Testicular hypotrophy does not correlate with grade of adolescent varicocele.
Testicular hypotrophy is the most widely accepted indication for correcting adolescent varicocele. Previous studies in adolescents have shown a relationship between increasing grade of varicocele and the likelihood of testicular hypotrophy. As this relationship has significant clinical implications, we studied the correlation between grade and testicular volume disproportion in our adolescent varicocele population. We reviewed the adolescent varicocele database at our institution. A total of 168 patients 8 to 21 years old were studied. We routinely calculated testis volumes using scrotal ultrasound. Testicular disproportion was calculated using the equation [(size of unaffected testis) - (size of affected testis)]/(size of unaffected testis) x 100%. Disproportion was categorized as less than 10%, 10% to 20% and more than 20%. Varicoceles were graded by an attending urologist with the patient standing, using the system of Dubin and Amelar. Analysis of variance and Pearson chi-square indicated no significant differences in volume differential between varicocele grades. Mean +/- SD volume differential was 18% +/- 15% for grade I, 25% +/- 20% for grade II and 19% +/- 14% for grade III. ANOVA revealed no significant difference in mean volume differential between the 3 varicocele grades (p = 0.10). When categorizing patients into 3 levels of volume differential (less than 10%, 10% to 20%, more than 20%) no significant correlation was observed between varicocele and volume differential (p = 0.48, chi-square test). Grade of varicocele does not correlate with presence or severity of testicular disproportion in adolescent boys with varicocele as measured by scrotal ultrasound.


Multimorbidity and functional decline in community-dwelling adults: a systematic review.
Multimorbidity affects up to one quarter of primary care populations. It is associated with reduced quality of life, an increased risk of mental health difficulties and increased healthcare utilisation. Functional decline is defined as developing difficulties with activities of daily living and is independently associated with poorer health outcomes. The aim of this systematic review was to examine the association between multimorbidity and functional decline and to what extent multimorbidity predicts future functional decline. A systematic literature search (1990-2014) and narrative analysis was conducted. Population; Community-dwelling adults (≥18 years), Risk; Multimorbidity defined as the presence of ≥2 chronic medical conditions in an individual, Primary outcome; Physical functional decline measured using a validated instrument, Study design; cross-sectional or cohort studies. The following databases were included: PubMed, EMBASE, CINAHL, the Cochrane Library and the International Research Community on Multimorbidity (IRCMo) publication list. Methodological quality assessment of included studies was conducted with a suitable risk of bias tool. A total of 37 studies were eligible for inclusion (28 cross-sectional studies and 9 cohort studies). The majority of cross-sectional studies (n = 24/28) demonstrated a consistent association between multimorbidity and functional decline. Twelve of these studies reported that increasing numbers of chronic condition counts were associated with worsening functional decline. Nine cohort studies included 14,133 study participants with follow-up periods ranging from one to six years. The majority (n = 5) found that multimorbidity predicted functional decline. Of the five studies that reported the impact of increasing numbers of conditions, all reported greater functional decline with increasing numbers of conditions. One study examined disease severity and found that this also predicted greater functional decline. Overall, cohort studies were of good methodological quality but were mixed in terms of participants, multimorbidity definitions, follow-up duration, and outcome measures. The available evidence indicates that multimorbidity predicts future functional decline, with greater decline in patients with higher numbers of conditions and greater disease severity. This review highlights the importance of considering physical functioning when designing interventions and systems of care for patients with multimorbidity, particularly for patients with higher numbers of conditions and greater disease severity.


Aspirin's ability to induce intestinal injury in rats is dependent on bile and can be reversed if pre-associated with phosphatidylcholine.
Clinical evidence suggests that aspirin and particularly enteric-coated aspirin induce significant injury to the lower gut. We have reported that NSAID injury to the small bowel is exacerbated by bile acids and that phosphatidylcholine (PC) can protect against this damage. Using a recently described method, we intra-duodenally administered either: saline, aspirin or aspirin pre-associated with PC. The rats were euthanized 90 minutes later at which time we assessed: tissue injury morphologically, vascular permeability with i.v. administered Evan's blue and intestinal bleeding by measuring luminal hemoglobin. In a separate experiment, aspirin-induced injury was studied in rats whose bile duct was ligated either alone or in the presence of rat bile (collected from donor animals). Intra-duodenal administration of aspirin induced mucosal injury (observed histologically), an increase in vascular permeability and blood loss into the intestinal lumen, all of which could be attenuated if the NSAID was pre-associated with PC. Furthermore, using 100 mg/kg dose of aspirin it was determined that bile duct ligation (BDL) significantly reduced aspirin-induced intestinal bleeding which was not different from control rats. Lastly, it was determined that intestinal bleeding was significantly increased in rats with BDL if the aspirin was administered in rodent bile. Aspirin-induced intestinal injury and bleeding in the rat is dependent on the presence of luminal bile, which is likely attributable to it's constituent bile acids. Pre-association of aspirin with PC provides a novel therapeutic approach to significantly reduce aspirin-induced small intestinal injury and bleeding, as may occur with enteric-coated aspirin.


Outcome of extracranial cervicocephalic arterial dissections: a follow-up study.
Cervicocephalic arterial dissections (CCAD) are an increasingly recognized cause of ischemic stroke in young adults. Various treatments have been suggested but no controlled trial has ever been performed. Medical treatment has included anticoagulant or platelet antiaggregant therapy. Surgical correction has been proposed for selected patients who have failed medical therapy. Percutaneous balloon angioplasty and stenting have been increasingly used in some patients, although long-term results are unknown. The objective of the study was to review our recent experience with the management and outcome of extracranial CCAD. We identified 27 patients with extracranial CCAD who were evaluated, treated and/or followed by our Stroke Service from September 1995 to August 2001. Clinical presentation, diagnostic evaluation, management, and outcome were reviewed. There were 15 men (56%) and 12 women (44%) with mean ages of 38 and 43 years respectively. Diagnosis was made by cerebral angiography in 15 (56%) patients and by MRI/MRA only in 12 (44%) patients. Twenty-two patients had spontaneous and five had traumatic extracranial CCAD. Most common associated disorders were arterial hypertension (37%) and migraine (26%). One patient presented only with a painful post-ganglionic Horner syndrome, another patient with neck pain and post-ganglionic Horner syndrome, another patient solely with protracted unilateral headaches, three with transient ischemic attacks (TIA), and 21 with ischemic strokes. The internal carotid artery (ICA) was the most frequently involved vessel (63%), followed by the vertebral artery (30%, and multivessel involvement in two patients (7%). Eighteen patients received anticoagulant therapy and nine platelet anti-aggregants. Follow-up extended from 2 to 115 months, with a mean of 58 months. At the end of follow-up, 23 (85%) patients had either no disability or only minor sequelae (modified Rankin score: 0 to 1), and four (15%) patients had moderate limitations (modified Rankin score: 2 to 3). Two patients had a recurrent ischemic stroke, one unrelated to recurrent CCAD, and the other following percutaneous balloon angioplasty/stenting for treatment of a persistent vertebral artery pseudoaneurysm. Most CCAD involved the extracranial ICA. The clinical presentation is variable, most patients having an ischemic stroke or TIAs. The short- and long-term outcome are usually favorable with either anticoagulant or platelet antiaggregant therapy. A medical initial approach to the management of extracranial CCAD is recommended for most patients.


[Longitudinal study of aflatoxin exposure in the development of primary liver cancer in patients with chronic hepatitis].
To study the relationship between aflatoxin exposure and the development of primary liver cancer (PLC) in patients with chronic hepatitis. A 21-year longitudinal study was carried out in a large cohort of 515 PLC high-risk individuals with HBV infection in PLC high prevalence region. (1) The PLC year-incidence of cohort was 1437.25/100,000. And it was significantly higher than that of the same natural peoples (184. 53/100,000, P = 0.000, RR = 7.79). There was no significant difference in the incidence of other tumors between these two groups (P = 0.576). (2) The PLC patients in the cohort were diagnosed at an average age 1.4 year younger than those in the same natural peoples and had an average survival of 6.42 months longer than the latter. (3) The PLC year-incidence of those with the exposure to aflatoxin was significantly higher than that of unexposed people (2784. 96/100,000 vs. 1251.02/100,000, P = 0.008, RR = 2.23). There was no relationship between the incidence rate of other tumors and the aflatoxin exposure. (4) The PLC year-incidence of aflatoxin-exposing people increased with the rising urine excretion of AFM1. When the urine excretion of AFM1 was more than 100 ng during 24 hours, the PLC year-incidence was high as 4717.82/100,000. The urine excretion of AFM1 was also obviously related with the abnormal liver function (P = 0.035). There was no relationship with the positive rate of HBeAg (P = 0.812). (5) The PLC year-incidence of those with the exposure to aflatoxin were infected with HBV (2 784. 96/100 000) significantly higher than that of cohort people (P = 0.001) and the same natural peoples (P = 0.000, RR = 15.09). (6) It took an average time of 14.65 years (median 13.68) from hepatitis occurrence to PLC diagnosis and 7.38 years (median 6.40) from liver cirrhosis to PLC diagnosis. HBV infection is a main etiological factor of PLC and the aflatoxin exposure has obvious synergistic effect in the carcinogenesis of PLC. Regular observation in a PLC high-risk cohort is effective for an early diagnosis and treatment. Hepatitis control and aflatoxin de-pollution is effective to inhibit the occurrence of PLC.


Assessment of injury to the posterior ligamentous complex in thoracolumbar spine trauma.
Posterior ligamentous complex (PLC), consisting of supraspinous ligament (SSL), interspinous ligament (ISL), ligamentum flavum (LF), and the facet joint capsules is thought to contribute significantly to the stability of thoracolumbar spine. Currently, no consensus exists on radiographic imaging parameters that may indicate injury to the posterior ligamentous complex. To identify imaging parameters that may suggest a disruption of the PLC of the thoracolumbar spine. A survey analysis of members of the Spine Trauma Study Group. None. Compilation of survey results. An extensive review of the literature from 1949 to the present was performed to identify key radiographic elements that have been suggested as indicators of PLC injury. Twelve items identified as such were placed on a survey and sent to the members of the Spine Trauma Study Group. They were asked to rank the items from most important to least important, and the results were compiled for analysis. Twenty-eight surveys were returned for final analysis. Fifty-percent (14/28) of the members ranked "vertebral body translation" on plain radiographs as the most important factor in determining disruption of PLC. Plain radiographic signs were ranked higher than computed tomography or magnetic resonance imaging indicators, and history of the mechanism ranked lowest. The members were also given freedom to add other criteria that they felt were important in determining PLC integrity. "Interspinous spacing 7 mm greater than that of level above or below on antero posterior plain X-rays" was the only new category that was suggested. Plain radiographic findings were felt to be most helpful in determining PLC injury by the members of the Spine Trauma Study Group. Physical examination findings and history of the mechanism of injury were ranked lower than imaging studies. Future analysis should focus on indicators of PLC injury when plain radiographic findings are either subtle or not present.


