Formation of circular satellite tobacco ringspot virus RNA in protoplasts transiently expressing the linear RNA.
The most abundant form of the satellite RNA of tobacco ringspot virus (sTRSV RNA) is a linear, unit length molecule of 359 nucleotide residues, designated L-(+)M. A postulated replication scheme for the satellite RNA has as its first, and apparently virus-independent, step the ligation of L-(+)M into the corresponding circular form C-(+)M. We transiently expressed L-(+)M wild type and L-(+)M mutants in tobacco protoplasts using an African cassava mosaic geminivirus vector. Measured extents of C-(+)M accumulation were correlated with computer-predicted folding to suggest wild-type secondary structure elements that might be deleted without reducing ligation. A 127-nucleotide residue mutant L-(+)M was created by replacing, with 7 and 3 residues, respectively, nucleotide residues 53-211 and 268-350, each of which was predicted to form a set of three adjacent imperfect stem-loops in wild-type L-(+)M. The mutant L-(+)M was found to be extensively ligated to C-(+)M in protoplasts and to retain a calculated helix of the wild-type molecule that incorporates the 3' terminal sequence. A trinucleotide in the 3' region was mutated so as to disrupt and restore, respectively, the calculated helix, reducing and restoring, respectively, C-(+)M formation. These results suggest that the 3' stem contributes to the suitability of the small L-(+)M molecules as a substrate for a protoplast RNA ligase and that computed folding of sTRSV RNA may be predictive of sTRSV RNA structure in vivo.


NIPBL: a new player in myeloid cell differentiation.
The nucleophosmin 1 gene (NPM1) is the most frequently mutated gene in acute myeloid leukemia. Notably, NPM1 mutations are always accompanied by additional mutations such as those in cohesin genes RAD21, SMC1A, SMC3, and STAG2 but not in the cohesin regulator, nipped B-like (NIPBL). In this work, we analyzed a cohort of adult patients with acute myeloid leukemia and NPM1 mutation and observed a specific reduction in the expression of NIPBL but not in other cohesin genes. In our zebrafish model, overexpression of the mutated form of NPM1 also induced downregulation of nipblb, the zebrafish ortholog of human NIPBL To investigate the hematopoietic phenotype and the interaction between mutated NPM1 and nipblb, we generated a zebrafish model with nipblb downregulation which showed an increased number of myeloid progenitors. This phenotype was due to hyper-activation of the canonical Wnt pathway: myeloid cells blocked in an undifferentiated state could be rescued when the Wnt pathway was inhibited by dkk1b mRNA injection or indomethacin administration. Our results reveal, for the first time, a role for NIPBL during zebrafish hematopoiesis and suggest that an interplay between NIPBL/NPM1 may regulate myeloid differentiation in zebrafish and humans through the canonical Wnt pathway and that dysregulation of these interactions may drive leukemic transformation.


Focal cortical and subcortical atrophy in early Parkinson's disease.
Neurodegeneration in clinically manifest Parkinson's disease affects the substantia nigra pars compacta, and gradually spreads to the limbic cortices and the neocortex. We used MRI imaging coupled with automated surface reconstruction and segmentation methods to examine cortical thickness and subcortical volumes in nondemented, early-stage Parkinson's disease patients compared to matched healthy control participants. These methods, which have been previously used to document cortical thickness changes in patients with Alzheimer's disease and Huntington's disease but not Parkinson's disease, use MR signal intensity information and the geometric constraints of the cortical and subcortical structures for an accurate tissue classification. Parkinson's disease patients were matched to the control group in psychomotor processing speed and executive functioning, but showed higher anxiety state scores. Our results demonstrated focal cortical thinning in the Parkinson's disease group in the orbitofrontal cortex, ventrolateral prefrontal cortex, and occipito-parietal areas. Subcortically, striatal volume loss was noted. These results demonstrate that both cortical and subcortical structural changes occur at relatively early stages of the disease, and are discussed in terms of the emotional dysregulation that occurs early on in patients with Parkinson's disease.


Preclinical mouse model to monitor live Muc5b-producing conjunctival goblet cell density under pharmacological treatments.
Modification of mucous cell density and gel-forming mucin production are established hallmarks of mucosal diseases. Our aim was to develop and validate a mouse model to study live goblet cell density in pathological situations and under pharmacological treatments. We created a reporter mouse for the gel-forming mucin gene Muc5b. Muc5b-positive goblet cells were studied in the eye conjunctiva by immunohistochemistry and probe-based confocal laser endomicroscopy (pCLE) in living mice. Dry eye syndrome (DES) model was induced by topical application of benzalkonium chloride (BAK) and recombinant interleukine (rIL) 13 was administered to reverse the goblet cell loss in the DES model. Almost 50% of the total of conjunctival goblet cells are Muc5b+ in unchallenged mice. The decrease density of Muc5b+ conjunctival goblet cell population in the DES model reflects the whole conjunctival goblet cell loss. Ten days of BAK in one eye followed by 4 days without any treatment induced a -18.3% decrease in conjunctival goblet cell density. A four days of rIL13 application in the DES model restored the normal goblet cell density. Muc5b is a biological marker of DES mouse models. We bring the proof of concept that our model is unique and allows a better understanding of the mechanisms that regulate gel-forming mucin production/secretion and mucous cell differentiation in the conjunctiva of living mice and can be used to test treatment compounds in mucosal disease models.


Internal carbon source from sludge pretreated by microwave-H2O2 for nutrient removal in A2/O-membrane bioreactors.
To improve the nutrient removal, the feasibility was studied for the organics released from sludge pretreated by microwave-H2O2 process (MHP) to be used as internal carbon source in two A2/O-membrane bioreactors (MBRs). The experiments were conducted for the nutrient removal and the membrane fouling. The results showed that the removal efficiencies of TN and TP were improved by 11% and 28.34%, respectively, as C/N ratio was adjusted to 8 by adding the internal carbon source, and the ratio of soluble chemical oxygen demand (sCOD) consumed easily for denitrification was about 46% of the total sCOD in the internal carbon source. The addition of the internal carbon sources did not lead to severe membrane fouling in the experimental A2/O-MBR. It is implied that the organics released from sludge pretreated by MHP could be used as the internal carbon source to enhance the nutrient removal in A2/O-MBRs.


Ligand-mediated control of dislocation dynamics and resulting particle morphology of GdOCl nanocrystals.
While much progress has been achieved in the shape-controlled synthesis of nanocrystals, chemical strategies to define morphology remain primarily empirical. Here, a mechanistic study of the influence of different coordinating ligands on the kinetics and thermodynamics of crystal growth during the preparation of GdOCl by the non-hydrolytic condensation of GdCl3 and Gd(O (i) Pr)3 is reported. Growth using oleylamine, octadecylamine, trioctylamine, and didodecylamine yields 2D nanosheets with approximately square cross sections, whereas growth in trioctylphosphine oxide yields larger and thicker platelets. The nanostructures are characterized by the presence of spiral growth patterns and dislocations. Apart from preferential binding to specific crystallographic facets, the coordinating ligands are suggested to control the extent of supersaturation, thereby facilitating and tuning dislocation-mediated growth. Upon depletion of monomers, thermodynamic surface energy considerations become of paramount importance and the nanocrystals are reshaped via mass transport from edges to sides yielding their eventual equilibrium shapes. The mechanisms developed here are thought to be broadly generalizable to the ligand-directed growth of nanomaterials.


Performance of Bacticard Candida compared with the germ tube test for the presumptive identification of Candida albicans.
Bacticard Candida was compared with the germ tube test for the rapid, presumptive identification of Candida albicans. This test kit detects the enzymatic activities l-proline aminopeptidase and beta-galactosaminidase in yeast colonies grown on culture media. Candida albicans produces both enzymes whereas other yeasts produce only one or neither of the enzymes. We evaluated 536 isolates including eight genera and 33 species of medically important yeasts, including 228 C. albicans and 36 C. dubliniensis. Both tests did not discriminate between C. albicans and C. dubliniensis isolates. The sensitivity and specificity for the Bacticard Candida test were 97.8 and 96.5%, respectively. Bacticard Candida and germ tube tests detected 246 (93.2%), and 256 (97%) C. albicans plus C. dubliniensis isolates. There were eight false-positive results with BactiCard Candida kit and four false-positive results with the germ tube test. Positive and negative predictive values for Bacticard Candida enzymatic test were 95.3 and 98.4%, respectively, while 97.4 and 98.1% for the germ tube test, its specificity being 98.1% and efficiency 97% (97.7% for germ tube). We have observed slightly lower values of sensitivity and specificity than those reported by others using the BactiCard test kit. Bacticard Candida provides a rapid and accurate alternative to the germ tube test for the presumptive identification of C. albicans.


Child care arrangements and repeated ear infections in young children.
The purpose of this study was to assess the association between child care arrangements and repeated ear infections in young children. Data on 5818 children younger than 6 years of age were taken from the 1988 National Health Interview Survey of Child Health. Seventeen percent of the children had had repeated ear infections in the year preceding the interview. Multiple logistic regression analysis showed that factors significantly associated with repeated ear infections were age (1 to 2 years), race (White), sex (male), and medical history (repeated tonsillitis, enlarged adenoids, or asthma). When these factors were controlled for, children in a current child care arrangement still had a 50% higher chance of repeated ear infections than did children not in care. Children in day care centers were at higher risk than children cared for in homes. Care in settings with more than six children elevated the risk, but hours per week of care did not. Decreasing the risk of repeated ear infections in young children in out-of-home child care settings may require smaller group sizes. Given the increasing use of child care, research should continue to identify specific aspects of child care that contribute to this problem.


