Differentiated induction of cytochrome P450b/e and P450p mRNAs by dose of phenobarbital in primary cultures of adult rat hepatocytes.
Phenobarbital induces cytochromes P450 (P450s) of not only the class IIB gene subfamily (i.e., P450b and P450e) but also the class IIIA gene subfamily (P450p and P450pcn2). To determine whether coinduction of these structurally dissimilar gene products involves the same mechanism, we examined the dose dependency of phenobarbital induction of the mRNAs for these four P450s in a new responsive system for primary monolayer cultures of adult rat hepatocytes on Matrigel, a reconstituted basement membrane. Two-day treatments of the cultures with phenobarbital produced marked dose-dependent increases in the levels of P450b, P450e, and P450p mRNAs, which reached maximal inductions ranging from 11- to greater than 193-fold. Although the dose-response relationships for the inductions of P450b and P450e mRNAs by phenobarbital were similar (ED50 = 1.5 x 10(-5) and 5.7 x 10(-6) M, respectively), the dose-response curve for the induction of P450p mRNA was positioned distinctly to the right (ED50 = 3.0 x 10(-4) M). This difference reflects a potency ratio of 20-fold for P450p/P450b mRNA induction. Phenobarbital also produced a weak dose-dependent induction of P450pcn2 mRNA, with a potency (ED50 = 3.4 x 10(-5) M) intermediate between those for P450b/e and P450p. In a similar experiment using two phenobarbital-like inducers, (trans)nonachlor and clotrimazole, the relative inductions of P450b, P450e, P450p, and P450pcn2 mRNAs proved to be similar to those produced by phenobarbital (P450p/P450b potency ratios = 14- and 16-fold, respectively). These findings provide strong further evidence in support of the newly emerging concept that "phenobarbital" induction of the responsive class IIB and class IIIA P450 isozymes likely reflects multiple mechanisms.


Introduction to complementary, alternative, and traditional therapies.
The use of complementary, alternative, and traditional therapies is increasing in the United States, and patients and their families are bringing these practices into the acute care setting. Acute and critical care nurses are in a unique and trusted position to advocate for their patients and to promote safe incorporation of complementary, alternative, and traditional therapies into the plan of care.


Elemental analysis of glass fragments by ICP-MS as evidence of association: analysis of a case.
Inductively coupled plasma mass spectrometry (ICP-MS) was used to analyze glass evidence from a case in which a person broke a window in each of 15 vehicles in a parking lot in order to gain entry into the vehicles. The results of the analysis by traditional methods, which measure the properties of color, thickness, density, and refractive index, are also reported. A total of 15 known samples representing the windows on the cars and 42 questioned glass fragments recovered from the suspect and the police vehicle where the suspect was sitting were submitted for analysis. Density comparisons separated one of the known samples into three samples, increasing the number of known samples from 15 to 17. The concentrations of 16 elements were measured for all but three of the samples using an external calibration ICP-MS method with internal standardization. Color assessment (non-instrumental) separated the 17 known samples into two groups, and refractive index measurements resulted in six groups when the Emmons double variation method was used and ten groups when the Glass Refractive Index Measurement 2 (GRIM2) system was used. Elemental analysis, by itself, differentiated all of the known samples from each other and associated four of the known sample fragments with several of the questioned sample fragments. The informing power of RI, density, and elemental analysis comparisons is evaluated and a summary of the case results is reported.


Genetic properties of the MAGIC maize population: a new platform for high definition QTL mapping in Zea mays.
Maize (Zea mays) is a globally produced crop with broad genetic and phenotypic variation. New tools that improve our understanding of the genetic basis of quantitative traits are needed to guide predictive crop breeding. We have produced the first balanced multi-parental population in maize, a tool that provides high diversity and dense recombination events to allow routine quantitative trait loci (QTL) mapping in maize. We produced 1,636 MAGIC maize recombinant inbred lines derived from eight genetically diverse founder lines. The characterization of 529 MAGIC maize lines shows that the population is a balanced, evenly differentiated mosaic of the eight founders, with mapping power and resolution strengthened by high minor allele frequencies and a fast decay of linkage disequilibrium. We show how MAGIC maize may find strong candidate genes by incorporating genome sequencing and transcriptomics data. We discuss three QTL for grain yield and three for flowering time, reporting candidate genes. Power simulations show that subsets of MAGIC maize might achieve high-power and high-definition QTL mapping. We demonstrate MAGIC maize's value in identifying the genetic bases of complex traits of agronomic relevance. The design of MAGIC maize allows the accumulation of sequencing and transcriptomics layers to guide the identification of candidate genes for a number of maize traits at different developmental stages. The characterization of the full MAGIC maize population will lead to higher power and definition in QTL mapping, and lay the basis for improved understanding of maize phenotypes, heterosis included. MAGIC maize is available to researchers.


Ten-year prospective study of public health problems associated with early drinking.
To compare early nondrinkers, experimenters, and drinkers on the prevalence of problem behaviors at grades 7 and 12 and at age 23 (N = 6338, 4265, and 3369, respectively). Results are based on longitudinal self-report data from individuals who were originally recruited from 30 California and Oregon schools at grade 7 (1985) and assessed again at grade 12 (1990) and at age 23 (1995). Logistic regression was used to develop weighted estimates of the prevalence of academic difficulties, employment problems, substance use, and delinquent and violent behaviors within the 3 drinking status groups at grades 7, 12, and/or at age 23. Huber variance estimates, which adjust for weighting and clustering of observations, were used to assess the statistical significance of differences across groups. Early drinkers and experimenters were more likely than nondrinkers to report academic problems, substance use, and delinquent behavior in both middle school and high school. By young adulthood, early alcohol use was associated with employment problems, other substance abuse, and criminal and violent behavior. Early drinkers do not necessarily mature out of a problematic lifestyle as young adults. Interventions for these high-risk youth should start early and address their other public health problems, particularly their tendency to smoke and use other illicit drugs.


Assessment of in vitro cytotoxic and genotoxic activities of some trimethoprim conjugates.
Trimethoprim, a commonly used antibacterial agent, is widely applied in the treatment of variety of infections in human. A few studies have demonstrated an extensive exposure of man to antibiotics, but there is still a lack of data for cytotoxic effects including nephrotoxicity, gastrointestinal toxicity, hematotoxicity, neurotoxicity and ototoxicity. The main purpose behind this study was to determine cytotoxic and genotoxic activities of trimethoprim (1), trimethoprim with maleic acid (2) and trimethoprim in conjugation with oxalic acid dihydrate (3). The cytotoxic effects of these three conjugates were elucidated by employing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoium bromide (MTT) assay using embryonic rat fibroblast-like cell line (F2408) and H-ras oncogene activated embryonic rat fibroblast-like cancer cell line (5RP7). Additionally, determination of genotoxic activity of these three compounds were studied by using cytokinesis blocked micronucleus assay (CBMN) in human lymphocytes. The results demonstrated that trimethoprim alone and its combination with other compounds are able to induce both cytotoxic and genotoxic damage on cultured cells (F2408, 5RP7, human lymphocytes).


Menaquinone-7 supplementation improves arterial stiffness in healthy postmenopausal women. A double-blind randomised clinical trial.
Observational data suggest a link between menaquinone (MK, vitamin K2) intake and cardiovascular (CV) health. However, MK intervention trials with vascular endpoints are lacking. We investigated long-term effects of MK-7 (180 µg MenaQ7/day) supplementation on arterial stiffness in a double-blind, placebo-controlled trial. Healthy postmenopausal women (n=244) received either placebo (n=124) or MK-7 (n=120) for three years. Indices of local carotid stiffness (intima-media thickness IMT, Diameter end-diastole and Distension) were measured by echotracking. Regional aortic stiffness (carotid-femoral and carotid-radial Pulse Wave Velocity, cfPWV and crPWV, respectively) was measured using mechanotransducers. Circulating desphospho-uncarboxylated matrix Gla-protein (dp-ucMGP) as well as acute phase markers Interleukin-6 (IL-6), high-sensitive C-reactive protein (hsCRP), tumour necrosis factor-α (TNF-α) and markers for endothelial dysfunction Vascular Cell Adhesion Molecule (VCAM), E-selectin, and Advanced Glycation Endproducts (AGEs) were measured. At baseline dp-ucMGP was associated with IMT, Diameter, cfPWV and with the mean z-scores of acute phase markers (APMscore) and of markers for endothelial dysfunction (EDFscore). After three year MK-7 supplementation cfPWV and the Stiffness Index βsignificantly decreased in the total group, whereas distension, compliance, distensibility, Young's Modulus, and the local carotid PWV (cPWV) improved in women having a baseline Stiffness Index β above the median of 10.8. MK-7 decreased dp-ucMGP by 50 % compared to placebo, but did not influence the markers for acute phase and endothelial dysfunction. In conclusion, long-term use of MK-7 supplements improves arterial stiffness in healthy postmenopausal women, especially in women having a high arterial stiffness.


Ligand-dependent and -independent function of the transactivation regions of the human estrogen receptor in yeast.
The estrogen receptor (ER) is a transcription factor involved in steroid hormone signal transduction in higher eukaryotes. The receptor also functions as a ligand-dependent transcriptional activator when introduced into Saccharomyces cerevisiae (baker's yeast), which suggests that at least some of the components of the signal transduction pathway are conserved between yeast and mammalian cells, and, moreover, allows the possibility of using this simple eukaryotic organism to dissect receptor function. However, whether the ER actually activates transcription in a mechanistically similar fashion in yeast and mammalian cells is unclear, since it has been reported that the transactivation function within the hormone binding domain (TAF-2) does not function in yeast. In this report, we have characterized the activity of the transactivation functions of the ER in yeast. Our results indicate that both TAF-2 and the N-terminal transactivation region (TAF-1) are functional in yeast and contribute synergistically to the receptor's total activity. These results are consistent with those obtained in mammalian cells. Furthermore, we show that in yeast the antagonistic effects of the antiestrogen nafoxidine arise from a modulation of the synergistic interactions of TAF-1 and TAF-2, and not simply from an inactivation of TAF-2 by antihormone. Finally, we characterize the effect of ER deletion mutants on chromatin structure in yeast. Our data lend support to the view that the formation of competent transcriptional initiation complexes requires a precise disruption of chromatin structure.


