (CEACAM1, 5, and 6) to stably adhere to gastric epithelial cells, which is necessary for delivery of the H. pylori CagA oncoprotein into the epithelial cells via a type IV secretion system. In contrast to human CEACAMs, however, HopQ does not interact with Ceacam1 (mouse CEACAM1) in vitro or in CHO cells ectopically expressing Ceacam1. Since the mouse genome lacks Ceacam5 and Ceacam6, no significant HopQ-Ceacam interaction may occur in mouse gastric epithelial cells. Here, we found that the mouse stomach has a much lower expression level of Ceacam1 than the expression level of CEACAM1 in the human stomach. Consistently, mouse gastric epithelial cells resist CagA delivery by cagA-positive H. pylori, and the delivery is restored by ectopic expression of human CEACAM1 or CEACAM5 in mouse gastric epithelial cells. Thus, despite the fact that mice are routinely used for H. pylori infection studies, a low expression level of Ceacam1 in the mouse stomach together with the loss or greatly reduced interaction of HopQ with Ceacams make the mouse an inappropriate model for studying the role of H. pylori-delivered CagA in gastric pathogenesis, including the development of gastric cancer.


[Benedikt Syndrome Associated with Neck Clipping of Ruptured Basilar-Superior Cerebellar Artery Aneurysm:A Case Report].
Benedikt syndrome is caused by a rare type of brain damage to the red nucleus and nearby locations. Clinically, it presents as ipsilateral oculomotor palsy and specific contralateral involuntary movement. These involuntary movements may affect activities of daily living(ADLs)in affected patients. We treated a case of Benedikt syndrome associated with neck clipping of a ruptured basilar-superior cerebellar artery aneurysm. A 66-year-old female patient was admitted to our department with headache. Initial computed tomography imaging and angiography showed subarachnoid hemorrhage and left basilar-superior cerebellar artery aneurysm. We performed neck clipping of the aneurysm. Immediately after the surgery, left oculomotor nerve palsy occurred. Right involuntary movement gradually developed from the fifth day after the surgery. Postoperative magnetic resonance imaging(MRI)showed an infarction around the left red nucleus. This specific involuntary movement significantly impaired the ADLs of the patient and persisted for two months. From three months after the surgery, it gradually improved. One and a half years after the surgery, the ADLs of the patient improved notably. It is important to preserve perforators in the treatment of distal basilar artery aneurysm. However, while ischemia of the red nucleus(e.g., due to perforator damage)may cause specific involuntary movements, they could recover spontaneously after a period of months.


Movement disorders induced by gastrointestinal drugs: two paediatric cases.
A number of frequently prescribed gastrointestinal drugs can cause movement disorders in children, as well as in adults. In our centre for paediatric neurology, we saw a 3-year-old girl with abnormal movements mostly of the legs with an inner restlessness (akathisia) while using cisapride. Another patient, a 17-year-old male, developed a hemiballism (a fierce movement of one arm and shoulder with a hurling appearance) while using ranitidine for gastric distress. In both children, the movement disorder disappeared after discontinuing the drug. The pathophysiological mechanisms of these drug-induced movement disorders might be related to the central function of histamine in the developing brain. These cases illustrate the importance of being alert for possible drug-induced events early in the process of diagnosing abnormal movement disorders.