Dipole potential and head-group spacing are determinants for the membrane partitioning of pregnanolone.
The membrane interactions of pregnanolone, an intravenous general anesthetic steroid, were characterized using fluorescence spectroscopy and monolayer technique. di-8-ANEPPS [4-[2-[6-(dioctylamino)-2-naphthalenyl]ethenyl]-1-(3-sulfopropyl)-pyridinium], a membrane dipole potential (Psi)-sensitive probe, revealed pregnanolone to decrease Psi similarly as reported previously for other anesthetics. The decrement in Psi was approximately 16 and 10 mV in dipalmitoylphosphatidylcholine (DPPC) and DPPC/cholesterol (90:10, mol/mol) vesicles, respectively. Diphenylhexatriene anisotropy indicated pregnanolone to have a negligible effect on the acyl chain order. In contrast, substantial changes were observed for the fluorescent dye Prodan, thus suggesting pregnanolone to reside in the interfacial region of lipid bilayers. Langmuir balance studies indicated increased association of pregnanolone to DPPC monolayers containing cholesterol or 6-ketocholestanol at surface pressures pi > 20 mN/m as well as to monolayers of the unsaturated 1-palmitoyl-2-oleoylphosphatidylcholine. In the same surface pressure range, the addition of phloretin, which decreases Psi, reduced the penetration of pregnanolone into the monolayers. These results suggest that membrane partitioning of pregnanolone is influenced by the spacing of the phosphocholine head groups as well as by membrane dipole potential. The latter can be explained in terms of electrostatic dipole-dipole interactions between pregnanolone and the membrane lipids with their associated water molecules. Considering the universal nature of these interactions, they are likely to affect membrane partitioning of most, if not all, weakly amphiphilic drugs.


Elevated frequencies of circulating Th22 cell in addition to Th17 cell and Th17/Th1 cell in patients with acute coronary syndrome.
Atherosclerosis is a chronic inflammatory disease mediated by immune cells. Th22 cells are CD4(+) T cells that secret IL-22 but not IL-17 or IFN-γ and are implicated in the pathogenesis of inflammatory disease. The roles of Th22 cells in the pathophysiologic procedures of acute coronary syndrome (ACS) remain unclear. The purpose of this study is to investigate the profile of Th22, Th17 and Th17/Th1 cells in ACS patients, including unstable angina (UA) and acute myocardial infarction (AMI) patients. In this study, 26 AMI patients, 16 UA patients, 16 stable angina (SA) patients and 16 healthy controls were included. The frequencies of Th22, Th17 and Th17/Th1 cells in AMI, UA, SA patients and healthy controls were examined by flow cytometry. Plasma levels of IL-22, IL-17 and IFN-γ were measured by enzyme-linked immunosorbent assay (ELISA). Th22, Th17 and Th17/Th1 cells were significantly increased in AMI and UA patients compared with SA patients and healthy controls. Moreover, plasma IL-22 level was significantly elevated in AMI and UA patients. In addition, Th22 cells correlated positively with IL-22 as well as Th17 cells in AMI and UA patients. Our findings showed increased frequencies of both Th22 and Th17 cells in ACS patients, which suggest that Th22 and Th17 cells may play a potential role in plaque destabilization and the development of ACS.


Deep phylogeographic structure may indicate cryptic species within the Sparid genus Spondyliosoma.
Two geographically nonoverlapping species are currently described within the sparid genus Spondyliosoma: Spondyliosoma cantharus (Black Seabream) occurring across Mediterranean and eastern Atlantic waters from NW Europe to Angola and S. emarginatum (Steentjie) considered endemic to southern Africa. To address prominent knowledge gaps this study investigated range-wide phylogeographic structure across both species. Mitochondrial DNA sequences revealed deep phylogeographic structuring with four regionally partitioned reciprocally monophyletic clades, a Mediterranean clade and three more closely related Atlantic clades [NE Atlantic, Angola and South Africa (corresponding to S. emarginatum)]. Divergence and distribution of the lineages reflects survival in, and expansion from, disjunct glacial refuge areas. Cytonuclear differentiation of S. emarginatum supports its validity as a distinct species endemic to South African waters. However, the results also indicate that S. cantharus may be a cryptic species complex wherein the various regional lineages represent established/incipient species. A robust multilocus genetic assessment combining morphological data and detailing interactions among lineages is needed to determine the full diversity within Spondyliosoma and the most adequate biological and taxonomic status.


Network Analysis of the Participation and Activity Inventory for Children and Youth (PAI-CY) 7-12 Years with Visual Impairment.
Children with visual impairment often experience more difficulties regarding participation compared to sighted peers. The Participation and Activity Inventory for Children and Youth (PAI-CY) has recently been developed to assess their participation needs. A novel application in the field of questionnaires is the use of network analysis to explore interrelations between items in order to capture their complex interactions as a reflection of the overall construct of measurement. This study aimed to apply network modeling for the PAI-CY 7-12 from the perspectives of children and their parents. Children and their parents (n = 195) completed the 55-item PAI-CY via face-to-face interviews and a web-based survey, respectively. Internal consistency, test-retest reliability, and concordance between children and parents were investigated. Two networks were created, along with visualizations of shared and differential connections between children and parents. Eight items were deleted. Network structures were dissimilar; for children, connections evolved around social contacts and school items, whereas for parents, mobility, leisure time, acceptance, self-reliance, and communication items prevailed. In the children's network, playing imaginary games, inviting a friend to play at home, and estimating the distance from others were most connected to other items. This study uniquely identifies connections between items of the PAI-CY 7-12, highlighting the different perspectives parents and children have on what defines participation, possibly implying that they perceive the relevance of various rehabilitation programs differently. Rehabilitation programs aimed at improving the most connected items might positively affect other items in the network, possibly improving children's participation.


Phase I safety, pharmacokinetics, and inhibition of SRC activity study of saracatinib in patients with solid tumors.
This dose-escalation study evaluated the safety, tolerability, and pharmacokinetics (PK) of the oral Src inhibitor saracatinib (AZD0530) in patients with advanced solid malignancies. Tumor biopsy samples were taken to investigate the effect of saracatinib on Src activity in tumors. Part A of the study followed a multiple-ascending dose design to establish the maximum tolerated dose (MTD) of saracatinib. Part B was a randomized, parallel-group, cohort-expansion phase to further assess tolerated doses. Safety, tolerability, and Src activity (immunohistochemistry and lysate-based methodologies) were assessed after 21 days of once-daily oral dosing. PK was assessed after single and multiple dosing. In part A, 30 patients received once-daily saracatinib at doses of 60 to 250 mg; the MTD was established as 175 mg. In part B, 51 patients were randomized to receive 50 mg (n = 16), 125 mg (n = 16), or 175 mg (n = 19) of saracatinib. The most common grade ≥3 events considered to be treatment related were anemia, diarrhea, and asthenia. Tumor Src activity was reduced following saracatinib treatment. The area under the concentration-time curve and C(max) of saracatinib increased with increasing dose. Saracatinib accumulated 4- to 5-fold on once-daily dosing to reach steady-state exposure after 10 to 17 days of dosing. The half-life was ∼40 hours. Saracatinib was well tolerated in patients with advanced solid malignancies. A reduction in tumor Src activity was observed. PK data show that saracatinib is suitable for once-daily oral dosing. Based on this study, the recommended dose for the phase II studies was chosen to be 175 mg/d.


Differential vagal effects on antegrade vs. retrograde atrioventricular conduction.
The influence of postganglionic vagal stimulation (PGVS) on antegrade and retrograde atrioventricular nodal conduction was studied in 17 isolated rabbit heart tissue preparations by pacing at the crista terminalis or His bundle, respectively. The effect of short bursts of PGVS on prolongation of atrioventricular conduction was phase dependent with respect to the cardiac cycle. This phasic dependency was more pronounced during antegrade atrioventricular conduction. Although the control retrograde atrioventricular conduction time was longer than the antegrade (P less than 0.05) at or near the time in the cycle during which vagal stimulation caused maximal prolongation of conduction time (optimal phase), PGVS-induced maximal prolongation of the antegrade atrioventricular conduction time was significantly greater than that of the retrograde (P less than 0.02). Moreover, when PGVS was introduced at a fixed phase in the cycle, but with increasing amplitude, antegrade atrioventricular conduction time was progressively prolonged, and block was observed first in the antegrade direction, whereas retrograde atrioventricular conduction continued. Microelectrode recordings during these experiments showed consistently that PGVS-induced hyperpolarization in the N region of the atrioventricular node was greater during antegrade atrioventricular conduction. This suggests that vagal effects depended not only on the intensity and phase of stimulation, but also on electronic influences which apparently are different during antegrade and retrograde conduction.


Relationship between Translational and Rotational Dynamics of Alkyltriethylammonium-Based Ionic Liquids.
1H spin-lattice relaxation experiments have been performed for a series of ionic liquids including bis(trifluoromethanesulfonyl)imide anion and cations of a varying alkyl chain length: triethylhexylammonium, triethyloctylammonium, decyltriethylammonium, dodecyltriethylammonium, triethyltetradecylammonium, and hexadecyltriethylammonium. The relaxation studies were carried out in abroad frequency range covering three orders of magnitude, from 10 kHz to 10 MHz, versus temperature. On the basis of a thorough, quantitative analysis of this reach data set, parameters characterizing the relative, cation-cation, translation diffusion (relative diffusion coefficients and translational correlation times), and rotational motion of the cation (rotational correlation times) were determined. Relationships between these quantities and their dependence on the alkyl chain length were discussed in comparison to analogous properties of molecular liquids. It was shown, among other findings, that the ratio between the translational and rotational correlation times is smaller than for molecular liquids and considerably dependent on temperature. Moreover, a comparison of relative and self-diffusion coefficients indicate correlated translational dynamics of the cations.


The influence of programmed cell death ligand 2 (PD-L2) expression on survival outcome and tumor microenvironment in diffuse large B cell lymphoma.
The frequency and significance of programmed cell death ligand (PD-L) 2 expression in diffuse large B cell lymphoma (DLBCL) remain undefined. We described the expression pattern of PD-L/PD-1 in 88 DLBCL patients using immunohistochemistry. The association between PD-L expression and clinical characteristics/outcomes were analyzed. PD-L1 and PD-L2 were expressed in 14.8% and 68.2% of DLBCL patients with median positivity on tumor cells of 100% and 90%, respectively. PD-1 on tumor-infiltrating lymphocytes (TILs) was expressed in 12.5% of patients. Interestingly, 45.5% of patients had PD-L2 expressing TILs which were significantly associated with bulky disease (p = .046) and elevated lactate dehydrogenase (p = .048). PD-L1 and/or PD-L2 expression on lymphoma cells was associated with inferior progression-free survival (Hazard ratio [HR] 2.20; 95% Confidence Interval [CI] 1.004-4.84, p = .049) and overall survival (HR 2.27; 95%CI 1.03-4.98, p = .042), using multivariate analysis. In summary, PD-L2 expression on DLBCL is common and, together with PD-L1, were related to poor outcomes.