Natural history of cervical intraepithelial neoplasia: a critical review.
The literature dealing with the natural history of cervical intraepithelial neoplasia (CIN) since 1950 is reviewed, in particular from the viewpoint of regression, persistence, and progression. When stratified into the various grades of severity, the composite data indicate the approximate likelihood of regression of CIN 1 is 60%, persistence 30%, progression to CIN 3 10%, and progression to invasion 1%. The corresponding approximations for CIN 2 are 40%, 40%, 20%, and 5%, respectively. The likelihood of CIN 3 regressing is 33% and progressing to invasion greater than 12%. It is obvious from the above figures that the probability of an atypical epithelium becoming invasive increases with the severity of the atypia, but does not occur in every case. Even the higher degrees of atypia may regress in a significant proportion of cases. As morphology by itself does not predict which lesion will progress or regress, future efforts should seek factors other than morphological to determine the prognosis in individual patients.


Effect of nitrate on the synthesis and decay of nitrate reductase of Neurospora.
1. A method was developed to examine the turnover of nitrate reductase by the use of tungstate. 2. Evidence is presented which suggests that the disappearance of nitrate reductase activity from Neurospora mycelia exposed to non-inducing conditions is due to the disappearance of the enzyme protein(s) from the mycelia, and not merely due to the disappearance of its (their) catalytic power. 3. The presence of NO(3) (-) in the culture medium slows down the rate of degradation of nitrate reductase in Neurospora in vivo.


Is Kinesiotherapy Effective in Relieving Pain and Reducing Swelling after Shoulder Arthroscopy?
PURPOSE OF THE STUDY The purpose of this study is to investigate the acute and long term effectiveness of kinesio taping applied following shoulder arthroscopy in relieving pain and reducing swelling. MATERIAL AND METHODS This study included 50 patients undergone shoulder arthroscopy between June 2016 and December 2017 in our clinic. Patients were randomly assigned into two groups. Group I consisted of patients who had kinesiotherapy; while Group II consisted of control patients whom we applied sham taping with no effect. Pre and postoperative pain and swelling status of patients were recorded and groups were compared. RESULTS Comparing two groups, we found that kinesio taping significantly reduced pain levels in the early post-operative period. However, we didn't detect any significant difference in reduction of shoulder swelling between two groups. CONCLUSIONS Kinesio taping could be an alternative treatment in relieving pain after shoulder arthroscopy. However, we didn't find a significant reduction in swelling in shoulder. Key words:kinesio taping, arthroscopy, shoulder.


The Relationship between Alcohol Outlets, HIV Risk Behavior, and HSV-2 Infection among South African Young Women: A Cross-Sectional Study.
Alcohol consumption has a disinhibiting effect that may make sexual risk behaviors and disease transmission more likely. The characteristics of alcohol-serving outlets (e.g. music, dim lights, lack of condoms) may further encourage risky sexual activity. We hypothesize that frequenting alcohol outlets will be associated with HIV risk. In a sample of 2,533 school-attending young women in rural South Africa, we performed a cross-sectional analysis to examine the association between frequency of alcohol outlet visits in the last six months and four outcomes related to HIV risk: number of sex partners in the last three months, unprotected sex acts in the last three months, transactional sex with most recent partner, and HSV-2 infection. We also tested for interaction by alcohol consumption. Visiting alcohol outlets was associated with having more sex partners [adjusted odds ratio (aOR), one versus zero partners (95% confidence interval (CI)): 1.51 (1.21, 1.88)], more unprotected sex acts [aOR, one versus zero acts (95% CI): 2.28 (1.52, 3.42)], higher levels of transactional sex [aOR (95% CI): 1.63 (1.03, 2.59)], and HSV-2 infection [aOR (95% CI): 1.30 (0.88, 1.91)]. In combination with exposure to alcohol consumption, visits to alcohol outlets were more strongly associated with all four outcomes than with either risk factor alone. Statistical evidence of interaction between alcohol outlet visits and alcohol consumption was observed for all outcomes except transactional sex. Frequenting alcohol outlets was associated with increased sexual risk in rural South African young women, especially when they consumed alcohol. Sexual health interventions targeted at alcohol outlets may effectively reach adolescents at high risk for sexually transmitted infections like HIV and HSV-2. HIV Prevention Trials Network HPTN 068.


Electric field detection of phase-locked near-infrared pulses using photoconductive antenna.
We have demonstrated that a photoconductive antenna gated with 5-fs ultrashort laser pulses can detect electric field transients of near-infrared pulses at least up to 180 THz. Measured sensitivity spectrum of the antenna shows a good agreement with a simple calculation, demonstrating the promising capability of the antenna to near infrared spectroscopy. Using this setup, near-infrared time-domain spectroscopy and characterization of phase controlled near-infrared pulses are demonstrated. Observed absorption spectrum of a polystyrene film and complex refractive index dispersion of a fused silica plate both agree well with those obtained by the conventional methods.


Correlation of clinical and ultrasonographic findings after Achilles tenotomy in idiopathic club foot.
Achilles tenotomy is a recognised step in the Ponseti technique for the correction of idiopathic congenital talipes equinovarus in most percutaneous cases. Its use has been limited in part by concern that the subsequent natural history of the tendon is unknown. In a study of 11 tendons in eight infants, eight tendons were shown to be clinically intact and ten had ultrasonographic evidence of continuity three weeks after tenotomy. At six weeks after tenotomy all tendons had both clinical and ultrasonographic evidence of continuity.


Refractometric sensor based on a phase-shifted long-period fiber grating.
A refractometric sensor based on a phase-shifted long-period fiber grating written by electric-arc discharges is presented. Transmission and reflective configurations for refractive index measurements are studied. It is observed that the reflective topology permits better performance compared with the transmission one, which is the approach normally utilized in the context of long-period fiber sensing. The resolution achieved in the measurement of refractive index enables the application of this sensing head structure in demanding situations, such as the measurement of the level of salinity of water.


Distinguishing between impairments of working memory and inhibitory control in cases of early dementia.
Dementia (most notably, Alzheimer's Disease) is often associated with impairments of both working memory and inhibitory control. However, it is unclear whether these are functionally distinct impairments. We addressed the issue of whether working memory and inhibitory control can be dissociated, using data from a sample of patients who were recruited in a longitudinal study (Crawford et al., 2013, 2015). The first case revealed a preserved working memory capacity together with poor inhibitory control in the anti-saccade task. A longitudinal follow-up revealed that the defective inhibitory control emerged 12-months before the dementia was evident on the mini-mental state examination assessment. A second case revealed a poor working memory together with a well-preserved level of inhibitory control. The dissociation of working memory and inhibitory control was confirmed statistically in 7 additional cases. These findings yield converging evidence that working memory and inhibitory control are distinct cognitive operations and challenges the Kimberg and Farah (2000) cognitive model of working memory.


Amino acid templating mechanisms in selection of nucleotides opposite abasic sites by a family a DNA polymerase.
Cleavage of the N-glycosidic bond that connects the nucleobase to the backbone in DNA leads to abasic sites, the most frequent lesion under physiological conditions. Several DNA polymerases preferentially incorporate an A opposite this lesion, a phenomenon termed "A-rule." Accordingly, KlenTaq, the large fragment of Thermus aquaticus DNA polymerase I, incorporates a nucleotide opposite an abasic site with efficiencies of A > G > T > C. Here we provide structural insights into constraints of the active site during nucleotide selection opposite an abasic site. It appears that these confines govern the nucleotide selection mainly by interaction of the incoming nucleotide with Tyr-671. Depending on the nucleobase, the nucleotides are differently positioned opposite Tyr-671 resulting in different alignments of the functional groups that are required for bond formation. The distances between the α-phosphate and the 3'-primer terminus increases in the order A < G < T, which follows the order of incorporation efficiency. Additionally, a binary KlenTaq structure bound to DNA containing an abasic site indicates that binding of the nucleotide triggers a remarkable rearrangement of enzyme and DNA template. The ability to resolve the stacking arrangement might be dependent on the intrinsic properties of the respective nucleotide contributing to nucleotide selection. Furthermore, we studied the incorporation of a non-natural nucleotide opposite an abasic site. The nucleotide was often used in studying stacking effects in DNA polymerization. Here, no interaction with Tyr-761 as found for the natural nucleotides is observed, indicating a different reaction path for this non-natural nucleotide.


Differential ability of human cutaneous nociceptors to signal mechanical pain and to produce vasodilatation.
We investigated the ability of human nociceptive primary afferent neurons to encode mechanical pain and to produce vasodilatation. Pain was induced by shooting a light metal cylinder (0.3 g) at different velocities (6-18 m/sec) perpendicularly against the hairy skin of the hand. When single impact stimuli were applied, monotonically increasing stimulus-response functions were obtained in 10 psychophysical experiments using magnitude estimation techniques. In 35 microneurographic experiments nine unmyelinated afferents were recorded from the superficial radial nerve. All units responded readily to impact stimulation even at stimulus intensities that were not rated as painful. However, there was a close linear correlation between the number of action potentials evoked from the nociceptors and the psychophysical magnitude estimates of the perceived sensation or the stimulus intensity. This was also reflected by a corresponding increase of neurogenic vasodilatation. While two thin myelinated afferents displayed qualitatively similar responses 12 low-threshold mechanosensitive afferents (4 rapidly adapting, 5 slowly adapting type 1, 3 slowly adapting type II) failed to encode the intensity of the applied impact force and often became desensitized. This indicates that the total number of action potentials is the determinant of the magnitude of mechanical pain and the associated vasodilatation following single brief stimuli. By contrast, the close correlation between nociceptor activity and sensation changed when trains of mechanical impact stimuli (five stimuli of constant intensity, intratrain frequency of 1/32 to 2 Hz) were applied. Magnitude estimates of pain intensity were frequency dependent and stimuli with short interstimulus intervals were perceived as more painful than those delivered with long intervals. However, the total number of action potentials evoked from C-fibers was higher at longer interstimulus intervals than shorter intervals, thus yielding a negative correlation between the magnitude estimates of the perceived painful sensation and the number of action potentials elicited from nociceptive afferents. This suggests that temporal summation of the nociceptive discharge at central neurons becomes increasingly more important for the sensory discriminative experience of pain evoked by repetitive stimulation. We conclude that human nociceptive C-fibers signal brief noxious mechanical stimuli by the total number of action potentials evoked during a short period of time. However, with repetitive stimulation the total number of action potentials evoked from nociceptors is less important for evoking pain and temporal summation of the nociceptive primary afferent discharge becomes the crucial factor for signaling the magnitude of sensation.