Toward Bifunctional Chelators for Thallium-201 for Use in Nuclear Medicine.
Auger electron therapy exploits the cytotoxicity of low-energy electrons emitted during radioactive decay that travel very short distances (typically <1 μm). 201Tl, with a half-life of 73 h, emits ∼37 Auger and other secondary electrons per decay and can be tracked in vivo as its gamma emissions enable SPECT imaging. Despite the useful nuclear properties of 201Tl, satisfactory bifunctional chelators to incorporate it into bioconjugates for molecular targeting have not been developed. H4pypa, H5decapa, H4neunpa-NH2, and H4noneunpa are multidentate N- and O-donor chelators that have previously been shown to have high affinity for 111In, 177Lu, and 89Zr. Herein, we report the synthesis and serum stability of [nat/201Tl]Tl3+ complexes with H4pypa, H5decapa, H4neunpa-NH2, and H4noneunpa. All ligands quickly and efficiently formed complexes with [201Tl]Tl3+ that gave simple single-peak radiochromatograms and showed greatly improved serum stability compared to DOTA and DTPA. [natTl]Tl-pypa was further characterized using nuclear magnetic resonance spectroscopy (NMR), mass spectroscopy (MS), and X-ray crystallography, showing evidence of the proton-dependent presence of a nine-coordinate complex and an eight-coordinate complex with a pendant carboxylic acid group. A prostate-specific membrane antigen (PSMA)-targeting bioconjugate of H4pypa was synthesized and radiolabeled. The uptake of [201Tl]Tl-pypa-PSMA in DU145 PSMA-positive and PSMA-negative prostate cancer cells was evaluated in vitro and showed evidence of bioreductive release of 201Tl and cellular uptake characteristic of unchelated [201Tl]TlCl. SPECT/CT imaging was used to probe the in vivo biodistribution and stability of [201Tl]Tl-pypa-PSMA. In healthy animals, [201Tl]Tl-pypa-PSMA did not show the myocardial uptake that is characteristic of unchelated 201Tl. In mice bearing DU145 PSMA-positive and PSMA-negative prostate cancer xenografts, the uptake of [201Tl]Tl-pypa-PSMA in DU145 PSMA-positive tumors was higher than that in DU145 PSMA-negative tumors but insufficient for useful tumor targeting. We conclude that H4pypa and related ligands represent an advance compared to conventional radiometal chelators such as DOTA and DTPA for Tl3+ chelation but do not resist dissociation for long periods in the biological environment due to vulnerability to reduction of Tl3+ and subsequent release of Tl+. However, this is the first report describing the incorporation of [201Tl]Tl3+ into a chelator-peptide bioconjugate and represents a significant advance in the field of 201Tl-based radiopharmaceuticals. The design of the next generation of chelators must include features to mitigate this susceptibility to bioreduction, which does not arise for other trivalent heavy radiometals.


Incidence and factors associated with medication nonadherence in patients with mental illness: a cross-sectional study.
In spite of the progress made in the treatment of psychiatric disorders during the last few decades, nonadherence continues to be a frequent phenomenon, often associated with potentially severe clinical consequences and increased health-care costs. There are numerous factors associated with medication nonadherence in patients with mental illness. The aim of the study was to determine the incidence and factors associated with medication nonadherence among psychiatric outpatients. A cross-sectional study was carried out in the outpatient psychiatric department of an Indian tertiary care private hospital over a period of 1 year. Patients aged 18 years and above who presented with mental illness as diagnosed by the International Classification of Diseases (ICD)-10 and who were receiving at least one psychotropic medication for at least 1 month were included in the study. Medication adherence was assessed using the Medication Adherence Rating Scale (MARS). Of the 400 patients, 172 (43%) were nonadherent to their prescribed medications. There is a statistically significant association between the education (P = 0.001), number of drugs (P = 0.002), family income (P = 0.013), and nonadherence. Among the 172 patients, 33.5 % were nonadherent to their therapy due to patient-related factors followed by drug-related factors (32%) and disease-related factors (31%). The overall incidence of medication nonadherence in patients with mental illness was 43%. Numerous factors contributed to medication nonadherence. Strategies need to be developed and implemented to enhance medication adherence, and thereby achieve a better therapeutic outcome in patients with mental illness.


Mossy fibre reorganization in the hippocampus of prion protein null mice.
Mice lacking prion protein (PrP-null) are resistant to transmissible spongiform encephalopathies. However, the normal functions of this highly conserved protein remain controversial. This study examines whether PrP-null mice develop normal neuronal pathways, specifically the mossy fibre pathway, within the hippocampus. Timm stained hippocampal sections from the PrP-null group had more granules than the controls in: the granule cell layer, the inner molecular layer of the dentate gyrus, and the infrapyramidal region of CA3. This resembles the mossy fibre collateral and terminal sprouting seen in certain epilepsies. The abnormal connectivity might be predicted to promote epileptiform activity, but extracellular electrophysiological recordings from the granule cell layer revealed a reduced excitability in the PrP-null group, both with and without blockade of GABA(A) receptor-mediated inhibition. We propose that reorganization of neuronal circuity is a feature of PrP-null mice.


Competency-based assessment for clinical supervisors: design-based research on a web-delivered program.
Clinicians need to be supported by universities to use credible and defensible assessment practices during student placements. Web-based delivery of clinical education in student assessment offers professional development regardless of the geographical location of placement sites. This paper explores the potential for a video-based constructivist Web-based program to support site supervisors in their assessments of student dietitians during clinical placements. This project was undertaken as design-based research in two stages. Stage 1 describes the research consultation, development of the prototype, and formative feedback. In Stage 2, the program was pilot-tested and evaluated by a purposeful sample of nine clinical supervisors. Data generated as a result of user participation during the pilot test is reported. Users' experiences with the program were also explored via interviews (six in a focus group and three individually). The interviews were transcribed verbatim and thematic analysis conducted from a pedagogical perspective using van Manen's highlighting approach. This research succeeded in developing a Web-based program, "Feed our Future", that increased supervisors' confidence with their competency-based assessments of students on clinical placements. Three pedagogical themes emerged: constructivist design supports transformative Web-based learning; videos make abstract concepts tangible; and accessibility, usability, and pedagogy are interdependent. Web-based programs, such as Feed our Future, offer a viable means for universities to support clinical supervisors in their assessment practices during clinical placements. A design-based research approach offers a practical process for such Web-based tool development, highlighting pedagogical barriers for planning purposes.


The role of calorie restriction and SIRT1 in prion-mediated neurodegeneration.
A central focus of aging research is to determine how calorie restriction (CR) extends lifespan and delays diseases of aging. SIRT1, the mammalian ortholog of Sir2 in yeast, is a longevity factor which mediates dietary restriction in diverse species. In addition, SIRT1 plays a protective role in several models of neurodegenerative disease. We tested the role of SIRT1 in mediating the effects of CR in a mouse model of prion disease. Prion diseases are protein misfolding disorders of the central nervous system with many similarities to other neurodegenerative diseases, including deposition of aggregated protein, gliosis, and loss of synapses and neurons. We report that the onset of prion disease is delayed by CR and in the SIRT1 KO mice fed ad libitum. CR exerts no further effect on the SIRT1 KO strain, suggesting the effects of CR and SIRT1 deletion are mechanistically coupled. In conjunction, SIRT1 is downregulated in certain brain regions of CR mice. The expression of PrP mRNA and protein is reduced in the brains of CR mice and in SIRT1 knockout mice, suggesting a possible mechanism for the delayed onset of disease, as PrP levels are a critical determinant of how quickly mice succumb to prion disease. Surprisingly, CR greatly shortens the duration of clinical symptoms of prion disease and ultimately shortens lifespan of prion-inoculated mice in a manner that is independent of SIRT1. Taken together, our results suggest a more complex interplay between CR, SIRT1, and neurodegenerative diseases than previously appreciated.


Repeated inseminations required for natural fertility in a wild bird population.
In most bird species, pairs copulate many times before egg laying. The exact function of repeated inseminations (i.e. successful copulations) is unknown, but several suggestions have been made. We tested the hypothesis that repeated inseminations are required to ensure fertilization of eggs, by using an experimental method where free-ranging male collared flycatchers (Ficedula albicollis) were prevented from inseminating their mates. We show that egg fertility was lower when females had not copulated during the studied part of their fertile period. By counting sperm on the inner perivitelline layer of eggs, we estimated that a minimum of 86 sperm must reach the site of fertilization to ensure average fertility. Using the timing of inseminations and the numbers of sperm on successive eggs, we show that repeated copulations are necessary to achieve an average rate of fertilization of a single clutch. Our results thus provide evidence that repeated inseminations function to ensure fertilization success. We discuss possible constraints on sperm production and utilization that may have contributed to this pattern.