[Psychomotor development of newborn infants at risk with reference to the neuron-specific enolase].
In a prospective study the psychomotor development of an unselected collective of risk newborn infants up to the end of the second year of life was examined. 199 children have developed normal, 21 showed developmental abnormalities, 84 light to moderate disturbances, and 30 severe disabilities. Already in the newborn period the concentration of neuron-specific enolase in the serum was evaluated. The NSE could be an indicator of brain damage. The determined values of NSE showed a log-normal-distribution at the three times (cord-blood, first or second day of life, third or fourth day of life, and ninth to eleventh day of life). Significant relationship between the psychomotor development during the first two years of life and of NSE-concentration was not found. Nevertheless the diagnostic value of the NSE is better with a specificity of 47.7% and a sensitivity of 80.0% than those of the results of intracranial ultrasound examination or of the erythrocytic-density-test. In contrast to these results the use of the neuron-specific enolase alone for the prediction of individual prognosis of children is not be recommended.


Conventional and functional MR imaging of peripheral nerve sheath tumors: initial experience.
Differentiating benign from malignant peripheral nerve sheath tumors can be very challenging using conventional MR imaging. Our aim was to test the hypothesis that conventional and functional MR imaging can accurately diagnose malignancy in patients with indeterminate peripheral nerve sheath tumors. This institutional review board-approved, Health Insurance Portability and Accountability Act-compliant study retrospectively reviewed 61 consecutive patients with 80 indeterminate peripheral nerve sheath tumors. Of these, 31 histologically proved peripheral nerve sheath tumors imaged with conventional (unenhanced T1, fluid-sensitive, contrast-enhanced T1-weighted sequences) and functional MR imaging (DWI/apparent diffusion coefficient mapping, dynamic contrast-enhanced MR imaging) were included. Two observers independently assessed anatomic (size, morphology, signal) and functional (ADC values, early arterial enhancement by dynamic contrast-enhanced MR) features to determine interobserver agreement. The accuracy of MR imaging for differentiating malignant from benign was also determined by receiver operating characteristic analysis. Of 31 peripheral nerve sheath tumors, there were 9 malignant (9%) and 22 benign ones (81%). With anatomic sequences, average tumor diameter (6.3 ± 1.8 versus 3.9 ± 2.3 mm, P = .009), ill-defined/infiltrative margins (77% versus 32%; P = .04), and the presence of peritumoral edema (66% versus 23%, P = .01) were different for malignant peripheral nerve sheath tumors and benign peripheral nerve sheath tumors. With functional sequences, minimum ADC (0.47 ± 0.32 × 10(-3) mm(2)/s versus 1.08 ± 0.26 × 10(-3) mm(2)/s; P < .0001) and the presence of early arterial enhancement (50% versus 11%; P = .03) were different for malignant peripheral nerve sheath tumors and benign peripheral nerve sheath tumors. The minimum ADC (area under receiver operating characteristic curve was 0.89; 95% confidence interval, 0.73-0.97) and the average tumor diameter (area under the curve = 0.8; 95% CI, 0.66-0.94) were accurate in differentiating malignant peripheral nerve sheath tumors from benign peripheral nerve sheath tumors. With threshold values for minimum ADC ≤ 1.0 × 10(-3) mm(2)/s and an average diameter of ≥4.2 cm, malignancy could be diagnosed with 100% sensitivity (95% CI, 66.4%-100%). Average tumor diameter and minimum ADC values are potentially important parameters that may be used to distinguish malignant peripheral nerve sheath tumors from benign peripheral nerve sheath tumors.


Prevention of Sudden Cardiac Death in Hypertrophic Cardiomyopathy: What has Changed in The Guidelines?
The new European Society of Cardiology guidelines for hypertrophic cardiomyopathy (HCM) define the estimation of sudden cardiac death (SCD) risk as an integral part of clinical management. An implantable cardioverter defibrillator (ICD) is recommended (class IIa) when the risk is ≥ 6%. To compare the SCD risk stratification according to the 2011 and 2014 recommendations for ICD implantation in patients with HCM. Retrospective study including 105 patients diagnosed with HCM. The indication for ICD was assessed using the 2011 and 2014 guidelines. Statistical analysis was performed using SPSS software version 188.8.131.52®. The tests performed were bilateral, considering the significance level of 5% (p < 0.05). Regarding primary prevention, according to the 2011 ACCF/AHA recommendations, 39.0% of the patients had indication for ICD implantation (level of evidence IIa). Using the 2014 guidelines, only 12.4% of the patients had an indication for ICD implantation. Comparing the two risk stratification models for patients with HCM, we detected a significant reduction in the number of indications for ICD implantation (p < 0.001). Of the 41 patients classified as IIa according to the 2011 recommendations, 68.3% received a different classification according to the 2014 guidelines. Significant differences were found when comparing the SCD risk stratification for ICD implantation in the two guidelines. The current SCD risk score seems to identify many low-risk patients who are not candidates for ICD implantation. The use of this new score results in a significant reduction in the number of ICD implanted.


The Contribution of Plasma and Brain Vitamin C on Age and Gender-Related Cognitive Differences: A Mini-Review of the Literature.
There is increasing evidence that sex differences in the brain may contribute to gender-related behavioral differences, including cognitive function. Literature has revealed gender dimorphisms in cognitive function between males and females. Additionally, several risk factors associated with cognitive decline depend on chronological age. It is well recognized that the process of aging is associated with a decline in cognitive ability and brain function. Various explanations may account for these gender-related cognitive differences and age-associated cognitive changes. Recent investigations have highlighted the importance of vitamin C in maintaining brain health and its association with cognitive function in both cognitively intact and impaired cohorts. The present review explores previous literature that has evaluated differences in plasma/brain vitamin C between genders and during aging. It then assesses whether these age and gender-related differences may affect the relationship between plasma/brain vitamin C and cognition. The purpose of this review was to examine the evidence for a link between plasma/brain vitamin C and cognition and the impact of gender and age on this relationship. Epidemiological studies have frequently shown higher vitamin C plasma concentrations in women. Similarly, aging has been systematically associated with reductions in plasma vitamin C levels. A range of animal studies has demonstrated potential gender and age-related differences in vitamin C brain distribution and utilization. The reviewed literature suggests that gender differences in plasma and brain vitamin C may potentially contribute to differences in gender-associated cognitive ability, particularly while females are pre-menopausal. Additionally, we can propose that age-associated differences in plasma and brain vitamin C may be potentially linked to age-associated cognitive differences, with older cohorts appearing more vulnerable to experience declines in plasma vitamin C concentrations alongside compromised vitamin C brain regulation. This review encourages future investigations to take into account both gender and age when assessing the link between plasma vitamin C concentrations and cognitive function. Further large scale investigations are required to assess whether differences in cognitive function between genders and age groups may be causally attributed to plasma vitamin C status and brain distribution and utilization.


[DROSOPHILA MELANOGASTER GASTROINTESTINAL MYIASIS: ABOUT A MOROCCAN CASE REPORT].
Myiasis is an infestation of humans or animals by larval forms of brachycetes. Commonly observed locations are otolaryngological and cutaneous. Gastrointestinal localization remains exceptional. In this work, we report the observation of a Moroccan case of gastrointestinal myiasis caused by Drosophila melanogaster. Observation. The patient was a 56-year-old man living in a rural region of northwest Morocco. He was being followed at the National Oncology Institute in Rabat for pulmonary and renal adenocarcinoma and was put on neo-adjuvant chemotherapy with concomitant radiochemotherapy. The patient presented vomiting twice containing about forty small white, circular and hairless worms measuring 4 mm in length. This sample was sent to the parasitology and mycology laboratory for identification. Microscopic study of the larvae and adult flies obtained after rearing in the laboratory of parasitology allowed the diagnosis of myiasis caused by Drosophila melanogaster. This observation is noteworthy for both its exceptional anatomical localization and the implicated parasite species.


Insulin action in human thighs after one-legged immobilization.
Insulin action was assessed in thighs of five healthy young males who had one knee immobilized for 7 days by a splint. The splint was not worn in bed. Subjects also used crutches to prevent weight bearing of the immobilized leg. Immobilization decreased the activity of citrate synthase and 3-OH-acyl-CoA-dehydrogenase in the vastus lateralis muscle by 9 and 14%, respectively, and thigh volume by 5%. After 7 days of immobilization, a two-step euglycemic hyperinsulinemic clamp procedure combined with arterial and bilateral femoral venous catheterization was performed. Insulin action on glucose uptake and tyrosine release of the thighs at mean plasma insulin concentrations of 67 (clamp step I) and 447 microU/ml (clamp step II) was decreased by immobilization, whereas immobilization did not affect insulin action on thigh exchange of free fatty acids, glycerol, O2, or potassium. Before and during the clamp step I, lactate release was significantly higher in the immobilized than in the control thigh. Seven days of one-legged immobilization causes local decreased insulin action on thigh glucose uptake and net protein degradation.


Aesthetic evaluation of the labiolingual position of maxillary lateral incisors by orthodontists and laypersons.
The maxillary anterior teeth play a crucial role in smile aesthetics. Previous studies regarding the importance of maxillary lateral incisors for smile aesthetics concentrated on their size, incisor edge level, and inclination, etc. However, the aesthetic effect of lateral incisor movement in the spatial position has not been studied yet. Therefore, the purpose of this study was to explore the influence of the labiolingual position of maxillary lateral incisors on the aesthetic perception of smiles by orthodontists and laypersons, as well as analyze differences in this perception between male and female raters. A three-dimensional (3D) dental model was generated from the photograph of a man's smile using iOrtho7.0 software (Time Angel, Wuxi, China). Based on this model, seven images were generated with different labiolingual positions of the maxillary lateral incisors in 0.5 mm increments (+ indicating labial translation, and-indicating lingual translation). The images were evaluated by 86 orthodontists and 161 laypersons using a visual analog scale, with lower scores indicating less attractiveness. Data were analyzed using Student's t test and one-way analysis of variance with post hoc test. There was no significant difference in smile ratings by males and females. Orthodontists assigned lower scores to all images than laypersons. The smile at + 1.5 mm was considered the least attractive by orthodontists, while smiles at + 1.5 mm and - 1.5 mm were regarded as the least attractive by laypersons. The smile at 0 mm was evaluated as the most attractive by all raters. Laypersons gave different scores to smiles at 0 or - 0.5 mm, but orthodontists did not. The labiolingual position of maxillary lateral incisors does affect the perception of smile aesthetics. Orthodontists may rate smile aesthetics more critically than laypersons. Therefore, communication and discussion between orthodontists and patients is needed to achieve better therapeutic and aesthetic outcomes.