The effect of dose, gender, and non-H-2 genes in murine mercury-induced autoimmunity.
We have studied the effect of dose, treatment time, gender and non-H-2 genes on immune parameters and toxicokinetics in murine mercury-induced autoimmunity (HgAI). The partly-proven mechanism for HgAI is the modification of the autoantigen fibrillarin by mercury, followed by a T cell-dependent immune response driven by the modified fibrillarin. In the H-2 congenic (H-2(S)) mouse strains A.SW and B10.S given (203)HgCl(2) in a dose of 0.25-8 mg Hg/l drinking water for up to 10 weeks, the internal dose measured as the whole-body retention of mercury reached steady state within 5 weeks. Fifty percent of the steady state level was reached already after 2 days. Conditions therefore exist for a rapid modification of fibrillarin, followed by a T cell-dependent immune response, which is consistent with the presence of anti-fibrillarin antibodies (AFA) in serum after 2 weeks. AFA developed in a dose-dependent pattern. Serum IgE showed a dose-dependent increase with a maximum after 1-2.5 weeks followed by a distinct decline towards the baseline level. Substantial polyclonal B-cell activation (PBA) developed in the highest dose groups only. Since AFA developed using lower doses too, PBA can be excluded as a general mechanism for induction of AFA. Tissue immune-complex (IC) deposits were present in the highest dose groups only, indicating a possible causality between PBA and IC deposits. The substantially lower whole body and organ mercury level needed to induce AFA in the A.SW strain as compared with the H-2 congenic B10.S strain, demonstrates that genetic factors outside the H-2 region, and not related to toxicokinetics, modifies the autoimmune response. In contrast, the difference in mercury thresholds for induction of IgE was only slight between A.SW and B10.S mice, indicating basically different mechanisms for induction of AFA and serum IgE.


DNA polymerase eta reduces the gamma-H2AX response to psoralen interstrand crosslinks in human cells.
DNA interstrand crosslinks are processed by multiple mechanisms whose relationships to each other are unclear. Xeroderma pigmentosum-variant (XP-V) cells lacking DNA polymerase eta are sensitive to psoralen photoadducts created under conditions favoring crosslink formation, suggesting a role for translesion synthesis in crosslink repair. Because crosslinks can lead to double-strand breaks, we monitored phosphorylated H2AX (gamma-H2AX), which is typically generated near double-strand breaks but also in response to single-stranded DNA, following psoralen photoadduct formation in XP-V fibroblasts to assess whether polymerase eta is involved in processing crosslinks. In contrast to conditions favoring monoadducts, conditions favoring psoralen crosslinks induced gamma-H2AX levels in both XP-V and nucleotide excision repair-deficient XP-A cells relative to control repair-proficient cells; ectopic expression of polymerase eta in XP-V cells normalized the gamma-H2AX response. In response to psoralen crosslinking, gamma-H2AX as well as 53BP1 formed coincident foci that were more numerous and intense in XP-V and XP-A cells than in controls. Psoralen photoadducts induced gamma-H2AX throughout the cell cycle in XP-V cells. These results indicate that polymerase eta is important in responding to psoralen crosslinks, and are consistent with a model in which nucleotide excision repair and polymerase eta are involved in processing crosslinks and avoiding gamma-H2AX associated with double-strand breaks and single-stranded DNA in human cells.


Phytophthora aquimorbida sp. nov. and Phytophthora taxon 'aquatilis' recovered from irrigation reservoirs and a stream in Virginia, USA.
Two distinct subgroups (L2 and A(-2)) were recovered from irrigation reservoirs and a stream in Virginia, USA. After molecular, morphological and physiological examinations, the L2 subgroup was named Phytophthora aquimorbida and the A(-2) designated as Phytophthora taxon 'aquatilis'. Both taxa are homothallic. P. aquimorbida is characterized by its noncaducous and nonpapillate sporangia, catenulate and radiating hyphal swellings and thick-walled plerotic oospores formed in globose oogonia mostly in the absence of an antheridium. P. taxon 'aquatilis' produces plerotic oospores in globose oogonia mostly with a paragynous antheridium. It has semi-papillate, caducous sporangia with variable pedicels, but it does not have hyphal swelling. Analyses of ITS, CO1, β-tubulin and NADH1 sequences revealed that P. aquimorbida is closely related to P. hydropathica, P. irrigata and P. parsiana, and P. taxon 'aquatilis' is related to P. multivesiculata. The optimum temperature for culture growth is 30 and 20 C for P. aquimorbida and P. taxon 'aquatilis' respectively. Both taxa were pathogenic to rhododendron plants and caused root discoloration, pale leaves, wilting, tip necrosis and dieback. Their plant biosecurity risk also is discussed.


Antipersonnel mines: who are the victims?
The International Committee of the Red Cross (ICRC) has surgically treated 49 946 war wounded from all sides of the Afghan conflict. Two hospitals were established in Peshawar (1981) and Quetta (1983) on the Afghan border of Pakistan and inside Afghanistan in Kabul (1988). One quarter of all war wounded were injured as a result of antipersonnel mines. In 1980 a UN weapons convention adopted specific rules on the use of land mines, yet despite this, mines are still present and causing casualties long after the ceasefire. In the ICRC hospital Peshawar, 48% of all war wounded in the last year were injured as a result of mines. Non-combatants accounted for 34%, with the majority being children < 16 years old (25%); 78% of all mine injured people claimed to be returning refugees, of whom 37% had returned within three months. A significant increase in injuries occurred in children, from 14% in 1990 to 25% in 1992. For a country recovering from war, the presence of mines causes a serious environmental, social, and economic burden, and for the victims, continued tragedy not only for their families but also the whole country for many years to come.


Factors influencing radiation treatment recommendations in early-stage Merkel cell carcinoma: a survey of US-based radiation oncologists.
It is unclear which subgroups of Merkel cell carcinoma (MCC) patients benefit the most from radiation. We surveyed radiation oncologists (RO) that regularly see and treat MCC to understand how they approach the treatment of early-stage MCC. A web-based survey was emailed to 63 ROs, who were identified through publications, guideline panel membership, and/or affiliation with institutions of high MCC volume. ROs provided treatment recommendations for two hypothetical case scenarios of early stage MCC. Among 28 respondents (45%), 26 regularly saw and treated MCC patients for a median 7 years (range, 0-30). While all ROs recommended adjuvant radiation for a stage II upper extremity MCC, only 48% recommended adjuvant radiation for stage I head and neck MCC. Among US-based MCC RO 'specialists,' there is equipoise on the role of radiotherapy for stage I MCC. Ideally, prospective studies are needed to identify which patients may benefit from radiation.


Long-term mutagenicity studies with chloroform and dimethylnitrosamine in female lacI transgenic B6C3F1 mice.
The weight of evidence indicates that chloroform induces cancer in the female B6C3F1 mouse liver via a nongenotoxic-cytotoxic mode of action. However, it is probable that DNA damage occurs secondary to events associated with cytolethality and regenerative cell proliferation. The purpose of the present study was to evaluate the potential mutagenic activity of chloroform in the B6C3F1 lacI transgenic mouse liver mutagenesis assay including mutagenic events that might occur secondary to cytolethality. The positive control, dimethylnitrosamine (DMN) is a DNA-reactive mutagen and carcinogen. DMN-induced mutations were anticipated to require only a brief exposure and without further treatment were predicted to remain unchanged over time at those frequencies. Chloroform-induced mutations secondary to toxicity were anticipated to require longer exposure periods and to occur only under conditions that produced sustained cytolethality and regenerative cell proliferation. Female B6C3F1 lacI transgenic mice were treated with daily doses of 2, 4, or 8 mg/kg of DMN by gavage for 4 days and then held until analysis 10, 30, 90, and 180 days postexposure. Livers from DMN-treated mice exhibited a dose-related 2- to 5-fold increase over control mutant frequencies and remained at those levels for 10 through 180 days postexposure. Thus, following the initial induction by DMN no selective mutation amplification or loss was seen for this extended period of time. Female B6C3F1 lacI mice were exposed daily for 6 hr/day 7 days/week to 0, 10, 30, or 90 ppm chloroform by inhalation, representing nonhepatotoxic, borderline, or overtly hepatotoxic chloroform exposures. Timepoints for determination of lacI mutant frequency were 10, 30, 90, and 180 days of exposure. No increase in lacI mutant frequency in the liver was observed at any dose or timepoint with chloroform, indicating a lack of DNA reactivity. DNA alterations secondary to toxicity either did not occur or were of a type not detectable by lacI mutant frequency analysis, such as large deletions.


Increasing macrolide resistance among Streptococcus agalactiae causing invasive disease in non-pregnant adults was driven by a single capsular-transformed lineage, Portugal, 2009 to 2015.
We characterised Lancefield group B streptococcal (GBS) isolates causing invasive disease among non-pregnant adults in Portugal between 2009 and 2015. All isolates (n = 555) were serotyped, assigned to clonal complexes (CCs) by multilocus sequence typing and characterised by surface protein and pilus island gene profiling. Antimicrobial susceptibility was tested by disk diffusion and resistance genotypes identified by PCR. Overall, serotype Ia was most frequent in the population (31%), followed by serotypes Ib (24%) and V (18%). Serotype Ib increased significantly throughout the study period (p < 0.001) to become the most frequent serotype after 2013. More than 40% of isolates clustered in the CC1/alp3/PI-1+PI-2a genetic lineage, including most isolates of serotypes Ib (n = 110) and V (n = 65). Erythromycin and clindamycin resistance rates were 35% and 34%, respectively, both increasing from 2009 to 2015 (p < 0.010) and associated with CC1 and serotype Ib (p < 0.001). The Ib/CC1 lineage probably resulted from acquisition of the type Ib capsular operon in a single recombination event by a representative of the V/CC1 macrolide-resistant lineage. Expansion of the new serotype Ib/CC1 lineage resulted in increased macrolide resistance in GBS, causing invasive disease among adults in Portugal. The presence of this clone elsewhere may predict more widespread increase in resistance.


