Peripherally administered cannabinoid receptor 2 (CB2R) agonists lose anti-allodynic effects in TRPV1 knockout mice, while intrathecal administration leads to anti-allodynia and reduced GFAP, CCL2 and TRPV1 expression in the dorsal spinal cord and DRG.
The transient receptor potential (TRP) superfamily of cation channels, of which the TRP vanilloid type 1 (TRPV1) receptor plays a critical role in inflammatory and neuropathic pain, is expressed on nociceptors and spinal cord dorsal horn neurons. TRPV1 is also expressed on spinal astrocytes and dorsal root ganglia (DRG) satellite cells. Agonists of the cannabinoid type 2 receptor (CB2R) suppress allodynia, with some that can bind TRPV1. The neuroimmune C-C class chemokine-2 (CCL2) expressed on injured DRG nociceptor cell bodies, Schwann cells and spinal astrocytes, stimulates immune cell accumulation in DRG and spinal cord, a known critical element in chronic allodynia. The current report examined whether two CB2R agonists, AM1710 and AM1241, previously shown to reverse light touch mechanical allodynia in rodent models of sciatic neuropathy, require TRPV1 activation that leads to receptor insensitivity resulting in reversal of allodynia. Global TRPV1 knockout (KO) mice with sciatic neuropathy given intrathecal or intraperitoneal AM1710 were examined for anti-allodynia followed by immunofluorescent microscopy analysis of lumbar spinal cord and DRG of astrocyte and CCL2 markers. Additionally, immunofluorescent analysis following intrathecal AM1710 and AM1241 in rat was performed. Data reveal that intrathecal AM1710 resulted in mouse anti-allodynia, reduced spinal astrocyte activation and CCL2 expression independent of TRPV1 gene deletion. Conversely, peripheral AM1710 in TRPV1-KO mice failed to reverse allodynia. In rat, intrathecal AM1710 and AM1241 reduced spinal and DRG TRPV1 expression, with CCL2-astrocyte and -microglial co-expression. These data support that CB2R agonists can impact spinal and DRG TRPV1 expression critical for anti-allodynia.


Nonlinear growth curve analysis: estimating the population parameters.
A model for analysing longitudinal growth data with covariates is proposed. It assumes that the data for each individual follow a nonlinear growth model, with parameters unique to that individual. The parameters of individuals are assumed to vary in the population, with the population mean of each parameter dependent upon covariates. Several simple, two-stage methods of estimation are described and compared for making inferences about the effects of covariates upon growth. We also present a method of model validation for nonlinear models. These two-stage methods solve the inference problem we encounter when performing multiple cross-sectional analyses of the effects of covariates along the age scale. In addition, these two-stage methods permit us to study the effects of covariates upon growth rate, or other, possibly nonlinear, growth parameters. We find that some methods are better than others at reproducing the covariate effects observed in a series of age-specific analyses of the data.


The phagocytosis of crystalline silica particles by macrophages.
Silicosis is a chronic lung disease induced by the inhalation of crystalline silica. Exposure of cultured macrophages to crystalline silica leads to cell death; however, the mechanism of cell-particle interaction, the fate of particles, and the cause of death are unknown. Time-lapse imaging shows that mouse macrophages avidly bind particles that settle onto the cell surface and that cells also extend protrusions to capture distant particles. Using confocal optical sectioning, silica particles were shown to be present within the cytoplasmic volume of live cells. In addition, electron microscopy and elemental analysis showed silica in internal cellular sections. To further examine the phagocytosis process, the kinetics of particle uptake was quantified using an assay in which cells were exposed to ovalbumin (OVA)-coated particles, and an anti-OVA antibody was used to distinguish surface-bound from internalized particles. Fc receptor-mediated uptake of antibody-coated silica particles was nearly complete within 5 minutes. In contrast, no OVA-coated particles were internalized at this time. After 30 minutes, 30% of bound silica was internalized and uptake continued slowly thereafter. OVA-coated latex beads, regardless of surface charge, were internalized at a similarly slow rate. These results demonstrate that macrophages internalize silica and that nonopsonized phagocytosis occurs by a temporally, and possibly mechanistically, distinct pathway from Fc receptor-mediated phagocytosis. Eighty percent of macrophages die within 12 hours of silica exposure. Neither OVA coating nor tetramethylrhodamine isothiocyanate labeling has any effect on cell death. Interestingly, antibody coating dramatically reduces silica toxicity. We hypothesize that the route of particle entry and subsequent phagosome trafficking affects the toxicity of internalized particles.