Research Resource: Correlating Human Cytochrome P450 21A2 Crystal Structure and Phenotypes of Mutations in Congenital Adrenal Hyperplasia.
Cytochrome P450 21A2 is a key player in steroid 21-hydroxylation and converts progesterone to 11-deoxycorticosterone and 17α-hydroxy progesterone to 11-deoxycortisol. More than 100 mutations in P450 21A2 have been established in patients thus far; these account for the vast majority of occurrences of congenital adrenal hyperplasia (CAH), which is among the most common heritable metabolic diseases in humans. CAH phenotypes range from the most severe, salt-wasting (SW), to the simple virilizing (SV), and nonclassical (NC) CAH forms. We recently determined the crystal structure of human P450 21A2 in complex with progesterone. To gain more insight into the structural and stability changes underlying the phenotypes of individual mutations, we analyzed 24 SW, SV, and NC mutants in the context of the crystal structure of the human enzyme. Our analysis reveals clear differences in the localization of SW, SV, and NC mutations, with many of the first type mapping to the active site and near the heme and/or substrate and mostly resulting in complete loss of enzyme activity. Conversely, NC mutations are often found near the periphery and close to the surface of the protein, and mutant enzymes retain partial activity. The main conclusion from the mutation-structure-activity study is that the severity of the CAH clinical manifestations can be directly correlated with the degree of mutation-induced damage in terms of protein fold stability and active site changes in the structural model. Thus, the NC phenotype is typically associated with mutations that have a compensatory effect, ie, H-bonding replacing hydrophobic interactions and vice versa.


Comparison of nerve conduction blocks by an opioid and a local anesthetic.
The experiments were done on frog sciatic nerves, using a sucrose-gap recording technique. The aim of our study was to investigate and to compare the tonic and phasic conduction blocking potency of tramadol and lidocaine on whole nerve and their interactions with Ca(2+). The concentration of a tramadol solution producing the same amount of tonic and phasic conduction blocks was three and six times higher than that needed for lidocaine, respectively. Increasing the Ca(2+) concentration in the test solution enhanced the conduction blocking potency of tramadol, but decreased that of lidocaine. It is concluded that tramadol blocks nerve conduction like a local anesthetic but with a weaker effect than that of lidocaine. Interactions of Ca(2+) and these drugs suggested that these drugs might have either different binding sites or different action mechanisms.


The Suicide Rate in the Elderly Population of Iran between 2008 and 2014.
We aimed to investigate the suicide rate led to death in the elderly population of Iran between 2008 and 2014. A cross-sectional study. The present study was conducted on all suicide-related deaths in elderly people (≥65 yr) during the years 2008 to 2014 reported to the Iranian Legal Medicine Organization. For data collection, legal medicine standard form was used and the cases were classified by age, gender, suicide way and time (year). The incidence of death from suicide was calculated by age and sex. Statistical soft-ware stata12 was used to analyze data. The significance level has been considered to be 0.05. Overall, 1,601 suicide-related deaths were investigated throughout the country. The mean age was 70.36 ± 0.17 years. The incidence trend (per 100,000 people) of the elderly suicides in Iran indicates that successful suicides have been on the rise, rising from 3.7 in 2008 to 4.37 per 100,000 people in 2014. It is necessary to identify and treat suicidal important predisposing factors of suicide such as psycho-social illnesses including depression and also implement prevention programs and policies for this fast-rising population age-group.


Comparative Analysis of Root Microbiomes of Rice Cultivars with High and Low Methane Emissions Reveals Differences in Abundance of Methanogenic Archaea and Putative Upstream Fermenters.
Rice cultivation worldwide accounts for ∼7 to 17% of global methane emissions. Methane cycling in rice paddies is a microbial process not only involving methane producers (methanogens) and methane metabolizers (methanotrophs) but also other microbial taxa that affect upstream processes related to methane metabolism. Rice cultivars vary in their rates of methane emissions, but the influence of rice genotypes on methane cycling microbiota has been poorly characterized. Here, we profiled the rhizosphere, rhizoplane, and endosphere microbiomes of a high-methane-emitting cultivar (Sabine) and a low-methane-emitting cultivar (CLXL745) throughout the growing season to identify variations in the archaeal and bacterial communities relating to methane emissions. The rhizosphere of the high-emitting cultivar was enriched in methanogens compared to that in the low emitter, whereas the relative abundances of methanotrophs between the cultivars were not significantly different. Further analysis of cultivar-sensitive taxa identified families enriched in the high emitter that are associated with methanogenesis-related processes. The high emitter had greater relative abundances of sulfate-reducing and iron-reducing taxa which peak earlier in the season than methanogens and are necessary to lower soil oxidation reduction potential before methanogenesis can occur. The high emitter also had a greater abundance of fermentative taxa which produce methanogenesis precursors (acetate, CO2, and H2). Furthermore, the high emitter was enriched in taxa related to acetogenesis which compete with methanogens for CO2 and H2 These taxa were enriched in a spatio-specific manner and reveal a complex network of microbial interactions on which plant genotype-dependent factors can act to affect methanogenesis and methane emissions.IMPORTANCE Rice cultivation is a major source of anthropogenic emissions of methane, a greenhouse gas with a potentially severe impact on climate change. Emission variation between rice cultivars suggests the feasibility of breeding low-emission rice, but there is a limited understanding of how genotypes affect the microbiota involved in methane cycling. Here, we show that the root microbiome of the high-emitting cultivar is enriched both in methanogens and in taxa associated with fermentation, iron, and sulfate reduction and acetogenesis, processes that support methanogenesis. Understanding how cultivars affect microbes with methanogenesis-related functions is vital for understanding the genetic basis for methane emission in rice and can aid in the development of breeding programs that reduce the environmental impact of rice cultivation.


[Chronic hepatitis B virus infection and the methylation status of p16INK4A promoter].
To explore the relationship among HBV-associated histopathological indexes, x gene mutations and the methylation status of p16INK4A promoter in liver with chronic hepatitis B virus infection, in order to illustrate their role in p16INK4A hypermethylation and HCC progression. Twenty-three cases of surgically resected HBV-associated hepatocellular carcinoma and twenty-five fine needle aspiration biopsy cases of chronic hepatitis B were chosen for this study. The methylation status of the p16INK4A promoter in tumors, their corresponding peritumoral samples and chronic hepatitis B cases was determined by methylation-specific polymerase chain reaction (MSP). EnVision two-step immunohistochemical staining showed the expression of viral antigens in situ. Tissue HBV DNA levels were determined by real-time fluorescence quantitative PCR. Polymerase chain reaction and the direct sequencing method was used for mutation analysis of HBV x gene. In peritumoral samples (P = 0. 025) and chronic hepatitis B cases (P = 0.029), the expression of HBx protein in methylated groups was all significantly higher than that in unmethylated groups of p16INK4A gene. But in tumors, there was no such significant difference. Other HBV antigens including HBsAg and HBcAg, tissue HBV DNA levels and point mutations of HBV x gene did not show a relationship with the methylation status of p16INK4A gene. The data suggest that p16INK4A hypermethylation correlated closely with higher HBx expression in precancerous lesions. HBx may play an important role in the early stage of HBV-associated hepatocarcinogenesis via induction of hepermethylation of p16INK4A promoter.


Multiple trait measurements in 43 inbred mouse strains capture the phenotypic diversity characteristic of human populations.
The breadth of genetic and phenotypic variation among inbred strains is often underappreciated because assessments include only a limited number of strains. Evaluation of a larger collection of inbred strains provides not only a greater understanding of this variation but collectively mimics much of the variation observed in human populations. We used a high-throughput phenotyping protocol to measure females and males of 43 inbred strains for body composition (weight, fat, lean tissue mass, and bone mineral density), plasma triglycerides, high-density lipoprotein and total cholesterol, glucose, insulin, and leptin levels while mice consumed a high-fat, high-cholesterol diet. Mice were fed a chow diet until they were 6-8 wk old and then fed the high-fat diet for an additional 18 wk. As expected, broad phenotypic diversity was observed among these strains. Significant variation between the sexes was also observed for most traits measured. Additionally, the response to the high-fat diet differed considerably among many strains. By the testing of such a large set of inbred strains for many traits, multiple phenotypes can be considered simultaneously and thereby aid in the selection of certain inbred strains as models for complex human diseases. These data are publicly available in the web-accessible Mouse Phenome Database (http://www.jax.org/phenome), an effort established to promote systematic characterization of biochemical and behavioral phenotypes of commonly used and genetically diverse inbred mouse strains. Data generated by this effort builds on the value of inbred mouse strains as a powerful tool for biomedical research.


Monocyte chemotactic peptide receptor. Functional characteristics and ligand-induced regulation.
Monocytes, macrophages, and neutrophils will demonstrate several important cellular functions in response to synthetic formylated oligopeptides. N-formyl-norleucyl-leucyl-phenylalanyl-norleucyl-tyrosyl-lysine (fNLPNTL) was a potent chemoattractant for human blood monocytes; a 1.0-nM concentration induced a maximal chemotactic response. Binding of 125I-labeled fNLPNTL to the monocyte formyl peptide receptor was rapid, specific, and saturable at 4, 24, or 37 degrees C. At 4 degrees C, monocytes from several different donors demonstrated between 10,000 and 18,000 receptors/cell with a dissociation constant (Kd) of 1.7-2.7 nM. The association of the 125I peptide with the cells was irreversible at the elevated temperatures and exceeded the amount of surface receptor by approximately four-fold, suggesting receptor-mediated peptide endocytosis. Processing of rhodamine-labeled fNLPNTL by monocytes was observed directly by video intensification microscopy. At 37 degrees C, diffuse membrane fluorescence was seen initially, followed by rapid aggregation and internalization of the peptide. Monocytes incubated with fNLPNTL displayed a temperature dependent loss of surface binding capacity (receptor down-regulation). This decrease was due to a decrease in surface receptor number rather than to a decrease in receptor affinity. A dose-response curve for peptide-induced receptor down-regulation correlated with a dose-response curve for 125I-labeled fNLPNTL uptake, suggesting that each uptake event led to the loss of one surface receptor. Surface receptor replenishment following down-regulation was rapid and not dependent on new protein synthesis, but was inversely related to both the time and peptide concentration used to induce down-regulation. An exact correlation between receptor down-regulation and functional deactivation of the chemotactic response could not be demonstrated.