Effects of dietary nitrate supplementation on microvascular physiology at 4559 m altitude - A randomised controlled trial (Xtreme Alps).
Native highlanders (e.g. Sherpa) demonstrate remarkable hypoxic tolerance, possibly secondary to higher levels of circulating nitric oxide (NO) and increased microcirculatory blood flow. As part of the Xtreme Alps study (a randomised placebo-controlled trial of dietary nitrate supplementation under field conditions of hypobaric hypoxia), we investigated whether dietary supplementation with nitrate could improve NO availability and microvascular blood flow in lowlanders. Plasma measurements of nitrate, nitrite and nitroso species were performed together with measurements of sublingual (sidestream dark-field camera) and forearm blood flow (venous occlusion plethysmography) in 28 healthy adult volunteers resident at 4559 m for 1 week; half receiving a beetroot-based high-nitrate supplement and half receiving an identically-tasting low nitrate 'placebo'. Dietary supplementation increased plasma nitrate concentrations 4-fold compared to the placebo group, both at sea level (SL; 19.2 vs 76.9 μM) and at day 5 (D5) of high altitude (22.9 vs 84.3 μM, p < 0.001). Dietary nitrate supplementation also significantly increased both plasma nitrite (0.78 vs. 0.86 μM SL, 0.31 vs. 0.41 μM D5, p = 0.03) and total nitroso product (11.3 vs. 19.7 nM SL, 9.7 vs. 12.3 nM D5, p < 0.001) levels both at sea level and at 4559 m. However, plasma nitrite concentrations were more than 50% lower at 4559 m compared to sea level in both treatment groups. Despite these significant changes, dietary nitrate supplementation had no effect on any measured read-outs of sublingual or forearm blood flow, even when environmental hypoxia was experimentally reversed using supplemental oxygen. In conclusion, dietary nitrate supplementation does not improve microcirculatory function at 4559 m.


Inhibiting miRNA in Caenorhabditis elegans using a potent and selective antisense reagent.
Antisense reagents can serve as efficient and versatile tools for studying gene function by inhibiting nucleic acids in vivo. Antisense reagents have particular utility for the experimental manipulation of the activity of microRNAs (miRNAs), which are involved in the regulation of diverse developmental and physiological pathways in animals. Even in traditional genetic systems, such as the nematode Caenorhabditis elegans, antisense reagents can provide experimental strategies complementary to mutational approaches. Presently no antisense reagents are available for inhibiting miRNAs in the nematode C. elegans. We have developed a new class of fluorescently labelled antisense reagents to inhibit miRNAs in developing worms. These reagents were synthesized by conjugating dextran with 2'-O-methyl oligoribonucleotide. The dextran-conjugated antisense reagents can be conveniently introduced into the germline of adult hermaphrodites and are transmitted to their progeny, where they efficiently and specifically inhibit a targeted miRNA in different tissues, including the hypodermis, the vulva and the nervous system. We show that these reagents can be used combinatorially to inhibit more than one miRNA in the same animal. This class of antisense reagents represents a new addition to the toolkit for studying miRNA in C. elegans. Combined with numerous mutants or reporter stains available, these reagents should provide a convenient approach to examine genetic interactions that involve miRNA, and may facilitate studying functions of miRNAs, especially ones whose deletion strains are difficult to generate.See related research article: http://jbiol.com/content/9/3/20.


A retrospective, case-control study on traditional environmental risk factors in inflammatory bowel disease in Vukovar-Srijem County, north-eastern Croatia, 2010.
Traditional environmental risk factors in inflammatory bowel disease (IBD), ulcerative colitis (UC) and Crohn's disease (CD), were examined as part of the retrospective epidemiologic study conducted in Vukovar-Srijem County, north-eastern Croatia in 2010. The geographical variations in the frequency of IBD in Croatia have been observed, which is also the trend in the Central Eastern European region and Europe as a whole, indicating the influence of environmental and lifestyle factors. However, the data on the spread of environmental IBD risk factors are still limited. The purpose of this study was to analyse the traditional environmental risk factors in IBD on our cohort sample, including measles virus infection and vaccination (MMR vaccine-Mumps, Measles, Rubella), tonsillectomy, appendectomy, current and former cigarette smoking and use of oral contraceptives in women. This retrospective, case-control study was performed as a part of a wider epidemiologic study aimed at assessing the incidence, prevalence and clinical expression of IBD, in Vukovar-Srijem County (population: 204,768; 2001), which is a lesser developed part of the continental Croatia that experienced deep demographic changes in the recent past. IBD patients were identified according to the hospital's patient records. There were 119 UC patients and 31 CD patients of a total of 150 patients in the cohort. A total of 150 individuals, volunteers, not having a diagnosis of IBD, age- and sex-matched, were used as the control group. Information on examined risk factors were obtained from all subjects in a previously conducted interview. Patients were contacted personally or by phone and interviewed by a gastroenterologist. There were no differences in the number of smokers, former smokers and non-smokers, between UC and CD patients and the controls, nor in the duration of smoking (years), in current smokers and ex-smokers. Only marginally significant longer time of non-smoking, in ex-smokers was found in IBD patients, compared to the controls, more pronounced in CD patients (p = 0.05). No difference was found in relation to tonsillectomy and risk of IBD. There was no difference in the number of female IBD patients and women from the controls using oral contraceptives. Duration (years) of oral contraceptives use was longer in women from the controls than in female IBD patients (p < 0.001). Frequency of appendectomy was the lowest in UC patients, compared to the controls and CD patients (3.4, 12.0 and 38.7%, respectively) (p < 0.001). No difference was found in relation to measles virus infection and risk of IBD. MMR vaccination rates were higher in CD patients (90.3%), compared to UC patients and the controls (74.8 and 67.3%, respectively) (p = 0.026). No association was found between smoking and tonsillectomy and risk of IBD. Our results do not support the idea of oral contraceptives use as a risk factor for IBD. Frequency of appendectomy was the lowest in UC patients, suggesting that appendectomy decreases the risk of UC, contributing the earlier results. MMR vaccination seemed to be associated with Crohn's disease. These results can add value to our understanding of the increasing incidence of IBD in Croatia and other Central Eastern European countries and can be introductory to future large-scale research.


The effect of dosing regimens on the antimalarial efficacy of dihydroartemisinin-piperaquine: a pooled analysis of individual patient data.
Dihydroartemisinin-piperaquine (DP) is increasingly recommended for antimalarial treatment in many endemic countries; however, concerns have been raised over its potential under dosing in young children. We investigated the influence of different dosing schedules on DP's clinical efficacy. A systematic search of the literature was conducted to identify all studies published between 1960 and February 2013, in which patients were enrolled and treated with DP. Principal investigators were approached and invited to share individual patient data with the WorldWide Antimalarial Resistance Network (WWARN). Data were pooled using a standardised methodology. Univariable and multivariable risk factors for parasite recrudescence were identified using a Cox's regression model with shared frailty across the study sites. Twenty-four published and two unpublished studies (n = 7,072 patients) were included in the analysis. After correcting for reinfection by parasite genotyping, Kaplan-Meier survival estimates were 97.7% (95% CI 97.3%-98.1%) at day 42 and 97.2% (95% CI 96.7%-97.7%) at day 63. Overall 28.6% (979/3,429) of children aged 1 to 5 years received a total dose of piperaquine below 48 mg/kg (the lower limit recommended by WHO); this risk was 2.3-2.9-fold greater compared to that in the other age groups and was associated with reduced efficacy at day 63 (94.4% [95% CI 92.6%-96.2%], p<0.001). After adjusting for confounding factors, the mg/kg dose of piperaquine was found to be a significant predictor for recrudescence, the risk increasing by 13% (95% CI 5.0%-21%) for every 5 mg/kg decrease in dose; p = 0.002. In a multivariable model increasing the target minimum total dose of piperaquine in children aged 1 to 5 years old from 48 mg/kg to 59 mg/kg would halve the risk of treatment failure and cure at least 95% of patients; such an increment was not associated with gastrointestinal toxicity in the ten studies in which this could be assessed. DP demonstrates excellent efficacy in a wide range of transmission settings; however, treatment failure is associated with a lower dose of piperaquine, particularly in young children, suggesting potential for further dose optimisation.


Reverse transcriptase: correlation of zinc content with activity.
Evidence is presented that DNA polymerase of avian myeloblastosis virus has an obligatory zinc requirement for activity. Previous studies indicate that the purified polymerase contains zinc in a stoichiometry of about 1 g-atom/mole. We now find that the enzyme-bound zinc is exchangeable with radioactive (65)Zn; after isoelectric focusing, the radioactive (65)Zn is coincident with polymerase activity. Dialysis of the (65)Zn-labeled polymerase against the chelator, 1,10-phenanthroline, results in a progressive loss of radioactive (65)Zn and polymerase activity. Thereupon, incubation of the inactivated enzyme with Zn(2+) fully restores activity. Thus, the DNA polymerase present in an oncogenic RNA virus, like animal DNA polymerases, can be rigorously classified as a zinc metalloenzyme. DNA polymerase of avian myeloblastosis virus is inactivated by 1,10-phenanthroline at a much faster rate than the bacterial and animal DNA polymerases that have been tested. It may, therefore, be possible to inactivate selectively DNA polymerases from animal tumor viruses by brief exposure to appropriate metal chelators.