Serologic response and safety to vaccination against avian influenza using inactivated H5N2 vaccine in zoo birds.
Due to the spread of the H5N1 highly pathogenic strain of avian influenza virus across Europe, a preventive vaccination occurred in early 2006 among 135 French zoologic institutions. Approximately 25,000 birds were vaccinated with a H5N2 inactivated vaccine. Among them, 4,369 birds were monitored by members of Association Francophone des Vétérinaires de Parc Zoologique regarding safety issues of the vaccination protocol. A total of 1,686 blood samples were collected before the first injection (n = 255), at the time of booster (n = 463), 60 day after the booster (n = 514), and 180 day (n = 229) and 330 day (n = 217) after the initial injection. Thus, sera of 126 species representing 15 different avian orders were tested using the hemagglutinin inhibition assay to evaluate seroconversion and the long-term serologic profile of selected anti-H5 antibody. Safety was considered satisfactory in all orders, and there were no deleterious effects on large-volume injection/body weight ratio. After the second injection, 71% of the birds developed a titer > or =32, with a mean titer of 558. Titers then decreased in all birds, with 42% of the remaining birds having a titer > or =32 at day 180 and only 26% at day 330. Results demonstrated that a booster 42 days after initial vaccination was mandatory to raise the titer above 32, considered to be the protective level in poultry, and to increase the number of seroconverted birds. Differences in the serologic responses among the orders and species of birds were detected and could be linked with the variation of vaccine dose injected per body weight or with species-specific immune response. The protocol for additional campaigns will be adjusted for some bird orders through the increase of injected dose or a half yearly booster to sustain better titers over the year. Vaccination is a useful tool, together with biosecurity, that should always be used as a primary method of preventing and controlling avian influenza outbreaks.


The missed pandemic: Intimate partner violence in female mental-health-care-users during the COVID-19 pandemic.
Adverse childhood experiences (ACEs) and interpersonal violence (IPV) in mentally ill women are often neglected and need to be reviewed in light of the suggested increase in IPV during the COVID-19 pandemic.We investigated the prevalence of ACEs and IPV in women living with severe mental illness (SMI) attending an outpatient psychiatry service at a public hospital in KwaZulu-Natal, South Africa, during the COVID-19 pandemic. We also described the association of ACEs with later IPV.A written survey comprising socio-demographic and clinical questionnaire, WHO Adverse Childhood Experiences International Questionnaire (ACE-IQ) for ACEs and the Women abuse screening tool (WAST) for IPV, was completed by the 154 women with SMI.141 (91.6%) participants scored positive for ACEs and 104 (67.5%) had experienced three or more ACEs. The most prevalent forms of ACEs were emotional neglect 72 (46.8%), one or no parents, parental separation, or divorce 104 (67.5%), contact sexual abuse 67 (43.5%) and witnessing a household member treated violently 67 (43.5%). Sixty-one (46.6%) participants reported IPV with scores 13 (indicative of abuse). On logistic regression, experience of three or more ACEs was significantly associated with IPV in adulthood (aOR 3.3, 95% CI: 1.2-9.6).The high prevalence of IPV and association of IPV with cumulative ACEs reflect firstly the hidden epidemic of domestic violence and secondly the vulnerability of those with ACEs to become victims of abuse later which is often missed in the care of women with SMI.


The dynamics of DNA methylation in maize roots under Pb stress.
Plants adapt to adverse conditions through a series of physiological, cellular, and molecular processes, culminating in stress tolerance. However, little is known about the associated regulatory mechanisms at the epigenetic level in maize under lead (Pb) stress. Therefore, in this study, we aimed to compare DNA methylation profiles during the dynamic development of maize roots following Pb treatment to identify candidate genes involved in the response to Pb stress. Methylated DNA immunoprecipitation-sequencing (MeDIP-seq) was used to investigate the genome-wide DNA methylation patterns in maize roots under normal condition (A1) and 3 mM Pb(NO3)2 stress for 12 h (K2), 24 h (K3) and 48 h (K4). The results showed that the average methylation density was the highest in CpG islands (CGIs), followed by the intergenic regions. Within the gene body, the methylation density of the introns was higher than those of the UTRs and exons. In total, 3857 methylated genes were found in 4 tested samples, including 1805 differentially methylated genes for K2 versus A1, 1508 for K3 versus A1, and 1660 for K4 versus A1. Further analysis showed that 140 genes exhibited altered DNA methylation in all three comparisons, including some well-known stress-responsive transcription factors and proteins, such as MYB, AP2/ERF, bZIP, serine-threonine/tyrosine-proteins, pentatricopeptide repeat proteins, RING zinc finger proteins, F-box proteins, leucine-rich repeat proteins and tetratricopeptide repeat proteins. This study revealed the genome-scale DNA methylation patterns of maize roots in response to Pb exposure and identified candidate genes that potentially regulate root dynamic development under Pb stress at the methylation level.