Mental health, depression, and anxiety in patients on maintenance dialysis.
Depression and anxiety are among the most common comorbid illnesses in people with end-stage renal disease (ESRD). Patients with ESRD face many challenges which increase the likelihood that they will develop depression or anxiety or worsen these conditions. These include a general feeling of unwellness; specific symptoms caused by ESRD or the patient's treatment; major disruptions in lifestyle; the need to comply with treatment regimens, including dialysis schedules, diet prescription, and water restriction; ancillary treatments and hospitalizations; and the fear of disability, morbidity, and shortened lifespan. Depression has been studied extensively in patients on maintenance dialysis, and much effort has been done to validate the proper screening tools to diagnose depression and to define the treatment options for patients on maintenance dialysis with depression. Anxiety is less well studied in this population of patients. Evidence indicates that anxiety is also common in maintenance dialysis. More attention should be paid to measuring the incidence and prevalence and developing methods of diagnosis and treatment approaches for anxiety in patients with ESRD. In this review, we attempted to underscore those aspects of depression and anxiety that have not been investigated extensively, especially with regard to anxiety. The interaction between racial/ethnic characteristics of patients on maintenance dialysis with depression and anxiety needs to be studied more extensively, in order to assess better approaches to healthcare for these individuals.


A blueprint for a multi-disease, multi-domain Bayesian adaptive platform trial incorporating adult and paediatric subgroups: the Staphylococcus aureus Network Adaptive Platform trial.
The Staphylococcus aureus Network Adaptive Platform (SNAP) trial is a multifactorial Bayesian adaptive platform trial that aims to improve the way that S. aureus bloodstream infection, a globally common and severe infectious disease, is treated. In a world first, the SNAP trial will simultaneously investigate the effects of multiple intervention modalities within multiple groups of participants with different forms of S. aureus bloodstream infection. Here, we formalise the trial structure, modelling approach, and decision rules that will be used for the SNAP trial. By summarising the statistical principles governing the design, our hope is that the SNAP trial will serve as an adaptable template that can be used to improve comparative effectiveness research efficiency in other disease areas.Trial registration NCT05137119 . Registered on 30 November 2021.


[Interleukins in chronic diseases of digestive organs].
To characterize quantitatively and functionally interleukines (IL) in chronic gastrointestinal diseases (CGD) depending on the affected organ, etiology, activity and stage of the disease. Levels of IL-1 beta, IL-4, IL-6, IL-8, tumor necrosis factor alpha (TNF-alpha), gamma interferon (Inf-gamma) in peripheral blood of 300 patients with chronic hepatitis, gastroduodenal ulcer, cholelithiasis, glutenic enteropathy and other CGD were measured by enzyme immunoassay. CGD are accompanied by cytokine imbalance the severity of which depended on the etiological factor (virus, alcohol, etc.), the disease activity, stage and kind of IL. A maximal IL rise was seen in exacerbations of ulcer, cholelithiasis, chronic viral, autoimmune diseases. At early disease stages higher were the levels of Inf-gamma, IL-1 beta, IL-8, IL-6; at late stages--of TNF-alpha, IL-4. In patients with highly and moderately active CGD content of IL reached 250-750 pg/ml that is 3-5 times higher than in inactive CGD. In alcohol-induced CGD and metabolic disturbances IL level was under 150 pg/ml. Assessment of the cytokine spectrum in CGD holds prognostic importance as this spectrum allows conclusion on the disease activity, its progression.


Prologue: 2019 Annual Meeting of the Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA).
The 2019 Annual Meeting of the Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA) was held in Paris, France, and was attended by rheumatologists, dermatologists, representatives of biopharmaceutical companies, and patients. As in previous years, GRAPPA members held a symposium for trainees to discuss their research in psoriatic disease with experts in the field. Other subjects featured during the annual meeting included a composites workshop to review continuous composite measures; the GRAPPA-Collaborative Research Network's third annual meeting; the need for a precision medicine approach to the treatment of psoriatic disease; updates from working groups in International Dermatology Outcome Measures and Outcome Measures in Rheumatology; a debate on the effectiveness of methotrexate in the treatment of psoriatic arthritis (PsA); updating recommendations for optimal treatment approaches for patients with PsA; an update on GRAPPA's research and educational projects; and the GRAPPA ultrasound (US) working group's goal to optimize the evaluation of enthesitis in patients with PsA using US through the development of a diagnostic US enthesitis tool. In this Prologue, we introduce the papers that summarize that meeting.


Phonetic perceptual identification by native- and second-language speakers differentially activates brain regions involved with acoustic phonetic processing and those involved with articulatory-auditory/orosensory internal models.
This experiment investigates neural processes underlying perceptual identification of the same phonemes for native- and second-language speakers. A model is proposed implicating the use of articulatory-auditory and articulatory-orosensory mappings to facilitate perceptual identification under conditions in which the phonetic contrast is ambiguous, as in the case of second-language speakers. In contrast, native-language speakers are predicted to use auditory-based phonetic representations to a greater extent for perceptual identification than second-language speakers. The English /r-l/ phonetic contrast, although easy for native English speakers, is extremely difficult for native Japanese speakers who learned English as a second language after childhood. Twenty-two native English and twenty-two native Japanese speakers participated in this study. While undergoing event-related fMRI, subjects were aurally presented with syllables starting with a /r/, /l/, or a vowel and were required to rapidly identify the phoneme perceived by pushing one of three buttons with the left thumb. Consistent with the proposed model, the results show greater activity for second- over native-language speakers during perceptual identification of /r/ and /l/ relative to vowels in brain regions implicated with instantiating forward and inverse articulatory-auditory articulatory-orosensory models [Broca's area, anterior insula, anterior superior temporal sulcus/gyrus (STS/G), planum temporale (PT), superior temporal parietal area (Stp), SMG, and cerebellum]. The results further show that activity in brain regions implicated with instantiating these internal models is correlated with better /r/ and /l/ identification performance for second-language speakers. Greater activity found for native-language speakers especially in the anterior STG/S for /r/ and /l/ perceptual identification is consistent with the hypothesis that native-language speakers use auditory phonetic representations more extensively than second-language speakers.


Behavior and neuropsychiatric manifestations in Angelman syndrome.
Angelman syndrome has been suggested as a disease model of neurogenetic developmental condition with a specific behavioral phenotype. It is due to lack of expression of the UBE3A gene, an imprinted gene located on chromosome 15q. Here we review the main features of this phenotype, characterized by happy demeanor with prominent smiling, poorly specific laughing and general exuberance, associated with hypermotor behavior, stereotypies, and reduced behavioral adaptive skills despite proactive social contact. All these phenotypic characteristics are currently difficult to quantify and have been subject to some differences in interpretation. For example, prevalence of autistic disorder is still debated. Many of these features may occur in other syndromic or nonsyndromic forms of severe intellectual disability, but their combination, with particularly prominent laughter and smiling may be specific of Angelman syndrome. Management of problematic behaviors is primarily based on behavioral approaches, though psychoactive medication (eg, neuroleptics or antidepressants) may be required.


Film coating of nifedipine extended release pellets in a fluid bed coater with a Wurster insert.
The objective of this work was to study the coating process of nifedipine extended release pellets using Opadry and Opadry II, in a fluid bed coater with a Wurster insert. The coating process was studied using a complete experimental design of two factors at two levels for each polymer. The variables studied were the inlet air temperature and the coating suspension flow rate. The agglomerate fraction and coating efficiency were the analyzed response variables. The air temperature was the variable that most influenced the coating efficiency for both polymers. In addition, a study of the dissolution profiles of coated and uncoated pellets using 0.5% sodium lauryl sulfate in simulated gastric fluid without enzymes (pH 1.2) was conducted. The results showed a prolonged release profile for the coated and uncoated pellets that was very similar to the standards established by the U.S. Pharmacopoeia. The drug content and the release profiles were not significantly affected by storage at 40 °C and 75% relative humidity. However, when exposed to direct sunlight and fluorescent light (light from fluorescent bulbs), the coated pellets lost only 5% of the drug content, while the uncoated ones lost more than 35%; furthermore, the dissolution profile of the uncoated pellets was faster.


The role of ceroid lipofuscinosis neuronal protein 5 (CLN5) in endosomal sorting.
Mutations in the gene encoding CLN5 are the cause of Finnish variant late infantile Neuronal Ceroid Lipofuscinosis (NCL), and the gene encoding CLN5 is 1 of 10 genes (encoding CLN1 to CLN9 and cathepsin D) whose germ line mutations result in a group of recessive disorders of childhood. Although CLN5 localizes to the lysosomal compartment, its function remains unknown. We have uncovered an interaction between CLN5 and sortilin, the lysosomal sorting receptor. However, CLN5, unlike prosaposin, does not require sortilin to localize to the lysosomal compartment. We demonstrate that in CLN5-depleted HeLa cells, the lysosomal sorting receptors sortilin and cation-independent mannose 6-phosphate receptor (CI-MPR) are degraded in lysosomes due to a defect in recruitment of the retromer (an endosome-to-Golgi compartment trafficking component). In addition, we show that the retromer recruitment machinery is also affected by CLN5 depletion, as we found less loaded Rab7, which is required to recruit retromer. Taken together, our results support a role for CLN5 in controlling the itinerary of the lysosomal sorting receptors by regulating retromer recruitment at the endosome.