THE ABSORPTION OF ADRENALIN AFTER INTRATRACHEAL INJECTION.
In the preceding pages we have submitted evidence which shows that a simple intratracheal injection of a solution in a normally breathing rabbit penetrates within a few seconds to the alveoli, chiefly those of the left lower lobe; that absorption is rapid and well maintained; and that the procedure may be repeated effectively a number of times even with a substance like adrenalin which decreases absorption. It was also shown that absorption of adrenalin from the lung could be obtained at a time when double the dose given intramuscularly exerted no blood pressure effect whatever, and that absorption could still take place after the development of pulmonary edema, when there was an undoubted dilution of the injected solution with a serum-containing liquid and when a diminution of the absorptive field had occurred. The solution injected, after reaching the alveoli, is probably largely taken up by the capillaries of the pulmonary veins. This is indicated by the great rapidity with which an intratracheal injection of adrenalin may cause a rise of blood pressure. In numerous instances, for example, the pressure began to rise less than 5 seconds after the completion of an injection, equaling and even surpassing in rapidity of effect an intramuscular injection. Absorption by the lymphatics probably plays a secondary part, an assumption rendered all themore likely if we consider that lymph nodes are interpolated in the lymphatic pulmonary path, where the bed of the lymph stream becomes greatly widened and the current slowed. Injection into the lungs, however, offers another advantage due to the vascular arrangement of the absorbing field which could be of value therapeutically. Absorption of liquids injected into the lung probably takes place largely through the capillaries of the pulmonary veins; to a slight extent possibly through the capillaries of the bronchial veins which empty partly into the pulmonary veins, partly into the azygos veins; and probably some absorption occurs also through the lymphatics. By far the larger proportion of the absorbed material will thus be rapidly delivered to the left auricle and then to the left ventricle. At each succeeding systole, as long as absorption continues, a fraction of the drug will be driven into the coronary arteries and be able to affect the musculature of the cardiac pump. This fact ought to render the procedure of intratracheal injection a valuable method when it becomes imperative to stimulate a suddenly failing heart as promptly as possible by drugs of the digitalis group. Intratracheal injection is perhaps better under the conditions mentioned than the intravenous route, for the surface veins cannot always be entered with promptness and certainty even under fairly normal conditions, and in cases of cardiac weakness the difficulties will be measurably increased, while an intratracheal injection can be carried out with ease. Moreover, it is legitimate to expect that some absorption will take place from the lung alveoli as long as the heart-lung circulation persists, no matter how feebly, and that thus some of the drug will reach the heart to act on this structure itself more promptly perhaps than when the drug is administered successfully through surface veins. As far as the intramuscular route is concerned, we have shown that the intratracheal injection of adrenalin gives prompt though diminished absorption at a time when double the dose intramuscularly exerts no blood pressure effect whatever. The technical difficulties of giving an intratracheal injection in animals are slight. Tracheotomy as practised by us in the present series of experiments is not necessary, for the injection may be given into the intact trachea without exposure of the trachea. The hypodermic needle is inserted through the skin about 1 cm. below the larynx in a slanting caudad direction; the entrance of the needle into the trachea is readily felt. The injection should not be so rapid that the injected solution fills the entire tracheal lumen, but it should flow down the sides of the trachea. If the lumen is entirely filled, an expiration may drive some of the injected liquid into the larynx causing cough. In our experiments each injection of about 0.5 cc. consumed approximately 5 seconds. In the human subject no data are available as far as our knowledge goes, but a priori it would seem that an intratracheal injection is almost as simple as in the lower animals. The free hypodermic needle could be inserted into the tracheal lumen immediately below the cricoid cartilage. The needle itself should preferably be connected with the syringe by a short length of rubber tubing to minimize the danger of breaking the needle by a sudden move of the patient. The amount of the solution should not be too small, so that at least a fraction of it may reach the alveoli as promptly as possible; 3 to 5 cc. probably would suffice. Insertion of the needle in the locality mentioned would puncture the isthmus of the thyroid, but this is of no significance, especially when the procedure is employed in cases of cardiac failure where the gravity of the condition would warrant incurring much heavier risks than a slight bleeding from the thyroidal isthmus. In conclusion it may be said that the incorporation of drugs by intratracheal injection, while not as generally applicable as other methods, nevertheless has advantages which warrant its use also in human therapeutics.


How urban characteristics affect vulnerability to heat and cold: a multi-country analysis.
The health burden associated with temperature is expected to increase due to a warming climate. Populations living in cities are likely to be particularly at risk, but the role of urban characteristics in modifying the direct effects of temperature on health is still unclear. In this contribution, we used a multi-country dataset to study effect modification of temperature-mortality relationships by a range of city-specific indicators. We collected ambient temperature and mortality daily time-series data for 340 cities in 22 countries, in periods between 1985 and 2014. Standardized measures of demographic, socio-economic, infrastructural and environmental indicators were derived from the Organisation for Economic Co-operation and Development (OECD) Regional and Metropolitan Database. We used distributed lag non-linear and multivariate meta-regression models to estimate fractions of mortality attributable to heat and cold (AF%) in each city, and to evaluate the effect modification of each indicator across cities. Heat- and cold-related deaths amounted to 0.54% (95% confidence interval: 0.49 to 0.58%) and 6.05% (5.59 to 6.36%) of total deaths, respectively. Several city indicators modify the effect of heat, with a higher mortality impact associated with increases in population density, fine particles (PM2.5), gross domestic product (GDP) and Gini index (a measure of income inequality), whereas higher levels of green spaces were linked with a decreased effect of heat. This represents the largest study to date assessing the effect modification of temperature-mortality relationships. Evidence from this study can inform public-health interventions and urban planning under various climate-change and urban-development scenarios.


Antidiabetic effect of Euterpe oleracea Mart. (açaí) extract and exercise training on high-fat diet and streptozotocin-induced diabetic rats: A positive interaction.
A growing body of evidence suggests a protective role of polyphenols and exercise training on the disorders of type 2 diabetes mellitus (T2DM). We aimed to assess the effect of the açaí seed extract (ASE) associated with exercise training on diabetic complications induced by high-fat (HF) diet plus streptozotocin (STZ) in rats. Type 2 diabetes was induced by feeding rats with HF diet (55% fat) for 5 weeks and a single dose of STZ (35 mg/kg i.p.). Control (C) and Diabetic (D) animals were subdivided into four groups each: Sedentary, Training, ASE Sedentary, and ASE Training. ASE (200 mg/kg/day) was administered by gavage and the exercise training was performed on a treadmill (30min/day; 5 days/week) for 4 weeks after the diabetes induction. In type 2 diabetic rats, the treatment with ASE reduced blood glucose, insulin resistance, leptin and IL-6 levels, lipid profile, and vascular dysfunction. ASE increased the expression of insulin signaling proteins in skeletal muscle and adipose tissue and plasma GLP-1 levels. ASE associated with exercise training potentiated the reduction of glycemia by decreasing TNF-α levels, increasing pAKT and adiponectin expressions in adipose tissue, and IR and pAMPK expressions in skeletal muscle of type 2 diabetic rats. In conclusion, ASE treatment has an antidiabetic effect in type 2 diabetic rats by activating the insulin-signaling pathway in muscle and adipose tissue, increasing GLP-1 levels, and an anti-inflammatory action. Exercise training potentiates the glucose-lowering effect of ASE by activating adiponectin-AMPK pathway and increasing IR expression.


Multimodal Imaging of Repetitive Transcranial Magnetic Stimulation Effect on Brain Network: A Combined Electroencephalogram and Functional Magnetic Resonance Imaging Study.
Repetitive transcranial magnetic stimulation (rTMS) has been increasingly used to treat many neurological and neuropsychiatric disorders. However, the clinical response is heterogeneous mainly due to our inability to predict the effect of rTMS on the human brain. Our previous investigation based on functional magnetic resonance imaging (fMRI) suggested that neuroimaging-guided navigation for rTMS could be informed by understanding connectivity patterns that correlate with treatment response. In this study, 20 individuals with a balance disorder called Mal de Debarquement Syndrome completed high-density resting-state electroencephalogram (EEG) and fMRI recordings before and after 5 days of rTMS stimulation over both dorsolateral prefrontal cortices. Based on temporal independent component analysis of source-level EEG data, large-scale electrophysiological resting-state networks were reconstructed and connectivity values in each individual were quantified both before and after treatment. Our results show that high-density, resting-state EEG can reveal connectivity changes in brain networks after rTMS that correlate with symptom changes. The connectivity changes measured by EEG were primarily superficial cortical areas that correlate with previously shown default mode network changes revealed by fMRI. Further, higher baseline EEG connectivity values in the primary visual cortex were predictive of symptom reduction after rTMS. Our findings suggest that multimodal EEG and fMRI measures of brain networks can be biomarkers that correlate with the treatment effect of rTMS. Since EEG is compatible with rTMS, real-time navigation based on an EEG neuroimaging marker may augment rTMS optimization.


[Caregiver burden in relatives of persons with schizophrenia: an overview of measure instruments].
The high prevalence and chronic evolution of schizophrenia are responsible for a major social cost. The adverse consequences of such psychiatric disorders for relatives have been studied since the early 1950s, when psychiatric institutions began discharging patients into the community. According to Treudley (1946) "burden on the family" refers to the consequences for those in close contact with a severely disturbed psychiatric patient. Grad and Sainsbury (1963) and Hoenig and Hamilton (1966) developed the first burden scales for caregivers of severely mentally ill patients, and a number of authors further developed instruments trying to distinguish between "objective" and "subjective" burden. Objective burden concerns the patient's symptoms, behaviour and socio-demographic characteristics, but also the changes in household routine, family or social relations, work, leisure time, physical health.... Subjective burden is the mental health and subjective distress among family members. While the first authors referred to those problems which are deemed to be related to, or caused by the patient, Platt et al. (1983) tried to distinguish between the occurrence of a problem, its alleged aetiology, and the perceived distress, when developing the SBAS questionnaire. These authors also proposed separate evaluations of behavioral disturbance and social performance by relatives, and a report of extra-disease stressors in family life. The SBAS is actually the most complete, but also complex instrument for evaluating burden in caregivers. Since 1967 Pasamanick and others proposed questionnaires for burden evaluation in relatives of schizophrenic patients. Relatives may be included in specific psychoeducational programs, but few of these programs have been evaluated in terms of caregiver burden. The theoretical approach in schizophrenia was not different from that one adopted in mentally ill population. Some instruments were validated first in a mentally ill group and then adapated for schizophrenic population. This paper describes the available data about intruments measuring caregiver burden in relatives of schizophrenic patients. Measures are summarized according to purpose, content and psychometric properties. Sixteen instruments have been collected from the litterature (1955-2001), and 2 instruments developed for relatives of mentally ill have also been taken into account. A group of 5 instruments focuses on the measurement of behavioural disturbance in persons with schizophrenia as perceived by their family members. Eleven scales include behavioural disturbance in a more complete decription of objective burden. Thirteen questionnaires also report the subjective distress in caregivers. One instrument has been developed in french language. Few of these instruments have been developed from a verbatim and really describe the caregiver's point of view. Most of them rely on experts point of view or on previously published studies. The content and domains explored by these instruments are variable. The psychometric properties are poorly documented for a number of them and no information is published about responsiveness. Some validated instruments are the Perceived Family Burden Scale (PFBS) the Involvement Evaluation Questionnaire (IEQ) and the Experience of Caregiving Inventory (ECI). In past studies, researchers more or less agreed about the dimensions that comprise the family burden. There was less agreement with regard to the definition of objective and subjective burden, and quite no agreement about the theoretical approach to the concept of burden. The evaluation of behavioural disturbance should now be excluded from the objective burden dimension. It is a specific domain, both objective and subjective, which can be described as a stressor in the stress-appraisal-coping model. A good approach of this domain can be found in the PFBS. It comprises 24 items and the principal components analysis produces 2 factors ("active" and "passive"), explaining 35% of the variance, with good consitency and acceptable test-retest reliability. The evaluation is both objective (presence or absence) and subjective (induced distress). The Behavior Disturbance Scale (BDS) may also be taken into account, although it is less validated. This scale derives from the SBAS, modified as a self-questionnaire, with both objective and subjective evaluations of all items. The concept of burden was recently modified in a new theoretical approach by Schene, when developing the IEQ. According to this author, the burden scale is supposed to exclude stressors (patient's behaviors), as well as outcome variables (distress or psychological impairment in caregiver). The "caregiving consequences" section comprises 36 items, which focus on the subjective aspects of the caregiver's experience. Principal component factor analysis generates 4 factors which explain 45% of the variance: tension, supervision, worrying, urging. The overall caregiving score substantially explains the connection between patient, caregiver, relationship variables and the caregiver's distress. This scale is a valid and simple instrument for caregiving eveluation The ECI also introduces a new approach of caregiving and rejects the notion of burden. The 66 item version is composed of 10 factors (8 "negative" and 2 "positive") with good internal consistency. The introduction of two positive factors (rewarding personal experiences, good aspects of the relationship with the patient) might be the basis of a useful outcome measure for intervention aimed at promoting caregiver well-being. Nevertheless, the authors fail to develop an overall score that includes these factors, and focus on the negative dimensions as predictors of morbidity and well-being. None of the variables included in the regression model explain a significant percent of the variance of the ECI positive score. None of these instruments was employed for evaluating programs or treatments, even psychoeducational programs for caregivers. This may be partly due to the lack of data about sensitivity to change. No instrument is now available for evaluating therapeutics from the caregiver's point of view. Developing such an instrument is necessary, in view of the increasing role of families in care for schizophrenic patients. These data and the review of the literature leeds us to propose the development of a self-administered questionnaire for evaluating subjective health-related quality of life in caregivers of schizophrenic patients. The instrument should be developed from the caregiver's point of view and be derived from qualitative interviews with relatives of patients suffering from schizophrenia. It's responsiveness should be documented before inclusion in clinical trials or evaluation of psychoeducational programs. We are now working with the National Union of Friends and Families of Patients to validate an instrument in french language.