Enhancing the "What" and "Why" of the Pharmacists' Patient Care Process With the "How" of Clinical Reasoning.
Objectives. Clinical reasoning is integral to the provision of patient-centered care as outlined in the Pharmacists' Patient Care Process (PPCP). However, the PPCP was not created to foster clinical reasoning in student pharmacists and cannot be the sole tool used to characterize or cultivate these skills. This article describes elements of clinical reasoning, the relationship between clinical reasoning and PPCP, and concepts from the clinical reasoning literature that should inform the teaching of clinical reasoning skills.Findings. Key elements of the PPCP were identified in clinical reasoning definitions, but differences emerged. The literature supports clinical reasoning as a bidirectional, fluid process that is highly collaborative. Effective clinical reasoning requires multiple types of "thinking," interaction with others and the environment, self-assessment, and a tolerance for nuance or ambiguity. Teaching strategies can be used in the didactic and experiential setting to target the cognitive and contextual factors associated with clinical reasoning.Summary. Educators should consult the literature to enhance our understanding of clinical reasoning in seeking to teach, model, and foster these skills in our students. Future scholarship should include the development of models to support clinical reasoning within the profession of pharmacy, adoption and experimentation with clinical reasoning teaching techniques, and valuation of the utility of various assessment tools and processes.


Intanza(®) 15 mcg intradermal influenza vaccine elicits cross-reactive antibody responses against heterologous A(H3N2) influenza viruses.
The aim of the present study was to explore the ability of Intanza(®) 15 μg, the intradermal (ID) trivalent inactivated split-virion influenza vaccine containing 15 μg hemagglutinin per strain, to enhance the antibody responses against heterologous circulating H3N2 strains in adults 60 years and older. During the 2006-2007 influenza season, subjects aged 60 years or older were randomly assigned to receive one dose of ID or an intramuscular (IM, Vaxigrip(®)) influenza vaccine, which contained the reassortant A/Wisconsin/67/05(H3N2) strain as the H3N2 component. Antibody responses were assessed against the homologous vaccine strain, against the A/Brisbane/10/07(H3N2) reassortant strain and against four heterologous H3N2 field isolates (A/Genoa/62/05(H3N2), A/Genoa/3/07(H3N2), A/Genoa/2/07(H3N2), A/Genoa/3/06(H3N2)). The viruses tested belonged to three different clades that were closely related antigenically to A/California/7/04(H3N2), A/Nepal/921/06(H3N2) and A/Brisbane/10/07(H3N2). Antibody responses to these viruses were measured in 25 subjects per group using both haemagglutination inhibition (HI) and neutralization (NT) assays. At least one Committee for Medicinal Products for Human Use (CHMP) immunogenicity criteria for vaccine approval in the elderly was reached by both vaccines against all the viruses used in the study. All three CHMP criteria were reached against A/California/7/04(H3N2)-like, A/Nepal/921/06(H3N2)-like and A/Brisbane/10/07(H3N2)-like viruses by Intanza(®) 15 μg ID vaccine, while IM vaccination did not meet seroprotection criteria against circulating A/Nepal/921/06(H3N2)-like and A/Brisbane/10/07(H3N2)-like viruses or seroconversion criteria against A/Brisbane/10/07(H3N2)-like viruses. Post-vaccination HI titer, seroconversion, and seroprotection rates were higher against all viruses in subjects who received Intanza(®) 15 μg. The superiority of the seroprotection rate against the A/Nepal/921/06(H3N2)-like strain attained statistical significance despite the small sample size. Upon Beyer correction for pre-vaccination status, post-immunization HI titers against A/California/7/04(H3N2)-like and A/Brisbane/10/07(H3N2)-like strains and NT post-immunization titers against A/Wisconsin/67/05(H3N2), A/California/7/04(H3N2)-like, A/Brisbane/10/07(H3N2)-like strains were significantly higher in subjects immunized with Intanza(®) 15 μg than in individuals receiving IM vaccine. This study, although limited in the size of study population, demonstrated the broader immune response elicited by an ID influenza vaccine vs. a standard IM influenza vaccine against heterologous viruses including field isolates.