Lactobacillus rhamnosus GG attenuates interferon-{gamma} and tumour necrosis factor-alpha-induced barrier dysfunction and pro-inflammatory signalling.
The intestinal epithelium forms a protective barrier against luminal contents and the external environment, mediated via intercellular tight junctions (TJs). The TJ can be disrupted via cell signalling induced by either enteric pathogens or pro-inflammatory cytokines, thereby contributing to various intestinal disorders ranging from acute infectious diarrhoea to chronic inflammatory bowel diseases. Probiotics, such as Lactobacillus rhamnosus GG (LGG), are reported to confer beneficial effects on epithelial cells, including antagonizing infections and reducing overt pro-inflammatory responses, but the underlying mechanisms of these observed effects require further characterization. We hypothesized that probiotics preserve barrier function by interfering with pro-inflammatory cytokine signalling. Caco-2bbe cells were seeded into Transwells to attain polarized monolayers with intercellular TJs. Monolayers were inoculated apically with the probiotic LGG 3 h prior to the addition of IFN-γ (100 ng ml(-1)) to the basolateral medium overnight. The monolayers were then placed in fresh basal medium±TNF-α (10 ng ml(-1)) and transepithelial electrical resistance (TER) measurements were taken over the time-course of TNF-α stimulation. To complement the TER findings, cells were processed for zona occludens-1 (ZO-1) immunofluorescence staining. As a measure of TNF-α downstream signalling, cells were immunofluorescently stained for NF-κB p65 subunit and CXCL-8 mRNA was quantified by qRT-PCR. Basal cell culture medium was collected after overnight TNF-α stimulation to measure secreted chemokines, including CXCL-8 (interleukin-8) and CCL-11 (eotaxin). Following LGG inoculation, IFN-γ priming and 24 h TNF-α stimulation, epithelial cells maintained TER and ZO-1 distribution. LGG diminished the nuclear translocation of p65, demonstrated by both immunofluorescence and CXCL-8 mRNA expression. CXCL-8 and CCL-11 protein levels were decreased in LGG-inoculated, cytokine-challenged cells. These findings indicate that LGG alleviates the effects of pro-inflammatory cytokines on epithelial barrier integrity and inflammation, mediated, at least in part, through inhibition of NF-κB signalling.


Molecular alterations associated with LNCaP cell progression to androgen independence.
There is no effective therapy currently available for androgen-independent (AI) prostate cancer (CaP). This is largely due to lack of information about the molecular mechanisms by which androgen-dependent tumor cells progress to androgen independence. In this study, we investigated molecular alterations occurring in AI LNCaP cells. We established and characterized three AI LNCaP sublines that exhibited a wide range of cytogenetic alterations. In order to understand why androgen-sensitive LNCaP cells can survive in an androgen-deprived environment, we analyzed the expression of signaling proteins associated with proliferation and survival of AI cells. In addition, gene expression profiling was performed to gain insight into molecular alterations in these LNCaP sublines. These LNCaP sublines exhibited heightened levels of androgen receptor (AR), HER2, MAPK, serine 473-phosphorylated Akt, and Bcl-2, implicating these proteins as mediators of AI growth. Gene expression profiling identified a common set of 66 genes that were differentially expressed in all three sublines compared to the parental LNCaP cells. Of these, 32 were apparently androgen regulated, while the remainder comprised an expression signature specific for androgen independence. We have developed AI LNCaP cell models and identified several genes that are specifically expressed in these models. Elucidating the relative importance of these genetic changes will help define the molecular mechanism by which CaP progresses to androgen independence.


Factors related to knowledge and perception of women about smoking: a cross sectional study from a developing country.
Smoking rates among women are currently low, but they are the fastest growing segment of cigarette smoking population in developing countries. We aimed to assess the knowledge and perceptions towards smoking and to identify the factors related with level of knowledge and perceptions among adult women in urban slums. This was a cross sectional study conducted on 250 adult (≥18 years of age) women attending primary care clinics in three slums of Karachi, Pakistan. A pre-tested and structured, interviewer administered questionnaire was used for data collection. Factors associated with level of understanding about smoking were analyzed with chi-square test. Most of the women knew that smoking has adverse effects on women and children's health but the knowledge of specific health effects was limited. About one third of the women knew that active smoking can cause lung disease, but only a small percentage (7%) knew that it could lead to heart disease. None of the women were aware that smoking contributes to infertility and osteoporosis. A small proportion of women were aware that smoking can lead to low birth weight (7%), congenital anomalies (5%) and less than 1% of women knew that it contributes to pregnancy loss, still birth and preterm delivery. The understanding of passive smoking affecting children's lung was low (20%) and a similar proportion voiced concern about the bad influence of maternal smoking on children. Educated women had better knowledge of health effects of smoking. Education was associated with having better knowledge about effects on women health in general (p = 0.02) and specific effects like lung (p = 0.03) and reproductive health effects (p < 0.001). Education was also associated with knowledge regarding effects on fetus (p < 0.001) and children (p < 0.005). Although most of the women disliked being around smokers, more than one third thought that smoking decreases boredom (39%), tension (38%) and also helps to relax (40%). A large proportion (48%) of women had the misconception that smoking helps to reduce weight. This study reveals that women are aware of the general ill effects of smoking but fail to identify smoking to be associated with female maladies particularly those who were illiterate and had lower levels of education. Understanding and attitudes needs to be improved by increasing health awareness and education of women in these urban communities with special emphasis on the effects of smoking on women's health.


The use of an integrative approach to improve accuracy of species identification and detection of new species in studies of stream fish diversity.
In this study, we made an inventory of the stream and headwater ichthyofauna of the left bank of the Itaipu Dam Reservoir, located in the lower part of the Upper Paraná River basin, using an integrative approach of molecular and morphological data. The area is located in the western portion of the Paraná state in Brazil, in an area of about 8,000 km2 highly impacted by deforestation and intensive agriculture. For taxonomic identification of species, we used an identification key combined with the DNA barcoding approach. We found 48 species belonging to six orders, 13 families, and 37 genera. The Siluriformes and Characiformes were the most representative orders (75%) and the Characidae was the most representative family (20.8%). Nine species prevailed in this region, making up to 86% of all specimens collected. The integrative approach proved to be useful by allowing the unambiguous identification of all species, including those cases in which morphological characters were not conclusive for species identification, cases of cryptic species, and species with high morphological plasticity. In addition, the integrative approach highlighted six to 13 new putative species depending on the approach considered. Our study provides a relevant contribution to the knowledge of fish diversity in a poorly studied area of the Paraná River basin. We showed that the use of an integrative approach in inventory studies improves species identification and the discovery of new, cryptic, and overlooked species, being a powerful and necessary tool to quantify biodiversity.


Estimating the sample mean and standard deviation from the sample size, median, range and/or interquartile range.
In systematic reviews and meta-analysis, researchers often pool the results of the sample mean and standard deviation from a set of similar clinical trials. A number of the trials, however, reported the study using the median, the minimum and maximum values, and/or the first and third quartiles. Hence, in order to combine results, one may have to estimate the sample mean and standard deviation for such trials. In this paper, we propose to improve the existing literature in several directions. First, we show that the sample standard deviation estimation in Hozo et al.'s method (BMC Med Res Methodol 5:13, 2005) has some serious limitations and is always less satisfactory in practice. Inspired by this, we propose a new estimation method by incorporating the sample size. Second, we systematically study the sample mean and standard deviation estimation problem under several other interesting settings where the interquartile range is also available for the trials. We demonstrate the performance of the proposed methods through simulation studies for the three frequently encountered scenarios, respectively. For the first two scenarios, our method greatly improves existing methods and provides a nearly unbiased estimate of the true sample standard deviation for normal data and a slightly biased estimate for skewed data. For the third scenario, our method still performs very well for both normal data and skewed data. Furthermore, we compare the estimators of the sample mean and standard deviation under all three scenarios and present some suggestions on which scenario is preferred in real-world applications. In this paper, we discuss different approximation methods in the estimation of the sample mean and standard deviation and propose some new estimation methods to improve the existing literature. We conclude our work with a summary table (an Excel spread sheet including all formulas) that serves as a comprehensive guidance for performing meta-analysis in different situations.


Removal of sulfur compounds from petroleum refinery wastewater through adsorption on modified activated carbon.
The adsorption of sulfur compounds from petroleum refinery wastewater on a chemically modified activated carbon (MAC) was investigated. The modification technique (nitric acid, hydrogen peroxide and thermal modification) enhanced the removal capacity of carbon and therefore decreases cost-effective removal of sulfide from refinery wastewater. Adsorption equilibrium and kinetics data were determined for sulfur removal from real refinery wastewater. The data were evaluated according to several adsorption isotherm and kinetics models. The Freundlich isotherm fitted well with the equilibrium data of sulfur on different adsorbents, whereas the kinetics data were best fitted by the pseudo-second-order model. Insights of sulfide removal mechanisms indicated that the sorption was controlled through the intraparticle diffusion mechanism with a significant contribution of film diffusion. The MAC adsorbent was found to have an effective removal capacity of approximately 2.5 times that of non-modified carbon. Using different MAC, sulfides were eliminated with a removal capacity of 52 mg g(-1). Therefore, MAC can be utilized as an effective and less expensive adsorbent for the reduction of sulfur in refinery wastewater.


Hotspots of biased nucleotide substitutions in human genes.
Genes that have experienced accelerated evolutionary rates on the human lineage during recent evolution are candidates for involvement in human-specific adaptations. To determine the forces that cause increased evolutionary rates in certain genes, we analyzed alignments of 10,238 human genes to their orthologues in chimpanzee and macaque. Using a likelihood ratio test, we identified protein-coding sequences with an accelerated rate of base substitutions along the human lineage. Exons evolving at a fast rate in humans have a significant tendency to contain clusters of AT-to-GC (weak-to-strong) biased substitutions. This pattern is also observed in noncoding sequence flanking rapidly evolving exons. Accelerated exons occur in regions with elevated male recombination rates and exhibit an excess of nonsynonymous substitutions relative to the genomic average. We next analyzed genes with significantly elevated ratios of nonsynonymous to synonymous rates of base substitution (dN/dS) along the human lineage, and those with an excess of amino acid replacement substitutions relative to human polymorphism. These genes also show evidence of clusters of weak-to-strong biased substitutions. These findings indicate that a recombination-associated process, such as biased gene conversion (BGC), is driving fixation of GC alleles in the human genome. This process can lead to accelerated evolution in coding sequences and excess amino acid replacement substitutions, thereby generating significant results for tests of positive selection.