Parental dietary seleno-L-methionine exposure and resultant offspring developmental toxicity.
Selenium (Se) leaches into water from agricultural soils and from storage sites for coal fly ash. Se toxicity causes population and community level effects in fishes and birds. We used the laboratory aquarium model fish, Japanese medaka (Oryzias latipes), an asynchronous breeder, to determine aspects of uptake in adults and resultant developmental toxicity in their offspring. The superior imaging properties of the model enabled detailed descriptions of phenotypic alterations not commonly reported in the existing Se literature. Adult males and females in treatment groups were exposed, separately and together, to a dry diet spiked with 0, 12.5, 25, or 50 μg/g (dry weight) seleno-L-methionine (SeMet) for 6 days, and their embryo progeny collected for 5 days, maintained under controlled conditions and observed daily for hatchability, mortality and/or developmental toxicity. Sites of alteration included: craniofacial, pericardium and abdomen (Pc/Ab), notochord, gall bladder, spleen, blood, and swim bladder. Next, adult tissue Se concentrations (liver, skeletal muscle, ovary and testis) were determined and compared in treatment groups of bred and unbred individuals. No significant difference was found across treatment groups at the various SeMet concentrations; and, subsequent analysis compared exposed vs. control in each of the treatment groups at 10 dpf. Increased embryo mortality was observed in all treatment groups, compared to controls, and embryos had a decreased hatching rate when both parents were exposed. Exposure resulted in significantly more total altered phenotypes than controls. When altered phenotypes following exposure of both parents were higher than maternal only exposure, a male role was suggested. The comparisons between treatment groups revealed that particular types of phenotypic change may be driven by the sex of the exposed parent. Additionally, breeding reduced Se concentrations in some adult tissues, specifically the liver of exposed females and skeletal muscle of exposed males. Detailed phenotypic analysis of progeny from SeMet exposed parents should inform investigations of later life stages in an effort to determine consequences of early life exposure.


Methodological challenges in the application of the glycemic index in epidemiological studies using data from the European Prospective Investigation into Cancer and Nutrition.
Associations between the glycemic index (GI) or glycemic load (GL) and diseases are heterogeneous in epidemiological studies. Differences in assigning GI values to food items may contribute to this inconsistency. Our objective was to address methodological issues related to the use of current GI and GL values in epidemiological studies. We performed ecological comparison and correlation studies by calculating dietary GI and GL from country-specific dietary questionnaires (DQ) from 422,837 participants from 9 countries participating in the European Prospective Investigation into Cancer and Nutrition study and single standardized 24-h dietary recalls (24-HDR) obtained from a representative sample (n = 33,404) using mainly Foster Powell's international table as a reference source. Further, 2 inter-rater and 1 inter-method comparison were conducted, comparing DQ GI values assigned by independent groups with values linked by us. The ecological correlation between DQ and 24-HDR was good for GL (overall r = 0.76; P < 0.005) and moderate for GI (r = 0.57; P < 0.05). Mean GI/GL differences between DQ and 24-HDR were significant for most centers. GL but not GI from DQ was highly correlated with total carbohydrate (r = 0.98 and 0.15, respectively; P < 0.0001) and this was higher for starch (r = 0.72; P < 0.0001) than for sugars (r = 0.36; P < 0.0001). The inter-rater and inter-method variations were considerable for GI (weighted kappa coefficients of 0.49 and 0.65 for inter-rater and 0.25 for inter-method variation, respectively) but only mild for GL (weighted kappa coefficients > 0.80). A more consistent methodology to attribute GI values to foods and validated DQ is needed to derive meaningful GI/GL estimates for nutritional epidemiology.


Voice Disorders in Teacher Students-A Prospective Study and a Randomized Controlled Trial.
Teachers are at risk of developing voice disorders, but longitudinal studies on voice problems among teachers are lacking. The aim of this randomized trial was to investigate long-term effects of voice education for teacher students with mild voice problems. In addition, vocal health was examined prospectively in a group of students without voice problems. First-semester students answered three questionnaires: one about background factors, one about voice symptoms (Screen6), and the Voice Handicap Index. Students with voice problems according to the questionnaire results were randomized to a voice training group or a control group. At follow-up in the sixth semester, all students answered Screen6 again together with four questions about factors that could have affected vocal health during their teacher education. The training group and the control group also answered the Voice Handicap Index a second time. At follow-up, 400 students remained in the study: 27 in the training group, 54 in the control group, and 319 without voice problems at baseline. Voice problems had decreased somewhat more in the training group than in the control group, but the difference was not statistically significant (P = 0.1). However, subgroup analyses showed significantly larger improvement among the students in the group with complete participation in the training program compared with the group with incomplete participation. Of the 319 students without voice problems at baseline, 14% had developed voice problems. Voice problems often develop in teacher students. Despite extensive dropout, our results support the hypothesis that voice education for teacher students has a preventive effect.


Chitosan monomer accelerates alkaline phosphatase activity on human osteoblastic cells under hypofunctional conditions.
Chitosan is a natural polyaminosaccharide that is extensively applied as an antitumor and antirheumatic drug. However, there are few reports about its effects on hypofunctional osteoblasts in vitro. We investigated the biological characteristics of a human osteoblastic cell line (NOS-1 cells) that was cultured with a chitosan monomer-containing medium under simulated microgravity conditions. After 7 days of cell incubation under the conventional conditions, the flasks were transferred to a microgravity simulator for 3 days. In the 0.005% chitosan monomer supplemented group, the marker enzyme of biological mineralization, the alkaline phosphatase (ALP) activity, was significantly higher compared with the control group (p<0.05). A cDNA microarray was performed to investigate the effects on the mRNA level by chitosan monomer, and the fluorescent signal was analyzed. The interferon gamma (IFN-gamma) receptor gene was detected with a signal ration of 2.2. The slight increase of IFN-gamma receptor expression was confirmed after 3 days of incubation according to RT-PCR analysis. Western blot analysis also showed the increased expression of IFN-gamma receptor. These results suggest that a supra-low concentration of chitosan monomer may increase the ALP activity of osteoblastic cells through the IFN-gamma receptor at the early phase of cell culture and recover the activity for biological mineralization under the hypofunctional condition.


Isolation and characterization of a bacteriophage infectious to an extreme thermophile, Thermus thermophilus HB8.
A bacteriophage (phiYS40) infectious to an extreme thermophile, Thermus thermophilus HB8, was isolated and characterized. phiYS40 grows over the temperature range of 56 to 78 C, and the optimum growth temperature is about 65 C. The phage had a latent period of 80 min and a burst size of about 80 at 65 C. The phage has a hexagonal head 0.125 mum in diameter, a tail 0.178 mum long and 0.027 mum wide, a base plate and tail fibers. The phage is thermostable in broth but rather unstable in a buffer containing 10 mM Tris, 10 mM MgCl2, pH 7.5. The addition of Casamino Acids (1 percent), polypeptone (0.8 percent), yeast extract (0.4 percent), NaCl (0.1 M) or spermidine (1 mM) to the buffer restores the thermostability of phiYS40 to the same degree as in broth. The phage is also thermostable in water of the hot spring from which this phage was isolated. The nucleic acid of PhiYS40 is a double-stranded DNA and has a molecular weight of 1.36 X 10-8. The guanine plus cytosine content of the DNA was determined to be about 35 percent from chemical determinations, buoyant density (1.693 g/cm-3 in CsCl), and melting temperature (83.5 C in 0.15 M NaCl plus 0.015 M sodium citrate).