Hounsfield unit value and clot length in the acutely occluded vessel and time required to achieve thrombectomy, complications and outcome.
Intra-arterial therapy for acute ischemic stroke (AIS) now has an established role. We investigated if Hounsfield Units (HU) quantification on non-contrast CT is associated with ease and efficacy of mechanical thrombectomy and outcomes. We retrospectively studied a prospectively maintained database of cases of AIS given intra-arterial therapy between May 2008 and August 2012. Functional outcome was assessed by 90-day follow-up modified Rankin Scale (mRS). Patients were dichotomized based on time to recanalization. HU were calculated on head CT and thrombus location and length were determined on CT angiography. Simple linear regression was used to analyze the association between clot length, average HU and other clinical variables. 141 patients were included. There was no difference in clot length or average HU among patients with good recanalization achieved within 1 h and those in which the procedures extended beyond 1 h. There was no relationship between clot length or density and recanalization. The thrombus length and density were not significantly different between patients with procedural complications and those without. Neither the presence of post-procedure intracranial hemorrhage nor the 90-day mRS was associated with thrombus length or density. We have not found any significant associations between either thrombus length or density and likelihood of recanalization, time to achieve recanalization, intraprocedural complications, postprocedural hemorrhage or functional outcome at 90 days. These results do not support a predictive value for thrombus quantification in the evaluation of AIS.


GIV is a nonreceptor GEF for G alpha i with a unique motif that regulates Akt signaling.
Heterotrimeric G proteins are molecular switches that control signal transduction. Ligand-occupied, G protein-coupled receptors serve as the canonical guanine nucleotide exchange factors (GEFs) that activate heterotrimeric G proteins. A few unrelated nonreceptor GEFs have also been described, but little or nothing is known about their structure, mechanism of action, or cellular functions in mammals. We have discovered that GIV/Girdin serves as a nonreceptor GEF for G alpha i through an evolutionarily conserved motif that shares sequence homology with the synthetic GEF peptide KB-752. Using the available structure of the KB-752 x G alpha i1 complex as a template, we modeled the G alpha i-GIV interface and identified the key residues that are required to form it. Mutation of these key residues disrupts the interaction and impairs Akt enhancement, actin remodeling, and cell migration in cancer cells. Mechanistically, we demonstrate that the GEF motif is capable of activating as well as sequestering the G alpha-subunit, thereby enhancing Akt signaling via the G betagamma-PI3K pathway. Recently, GIV has been implicated in cancer metastasis by virtue of its ability to enhance Akt activity and remodel the actin cytoskeleton during cancer invasion. Thus, the novel regulatory motif described here provides the structural and biochemical basis for the prometastatic features of GIV, making the functional disruption of this unique G alpha i-GIV interface a promising target for therapy against cancer metastasis.


Diffusion tensor imaging and disability progression in multiple sclerosis: A 4-year follow-up study.
Diffusion tensor imaging (DTI) is sensitive technique to detect widespread changes in water diffusivity in the normal-appearing white matter (NAWM) that appears unaffected in conventional magnetic resonance imaging. We aimed to investigate the prognostic value and stability of DTI indices in the NAWM of the brain in an assessment of disability progression in patients with a relapsing-onset multiple sclerosis (MS). Forty-six MS patients were studied for DTI indices (fractional anisotropy (FA), mean diffusivity (MD), radial (RD), and axial (AD) diffusivity) in the NAWM of the corpus callosum (CC) and the internal capsule at baseline and at 1 year after. DTI analysis for 10 healthy controls was also performed at baseline. Simultaneously, focal brain lesion volume and atrophy measurements were done at baseline for MS patients. Associations between DTI indices, volumetric measurements, and disability progression over 4 years were studied by multivariate logistic regression analysis. At baseline, most DTI metrics differed significantly between MS patients and healthy controls. There was tendency for associations between baseline DTI indices in the CC and disability progression (p < 0.05). Changes in DTI indices over 1 year were observed only in the CC (p < 0.008), and those changes were not found to predict clinical worsening over 4 years. Clear-cut association with disability progression was not detected for baseline volumetric measurements. Aberrant diffusivity measures in the NAWM of the CC may provide additional information for individual disability progression over 4 years in MS with the relapsing-onset disease. CC may be a good target for DTI measurements in monitoring disease activity in MS, and more studies are needed to assess the related prognostic potential.


Supplemental Nutrition Assistance Program Data: Why Disclosure Is Needed.
The Supplemental Nutrition Assistance Program (SNAP) provides funding to low-income households to purchase food at participating stores. The goals of the program include reducing hunger, improving nutrition, and strengthening the US food system. These are interrelated, as food access and choice depend on availability.SNAP generates data that could be useful for program evaluation and evidence-based policymaking to reach public health goals. However, the US Department of Agriculture (USDA) does not collect or disclose all SNAP-related data. In particular, the USDA does not systematically collect food expenditure data, and although it does collect transaction (sales) and redemption data (the amount retailers are reimbursed through SNAP), it does not release these data at the store level.In 2018, Congress quietly changed the law to prohibit the USDA from disclosing store-level transaction and redemption data, and in 2019, the US Supreme Court blocked disclosure of these data. These federal proceedings can inform the outcome of additional efforts to disclose SNAP-related data, as well as future research and policy evaluation to support improved public health outcomes for SNAP beneficiaries.


The effects of cognitive-behavioral therapy on depression, anger, and self-control for Korean soldiers.
This quasi-experimental study examined the effects of cognitive-behavioral therapy (CBT) to control depression, anger, and self-control in soldiers from South Korea. Using a pretest-posttest design, the sample was composed of an experimental group (n = 16) and a control group (n = 12). The experimental group participated in four sessions of CBT. No significant differences were found between the groups with regard to demographic characteristics. Changes in the variables after the intervention were analyzed using Wilcoxon's signed-rank sum test. The depression scores decreased significantly after the intervention (z = -3.05, p = 0.002); whereas, the scores of state-trait anger and self-control did not change. In the control group, none of the outcome variable scores changed significantly. The results indicate that the developed CBT program might be an effective modality to decrease the depression of soldiers who are in military service.


Transcriptional response of bronchial epithelial cells to Pseudomonas aeruginosa: identification of early mediators of host defense.
The airway epithelium responds to microbial exposure by altering expression of a variety of genes to increase innate host defense. We aimed to delineate the early transcriptional response in human primary bronchial epithelial cells exposed for 6 h to a mixture of IL-1beta and TNF-alpha or heat-inactivated Pseudomonas aeruginosa. Because molecular mechanisms of epithelial innate host defense are not fully understood, the open-ended expression-profiling technique SAGE was applied to construct gene expression profiles covering 30,000 genes: 292 genes were found to be differentially expressed. Expression of seven genes was confirmed by real-time qPCR. Among differentially expressed genes, four classes or families were identified: keratins, proteinase inhibitors, S100 calcium-binding proteins, and IL-1 family members. Marked transcriptional changes were observed for keratins that form a key component of the cytoskeleton in epithelial cells. Expression of antimicrobial proteinase inhibitors SLPI and elafin was elevated after microbial or cytokine exposure. Interestingly, expression of numerous S100 family members was observed, and eight members, including S100A8 and S100A9, were among the most differentially expressed genes. Differential expression was also observed for the IL-1 family members IL-1beta, IL-1 receptor antagonist, and IL-1F9, a newly discovered IL-1 family member. Clustering of differentially expressed genes into biological processes revealed that the early inflammatory response in airway epithelial cells to IL-1beta-TNF-alpha and P. aeruginosa is characterized by expression of genes involved in epithelial barrier formation and host defense.


Overview and new developments in softer X-ray (2A < lambda < 5A) protein crystallography.
New methodologies with synchrotron radiation and X-ray free electron lasers (XFELs) in structural biology are being developed. Recent trends in harnessing softer X-rays in protein crystallography for phase determination are described. These include reference to a data-collection test at 2.6 A wavelength with a lysozyme crystal on SRS station 7.2 (Helliwell, 1983) and also use of softer X-rays (2 A wavelength) to optimise f " at the xenon L1 absorption edge in the Single Isomorphous Replacement Optimised Anomalous Scattering ('SIROAS') structure determination of apocrustacyanin A1 with four, partially occupied, xenon atoms (Cianci et al., 2001; Chayen et al., 2000). The hand of the protein was determined using the f " enhanced sulphur anomalous signal from six disulphides in the protein dimer of 40 kDa. In a follow-up study the single wavelength xenon L1-edge f " optimised data set alone was used for phase determination and phase improvement by solvent flattening etc. (CCP4 DM) (Olczak et al., 2003). Auto-tracing of the protein was feasible but required additional diffraction data at higher resolution. This latter could be avoided in future by using improved tilted detector settings during use of softer X-rays, i.e. towards back-scattering recording (Helliwell, 2002). The Olczak et al. study has already led to optimisation of the new SRS beamline MPW MAD 10 (see www.nwsgc.ac.uk) firstly involving the thinning of the beryllium windows as much as possible and planning for a MAR Research tilted detector 'desk top beamline' geometry. Thus the use of softer, i.e. 2 to 3 A wavelength range, X-rays will allow optimisation of xenon and iodine L-edge f " and enhancing of sulphur f " signals for higher throughput protein crystallography. Softer X-rays utilisation in protein crystallography includes work done on SRS bending-magnet station 7.2 in the early 1980s by the author as station scientist (Helliwell, 1984). In the future development of XFELs these softer X-ray wavelengths could also be harnessed and relax the demands to some extent on the complexity and cost of an XFEL. Thus, by use of say 4 A XFEL radiation and use of a back-scattering geometry area detector the single molecule molecular transform could be sampled to a spatial resolution of 2 A, sufficient, in principle, for protein model refinement (Miao et al., 1999). Meanwhile, Miao et al. (2003) report the first experimental recording of the diffraction pattern from intact Escherichia coli bacteria using coherent X-rays, with a wavelength of 2 A, at a resolution of 30 nm and a real-space image constructed. The new single-particle X-ray diffraction-imaging era has commenced.


Association between seropositivity for Neospora caninum and reproductive performance of beef heifers in the Pantanal of Mato Grosso do Sul, Brazil.
Neospora caninum is an obligate intracellular parasite that can infect domestic and wild canids, ruminants, and horses. The purpose of this study was to evaluate the association between the presence of antibodies to N. caninum and reproductive loss in beef heifers in the South Pantanal region of Mato Grosso, Brazil. A total of 1098 heifers were evaluated from breeding to calving with regard to reproductive performance, and serology for neosporosis was assayed by indirect immunofluorescence reaction. The prevalence of N. caninum in cows that had failed to conceive was 22.13% higher than in those that had conceived (p > 0.0001), indicating a significant correlation between non-conception and presence of anti-N. caninum antibodies. The extraction rates for seropositive and seronegative heifers were 28.24 and 50.12%, respectively, with a statistically significant value (p > 0.0001) that showed a decrease of 44% in the extraction rate of seropositive animals. The results emphasize the importance of diagnosis for monitoring of neosporosis in primiparous cows, and indicators of seropositive females as a factor in strategies for culling the beef cattle herd.