Thermosensitive hydrogels composed of hyaluronic acid and gelatin as carriers for the intravesical administration of cisplatin.
The aim of this work was to evaluate the use of thermosensitive hydrogels for intravesical cisplatin delivery into the bladder. Poly(N-isopropylacrylamide) (PNIPAM) was grafted onto hyaluronic acid (HA) to synthesize an HPN copolymer, which was further grafted with gelatin to form an HPNG copolymer. A 3% concentration of HPN and HPNG was sufficient to exert a thermosensitive response, whereas a concentration of 8% was needed for PNIPAM to form the hydrogel. The physicochemical and drug delivery properties were examined by scanning electron microscopy (SEM), the lower critical solution temperature (LCST), hydration ratio, and in vitro cisplatin release. The incorporation of HA and gelatin produced a different microstructure compared to the parent PNIPAM hydrogel. Gelatin conjugation increased the fibrous structure in the matrix. The LCSTs of PNIPAM, HPN, and HPNG were 32.3, 32.0, and 30.7°C, respectively. The copolymers showed an eightfold increase in the hydration capacity compared to PNIPAM, with no significant difference in values between HPN and HPNG. The release of cisplatin from an aqueous solution (control) was nearly complete after 8 h, compared to 85, 80, and 52% release from PNIPAM, HPN, and HPNG, respectively. In vivo evaluation of cisplatin levels in bladder tissues was performed following intravesical instillation in rats. When the dwell time was extended to 6 h, PNIPAM showed a sevenfold enhancement in the drug concentration in the bladder wall. HPNG also showed a twofold increase in the drug concentration. The administration of cisplatin by the HPN carrier did not change the drug accumulation compared to the control. Confocal laser scanning microscopic results confirmed the trend of drug absorption from various systems. A histological examination showed no adverse change in the urothelium with HPN or HPNG application. PNIPAM caused partial desquamation of umbrella cells. The thermosensitive hydrogels prepared in this study may be promising carriers for targeted drug delivery to the bladder.


Pathogen-specific CD8 T cell responses are directly inhibited by IL-10.
Regulation of CD8 T cell expansion and contraction is essential for successful immune defense against intracellular pathogens. IL-10 is a regulatory cytokine that can restrict T cell responses by inhibiting APC functions. IL-10, however, can also have direct effects on T cells. Although blockade or genetic deletion of IL-10 enhances T cell-mediated resistance to infections, the extent to which IL-10 limits in vivo APC function or T cell activation/proliferation remains unknown. Herein, we demonstrate that primary and memory CD8 T cell responses following Listeria monocytogenes infection are enhanced by the absence of IL-10. Surface expression of the IL-10R is transiently up-regulated on CD8 T cells following activation, suggesting that activated T cells can respond to IL-10 directly. Consistent with this notion, CD8 T cells lacking IL-10R2 underwent greater expansion than wild-type T cells upon L. monocytogenes infection. The absence of IL-10R2 on APCs, in contrast, did not enhance T cell responses following infection. Our studies demonstrate that IL-10 produced during bacterial infection directly limits expansion of pathogen-specific CD8 T cells and reveal an extrinsic regulatory mechanism that modulates the magnitude of memory T cell responses.


Carotid endarterectomy versus carotid angioplasty with or without stenting for treatment of carotid artery stenosis: an updated meta-analysis of randomized controlled trials.
Carotid angioplasty with or without stent placement (CAS) has been proposed as an alternative to carotid endarterectomy (CEA) for the treatment of carotid artery stenosis. We performed a systematic review and meta-analysis of randomized controlled trials to compare the safety and efficacy of endovascular techniques with surgery for carotid stenosis. We searched MEDLINE, PubMed and Cochrane databases to identify randomized controlled trials comparing CAS with CEA. Both random and fixed effects models were used to calculate the pooled odds ratios (OR) and their confidence intervals, with values lower than one indicating a benefit from the endovascular approach. Continuity correction was used for studies with zero events in one arm. We identified 11 trials randomizing a total of 3 258 patients; 1 623 to CEA and 1 635 to CAS. By random effects model, there was no significant difference between the treatments for any stroke (OR, 1.28; 95% CI, 0.82-2.02), or death or any stroke at 30-day (OR, 1.30; 95% CI, 0.92-1.84) and death or any stroke at 6 months (OR, 1.34; 95% CI, 0.86-2.09) or 1 year (OR, 1.41; 95% CI, 0.24-8.27). However there was a significantly higher risk of 30-day death or any stroke (OR, 1.33; 95% CI, 1.01-1.75) after CAS by fixed effects model. Endovascular treatment significantly reduced the risk of 30-day cranial nerve injury (OR, 0.13; 95% CI, 0.04-0.44). In conclusion treating carotid artery stenosis with CAS offers lower rates of cranial nerve injury compared with CEA. CAS could not be proved to be as safe as CEA in treating carotid artery stenosis. The results of ongoing randomized trials comparing CAS with CEA are easily awaited because they may provide sufficient evidence for a change in clinical practice.


Erythrocytes-based quartz crystal microbalance cytosensor for in situ detection of cell surface sialic acid.
An erythrocytes (RBCs) cytosensor was first fabricated for in situ analysis of sialic acid (SA) on the cell surface based on a quartz crystal microbalance (QCM). RBCs, as a recognition element, were immobilized on a concanavalin A (ConA)-modified gold chip through the specific recognition between ConA and mannose on the cell surface. 4-Aminobenzeneboronic acid (APBA)-functionalized gold nanoparticles (AuNPs/APBA) were used as a signal amplification nanoprobe for labeling SA on the surface of RBCs. Compared to that of APBA, the frequency response of the cytosensor could be significantly enhanced 18-fold by using a AuNPs/APBA nanoprobe. RBCs can be detected in the range of 2.6 × 103 to 7.2 × 106 cells per mL with a detection limit of 1.1 × 103 cells per mL. The proposed cytosensor was further applied to detect the expression level of SA on normal and diabetic RBCs in situ, which showed that the average number of SA expressed on single normal and diabetic RBC surfaces were 2.1 ± 0.2 × 108 and 8.2 ± 0.7 × 107 with a relative standard deviation of 4.3% and 3.6%, respectively. The strategy shows an in situ and high sensitivity method for the quantitative evaluation of SA expression on RBC surface and provides a new alternative methodology to analyze glycan expression at the cell surface.


The permanence of the visual recovery that follows reverse occlusion of monocularly deprived kittens.
While the behavioral and physiologic effects of an early period of monocular deprivation can be extremely severe, they are not necessarily irreversible. Considerable recovery can occur if visual input is restored to the deprived eye sufficiently early, particularly if the nondeprived eye is occluded at the same time (reverse occlusion). This study examines the permanence of the visual recovery promoted by a period of reverse occlusion in kittens that were monocularly deprived from near birth for periods ranging from 3 to 18 weeks. During the period of reverse occlusion, the vision of the initially deprived eye improved from apparent blindness to good levels of acuity. However, upon restoring visual input to the formerly nondeprived eye a surprisingly rapid and reciprocal change occurred in the visual acuity of both eyes. Much of the substantial gain in the vision of the initially deprived eye that occurred during reverse occlusion was lost within 3 weeks, while at the same time the vision of the initially nondeprived eye improved substantially. Nevertheless, in many animals the acuity of the initially nondeprived eye did not recover to levels it had reached prior to reverse occlusion. These results hold important implications for the nature of the mechanisms responsible for the dramatic physiologic effects of monocular occlusion and reverse occlusion on the visual cortex. The results also may help elucidate recent observations on patching therapy in human amblyopia.


Nicotine reinforcement and cognition restored by targeted expression of nicotinic receptors.
Worldwide, 100 million people are expected to die this century from the consequences of nicotine addiction, but nicotine is also known to enhance cognitive performance. Identifying the molecular mechanisms involved in nicotine reinforcement and cognition is a priority and requires the development of new in vivo experimental paradigms. The ventral tegmental area (VTA) of the midbrain is thought to mediate the reinforcement properties of many drugs of abuse. Here we specifically re-expressed the beta2-subunit of the nicotinic acetylcholine receptor (nAChR) by stereotaxically injecting a lentiviral vector into the VTA of mice carrying beta2-subunit deletions. We demonstrate the efficient re-expression of electrophysiologically responsive, ligand-binding nicotinic acetylcholine receptors in dopamine-containing neurons of the VTA, together with the recovery of nicotine-elicited dopamine release and nicotine self-administration. We also quantified exploratory behaviours of the mice, and showed that beta2-subunit re-expression restored slow exploratory behaviour (a measure of cognitive function) to wild-type levels, but did not affect fast navigation behaviour. We thus demonstrate the sufficient role of the VTA in both nicotine reinforcement and endogenous cholinergic regulation of cognitive functions.


Dynamical basis for drug resistance of HIV-1 protease.
Protease inhibitors designed to bind to protease have become major anti-AIDS drugs. Unfortunately, the emergence of viral mutations severely limits the long-term efficiency of the inhibitors. The resistance mechanism of these diversely located mutations remains unclear. Here I use an elastic network model to probe the connection between the global dynamics of HIV-1 protease and the structural distribution of drug-resistance mutations. The models for study are the crystal structures of unbounded and bound (with the substrate and nine FDA approved inhibitors) forms of HIV-1 protease. Coarse-grained modeling uncovers two groups that couple either with the active site or the flap. These two groups constitute a majority of the drug-resistance residues. In addition, the significance of residues is found to be correlated with their dynamical changes in binding and the results agree well with the complete mutagenesis experiment of HIV-1 protease. The dynamic study of HIV-1 protease elucidates the functional importance of common drug-resistance mutations and suggests a unifying mechanism for drug-resistance residues based on their dynamical properties. The results support the robustness of the elastic network model as a potential predictive tool for drug resistance.


Promotion by prolactin of the growth of human breast neoplasms cultured in vitro in the soft agar clonogenic assay.
The role of prolactin (PRL) in supporting the growth of human breast cancer is still unclear. The ability to grow primary breast cancer specimens in the soft agar clonogenic assay in the absence of serum gave us the opportunity to evaluate the growth-promoting effect of PRL and to compare it to that of estradiol in the same tumor samples. PRL was tested both at physiological concentrations (20 ng/ml) as well as in pharmacological amounts (200 ng/ml) comparable to circulating blood levels in hyperprolactinemic states. Estradiol was simultaneously tested in physiological amounts (10(-8)M). In 17 infiltrating ductal carcinomas, the lower dose of PRL stimulated colony formation to 126 +/- 5.2% (SE) of control, while the higher dose increased colony number to 159 +/- 10.4% of control. This latter effect was comparable to that observed with estradiol (159 +/- 8.5% of control). The effect of PRL was more pronounced in estrogen receptor-positive tumors. Nine of ten estrogen receptor-positive tumors were PRL sensitive, while three of seven estrogen receptor-negative tumors exhibited a clear response to PRL administration. PRL did not stimulate colony formation in a malignant cystosarcoma phylloides and in two benign lesions (fibroadenoma and fibrocystic disease). We conclude that, at least under the conditions of the soft agar clonogenic assay, PRL exerts a dose-dependent growth-promoting effect on human breast cancer. Such effect is comparable to that of estradiol when PRL is added in concentrations similar to circulating blood levels in hyperprolactinemic patients.