"Show me the money": vulnerability to gambling moderates the attractiveness of money versus suspense.
Do people take risks to obtain rewards or experience suspense? We hypothesized that people vulnerable to gambling are motivated more by the allure of winning money whereas people less vulnerable to gambling are motivated more by the allure of suspense. Consistent with this hypothesis, participants with high scores on a subscale of the Gambling Attitudes and Beliefs Survey--a measure of vulnerability to gambling--reported more of a motivation to earn money (pilot study), were more likely to accept a certain or near-certain amount of money than to gamble for that same amount (Studies 1-2), and worked harder to earn money (Study 3). People vulnerable to gambling also made more accurate predictions about how much they would gamble. People less vulnerable to gambling, in contrast, gambled more than people vulnerable to gambling, but did not know that they would.


Simulated pivot-shift testing with single and double-bundle anterior cruciate ligament reconstructions.
One of the principal rationales for performing a double-bundle reconstruction of the anterior cruciate ligament is the suggestion that it may be superior to a single-bundle reconstruction in restoring a normal pivot-shift sign. The purpose of this study was to measure the abilities of single-bundle and anatomic double-bundle reconstructions to restore normal knee kinematics and graft forces during a simulated pivot-shift test. Graft force and knee kinematics were recorded during a simulated pivot-shift event with and without the anterior cruciate ligament and after graft reconstructions. With a single bundle, the graft was tensioned to restore anterior-posterior laxity at 30 degrees of flexion. With double-bundle reconstructions, the anteromedial graft was first tensioned as above and then the posterolateral graft tension was set with use of one of four protocols: posterolateral tension = anteromedial tension at 10 degrees of flexion (DB1); posterolateral tension = anteromedial tension at 30 degrees (DB2); posterolateral tension = (anteromedial tension + 30 N) at 10 degrees (DB3); and posterolateral tension = (anteromedial tension + 30 N) at 30 degrees (DB4). A single-bundle reconstruction restored all displacements and rotations during the pivot shift to the intact knee levels. The mean tibial rotations and lateral plateau displacements during the pivot shift with DB2, DB3, and DB4 reconstructions were less than those in the intact knee and also less than those in a single-bundle reconstruction. Before the pivot shift, the mean graft forces with all reconstructions were greater than that of the intact knee; the mean graft forces with the DB3 and DB4 reconstructions were also greater than that of a single-bundle reconstruction. After the pivot shift, the mean graft forces for all reconstructions were less than the levels before the pivot shift with single-bundle forces lower than intact knee levels and DB4 forces higher than intact knee levels. Reduction or elimination of the pivot-shift sign is an important goal for anterior cruciate ligament reconstruction. In our model, the results show that a single-bundle reconstruction was sufficient to restore intact knee kinematics during a simulated pivot-shift event. The higher graft forces with some double-bundle graft-tensioning protocols reduced the coupled rotations and displacements from an applied valgus moment to less than the intact levels. This overcorrection should theoretically make the knee less likely to pivot but could have unknown clinical consequences.