Histiocytic sarcoma transdifferentiated from follicular lymphoma presenting as a cutaneous tumor.
Histiocytic sarcoma represents a rare and poorly understood tumor of histiocytic/dendritic cell lineage that can rarely present in the skin. Previously reported cases of histiocytic sarcoma after follicular lymphoma suggested that follicular lymphoma can transdifferentiate into histiocytic sarcoma. We describe another case involving a 40-year old male who developed histiocytic sarcoma in his right thigh 4 years after the diagnosis of grade 1 follicular lymphoma in the left neck. The two neoplasms were morphologically and immunophenotypically different but had identical immunoglobulin heavy chain gene and bcl-2 gene rearrangements, as demonstrated by polymerase chain gene reaction analysis, and the presence of t(14;18)(q32;q21) translocation was confirmed via fluorescence in situ hybridization (FISH) analysis. Because of spindle cell morphology and focal S-100 positivity, malignant peripheral nerve sheath tumor and melanoma diagnoses were made initially and extensive workup was required to discover the correct diagnosis. Lineage transdifferentiation can occur in mature lymphoid neoplasms and awareness of this phenomenon and appropriate workup is crucial for correct diagnosis, as different treatment protocols and prognosis may vary.


Assessment of acylation routes and structural characterisation by liquid chromatography/tandem mass spectrometry of semi-synthetic acyl ester analogues of lipophilic marine toxins.
Esterification is one of the most important metabolic routes of lipophilic marine toxins in shellfish. In this work we assessed several chemical acylation reactions aimed at obtaining acyl ester analogues via partial synthesis from the free toxins. The procedures developed including sensitive and selective methods based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) can be applied to obtain reference materials that may be used as analytical standards (internal/external) for method development and calibration, as well as to perform toxicological in vitro and in vivo studies. Acylation systems involved both anhydrous and non-anhydrous fatty acid or acid halides as a source of the acyl radical, and several catalysers of the reaction. A series of mass spectrometric experiments involving product ion scans and multiple reaction monitoring (MRM) were used to confirm the identity and to elucidate the fragmentation pathways of the synthesised products. Reaction yields regarding reaction time and temperature were examined at sub-nmol scale for the acylation system consisting of palmitic anhydride and 4-(dimethylamino)pyridine (DMAP) in anhydrous pyridine, showing the best conditions at 75 °C for 60 min, 75 °C for 120 min and 100 °C for 270 min for cyclic imines, azaspiracid-1 and pectenotoxin-2, respectively. The esterification approach was verified at a larger scale for the esterification of gymnodimine-A (GYM-A), which kept a good yield >90% for the synthesis of 10-O-palmitoyl-GYM-A. Acyl ester analogues of lipophilic marine toxins have been synthesised and their structure elucidated by LC/MS/MS. For acyl ester analogues identical to natural metabolites, the procedures developed have potential to be applied for the semi-synthesis of metabolites in a sustainable, scalable and controlled way, avoiding extensive and tedious isolation and purification procedures from naturally contaminated shellfish. For the semi-synthetic esters structurally different than those found in shellfish, they may have applicability as internal standards for accurate quantifications of natural metabolites present in complex matrices.


New muscle power test in neuromuscular disease. Feasibility and reliability.
We studied the feasibility, reliability, and reproducibility of an "all-out" 30-second cycling or arm cranking test of muscle power and muscle endurance (the Wingate Anaerobic Test) in 66 girls and boys aged 5 to 18 years old who had cerebral palsy or a myopathy. The arm and leg tests, given in duplicate, were feasible in 94% and 61% of subjects, respectively. Reliability coefficients exceeded .95 in patients with spastic cerebral palsy and myopathies and were somewhat lower in those with athetotic cerebral palsy. Means and SDs were similar in both trials. We conclude that the Wingate Anaerobic Test is feasible, highly reliable, and reproducible in these patients.