Casein kinase II phosphorylates DNA-polymerase-alpha--DNA-primase without affecting its basic enzymic properties.
Immunoaffinity-purified DNA-polymerase-alpha--DNA-primase complex from calf thymus was phosphorylated in vitro by highly purified casein kinase II from the same tissue. Specific phosphorylation of the DNA-polymerizing alpha subunit and the primase-associated gamma subunit was observed. About 1 mol phosphate/mol polymerase--primase was incorporated. Despite this effect, neither the DNA polymerase nor the DNA primase activity were changed after phosphorylation by casein kinase II. Furthermore, dephosphorylation of polymerase--primase with alkaline phosphatase did not change the polymerase or the primase activity to a significant extent. Moreover, both alkaline phosphatase and casein kinase II had no effect on the processivity of DNA synthesis and on the lengths and amounts of primers formed by the DNA primase. Because DNA polymerase alpha maintained all its basic properties even after extensive treatment with alkaline phosphatase, it is unlikely that phosphorylation has a direct influence on the activities of the DNA-polymerase-alpha--DNA-primase complex. The possible influence of post-translational phosphorylation on the formation of a complex of polymerase alpha and its accessory proteins is discussed.


An advanced algorithm for dispersion encoded full range frequency domain optical coherence tomography.
Dispersion encoded full range (DEFR) optical coherence tomography (OCT) has become highly attractive as it is a simple way to increase the measurement range of OCT systems. Full range OCT is especially favorable as it does not only increase the measurement range but also shifts the highest sensitivity into the center of the measurement range. While the early versions of DEFR were highly computational expensive, new versions reduce the number of necessary Fourier transforms. Recently it has been shown that a GPU based algorithm can perform DEFR with more than 20,000 A-lines per second. We present a new version of the DEFR algorithm that requires only one Fourier transform per A-scan and uses convolution in z-space instead of multiplication in k-space, therefore reducing the computational effort considerably. While dispersion encoding has so far only been used to suppress mirror artifacts, we show that, with dispersion encoding and only one more Fourier transform, autocorrelation terms can be removed likewise. Since very high values of dispersion reduce the effective measurement range in dispersion encoded OCT, we present an estimate for a sufficient amount of dispersion for a successful image recovery, which is depending on the thickness of the scattering layers. Furthermore, we demonstrate the usability of ZnSe as a new dispersive material with a very high dispersion and describe a simple method to extract the dispersive phase from the measurement of a single reflex of a glass surface. Using a standard consumer PC, an artifact-free recovery of 1000 - 2000 A-scans per second with 2048 depth values including autocorrelation removal was achieved. The dynamic range (sensitivity) is not reduced and the suppression ratio of mirror artifacts and autocorrelation signals is more than 50dB using ZnSe.


Development of a targeted transgenesis strategy in highly differentiated cells: a powerful tool for functional genomic analysis.
Functional genomic analysis is a challenging step in the so-called post-genomic field. Identification of potential targets using large-scale gene expression analysis requires functional validation to identify those that are physiologically relevant. Genetically modified cell models are often used for this purpose allowing up- or down-expression of selected targets in a well-defined and if possible highly differentiated cell type. However, the generation of such models remains time-consuming and expensive. In order to alleviate this step, we developed a strategy aimed at the rapid and efficient generation of genetically modified cell lines with conditional, inducible expression of various target genes. Efficient knock-in of various constructs, called targeted transgenesis, in a locus selected for its permissibility to the tet inducible system, was obtained through the stimulation of site-specific homologous recombination by the meganuclease I-SceI. Our results demonstrate that targeted transgenesis in a reference inducible locus greatly facilitated the functional analysis of the selected recombinant cells. The efficient screening strategy we have designed makes possible automation of the transfection and selection steps. Furthermore, this strategy could be applied to a variety of highly differentiated cells.


Scoping Review of Hospital Business Continuity Plans to Validate the Improvement after the 2011 Great East Japan Earthquake and Tsunami.
During a disaster, all hospitals are expected to function as "social critical institutions" that protect the lives and health of people. In recent disasters, numerous hospitals were damaged, and this hampered the recovery of the affected communities. Had these hospitals business continuity plans (BCPs) to recover quickly after the disaster, most of the damage could have been avoided. This study conducted a scoping review of the historical trend and regional differences in hospital BCPs to validate the improvement of the BCP concept based on our own experience at Tohoku University Hospital, which was affected by the 2011 Great East Japan Earthquake and Tsunami (GEJET). We searched PubMed by using keywords related to BCP and adapted 97 articles for our analysis. The number of articles on hospital BCPs has increased in the 2000s, especially after Hurricane Katrina in 2005. While there are regional specificity of hazards, there were many common topics and visions for BCP implementation, education, and drills. From our 2011 GEJET experience, we found that BCPs assuming region-specific disasters are applicable in various types of disasters. Thus, we suggest the following integral and universal components for hospital BCPs: (1) alternative methods and resources, (2) priority of operation, and (3) resource management. Even if the type and extent of disasters vary, the development of BCPs and business continuity management strategies that utilize the abovementioned integral components can help a hospital survive disasters in the future.


Influence of plant silicon in Busseola fusca (Lepidoptera: Noctuidae) larvae - Poaceae interactions.
The noctuid stem borer Busseola fusca is an important pest of maize and sorghum in Sub-Saharan Africa. The presence of this species occurred mostly on cultivated than on wild habitats. Busseola fusca is oligophagous having a narrow range of a wild grass species. This might be due, in part, to differences in silicon (Si) content in plant tissues between cultivated and wild grasses. In the present study, we have tested this hypothesis by studying the survival and the relative growth rate (RGR) expressed as daily weight gains of B. fusca larvae on maize and six wild host plants, mostly present in the natural habitat where B. fusca occurred, and correlated with their Si contents. Survival and RGR of B. fusca larvae were considerably higher on maize and wild sorghum than on the other grass species, and they were negatively related to plant Si content. This was corroborated with results on RGR from artificial diets amended with increasing levels of Si. In addition, if Si was added to maize growing substrate B. fusca larval growth was significantly reduced confirming the involvement of Si in B. fusca larvae - Poaceae interactions. The results provide insight into the possible mechanisms of oligophagy of B. fusca and provide a correlative support for a physical role of plant endogenous Si in impeding feeding of B. fusca larvae.


A Process Evaluation of the Alaska Native Colorectal Cancer Family Outreach Program.
This article presents the results of a process evaluation of the Alaska Native (AN) Colorectal Cancer (CRC) Family Outreach Program, which encourages CRC screening among AN first-degree relatives (i.e., parents, siblings, adult children; hereafter referred to as relatives) of CRC patients. Among AN people incidence and death rates from CRC are the highest of any ethnic/racial group in the United States. Relatives of CRC patients are at increased risk; however, CRC can be prevented and detected early through screening. The evaluation included key informant interviews (August to November 2012) with AN and non-AN stakeholders and program document review. Five key process evaluation components were identified: program formation, evolution, outreach responses, strengths, and barriers and challenges. Key themes included an incremental approach that led to a fully formed program and the need for dedicated, culturally competent patient navigation. Challenges included differing relatives' responses to screening outreach, health system data access and coordination, and the program impact of reliance on grant funding. This program evaluation indicated a need for more research into motivating patient screening behaviors, electronic medical records systems quality improvement projects, improved data-sharing protocols, and program sustainability planning to continue the dedicated efforts to promote screening in this increased risk population.


Significant variations in the seroprevalence of C6 ELISA antibodies in a highly endemic area for Lyme borreliosis: evaluation of age, sex and seasonal differences.
The purpose of this study was to assess the seroprevalence of C6 ELISA antibodies in healthy blood donors in Kalmar County, Sweden, in relation to age, sex and time of year (peak season vs off season). In addition, we wanted to assess serological status over time in a group of C6 ELISA seropositive blood donors. Sera were collected from 273 (131 women, 142 men) blood donors in autumn 2011 and 300 (144 women, 156 men) in winter 2014. All sera were analysed in the C6 ELISA and the results were interpreted according to the manufacturer's instructions. The seroprevalence was 22% (females 16%, males 28%) in 2011 and 24% (females 15%, males 33%) in 2014. The seroprevalence was significantly higher in males and increased with age. The highest seroprevalence was observed among elderly men, 60-70 years old (46% in 2011 and 52% in 2014). No significant difference was detected in seropositivity between the samples collected in winter and autumn. All (34/34) seropositive blood donors followed over time remained seropositive at follow-up after 22-29 months. C6 ELISA seroprevalence in healthy blood donors is high in Kalmar County, thereby reducing the specificity of a positive test result regarding the clinical diagnosis of Lyme borreliosis (LB). Although C6 seroprevalence appears not to be affected by seasonal sample time, it varies greatly with age and sex. A careful evaluation of pre-test probability is therefore of the utmost importance in the clinical diagnosis of LB, especially in elderly men. We suggest that colleagues in other endemic regions also consider initiating similar evaluations to optimize the laboratory and clinical diagnosis of LB in relation to age and sex.


Influence of stimulus intensity on the soleus H-reflex amplitude and modulation during locomotion.
Diverging results have been reported regarding the modulation and amplitude of the soleus H-reflex measured during human walking and running. A possible explanation to this could be the use of too high stimulus strength in some studies while not in others. During activities like walking and running it is necessary to use a small M-wave to control the effective stimulus strength during all phases of the movement. This implies that the descending part of the H-reflex recruitment curve is being used, which may lead to an unwanted suppression of the H-reflex due to limitations imbedded within the H-reflex methodology itself. Accordingly, the purpose of the present study was to study the effect on the soleus H-reflex during walking and running using stimulus intensities normally considered too high (up to 45% Mmax). Using M-waves of 25-45% Mmax as opposed to 5-25% Mmax showed a significant suppression of the peak H-reflex during the stance phase of walking, while no changes were observed during running. No differences were observed regarding modulation pattern. So a possible use of too high stimulus intensity cannot explain the differences mentioned. The surprising result in running may be explained by the much higher voluntary muscle activity, which implies the existence of a V-wave influencing the H-reflex amplitude in positive direction.