Facilitating the transition home after military deployment: a systematic literature review of post-deployment adaptation programmes.
Background: Many countries pay special attention to the transition of their military personnel from deployment to home via post-deployment adaptation programmes (PDAPs). Objective: This systematic review aims to provide a structured analysis of structure, process, and outcomes based on available empirical evidence for PDAPs. Methods: A systematic literature search was performed to identify original peer-reviewed studies on PDAP in six databases (MEDLINE, Embase, PsycINFO, Cochrane Central Register of Controlled Trials, PTSDPubs, and OpenGrey). The overall risk of bias of the articles was assessed using GRADE guidelines. The literature was analysed in its description of PDAP structure, process, and outcomes. In addition, when effects were reported, these were extracted. Results: The search resulted in 1535 unique records that were screened for eligibility; 16 articles were included, of which only three showed low risk of bias according to GRADE. Few studies clearly described and investigated PDAP structure elements (i.e. combat exposure), process elements (i.e. who delivered the intervention), and other outcome elements (i.e. adjustment issues). Most articles included some form of third location decompression (N = 10; psychoeducation and time to rest and relax on a location that is not in the mission theatre or at home) and Battlemind (N = 4; interactive training focusing on how to adapt skills and cognitions after deployment at home). The results suggest positive mental health effects and satisfaction of these two types of PDAP, although the evidence is poor. Conclusions: Empirical, high-quality evidence for PDAP is scarce. In addition, the existing literature reveals a lack of systematic method in describing the goals of PDAP and the ways of achieving these. This leads to incomparability between studies and hinders the building of a knowledge base on PDAPs. A structure is proposed for describing the structure, process, and outcomes of PDAPs with measured effects. High -quality evidence for post-deployment adaptation programmes is scarce.Programme structure, process, and aims with measured effects are not delineated.Positive mental health effects are reported for Battlemind and third location decompression.


Identification of a novel coronavirus possibly associated with acute respiratory syndrome in alpacas (Vicugna pacos) in California, 2007.
Alpaca respiratory syndrome (ARS) was first recognized in California in October 2007. This syndrome is characterized by acute respiratory signs, high fever, and occasional sudden death, and has mostly been observed in pregnant alpacas (Vicugna pacos), although all signalments have been affected. A similarity in clinical signs to cases located on the East Coast of the United States was observed; however, a causative agent had not been identified. Preliminary diagnostic submissions to the California Animal Health and Food Safety Laboratory System (CAHFS) were negative for known bacterial, parasitic, fungal, and viral pathogens, as well as for toxins, making the etiology of this disease unknown. However, based on pathologic findings, a viral or toxic etiology was strongly considered. A novel coronavirus was recovered from lung tissue of a clinical case submitted to CAHFS. The coronavirus identity was confirmed in tissue culture by transmission electron microscopy and by sequence analysis of a conserved region within the viral genome. Statistical analysis calculating a serologic association between the serum virus neutralization antibody titer and coronavirus, the presence of exposure history on 40 animals with a history of ARS, and 167 controls provided an odds ratio of 121 (95% confidence interval: 36.54 and 402.84; P < 0.0001). The findings indicate that the ARS-associated coronavirus described is distinct from the previously reported gastrointestinal-associated coronavirus identified in alpaca herds.


NKT cells in cardiovascular diseases.
Despite life-style advice and the prescription of cholesterol-lowering and anti-thrombotic drugs, cardiovascular diseases are still the leading cause of death worldwide. Therefore, there is an urgent need for new therapeutic strategies focussing on atherosclerosis, the major underlying pathology of cardiovascular diseases characterized by an accumulation of lipids in an inflamed arterial/vessel wall. CD1d-restricted lipid-sensing natural killer T (NKT) cells, bridging the innate and adaptive immunity, and CD1d-expressing antigen-presenting cells are detected in atherosclerotic lesions of mice and humans. In this review we will summarize studies that point to a critical role for NKT cells in the pathogenesis of atherosclerosis and other cardiovascular diseases by the secretion of pro-atherogenic cytokines and cytotoxins. These pro-atherogenic NKT cells are potential targets for new therapeutic strategies in the prevention and treatment of cardiovascular diseases. Additionally, proteins transferring lipids during atherosclerosis, which are also important in the loading of lipids onto CD1d and possible endogenous ligands responsible for the activation of NKT cells during atherosclerosis will be discussed.


Metabolic impact of pheochromocytoma/paraganglioma: targeted metabolomics in patients before and after tumor removal.
Excess catecholamine release by pheochromocytomas and paragangliomas (PPGL) leads to characteristic clinical features and increased morbidity and mortality. The influence of PPGLs on metabolism is ill described but may impact diagnosis and management. The objective of this study was to systematically and quantitatively study PPGL-induced metabolic changes at a systems level. Targeted metabolomics by liquid chromatography-tandem mass spectrometry of plasma specimens in a clinically well-characterized prospective cohort study. Analyses of metabolic profiles of plasma specimens from 56 prospectively enrolled and clinically well-characterized patients (23 males, 33 females) with catecholamine-producing PPGL before and after surgery, as well as measurement of 24-h urinary catecholamine using LC-MS/MS. From 127 analyzed metabolites, 15 were identified with significant changes before and after surgery: five amino acids/biogenic amines (creatinine, histidine, ornithine, sarcosine, tyrosine) and one glycerophospholipid (PCaeC34:2) with increased concentrations and six glycerophospholipids (PCaaC38:1, PCaaC42:0, PCaeC40:2, PCaeC42:5, PCaeC44:5, PCaeC44:6), two sphingomyelins (SMC24:1, SMC26:1) and hexose with decreased levels after surgery. Patients with a noradrenergic tumor phenotype had more pronounced alterations compared to those with an adrenergic tumor phenotype. Weak, but significant correlations for 8 of these 15 metabolites with total urine catecholamine levels were identified. This first large prospective metabolomics analysis of PPGL patients demonstrates broad metabolic consequences of catecholamine excess. Robust impact on lipid and amino acid metabolism may contribute to increased morbidity of PPGL patients.


Indiana family physician attitudes and practices concerning smoking cessation.
Most physicians are aware of the health benefits of smoking cessation and agree they have a responsibility to help smokers quit. Many physicians, however, do not regularly address smoking cessation with their patients. Questionnaires were sent to 2,095 family practice physicians in Indiana. Information obtained included: demographic data; office-based smoking cessation practices; counseling; and physicians' perceptions of intervention outcomes. Most physicians (86%) asked new patients if they smoked, and 23% questioned patients about their exposure to passive smoke. Younger physicians, female physicians and urban physicians were more likely to ask new patients if they smoked. A formal smoking cessation program was used by 28% of the responding physicians. Among those not using a program, 7% reported plans to implement one in the coming year, 40% were not planning to implement one, and 53% were unsure. Physician and practice characteristics were not correlated with the use of smoking cessation programs. Only 11% of physicians considered their smoking cessation counseling skills to be excellent; 27% indicated the need for improvement in skills. One-half (52%) believed their counseling efforts were effective; almost half (45%) believed that current reimbursement policies limited their involvement in smoking cessation interventions. Most respondents have not instituted smoking cessation programs in their practices. It is likely that a combination of strategies, including both undergraduate, graduate and continuing medical education programs and reform in reimbursement practices for cessation programs, will be required to achieve significant increases in long-term smoking abstinence rates.


Cobalt chloride, a hypoxia-mimicking agent, targets sterol synthesis in the pathogenic fungus Cryptococcus neoformans.
We investigated the effects of the hypoxia-mimetic CoCl2 in the pathogenic fungus Cryptococcus neoformans and demonstrated that CoCl2 leads to defects in several enzymatic steps in ergosterol biosynthesis. Sterol defects were amplified in cells lacking components of the Sre1p-mediated oxygen-sensing pathway. Consequently, Sre1p and its binding partner Scp1p were essential for growth in the presence of CoCl2. Interestingly, high copies of a single gene involved in ergosterol biosynthesis, ERG25, rescued this growth defect. We show that the inhibitory effect of CoCl2 on scp1Delta and sre1Delta cells likely resulted from either an accumulation of non-viable methylated sterols or a decrease in the amount of ergosterol. Similar findings were also observed in the ascomycetous yeast, Schizosaccharomyces pombe, suggesting that the effects of CoCl2 on the Sre1p-mediated response are conserved in fungi. In addition, gene expression analysis revealed limited overlap between Sre1p-dependant gene activation in the presence of CoCl2 and low oxygen. The majority of genes similarly affected by both CoCl2 and low oxygen were involved in ergosterol synthesis and in iron/copper transport. This article identifies the Sre1p pathway as a common mechanism by which yeast cells sense and adapt to changes in both CoCl2 concentrations and oxygen levels.


Development of a comprehensive nursing/toxicity assessment form.
To develop and implement a form that integrates nursing and clinical trials toxicity assessments and interventions. Published books, journal articles, and toxicity scales. The combined nursing and biometry department Documentation/Toxicity Committee at the London Regional Cancer Centre designed and implemented the comprehensive Patient Assessment Record (PAR) to provide consistent nursing and toxicity data related to patients enrolled in clinical trials research protocols. Nursing interventions and evaluations are documented on the Nursing Intervention Record (NIR). Initial evaluation suggested that the PAR has been an effective form. Audit results indicated that the form minimizes the number of individuals collecting the same data while maintaining the quality of toxicity data. The collection of toxicity data for patients enrolled in clinical trials protocols has been incorporated into primary nurses' job descriptions. This additional responsibility has enhanced their role. Both the PAR and the NIR have assisted the multidisciplinary teams in meeting their documentation needs.


[Malaria control in tropical Africa. 1956].
Although spraying with insecticides has caused a spectacular regression in malaria in many parts of Africa, it cannot be said that the transmission of the disease has been definitely interrupted in any large area on the mainland. There is therefore some uneasiness regarding the methods so far adopted and doubts as to whether the very considerable expenditure involved is justified. There is, moreover, acute anxiety about the possible development of resistance to insecticides by the vector anopheles. Fortunately, so far there does not appear to be any in Africa, and the author considers that logical aim should be complete eradication of malaria before resistance does appear to the powerful insecticides now available. Control measures must inevitably be continued indefinitely, but if the aim of eradication is achieved, operations could cease and it would be possible for anopheles to co-exist with man without transmitting malaria. The author points out that before any continent-wide campaign can be begun, there are, in addition to the problem of funds, a number of other problems which must be carefully studied. Among these are the lack of trained specialized staff at all levels, transport difficulties, shortage of water over vast areas in the dry season and floods during the rains, the differing habits of the various African populations, and the biology of the vector mosquitos. Despite these problems, however, the time for concerted action to achieve eradication in Africa draws nearer with each succeeding day.