The action spectrum for vitamin D3: initial skin reaction and prolonged exposure.
Vitamin D3 photosynthesis in the skin is formulated as a set of reaction equations, including side-reactions to lumisterol, tachysterol and toxisterols, and the accompanying reverse reactions, isomerisation of previtamin D3 to vitamin D3 and photodegradation of vitamin D3. The solution of this set is given for the stationary irradiance spectrum. The effective action spectrum for the instantaneous vitamin D3 production changes shape as a function of exposure, and therefore, no single action spectrum can be used. We assessed the action spectrum for unexposed skin and for skin that has been exposed to 7.5 Standard Erythemal Doses (SED). We constructed two new estimates: (1) the RIVM action spectrum, based on absorption spectra, quantum yields and skin transmission spectra, and (2) the modified QUT action spectrum, which is adjusted for self-absorption and skin transmission. For previously unexposed skin, the modified QUT action spectrum gives a qualitatively similar, but larger estimate than the RIVM action spectrum. We have not been able to solve the lack of quantitative agreement between the vitamin D production estimates from the three action spectrum estimates (RIVM, modified QUT and CIE). All new action spectra have stronger emphasis on the short wavelengths than the CIE action spectrum. We showed that, for wavelengths larger than 300 nm, the bandwidth that was used in the experiment that formed the basis of the CIE action spectrum, gives a red-shift of about 1 nm. Generally, with the formation of previtamin D3, the return reaction to provitamin D3 limits the production of vitamin D3. After some exposure, the new action spectrum has negative values for the longer wavelengths in the UVB. For the RIVM action spectrum, this happens after 7.5 SED, for the modified QUT action spectrum already after 1.25 SED, and after 7.5 SED the net production rate is largely cancelled. Thus prolonged exposure of previously unexposed skin saturates vitamin D3 formation. For maximum vitamin D production after 1.25 SED, sunscreens should block wavelengths larger than 310 nm. Sunscreens that block only UVB could result in reduction in vitamin D production after prolonged exposure, or even a destruction of vitamin D that has just been formed.


Clinical trials: comparing dressings.
This article describes results from one of the sites involved in a multi-centred comparative study designed to assess the performance of a collagen-alginate dressing used in the treatment of leg ulceration. The research methods used in the trial, as well as the results, will be of interest to practising nurses. Those involved in similar work are invited to submit articles to be considered for publication in future issues of this supplement.


Coronary artery disease in Africa and the Middle East.
Countries in Africa and the Middle East bear a heavy burden from cardiovascular disease. The prevalence of coronary heart disease is promoted in turn by a high prevalence of cardiovascular risk factors, particularly smoking, hypertension, dyslipidemia, diabetes, and sedentary lifestyles. Patients in Africa and the Middle East present with myocardial infarction at a younger age, on average, compared with patients elsewhere. The projected future burden of mortality from coronary heart disease in Africa and the Middle East is set to outstrip that observed in other geographical regions. Recent detailed nationally representative epidemiological data are lacking for many countries, and high proportions of transient expatriate workers in countries such as Saudi Arabia and the United Arab Emirates complicate the construction of such datasets. However, the development of national registries in some countries is beginning to reveal the nature of coronary heart disease. Improving lifestyles (reducing calorie intake and increasing physical activity) in patients in the region will be essential, although cultural and environmental barriers will render this difficult. Appropriate prescribing of pharmacologic treatments is essential in the prevention and management of cardiovascular disease. In particular, recent controversies relating to the therapeutic profile of beta-blockers may have reduced their use. The current evidence base suggests that beta-blockers are as effective as other therapies in preventing cardiovascular disease and that concerns relating to their use in hypertension and cardiovascular disease have been overstated.