A meta-analysis of health state valuations for people with diabetes: explaining the variation across methods and implications for economic evaluation.
To review published studies on the effect of diabetes and its complications on utility scores to establish whether there is systematic variation across studies and to examine the implications for the estimation of quality-adjusted life years (QALYs). A systematic review was performed using studies reporting QALY measures elicited from people with diabetes including those with a history of complications. Meta-analysis was used to obtain the average utility, and meta-regression was employed to examine the impact of study characteristics and elicitation methods on these values. The effect of different utility scores on QALYs was examined using diabetes simulation models. In the meta-analysis based on 45 studies reporting 66 values, the average utility score was 0.76 (95% CI 0.75-0.77). A meta-regression showed significant variation due to age, method of elicitation and the proportion of males. The average utility score for individual complications ranged from 0.48 (95% CI 0.25, 0.71) for chronic renal disease to 0.75 (95% CI 0.73, 0.78) for myocardial infarction, and these differences produced meaningful changes in simulated QALYs. There was significant heterogeneity between studies. We provide summary utility scores for diabetes and its major complications that could help inform economic evaluation and policy analysis.


Lipid peroxides, anti-oxidants and nitric oxide in patients with pre-eclampsia and essential hypertension.
The aim was to study the role of lipid peroxides, nitric oxide and anti-oxidants in pre-eclampsia and essential hypertension. The study was done in 28 patients with pre-eclampsia and 30 patients of essential hypertension. The plasma concentrations of lipid peroxides as a measure of pro-oxidant status, nitric oxide and RBC content of anti-oxidants were measured in these patients and compared with controls. It was observed that in patients with pre-eclampsia the plasma concentrations of nitric oxide and lipid peroxides were elevated with a decrease in those of anti-oxidants such as catalase and superoxide dismutase and a concomitant increase in that of glutathione peroxidase. On the other hand, in patients with essential hypertension, the concentrations of nitric oxide and superoxide dismutase were found to be low where as that of lipid peroxides were high. When the lipid peroxides/nitric oxide (LP/NO) ratio was calculated a far more significant rise in the concentrations of lipid peroxides compared to NO levels was noted especially in patients with pre-eclampsia and patients with uncontrolled essential hypertension. These results suggest that oxidants and anti-oxidants are altered in human essential hypertension and pre-eclampsia and that they may play a significant role in their pathophysiology.


Sperm economy between female mating frequency and male ejaculate allocation.
Why females of many species mate multiply is a major question in evolutionary biology. Furthermore, if females accept matings more than once, ejaculates from different males compete for fertilization (sperm competition), which confronts males with the decision of how to allocate their reproductive resources to each mating event. Although most existing models have examined either female mating frequency or male ejaculate allocation while assuming fixed levels of the opposite sex's strategies, these strategies are likely to coevolve. To investigate how the interaction of the two sexes' strategies is influenced by the level of sperm limitation in the population, we developed models in which females adjust their number of allowable matings and males allocate their ejaculate in each mating. Our model predicts that females mate only once or less than once at an even sex ratio or in an extremely female-biased condition, because of female resistance and sperm limitation in the population, respectively. However, in a moderately female-biased condition, males favor partitioning their reproductive budgets across many females, whereas females favor multiple matings to obtain sufficient sperm, which contradicts the predictions of most existing models. We discuss our model's predictions and relationships with the existing models and demonstrate applications for empirical findings.


Immunohistochemical changes in sigmoid colon after allogeneic and autologous bone marrow transplantation.
To determine whether there are characteristic immunohistological changes in the colonic mucosa in acute graft versus host disease (GvHD). Consecutive allogeneic (n = 11) and autologous (n = 11) bone marrow transplant recipients underwent endoscopic biopsy of sigmoid mucosa before transplant and on day 30 post-transplant. Immunohistochemical staining and quantitation of intraepithelial and lamina propria mononuclear cells were undertaken using a panel of monoclonal antibodies and a Streptavidin-biotin alkaline phosphatase staining technique. In the allogeneic group (nine of whom had clinical acute GvHD) there was a fivefold increase in lamina propria CD16+ mononuclear cells (3.1 +/- 4.3 to 17.0 +/- 12.2 per 100 lamina propria nucleated cells), compared with autologous transplant recipients in whom this rise was twofold (5.5 +/- 4.6 to 10.6 +/- 7.1 per 100 lamina propria nucleated cells). The CD16+ mononuclear cells had morphological appearances of tissue macrophages, but in neither the allogeneic nor autologous groups was there an increase in total macrophage numbers (CD14+). In patients with acute GvHD the lamina propria CD4+:CD8+ lymphocyte ratio fell (1.97 +/- 1.12 to 1.07 +/- 1.01), primarily because of a fall in the number of lamina propria CD4+ lymphocytes. In both allogeneic and autologous groups there was a fall in intraepithelial lymphocyte numbers, but there was no change in CD19+ (B cell), CD25+ (interleukin-2 receptor positive) or CD56+ (natural killer) cell numbers. Following bone marrow transplantation, there appears to be upregulation of lamina propria tissue macrophage CD16 (an Fc receptor for IgG), a change which is more noticeable after allogeneic transplantation and which may be related to the development of acute GvHD. In patients with acute GvHD there was a fall in the lamina propria CD4+:CD8+ lymphocyte ratio. If these changes are functionally important, they may have significant implications for understanding the pathogenesis of GvHD.


Facile promoter deletion in Escherichia coli in response to leaky expression of very robust and benign proteins from common expression vectors.
Overexpression of proteins in Escherichia coli is considered routine today, at least when the protein is soluble and not otherwise toxic for the host. We report here that the massive overproduction of even such "benign" proteins can cause surprisingly efficient promoter deletions in the expression plasmid, leading to the growth of only non-producers, when expression is not well repressed in the newly transformed bacterial cell. Because deletion is so facile, it might impact on high-throughput protein production, e.g. for structural genomics, where not every expression parameter will be monitored. We studied the high-level expression of several robust non-toxic proteins using a T5 promoter under lac operator control. Full induction leads to no significant growth retardation. We compared expression from almost identical plasmids with or without the lacI gene together in strains expressing different levels of LacI. Any combination without net overexpression of LacI led to an efficient promoter deletion in the plasmid, although the number of growing colonies and even the plasmid size - all antibiotic-resistant non-producers - was almost normal, and thus the problem not immediately recognizable. However, by assuring sufficient repression during the initial establishment phase of the plasmid, deletion was completely prevented. The deletions in the insufficiently repressed system are caused entirely by the burden of high-level translation. Since the E. coli Dps protein, known to protect DNA against stress in the stationary phase, is accumulated in the deletion mutants, the mutation may have taken place during a transient stationary phase. The cause of the deletion is thus distinct from the well known interference of high-level transcription with plasmid replication. The deletion can be entirely prevented by overexpressing LacI, a useful precaution even without any signs of stress caused by the protein.


From Molecular Descriptors to Intrinsic Fish Toxicity of Chemicals: An Alternative Approach to Chemical Prioritization.
The European and U.S. chemical agencies have listed approximately 800k chemicals about which knowledge of potential risks to human health and the environment is lacking. Filling these data gaps experimentally is impossible, so in silico approaches and prediction are essential. Many existing models are however limited by assumptions (e.g., linearity and continuity) and small training sets. In this study, we present a supervised direct classification model that connects molecular descriptors to toxicity. Categories can be driven by either data (using k-means clustering) or defined by regulation. This was tested via 907 experimentally defined 96 h LC50 values for acute fish toxicity. Our classification model explained ≈90% of the variance in our data for the training set and ≈80% for the test set. This strategy gave a 5-fold decrease in the frequency of incorrect categorization compared to a quantitative structure-activity relationship (QSAR) regression model. Our model was subsequently employed to predict the toxicity categories of ≈32k chemicals. A comparison between the model-based applicability domain (AD) and the training set AD was performed, suggesting that the training set-based AD is a more adequate way to avoid extrapolation when using such models. The better performance of our direct classification model compared to that of QSAR methods makes this approach a viable tool for assessing the hazards and risks of chemicals.


Effect of preventive messages tailored to family history on health behaviors: the Family Healthware Impact Trial.
We wanted to determine the impact of automated family history assessment and tailored messages for coronary heart disease, stroke, diabetes, colorectal, breast, and ovarian cancer on preventive behaviors compared with a standard preventive message. The study was a cluster-randomized clinical trial that included 41 primary care practices, the majority in the Midwest, using Family Healthware, a self-administered, Web-based tool that assesses familial risk for the diseases and provides personalized risk-tailored messages. Patients in the control group received an age- and sex-specific health message related to lifestyle and screening. Smoking cessation, fruit and vegetable intake, physical activity, aspirin use, blood pressure, and cholesterol and blood glucose screening were assessed at baseline and 6 months after the intervention. Of 4,248 participants, 3,344 (78%) completed the study. Participants were white (91%), female (70%), and insured (97%), and had a mean age of 50.6 years (range 35-65 years). Intervention participants were more likely to increase daily fruit and vegetable consumption from 5 or fewer servings a day to 5 or more servings a day (OR = 1.29; 95% confidence interval [CI], 1.05-1.58) and to increase physical activity (OR = 1.47; 95% CI, 1.08-1.98) to 5 to 6 times a week for 30 minutes or more a week. The absolute differences in proportion were 3% and 4%, respectively. Intervention participants were less likely to move from not having cholesterol screening in the last 5 years to having their cholesterol measured within 5 years (OR = 0.34; 95% CI, 0.17-0.67), with an absolute difference of 15%. Messages tailored to an individual's familial risk for 6 common diseases modestly increased self-reported physical activity and fruit and vegetable intake but reduced the likelihood of receiving cholesterol screening.