Chiari Type II malformation: past, present, and future.
The Chiari Type II malformation (CM II) is a unique hindbrain herniation found only in patients with myelomeningocele and is the leading cause of death in these individuals younger than 2 years of age. Several theories exist as to its embryological evolution and recently new theories are emerging as to its treatment and possible prevention. A thorough understanding of the embryology, anatomy, symptomatology, and surgical treatment is necessary to care optimally for children with myelomeningocele and prevent significant morbidity and mortality. A review of the literature was used to summarize the clinically pertinent features of the CM II, with particular attention to pitfalls in diagnosis and surgical treatment. Any child with CM II can present as a neurosurgical emergency. Expeditious and knowledgeable evaluation and prompt surgical decompression of the hindbrain can prevent serious morbidity and mortality in the patient with myelomeningocele, especially those younger than 2 years old. Symptomatic CM II in the older child often presents with more subtle findings but rarely in acute crisis. Understanding of CM II continues to change as innovative techniques are applied to this challenging patient population.


[Laboratory tests in The Child and Adolescent Health Survey].
Laboratory tests will be performed as part of the National Health Survey for Children and Adolescents to identify health risks and to determine the seroprevalence of disease markers. The parameters were selected from haematology, clinical chemistry, allergology, endocrinology and serology of infectious diseases as indicators of individual health risks, to validate information collected by means of the questionnaires and to identify age specific immunity gaps of vaccine-preventable diseases.


Evidence of selective processing of immunodominant epitopes in virally infected cells.
Recent advances in clarifying the molecular mechanisms involved in Ag processing and presentation have relied heavily on the use of somatic cell mutants deficient in proteasome subunits, TAP transporter, and cell surface expression of MHC class I molecules. Of particular interest currently are those mutants that lack specific protease activity involved in the generation of antigenic peptides. It is theoretically possible that deficiencies of this nature could selectively prevent the cleavage of certain peptide bonds and thus generate only a subset of antigenic peptides. Gro29/Kb cell line is derived from the wild-type murine Ltk- cell line. This cell line is one example of a mutant that lacks specific protease activities. This deficiency manifests itself in an inability to generate a subset of immunodominant peptide epitopes derived from vesicular stomatitis virus and herpes simplex virus. This in turn leads to a general inability to present these viral epitopes to cytotoxic T lymphocytes (CTL). These studies describe a unique Ag processing deficiency and provide new insight into the role of proteasome-independent proteases in MHC class I-restricted peptide generation.


Correlations between endotoxin, interferon-gamma, biopterin and serum phospholipase A2-activities during lethal gram negative sepsis in rats.
To establish a standardised reproducible animal model of intraperitoneal sepsis, and to investigate early immunoserological responses to find a mediator-based system for evaluation and grading of diffuse peritonitis in patients Prospective experimental study 4 Teaching hospitals, Germany and Austria 42 LEW. 1W rats, 12 of which acted as controls Gram negative sepsis was induced by intraperitoneal injection of 6 ml of a mixture of Escherichia coli (K1:H+) 10(10) organisms/ml, autogenous haemoglobin 2.9 ml (haemoglobin concentration 3%), 0.9% sodium chloride 3 ml, and suspension 0.1 ml. Control rats were given physiological saline 6 ml alone. Concentrations of endotoxin, interferon gamma (IFN-gamma), and biopterin, and serum phospholipase A2 (PLA2) activity. There were significant differences between the septic and control rats in concentrations of endotoxin (EU/ml) (median (interquartile range) 21.85 (2.02-159.5) compared with 0, p < 0.0001; IFN-gamma (pg/ml) 1263.0 (271.0-7575.0) compared with 101.0 (89.0-141.0), p < 0.0001; biopterin (nmol/L) 111.0 (66.4-156.3) compared with 53.7 (38.3-67.6), p < 0.001; and PLA2 (U/L) 163.0 (125.8-209.0) compared with 112.5 (88.5-126.5) p < 0.01. Measurements of concentrations of endotoxin, IFN-gamma, pteridines, and PLA2 activity may well be adequate markers for early recognition of sepsis, and perhaps for grading it during the first 6 hours after induction. The allow a clear distinction to be made between septic and non-septic disorders in 87% of cases.


National policies and local planning for health services.
The primary reason for health planning in this country is the numerous instances in which the interests of the individual, health-care institution and those of the community may diverge, as in the case of hospital staff appointments for physicians. From a technical standpoint, it is much more difficult to plan for health services at the local level than nationally. Notwithstanding, health services are mostly provided at the local level, and health planning should be geared to the solution of local problems. In performing health planning, the local area can benefit from outside assistance. In the past decade, local health planning has been hampered by unstable federal funding. The absence of national policies and guidelines has led to a constant quest for new ideas. In the absence of substantive concerns, requirements for consumer representation have led to a preoccupation with structure and organization. What is required, in addition to steadier funding, is a fostering of local capabilities for health planning. Health planning organizations will require a good deal of technical assistance in the form of concrete ideas on ways to enhance the flexibility and versatility of health facilities and personnel, monitoring natural experiments and learning their lessons, and elucidating the public policy implications of empirical research findings and even of opposite propositions from theory. In specified circumstances the federal government is expected to serve as the superseding decision maker.


Analysis of lap times in international swimming competitions.
Swimming performances were analysed for the top 16 finishers (semi-finalists, finalists) in nine international competitions over a 7-year period (1530 males, 1527 female). Total race time and intermediate lap times were log-transformed and analysed for effects of sex (male, female), stroke (freestyle, form strokes, individual medley), event (100, 200, and 400 m), and place (1-16). Between-athlete correlations characterized the relationship of each lap to final time, and within-athlete estimates quantified the effect of lap time on improvements in final time. Finalists exhibited very large correlations (r = 0.7-0.9) with final time in the second 50-m lap of 100-m events and the middle two 50-m and 100-m laps of 200-m and 400-m events respectively. For an individual swimmer, an achievable change in lap time was associated with an approximate 0.4-0.8% improvement in final time for finalists and an approximate 0.5-1.1% improvement in final time for semi-finalists, depending on sex, stroke, and event. The pattern of lap times was similar for the top 16 swimmers and between the best and worst swims for finalists. These findings indicate that substantial improvements can be made via the final lap in sprints and the middle two laps of 200- to 400-m events, but the overall pattern of lap times should not be changed.


A genome-scan method to identify selected loci appropriate for both dominant and codominant markers: a Bayesian perspective.
Identifying loci under natural selection from genomic surveys is of great interest in different research areas. Commonly used methods to separate neutral effects from adaptive effects are based on locus-specific population differentiation coefficients to identify outliers. Here we extend such an approach to estimate directly the probability that each locus is subject to selection using a Bayesian method. We also extend it to allow the use of dominant markers like AFLPs. It has been shown that this model is robust to complex demographic scenarios for neutral genetic differentiation. Here we show that the inclusion of isolated populations that underwent a strong bottleneck can lead to a high rate of false positives. Nevertheless, we demonstrate that it is possible to avoid them by carefully choosing the populations that should be included in the analysis. We analyze two previously published data sets: a human data set of codominant markers and a Littorina saxatilis data set of dominant markers. We also perform a detailed sensitivity study to compare the power of the method using amplified fragment length polymorphism (AFLP), SNP, and microsatellite markers. The method has been implemented in a new software available at our website (http://www-leca.ujf-grenoble.fr/logiciels.htm).


Production and control of live oral poliovirus vaccine in WI-38 human diploid cells.
Production of live poliomyelitis vaccine in human diploid cells WI-38 can be carried out successfully on a continuous basis. Starting at the 8th population doubling level it is possible to produce 128 ampoules, each containing 1 ml of cell suspension at the 16th level. Each of these cell seed ampoules will yield 85 bottles each with a surface area of 1400 sq. cm at the production level (P29) together with 85 Roux bottle cultures which act as controls. A further passage from the control cells yields a sufficient supply of cells for karyology, tumourigenicity and other tests. Virus yields of the Sabin strains are rather lower than those obtained in primary monkey kidney cells, but this is compensated for by the very much lower reject rate. Karyological characterisation is undoubtedly the most demanding of the testing procedures both in time and expertise. Other tests for contamination, other than bacteriological have yielded uniformly negative results. There is urgent need for standardization of the assay systems used in different laboratories and there is perhaps a case for the use of reference vaccine produced in homogeneic cells.


Sexual risk related behaviour among youth living with HIV in central Uganda: implications for HIV prevention.
As young people living with HIV grow their sexual behaviour and it's implication on HIV prevention is of concern. This study describes the sexual risk related-behaviours and factors associated with abstinence among Youth Living with HIV in central Uganda. We conducted a cross-sectional study among 338 unmarried youth between 15 and 24 years accessing HIV care in central Uganda. Data was collected using interviewer administered structured questionnaires. Adjusted prevalence proportion ratios (adj. PPRs) of factors associated with sexual abstinence for at least six months were determined by multivariable log-binomial regression. Overall, 79% (269/338) of respondents were abstaining from sexual intercourse for atleast six months, although, 45% (150/338) had ever been sexually active. Of the 283 respondents who desired to get married in future, 40% preferred negative marriage partners. Only 31% (39/126) of respondents in boy/girl relationships had disclosed their HIV status to their partners. Among those currently sexually active (n = 69), 57% did not consistently use condoms and 30% had more than one sexual partner in the past six months. The adj.PRR of abstinence was higher among youth between 15 and 19 years compared to those between 20 and 24 years (adj. PPR = 1.26, 95% CI; 1.08-1.46). The prevalence of abstinence was significantly lower among respondent who consumed alcohol (adj. PPR = 0.31, 95% CI 0.16-0.61). Tailored interventions promoting disclosure, consistent condoms use and discouraging alcohol consumption among sero-positive youth could reduce HIV transmission risk.