Risk factors for urinary incontinence in Taiwanese women aged 20-59 years.
To assess the risk factors for urinary incontinence among Taiwanese women. A sample of 4,549 women was selected using a multistage random sampling protocol. The women selected were interviewed face-to-face by well-trained interviewers. The usual risk factors, which included marital status, age, body mass index, menstrual status, alcohol intake, drug allergy, diabetes mellitus, hypertension and gynecologic events, were evaluated. The factors were assessed by frequency analysis and logistic regression analysis using a significance level of less than 0.05. A total of 3,537 women were successfully interviewed, producing a response rate of 77.8% (3,735/4,549). The prevalence of urinary incontinence increased significantly with marriage (21.7%; 95% confidence interval, CI, 20.2-23.2), alcohol intake (24.9%; 95% CI, 19.3-30.5), drug allergy (24.5%; 95% CI, 19.9-29.2), diabetes mellitus (40.3%; 95% CI, 29.3-51.2), hypertension (30.1%; 95% CI, 23.8-36.4), body mass index (odds ratio, 1.05 per unit increase; 95% CI, 1.02-1.09) and a previous gynecologic operation (25.5%; 95% CI, 19.9-31.2). Age was also a risk factor for urinary incontinence (odds ratio, 1.04; 95% CI, 1.03-1.05), but there was no relationship between urinary incontinence and parity, route of delivery, smoking or menstrual status. There is a high prevalence of urinary incontinence among women who suffer from diabetes or hypertension, or who have undergone a gynecologic operation, in particular hysterectomy. From a public health viewpoint, it is important to promote better health education in order to improve understanding of urinary incontinence and its risk factors and to increase the awareness of the availability of mainstream treatments.


Dopamine signaling is required for depolarization-induced slow current in cerebellar Purkinje cells.
Brief strong depolarization of cerebellar Purkinje cells produces a slow inward cation current. This current, called depolarization-induced slow current (DISC), is triggered by Ca influx in the Purkinje cell and is attenuated by a blocker of vesicular fusion. Previous work in other brain regions, such as the substantia nigra and ventral tegmental area, has shown that dopamine can be released from dendrites to produce paracrine and autocrine signaling. Here, we test the hypothesis that postsynaptic release of dopamine and autocrine activation of dopamine receptors is involved in DISC. Light immunohistochemistry showed that D(3) dopamine receptors, vesicular monoamine transporter type 2 (VMAT2), and dopamine plasma membrane transporters (DATs) were all expressed in cerebellar Purkinje cells. However, their expression was strongest in the gyrus region of cerebellar lobules IX and X. Comparison of DISC across lobules revealed that it was weak in the anterior portions of the cerebellum (lobules II, V, and VI) and strong in lobules IX and X. DISC was blocked by dopamine receptor antagonists (haloperidol, clozapine, eticlopride, and SCH23390). Likewise, DISC was strongly attenuated by inhibitors of VMAT (reserpine and tetrabenazine) and DAT (GBR12909 and rimcazole). These drugs did not produce DISC attenuation through blockade of depolarization-evoked Purkinje cell Ca transients. Purkinje cells in cerebellar slices derived from DAT-null mice expressed DISC, but this DISC ran down at a significantly higher rate than littermate controls. Together, these results suggest that strong Purkinje cell depolarization produces Ca-dependent release of vesicular postsynaptic dopamine that then excites Purkinje cells in an autocrine manner.


Escherichia coli O157:H7 infections associated with consumption of locally grown strawberries contaminated by deer.
An outbreak of Escherichia coli O157:H7 was identified in Oregon through an increase in Shiga toxin-producing E. coli cases with an indistinguishable, novel pulsed-field gel electrophoresis (PFGE) subtyping pattern. We defined confirmed cases as persons from whom E. coli O157:H7 with the outbreak PFGE pattern was cultured during July-August 2011, and presumptive cases as persons having a household relationship with a case testing positive for E. coli O157:H7 and coincident diarrheal illness. We conducted an investigation that included structured hypothesis-generating interviews, a matched case-control study, and environmental and traceback investigations. We identified 15 cases. Six cases were hospitalized, including 4 with hemolytic uremic syndrome (HUS). Two cases with HUS died. Illness was significantly associated with strawberry consumption from roadside stands or farmers' markets (matched odds ratio, 19.6; 95% confidence interval, 2.9-∞). A single farm was identified as the source of contaminated strawberries. Ten of 111 (9%) initial environmental samples from farm A were positive for E. coli O157:H7. All samples testing positive for E. coli O157:H7 contained deer feces, and 5 tested farm fields had ≥ 1 sample positive with the outbreak PFGE pattern. The investigation identified fresh strawberries as a novel vehicle for E. coli O157:H7 infection, implicated deer feces as the source of contamination, and highlights problems concerning produce contamination by wildlife and regulatory exemptions for locally grown produce. A comprehensive hypothesis-generating questionnaire enabled rapid identification of the implicated product. Good agricultural practices are key barriers to wildlife fecal contamination of produce.


Pharmacology of traumatic brain injury.
The intensity of experimental and clinical research to identify a neuroprotective drug for the treatment of traumatic brain injury is motivated by the devastating morbidity and mortality of this condition. Encouraging experimental work has led so far to disappointing clinical trials and the identification of new potential therapeutic targets is critically dependent on a better understanding of the chronic pathophysiology triggered by the initial insult. Future advances in the pharmacological treatment of traumatic brain injury are likely to include the evaluation of sequentially timed therapies combining multiple and targeted agents, and manipulation of the newly discovered neurogenic potential of the adult brain together with the refinement of traditional interventions to block specific cytotoxic cascades.


Therapeutic Drug Monitoring of Antiepileptic Drugs in Epilepsy: A 2018 Update.
Antiepileptic drugs (AEDs) are the mainstay of epilepsy treatment. Since 1989, 18 new AEDs have been licensed for clinical use and there are now 27 licensed AEDs in total for the treatment of patients with epilepsy. Furthermore, several AEDs are also used for the management of other medical conditions, for example, pain and bipolar disorder. This has led to an increasingly widespread application of therapeutic drug monitoring (TDM) of AEDs, making AEDs among the most common medications for which TDM is performed. The aim of this review is to provide an overview of the indications for AED TDM, to provide key information for each individual AED in terms of the drug's prescribing indications, key pharmacokinetic characteristics, associated drug-drug pharmacokinetic interactions, and the value and the intricacies of TDM for each AED. The concept of the reference range is discussed as well as practical issues such as choice of sample types (total versus free concentrations in blood versus saliva) and sample collection and processing. The present review is based on published articles and searches in PubMed and Google Scholar, last searched in March 2018, in addition to references from relevant articles. In total, 171 relevant references were identified and used to prepare this review. TDM provides a pragmatic approach to epilepsy care, in that bespoke dose adjustments are undertaken based on drug concentrations so as to optimize clinical outcome. For the older first-generation AEDs (carbamazepine, ethosuximide, phenobarbital, phenytoin, primidone, and valproic acid), much data have accumulated in this regard. However, this is occurring increasingly for the new AEDs (brivaracetam, eslicarbazepine acetate, felbamate, gabapentin, lacosamide, lamotrigine, levetiracetam, oxcarbazepine, perampanel, piracetam, pregabalin, rufinamide, stiripentol, sulthiame, tiagabine, topiramate, vigabatrin, and zonisamide).


Fertilizer from dried human urine added to ash and lime - a potential product from eco-sanitation system.
This research explored the possibility of making fertilizer at a laboratory from source separated and untreated human urine added to ash and lime by drying at low temperatures. A mixture of ash and lime (1:1) was used as drying agent and human urine was applied as undiluted and fresh. Ash and lime were chosen as drying agents for maintaining a pH > 10 during the drying process, which should inhibit urea hydrolysis in urine, and thereby urea should be retained in the drying agent. The drying technique was developed and drying capacity of the system was quantified; three specific temperatures (20 °, 35 °, 60 °C) and two airflow rates (1 L/min and 5 L/min) were used in the experiment. A mass balance for nitrogen in the system was obtained. It was evident from the experiment that urea can be retained by maintaining a high pH (>10). Urine drying at 20 °C was not a feasible option, since rate of evaporation was very low. The highest retention of inflow nitrogen at 35 °C and 60 °C were 74% and 54%, respectively, in the produced fertilizer. Reduced evaporation rate, flooding of urine over drying agent, and blockage in airflow influenced nitrogen loss and concentration of nitrogen in the final product.


A game-theoretic model of kleptoparasitic behavior in polymorphic populations.
Kleptoparasitism, the stealing of food by one animal from another, is a widespread biological phenomenon. In this paper we build upon earlier models to investigate a population of conspecifics involved in foraging and, potentially, kleptoparasitism. We assume that the population is composed of four types of individuals, according to their strategic choices when faced with an opportunity to steal and to resist an attack. The fitness of each type of individual depends upon various natural parameters, for example food density, the handling time of a food item and the probability of mounting a successful attack against resistance, as well as the choices that they make. We find the evolutionarily stable strategies (ESSs) for all parameter combinations and show that there are six possible ESSs, four pure and two mixtures of two strategies, that can occur. We show that there is always at least one ESS, and sometimes two or three. We further investigate the influence of the different parameters on when each type of solution occurs.


Engineering hyperthermostability into a GH11 xylanase is mediated by subtle changes to protein structure.
Understanding the structural basis for protein thermostability is of considerable biological and biotechnological importance as exemplified by the industrial use of xylanases at elevated temperatures in the paper pulp and animal feed sectors. Here we have used directed protein evolution to generate hyperthermostable variants of a thermophilic GH11 xylanase, EvXyn11. The Gene Site Saturation Mutagenesis (GSSM) methodology employed assesses the influence on thermostability of all possible amino acid substitutions at each position in the primary structure of the target protein. The 15 most thermostable mutants, which generally clustered in the N-terminal region of the enzyme, had melting temperatures (Tm) 1-8 degrees C higher than the parent protein. Screening of a combinatorial library of the single mutants identified a hyperthermostable variant, EvXyn11TS, containing seven mutations. EvXyn11TS had a Tm approximately 25 degrees C higher than the parent enzyme while displaying catalytic properties that were similar to EvXyn11. The crystal structures of EvXyn11 and EvXyn11TS revealed an absence of substantial changes to identifiable intramolecular interactions. The only explicable mutations are T13F, which increases hydrophobic interactions, and S9P that apparently locks the conformation of a surface loop. This report shows that the molecular basis for the increased thermostability is extraordinarily subtle and points to the requirement for new tools to interrogate protein folding at non-ambient temperatures.