Types of gambling and levels of harm: A UK study to assess severity of presentation in a treatment-seeking population.
Background and aim Previous international research emphasized that some forms of gambling are more "addictive" than others. More recently, research has shown that we should shift our attention from the type of gambling activity to the level of involvement in a number of different gambling activities. The aim of our study was to verify whether a higher Problem Gambling Severity Index (PGSI) score was associated with particular gambling activities and evaluate the impact of involvement on gambling behavior. Methods A total of 736 treatment-seeking individuals with gambling disorder were assessed at the National Problem Gambling Clinic in London. First, the independent two-sample t-test and the Mann-Whitney test were used to verify if the PGSI score changed significantly according to the gambling activity at a bivariate level. Second, we conducted a cluster analysis and finally, we fitted a linear regression model in order to verify if some variables are useful to predict gambling addiction severity. Results The PGSI score was significantly higher for lower stakes gaming machine gamblers (1% significance level) and for fixed-odds betting terminal (FOBT) gamblers (5% significance level) at a bivariate level. Moreover, such finding was confirmed by cluster and linear regression analyses. Conclusions The results of this study indicated that gambling addiction severity was related to gambling involvement and, for a given level of gambling involvement, gambling addiction severity may vary according to gambling type, with a particularly significant increase for FOBT and gaming machine gambling.


Curcumin, the active constituent of turmeric, inhibits amyloid peptide-induced cytochemokine gene expression and CCR5-mediated chemotaxis of THP-1 monocytes by modulating early growth response-1 transcription factor.
Epidemiological studies show reduced risk of Alzheimer's disease (AD) among patients using non-steroidal inflammatory drugs (NSAID) indicating the role of inflammation in AD. Studies have shown a chronic CNS inflammatory response associated with increased accumulation of amyloid peptide and activated microglia in AD. Our previous studies showed that interaction of Abeta1-40 or fibrilar Abeta1-42 caused activation of nuclear transcription factor, early growth response-1 (Egr-1), which resulted in increased expression of cytokines (TNF-alpha and IL-1beta) and chemokines (MIP-1beta, MCP-1 and IL-8) in monocytes. We determined whether curcumin, a natural product known to have anti-inflammatory properties, suppressed Egr-1 activation and concomitant expression of cytochemokines. We show that curcumin (12.5-25 microm) suppresses the activation of Egr-1 DNA-binding activity in THP-1 monocytic cells. Curcumin abrogated Abeta1-40-induced expression of cytokines (TNF-alpha and IL-1beta) and chemokines (MIP-1beta, MCP-1 and IL-8) in both peripheral blood monocytes and THP-1 cells. We found that curcumin inhibited Abeta1-40-induced MAP kinase activation and the phosphorylation of ERK-1/2 and its downstream target Elk-1. We observed that curcumin inhibited Abeta1-40-induced expression of CCR5 but not of CCR2b in THP-1 cells. This involved abrogation of Egr-1 DNA binding in the promoter of CCR5 by curcumin as determined by: (i) electrophoretic mobility shift assay, (ii) transfection studies with truncated CCR5 gene promoter constructs, and (iii) chromatin immunoprecipitation analysis. Finally, curcumin inhibited chemotaxis of THP-1 monocytes in response to chemoattractant. The inhibition of Egr-1 by curcumin may represent a potential therapeutic approach to ameliorate the inflammation and progression of AD.


Relationships between aromatase activity in the brain and gonads and behavioural deficits in homozygous and heterozygous aromatase knockout mice.
The present study was carried out to determine whether aromatase knockout (ArKO) mice are completely devoid of aromatase activity in their brain and gonads and to compare aromatase activity in wild-type and ArKO mice, as well as in heterozygous (HET) mice of both sexes that were previously shown to display a variety of reproductive behaviours at levels intermediate between wild-type and ArKO mice. Aromatase activity was extremely low, and undetectable by the tritiated water assay, in homogenates of the preoptic area-hypothalamus of adult wild-type mice, but was induced following a 12-day treatment with testosterone. The induction of aromatase activity by testosterone was significantly larger in males than in females. Even after 12 days exposure to testosterone, no aromatase activity was detected in the brain of ArKO mice of either sex whereas HET mice showed intermediate levels of activity between ArKO and wild-type. Aromatase activity was also undetectable in the ovary of adult ArKO females but was very high in the wild-type ovary and intermediate in the HET ovary. In wild-type mice, a high level of aromatase activity was detected on the day of birth even without pretreatment with testosterone. This neonatal activity was higher in males than in females, but females nevertheless appear to display a substantial level of oestrogen production in their brain. Aromatase activity was undetectable in the brain of newborn ArKO males and females and was intermediate between wild-type and ArKO in HET mice. In conclusion, the present study confirms that ArKO mice are unable to synthesize any oestrogens, thereby validating the ArKO mouse as a valuable tool in the study of the physiological roles of oestradiol. In addition, it demonstrates that the intermediate behaviour of HET mice presumably reflects the effect of gene dosage on aromatase expression and activity, that aromatase activity is sexually differentiated in mice during the neonatal period as well as in adulthood and, finally, that the neonatal female brain produces substantial amounts of oestrogens that could play a significant role in the sexual differentiation of the female brain early in life.


Measurement by fluorescence of interstitial adenosine levels in normoxic, hypoxic, and ischemic perfused rat hearts.
An improved assay was used to investigate the effects of hypoxia or ischemia on interstitial fluid and coronary venous effluent levels of adenosine in isolated perfused nonworking rat hearts. The adenosine in 5- to 10-microliter samples of left ventricular epicardial surface transudates and coronary effluents was reacted with chloroacetaldehyde, and the fluorescent derivative (1,N6-ethenoadenosine) was quantitated using high pressure liquid chromatography and fluorescence detection. Hearts responding to hypoxia could be separated into two groups. In one group of hearts, the control (normoxic) transudate and effluent adenosine concentrations were 94 +/- 24 and 41 +/- 6 pmol/ml, respectively. These values increased by 118 and 96%, respectively, with 5 minutes of hypoxia (30% O2), and returned to control levels 5 minutes after resumption of normoxia. In a second group of hearts, the normoxic control levels of adenosine in the transudates (42 +/- 7 pmol/ml) and coronary effluents (62 +/- 17 pmol/ml) were increased with hypoxia by 174 and 1,178%, respectively. However, the transudate levels continued to rise for 5 minutes after resumption of normoxic perfusion while effluent levels fell. In another series of hearts, global ischemia for 30 seconds elicited an elevation of transudate adenosine levels by 362 to 641% above control (58 +/- 15 pmol/ml) as determined 30 seconds after resumption of perfusion flow.(ABSTRACT TRUNCATED AT 250 WORDS)


An evaluation of enteral feeding practices in critically ill children.
Establishing and sustaining enteral feeding in critically ill children is challenging and has met with many problems. The aim of this study was to investigate (a) how actual calorie intake compared with estimated caloric requirements and (b) whether feeding guideline adherence resulted in improved nutritional intake. A prospective observational study was undertaken over 1 month in a tertiary referral paediatric intensive care unit (PICU) in the northwest of England. Forty-seven children were studied, with a wide range of diagnoses in a 1-month period. Only 47% of the children had enteral feeds started within our 6 h post-admission target. Over half (55%) of the children received less than half of their estimated calorie requirements, but if feeding guidelines were followed, this resulted in a significantly higher (p = 0.004) delivery of the child's estimated requirements. This study found that many children are not receiving adequate nutrition in PICU and that the use of feeding guidelines significantly improves calorie delivery in PICU patients. This paper highlights the dearth of research related to enteral feeding in critically ill children. We found that the use of feeding guidelines improved calorie delivery and so units should be encouraged to develop their own guidelines based on the best evidence available.


Appositional angle closure in Chinese with primary angle closure and primary angle closure glaucoma after laser peripheral iridotomy.
To determine the prevalence of appositional angle closure (AAC) after laser peripheral iridotomy (LPI) in the eyes of Chinese patients with primary angle closure (PAC) and primary angle closure glaucoma (PACG) and to evaluate its pathogenesis by investigating anatomic characteristics. This was a cross-sectional observational study. PAC and PACG subjects were consecutively enrolled after LPI. Ultrasound biomicroscopy (UBM) images, obtained in darkness, of each quadrant without peripheral anterior synechia (PAS) under gonioscopy were qualitatively assessed. Darkroom provocative test (DRPT) was also performed. A total of 134 eyes of 134 patients were enrolled. AAC was observed in ≥ 1 quadrant of UBM image in 85 subjects (63.4%). There were 116 randomly selected quadrants without PAS for 134 patients (86.6%). AAC existed in 38 quadrants (32.8%). Among these, plateau iris accounted for 44.7%, anteriorly inserted iris for 13.2%, thick iris for 13.2%, and anteriorly inserted iris combined with thick iris for 18.4% of the total. One hundred fifteen patients underwent DRPT and its positive rate of eyes with AAC ≥ 2 quadrants (37.5% [12 of 32 patients]) was significantly higher than those ≤ 1 quadrant (16.9% [14 of 83 patients]; P = 0.018). However, no significant differences were found between eyes with nonsynechia plateau iris ≥ 2 quadrants (36.4% [4 of 11 patients]) and those ≤ 1 quadrant (21.2% [22 of 104 patients]; P = 0.266). Approximately two-thirds of PAC and PACG eyes of Chinese patients after LPI had AAC. Plateau iris accounted for less than 50% of AAC. Other factors such as a thick peripheral iris and an anteriorly inserted iris also contributed to AAC. DRPT results suggested AAC might have more functional meaning than plateau iris.


Effectiveness of dry needling on the local pressure pain threshold in patients with masticatory myofascial pain. Systematic review and preliminary clinical trial.
Objective: To systematically review the literature to identify controlled clinical trials evaluating the effectiveness of deep dry needling (DDN) in the treatment of patients with chronic masticatory myofascial pain (MMP).Methods: The sample size for a clinical trial was calculated and involved five patients who consecutively presented for treatment of MMP. The percentage of change in the means of three consecutive measurements of the pressure pain threshold (PPT) of myofascial trigger points (MTPs) was calculated and the statistical significance of this difference evaluated using the Wilcoxon test.Results: Twenty-five studies were considered for inclusion based on title and abstract. Only 2 studies met the inclusion criteria and were used to calculate the sample size. DDN significantly increased (p = 0.04) the PPT in MTP (44.6%) compared with sham procedure (-5.5%).Conclusion: Patients with chronic MMP treated with DDN of MTPs showed an increase in PPT measurements on the experimental side.Abbreviations: CGRP: calcitonin gene-related peptide; DC/TMD: diagnostic criteria for temporomandibular disorders; DDN: deep dry needling; DN: dry needling; LILACS: Latin American and Caribbean Health Sciences; MMP: masticatory myofascial pain; MTP: myofascial trigger point; MTPs: myofascial trigger points; PPT: pressure pain threshold; RCTs: randomized clinical trials; SciELO: Scientific Electronic Library Online; SP: Substance P; TMD: temporomandibular disorders.