Temporal and latitudinal trends of p,p'-DDE in eggs and carcasses of North American birds from 1980 to 2005.
The use of 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (DDT) in agriculture in the United States and Canada was prohibited in the early 1970s; however, it continued to be used restrictively in Mexico until 2000. Forty years later, 1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (p,p'-DDE), continues to be detected in eggs and bird carcasses in North America. The use of DDE has been associated with reproductive failure of several avian species, primarily through eggshell thinning. To assess the temporal and latitudinal distribution of p,p'-DDE in North America, the authors examined DDE concentrations reported in bird tissues in the scientific literature published between 1980 and 2009. Overall, the majority of supported models suggested that DDE concentrations in birds were greater in the upper mid-latitudes (38°-48°) than in other parts of North America. However, spatial trends of DDE seemed to be influenced by regions with large amounts of data, such as the Great Lakes area. Concentrations of p,p'-DDE in eggs averaged 2.5 μg/g, 3.2 μg/g, and 29.5 μg/g wet weight in 1980 and decreased to 1.64 μg/g, 0.87 μg/g, and 1.01 μg/g wet weight by the mid-2000s for the central, eastern, and western North America regions, respectively. The results indicate that, over time, all DDE residues observed in birds have decreased significantly in North America. Environ Toxicol Chem 2016;35:1340-1348. © 2016 SETAC.


Random blood glucose testing in dental practice: a community-based feasibility study from The Dental Practice-Based Research Network.
The prevalence of diabetes mellitus (DM) has been increasing. Instances of patients' not having received a diagnosis have been reported widely, as have instances of poor control of DM or prediabetes among patient's who have the disease. These facts indicate that blood glucose screening is needed. As part of The Dental Practice-Based Research Network, the authors conducted a study in community dental practices to test the feasibility of screening patients for abnormal random blood glucose levels by means of glucometers and finger-stick testing. Practitioners and staff members were trained to use a glucometer, and they then screened consecutive patients older than 19 years at each practice until 15 patients qualified for the study and provided consent. Perceived barriers to and benefits of blood glucose testing (BGT) were reported by patients and dental office personnel on questionnaires. Twenty-eight practices screened 498 patients. A majority of the respondents from the 67 participating dental offices considered BGT useful and worth routine implementation. They did not consider duration of BGT or its cost to be significant barriers. Among patients, more than 80 percent thought BGT in dental practice was a good idea and found it easy to withstand; 62 percent were more likely to recommend their dentists to others if BGT was offered. BGT was well received by patients and practitioners. These results support the feasibility of implementation of BGT in community dental practices. Improved diagnosis and control of DM may be achieved through implementation of BGT in community dental practices.


Loss of function of FATTY ACID DESATURASE7 in tomato enhances basal aphid resistance in a salicylate-dependent manner.
We report here that disruption of function of the ω-3 FATTY ACID DESATURASE7 (FAD7) enhances plant defenses against aphids. The suppressor of prosystemin-mediated responses2 (spr2) mutation in tomato (Solanum lycopersicum), which eliminates the function of FAD7, reduces the settling behavior, survival, and fecundity of the potato aphid (Macrosiphum euphorbiae). Likewise, the antisense suppression of LeFAD7 expression in wild-type tomato plants reduces aphid infestations. Aphid resistance in the spr2 mutant is associated with enhanced levels of salicylic acid (SA) and mRNA encoding the pathogenesis-related protein P4. Introduction of the Naphthalene/salicylate hydroxylase transgene, which suppresses SA accumulation, restores wild-type levels of aphid susceptibility to spr2. Resistance in spr2 is also lost when we utilize virus-induced gene silencing to suppress the expression of NONEXPRESSOR OF PATHOGENESIS-RELATED PROTEINS1 (NPR1), a positive regulator of many SA-dependent defenses. These results indicate that FAD7 suppresses defenses against aphids that are mediated through SA and NPR1. Although loss of function of FAD7 also inhibits the synthesis of jasmonate (JA), the effects of this desaturase on aphid resistance are not dependent on JA; other mutants impaired in JA synthesis (acx1) or perception (jai1-1) show wild-type levels of aphid susceptibility, and spr2 retains aphid resistance when treated with methyl jasmonate. Thus, FAD7 may influence JA-dependent defenses against chewing insects and SA-dependent defenses against aphids through independent effects on JA synthesis and SA signaling. The Arabidopsis (Arabidopsis thaliana) mutants Atfad7-2 and Atfad7-1fad8 also show enhanced resistance to the green peach aphid (Myzus persicae) compared with wild-type controls, indicating that FAD7 influences plant-aphid interactions in at least two plant families.