Exploring the vascular smooth muscle receptor landscape in vivo: ultrasound Doppler versus near-infrared spectroscopy assessments.
Ultrasound Doppler and near-infrared spectroscopy (NIRS) are routinely used for noninvasive monitoring of peripheral hemodynamics in both clinical and experimental settings. However, the comparative ability of these methodologies to detect changes in microvascular and whole limb hemodynamics during pharmacological manipulation of vascular smooth muscle receptors located at varied locations within the arterial tree is unknown. Thus, in 10 healthy subjects (25 ± 2 yr), changes in resting leg blood flow (ultrasound Doppler; femoral artery) and muscle oxygenation (oxyhemoglobin + oxymyoglobin; vastus lateralis) were simultaneously evaluated in response to intra-arterial infusions of phenylephrine (PE, 0.025-0.8 μg·kg(-1)·min(-1)), BHT-933 (2.5-40 μg·kg(-1)·min(-1)), and angiotensin II (ANG II, 0.5-8 ng·kg(-1)·min(-1)). All drugs elicited significant dose-dependent reductions in leg blood flow and oxyhemoglobin + oxymyoglobin. Significant relationships were found between ultrasound Doppler and NIRS changes across doses of PE (r(2) = 0.37 ± 0.08), BHT-933 (r(2) = 0.74 ± 0.06), and ANG II (r(2) = 0.68 ± 0.13), with the strongest relationships evident with agonists for receptors located preferentially "downstream" in the leg microcirculation (BHT-933 and ANG II). Analyses of drug potency revealed similar EC50 between ultrasound Doppler and NIRS measurements for PE (0.06 ± 0.02 vs. 0.10 ± 0.01), BHT-933 (5.0 ± 0.9 vs. 4.5 ± 1.3), and ANG II (1.4 ± 0.8 vs. 1.3 ± 0.3). These data provide evidence that both ultrasound Doppler and NIRS track pharmacologically induced changes in peripheral hemodynamics and are equally capable of determining drug potency. However, considerable disparity was observed between agonist infusions targeting different levels of the arterial tree, suggesting that receptor landscape is an important consideration for proper interpretation of hemodynamic monitoring with these methodologies.


Effects of Choline, Betaine, and Wheat Floral Extracts on Growth of Fusarium graminearum.
Fusarium head blight has been more severe when infection occurs during anthesis, indicating that floral organs may be important infection courts. Choline acetate and glycinebetaine have been extracted from wheat and reported to be growth stimulants of Fusarium graminearum. They are hypothesized to enhance infection and tissue colonization. Growth of F. graminearum was examined on media amended with extracts from floral parts of nine wheat genotypes with various Fusarium head blight resistance levels. Results indicated no significant effect of anther, palea, or lemma extracts on radial growth when compared with unamended controls. Effects on spore germination and hyphal growth of F. graminearum by choline, betaine, and an equimolar mixture at concentrations ranging from 0.01 to 1,000 μM also were examined. Spore germination was not significantly (P ≤ 0.05) affected by choline, betaine, or a combination of the compounds compared with unamended controls. Radial hyphal growth also was not consistently affected (P ≤ 0.05) by choline or betaine when compared with controls. Equimolar mixtures of the two compounds showed significant slight reduction in growth rate at higher concentrations when compared with controls. The reduction in growth rate was due to higher concentrations of betaine. Results of this study indicate that endogenous compounds in floral parts may not be associated with wheat resistance to F. graminearum.


The inter-locus recombinant HLA-B*4601 has high selectivity in peptide binding and functions characteristic of HLA-C.
The vast majority of new human HLA class I alleles are formed by conversions between existing alleles of the same locus. A notable exception to this rule is HLA-B*4601 formed by replacement of residues 66-76 of the alpha 1 helix of B*1501 by the homologous segment of Cw*0102. This inter-locus recombination, which brings together characteristic elements of HLA-B and HLA-C structure, is shown here to influence function dramatically. Naturally processed peptides bound by B*4601 are distinct from those of its parental allotypes B*1501 and Cw*0102 and dominated by three high abundance peptides. Such increased peptide selectivity by B*4601 is unique among HLA-A,B,C allotypes. For other aspects of function, presence of the small segment of HLA-C-derived sequence in an otherwise HLA-B framework converts B*4601 to an HLA-C-like molecule. Alloreactive cytotoxic T lymphocytes (CTL), natural killer (NK) cells, and cellular glycosidases all recognize B*4601 as though it were an HLA-C allotype. These unusual properties are those of an allotype which has frequencies as high as 20% in south east Asian populations and is associated with predisposition to autoimmune diseases and nasopharyngeal carcinoma.


Phylogenetic analysis and real time PCR detection of a presumbably undescribed Peronospora species on sweet basil and sage.
Downy mildew of sweet basil (Ocimum basilicum) has become a serious disease issue for the producers of sweet basil in Switzerland since it was first recorded in 2001. Reported in Africa in Uganda as early as 1933, major outbreaks of this disease in Europe were first noted in Italy in 1999 and in the USA from 1993. Previous reports have named the pathogen as Peronospora lamii. Its preferential hosts belong to the Lamiaceae family including basils (Ocimum spp.), mints (Menta spp.), sages (Salvia spp.) and other aromatics. This study investigated the taxonomic status of the downy mildew pathogen, using both morphological characters and molecular analysis of the ITS region of the rDNA. The inherent variability of conidial dimensions made species differentiation difficult. Sequence homology and phylogenetic analysis of nine collections of the Peronospora on sweet basil showed unique ITS sequences distinct from those of P. lamii and any other sequenced Peronospora species. This paper describes and illustrates the morphology of this presumably undescribed species of Peronospora. Its taxonomic position and relationships with other related species in the same genus are presented and discussed. In addition to this work, PCR primers for real time PCR analysis have been developed for the specific detection of this downy mildew pathogen from infected tissues or seeds. It is shown that these primers can also be used in classic PCR.


Stair design in the United States and obesity: the need for a change.
Maintenance of healthy body weight is a function of total energy expenditure including household and workplace activity. Light to moderate physical activity has been found to be effective in motivating sedentary and obese individuals, and is at least as effective as structured exercise in lowering weight in some studies. Stair use offers a promising intervention for increasing physical activity, because it involves a lifestyle choice that must be made (people must get to their destination), and it requires no personal financial cost. Stairs in United States buildings are frequently hidden from entrances with small signs denoting their location, mainly in connection to fire exits. Using the stairs is usually seen as a way of escaping from fires rather than as a recommended daily activity. To comply with State Fire Marshal regulations, stairs are usually guarded by heavy, spring doors, without air conditioning, and are noncarpeted. In this article, several suggestions to change the architectural design in buildings to be more physical activity-friendly are discussed. Such changes would make stairs attractive, safe, and readily accessible. Local and state authorities may also allow incentives for such designs to compensate for their additional costs. Moreover, standard national building codes that incorporate health concerns should be devised. Stair use at work and in living places has the potential to increase physical activity and decrease obesity. Changing stair design to encourage their use requires a series of interventions both architecturally and legislatively to create physical environments that support active lifestyles.


Absolute risk of endometrial carcinoma during 20-year follow-up among women with endometrial hyperplasia.
PURPOSE The severity of endometrial hyperplasia (EH)-simple (SH), complex (CH), or atypical (AH)-influences clinical management, but valid estimates of absolute risk of clinical progression to carcinoma are lacking. Materials and METHODS We conducted a case-control study nested in a cohort of 7,947 women diagnosed with EH (1970-2002) at one prepaid health plan who remained at risk for at least 1 year. Patient cases (N = 138) were diagnosed with carcinoma, on average, 6 years later (range, 1 to 24 years). Patient controls (N = 241) were matched to patient cases on age at EH, date of EH, and duration of follow-up, and they were counter-matched to patient cases on EH severity. After we independently reviewed original slides and medical records of patient controls and patient cases, we combined progression relative risks (AH v SH, CH, or disordered proliferative endometrium [ie, equivocal EH]) from the case-control analysis with clinical censoring information (ie, hysterectomy, death, or left the health plan) on all cohort members to estimate interval-specific (ie, 1 to 4, 5 to 9, and 10 to 19 years) and cumulative (ie, through 4, 9, and 19 years) progression risks. Results For nonatypical EH, cumulative progression risk increased from 1.2% (95% CI, 0.6% to 1.9%) through 4 years to 1.9% (95% CI, 1.2% to 2.6%) through 9 years to 4.6% (95% CI, 3.3% to 5.8%) through 19 years after EH diagnosis. For AH, cumulative risk increased from 8.2% (95% CI, 1.3% to 14.6%) through 4 years to 12.4% (95% CI, 3.0% to 20.8%) through 9 years to 27.5% (95% CI, 8.6% to 42.5%) through 19 years after AH. CONCLUSION Cumulative 20-year progression risk among women who remain at risk for at least 1 year is less than 5% for nonatypical EH but is 28% for AH.


Restitution slope is principally determined by steady-state action potential duration.
The steepness of the action potential duration (APD) restitution curve and local tissue refractoriness are both thought to play important roles in arrhythmogenesis. Despite this, there has been little recognition of the apparent association between steady-state APD and the slope of the restitution curve. The objective of this study was to test the hypothesis that restitution slope is determined by APD and to examine the relationship between restitution slope, refractoriness and susceptibility to VF. Experiments were conducted in isolated hearts and ventricular myocytes from adult guinea pigs and rabbits. Restitution curves were measured under control conditions and following intervention to prolong (clofilium, veratridine, bretylium, low [Ca]e, chronic transverse aortic constriction) or shorten (catecholamines, rapid pacing) ventricular APD. Despite markedly differing mechanisms of action, all interventions that prolonged the action potential led to a steepening of the restitution curve (and vice versa). Normalizing the restitution curve as a % of steady-state APD abolished the difference in restitution curves with all interventions. Effects on restitution were preserved when APD was modulated by current injection in myocytes pre-treated with the calcium chelator BAPTA-AM - to abolish the intracellular calcium transient. The non-linear relation between APD and the rate of repolarization of the action potential is shown to underpin the common influence of APD on the slope of the restitution curve. Susceptibility to VF was found to parallel changes in APD/refractoriness, rather than restitution slope. Steady-state APD is the principal determinant of the slope of the ventricular electrical restitution curve. In the absence of post-repolarization refractoriness, factors that prolong the action potential would be expected to steepen the restitution curve. However, concomitant changes in tissue refractoriness act to reduce susceptibility to sustained VF. Dependence on steady-state APD may contribute to the failure of restitution slope to predict sudden cardiac death.