How our own speech rate influences our perception of others.
In conversation, our own speech and that of others follow each other in rapid succession. Effects of the surrounding context on speech perception are well documented but, despite the ubiquity of the sound of our own voice, it is unknown whether our own speech also influences our perception of other talkers. This study investigated context effects induced by our own speech through 6 experiments, specifically targeting rate normalization (i.e., perceiving phonetic segments relative to surrounding speech rate). Experiment 1 revealed that hearing prerecorded fast or slow context sentences altered the perception of ambiguous vowels, replicating earlier work. Experiment 2 demonstrated that talking at a fast or slow rate prior to target presentation also altered target perception, though the effect of preceding speech rate was reduced. Experiment 3 showed that silent talking (i.e., inner speech) at fast or slow rates did not modulate the perception of others, suggesting that the effect of self-produced speech rate in Experiment 2 arose through monitoring of the external speech signal. Experiment 4 demonstrated that, when participants were played back their own (fast/slow) speech, no reduction of the effect of preceding speech rate was observed, suggesting that the additional task of speech production may be responsible for the reduced effect in Experiment 2. Finally, Experiments 5 and 6 replicate Experiments 2 and 3 with new participant samples. Taken together, these results suggest that variation in speech production may induce variation in speech perception, thus carrying implications for our understanding of spoken communication in dialogue settings. (PsycINFO Database Record


Nucleic Acid Detection Using CRISPR/Cas Biosensing Technologies.
For infectious diseases, rapid and accurate identification of the pathogen is critical for effective management and treatment, but diagnosis remains challenging, particularly in resource-limited areas. Methods that accurately detect pathogen nucleic acids can provide robust, accurate, rapid, and ultrasensitive technologies for point-of-care diagnosis of pathogens, and thus yield information that is invaluable for disease management and treatment. Several technologies, mostly PCR-based, have been employed for pathogen detection; however, these require expensive reagents and equipment, and skilled personnel. CRISPR/Cas systems have been used for genome editing, based on their ability to accurately recognize and cleave specific DNA and RNA sequences. Moreover, following recognition of the target sequence, certain CRISPR/Cas systems including orthologues of Cas13, Cas12a, and Cas14 exhibit collateral nonspecific catalytic activities that can be employed for nucleic acid detection, for example by degradation of a labeled nucleic acid to produce a fluorescent signal. CRISPR/Cas systems are amenable to multiplexing, thereby enabling a single diagnostic test to identify multiple targets down to attomolar (10-18 mol/L) concentrations of target molecules. Developing devices that couple CRISPR/Cas with lateral flow systems may allow inexpensive, accurate, highly sensitive, in-field deployable diagnostics. These sensors have myriad applications, from human health to agriculture. In this review, we discuss the recent advances in the field of CRISPR-based biosensing technologies and highlight insights of their potential use in a myriad of applications.


Tolerability of effective high doses of paliperidone palmitate in patients with severe resistant schizophrenia.
The aim of the study was to evaluate the effectiveness and tolerability of doses of paliperidone palmitate (PP) of 175 mgEq and over/28 days in patients with severe schizophrenia [Clinical Global Impression-Severity scale (CGI-S)≥5] and their retention in treatment. A 36-month prospective, observational study was carried out of patients with severe schizophrenia who were treated with 175 mg and over every 28 days of PP to achieve clinical stabilization (N=30). Assessment included CGI-S, WHO Disability Assessment Schedule, Camberwell Assessment of Need, and Medication Adherence Report Scale. Laboratory tests, weight, side effects, reasons for discharge, and hospital admissions were measured. The average dose of PP was 228.7 (11.9) mgEq/28 days. There was one discharge because of side effects. Weight and prolactin levels decreased. After 3 years, CGI-S (P<0.01), Camberwell Assessment of Need (P<0.01), and WHO Disability Assessment Schedule in the four areas (P<0.05) decreased. The Medication Adherence Report Scale increased (P<0.001). There were fewer hospital admissions (P<0.001). Retention in treatment after 36 months was 90%. Tolerability of 175 mgEq/28 days and over of PP was very good, being useful in improving treatment adherence in severely ill patients and helping in this way to achieve clinical stabilization and better social functioning. These patients were clozapine candidates; thus, high doses of PP could be an alternative for them.


Collembola as alternative prey sustaining spiders in arable ecosystems: prey detection within predators using molecular markers.
Collembola comprise a major source of alternative prey to linyphiid spiders in arable fields, helping to sustain and retain these predators as aphid control agents within the crop. Polymerase chain reaction primers were developed for the amplification, from spider gut samples, of DNA from three of the most abundant species of Collembola in wheat crops in Europe, namely Isotoma anglicana, Lepidocyrtus cyaneus and Entomobrya multifasciata. The primers amplified fragments of the mitochondrial cytochrome oxidase subunit I (COI) gene and were designed following alignment of comparable sequences for a range of predator and prey species. Each of the primer pairs proved to be species-specific to a Collembola species, amplifying DNA fragments from 211 to 276 base pairs in length. Following consumption of a single collembolan, prey DNA was detectable in 100% of spiders after 24 h of digestion. We report the first use of DNA-based techniques to detect predation by arthropods on natural populations of prey in the field. All three species of Collembola were consumed by the spiders. By comparing the ratios of the Collembola species in the field with the numbers of spiders that gave positive results for each of those species, it was possible to demonstrate that the spiders were exercising prey choice. Overall, a single target species of Collembola was eaten by 48% of spiders while a further 16% of spiders contained DNA from two different species of Collembola. Preference was particularly evident for I. anglicana, the species most frequently found in spider guts yet the least numerous of the three target species in the field.


Mutational and functional analysis of Large in a novel CHO glycosylation mutant.
Inactivating mutations of Large reduce the functional glycosylation of alpha-dystroglycan (alpha-DG) and lead to muscular dystrophy in mouse and humans. The N-terminal domain of Large is most similar to UDP-glucose glucosyltransferases (UGGT), and the C-terminal domain is related to the human i blood group transferase beta1,3GlcNAcT-1. The amino acids at conserved motifs DQD+1 and DQD+3 in the UGGT domain are necessary for mammalian UGGT activity. When the corresponding residues were mutated to Ala in mouse Large, alpha-DG was not functionally glycosylated. A similar result was obtained when a DXD motif in the beta1,3GlcNAcT-1 domain was mutated to AIA. Therefore, the first putative glycosyltransferase domain of Large has properties of a UGGT and the second of a typical glycosyltransferase. Co-transfection of Large mutants affected in the different glycosyltransferase domains did not lead to complementation. While Large mutants were more localized to the endoplasmic reticulum than wild-type Large or revertants, all mutants were in the Golgi, and only very low levels of Golgi-localized Large were necessary to generate functional alpha-DG. When Large was overexpressed in ldlD.Lec1 mutant Chinese hamster ovary (CHO) cells which synthesize few, if any, mucin O-GalNAc glycans and no complex N-glycans, functional alpha-DG was produced, presumably by modifying O-mannose glycans. To investigate mucin O-GalNAc glycans as substrates of Large, a new CHO mutant Lec15.Lec1 that lacked O-mannose and complex N-glycans was isolated and characterized. Following transfection with Large, Lec15.Lec1 cells also generated functionally glycosylated alpha-DG. Thus, Large may act on the O-mannose, complex N-glycans and mucin O-GalNAc glycans of alpha-DG.


Identification of MUM1 as a prognostic immunohistochemical marker in follicular lymphoma using computerized image analysis.
Detection of MUM1+ cells in follicular lymphoma (FL) tissues was previously found to be associated with poor prognosis in a single report, whereas the usefulness of Ki-67 immunostaining remains debated. Our goal was to establish whether these markers have predictive value for patients with FL. We analyzed MUM1 and Ki-67 expression using immunohistochemistry in biopsy samples from 434 patients from the PRIMA randomized trial. The MUM1 prognostic value was then validated in a cohort of 138 patients from the FL2000 randomized trial, using the optimal cutoff value obtained from the PRIMA cohort. The surface of positive staining was quantified using computerized image analysis. In the PRIMA cohort, both high levels of MUM1 positivity (cutoff value of 0.80%) and high levels of Ki-67 positivity (cutoff value of 10.25%) were significantly associated with a shorter progression-free survival (PFS) (P = .004 and P = .007 for MUM1 and Ki-67, respectively). In a multivariate Cox proportional hazards regression model, only MUM1 retained a statistical significance (hazards ratio 1.56; 95% confidence interval, 1.02-2.37; P = .038) after adjustment for the maintenance arm of treatment and the follicular lymphoma international prognostic index score. In the FL2000 cohort, high levels of MUM1 positivity were significantly associated to a shorter PFS (P = .004) and to a trend toward a shorter overall survival (P = .043). This remained significant using a multivariate Cox regression model after adjustment for the follicular lymphoma international prognostic index and the treatment arm for PFS (P = .016). These results show that MUM1 is a strong and robust predictive immunohistochemical marker in patients with FL.


Kidney is in trouble with mediators.
The kidneys receive 20-25 % of cardiac output and play a main role in the control of cardiovascular homeostasis. It is an endocrine organ that regulates and produces many substances, scavenger particles and immune complexes. Cytokines, growth factors, reactive oxygen metabolites, bioactive lipids, proteases, vasoactive substances such as nitric oxide (NO), adrenomedullin (AM), urotensin-II (U-II), have been released in several diseases, and kidney is one of mostly affected organs in body. Some of these mediators act in a paracrine fashion while some act in autocrine. They play important roles in modulating the cardiovascular responses, renal hemodynamics, and probably in mediating the clinical and laboratory manifestations of several renal diseases. These mediators are like "a double edged sword". While small amounts of them mediate many physiological events, little excess may cause the damage to the healthy cells. Many investigators have searched the role(s) of mediators in several diseases. However, the findings are mostly like the model of "chicken and egg", and indistinguishable as to whether they are the causes of, or results of the diseases. We will discuss mainly the possible roles of NO, AM and U-II in children with several renal diseases and summarize what is known, and what must be known about these mediators.