Dexamethasone and Monophosphoryl Lipid A Induce a Distinctive Profile on Monocyte-Derived Dendritic Cells through Transcriptional Modulation of Genes Associated With Essential Processes of the Immune Response.
There is growing interest in the use of tolerogenic dendritic cells (tolDCs) as a potential target for immunotherapy. However, the molecular bases that drive the differentiation of monocyte-derived DCs (moDCs) toward a tolerogenic state are still poorly understood. Here, we studied the transcriptional profile of moDCs from healthy subjects, modulated with dexamethasone (Dex) and activated with monophosphoryl lipid A (MPLA), referred to as Dex-modulated and MPLA-activated DCs (DM-DCs), as an approach to identify molecular regulators and pathways associated with the induction of tolerogenic properties in tolDCs. We found that DM-DCs exhibit a distinctive transcriptional profile compared to untreated (DCs) and MPLA-matured DCs. Differentially expressed genes downregulated by DM included MMP12, CD1c, IL-1B, and FCER1A involved in DC maturation/inflammation and genes upregulated by DM included JAG1, MERTK, IL-10, and IDO1 involved in tolerance. Genes related to chemotactic responses, cell-to-cell signaling and interaction, fatty acid oxidation, metal homeostasis, and free radical scavenging were strongly enriched, predicting the activation of alternative metabolic processes than those driven by counterpart DCs. Furthermore, we identified a set of genes that were regulated exclusively by the combined action of Dex and MPLA, which are mainly involved in the control of zinc homeostasis and reactive oxygen species production. These data further support the important role of metabolic processes on the control of the DC-driven regulatory immune response. Thus, Dex and MPLA treatments modify gene expression in moDCs by inducing a particular transcriptional profile characterized by the activation of tolerance-associated genes and suppression of the expression of inflammatory genes, conferring the potential to exert regulatory functions and immune response modulation.


A novel approach to the synthesis of amino-sugars. Routes to selectively protected 3-amino-3-deoxy-aldopentoses based on pyridinium salt photochemistry.
A new approach for the synthesis of selectively blocked 3-amino-3-deoxyaldopentoses is presented. The strategy is based on employment of a pyridinium salt photocyclization-aziridine ring-opening sequence to prepare stereochemically defined, enantiomerically enriched aminocyclopentendiol derivatives. Ring-opening reactions transform these substances into terminally differentiated aminopolyols, which serve as precursors to the target amino-aldopentoses. The utility of this strategy is demonstrated by its application to the syntheses of protected derivatives of D- and L-3-amino-3-deoxyxylose, L-3-amino-3-deoxyarabinose, and a late-stage intermediate in a potential route to N-acetylneuraminic acid.


Rethinking childhood ependymoma: a retrospective, multi-center analysis reveals poor long-term overall survival.
Ependymoma is the third most common brain tumor in children, but there is a paucity of large studies with more than 10 years of follow-up examining the long-term survival and recurrence patterns of this disease. We conducted a retrospective chart review of 103 pediatric patients with WHO Grades II/III intracranial ependymoma, who were treated at Dana-Farber/Boston Children's Cancer and Blood Disorders Center and Chicago's Ann & Robert H. Lurie Children's Hospital between 1985 and 2008, and an additional 360 ependymoma patients identified from the Surveillance Epidemiology and End Results (SEER) database. For the institutional cohort, we evaluated clinical and histopathological prognostic factors of overall survival (OS) and progression-free survival (PFS) using the log-rank test, and univariate and multivariate Cox proportional-hazards models. Overall survival rates were compared to those of the SEER cohort. Median follow-up time was 11 years. Ten-year OS and PFS were 50 ± 5% and 29 ± 5%, respectively. Findings were validated in the independent SEER cohort, with 10-year OS rates of 52 ± 3%. GTR and grade II pathology were associated with significantly improved OS. However, GTR was not curative for all children. Ten-year OS for patients treated with a GTR was 61 ± 7% and PFS was 36 ± 6%. Pathological examination confirmed most recurrent tumors to be ependymoma, and 74% occurred at the primary tumor site. Current treatment paradigms are not sufficient to provide long-term cure for children with ependymoma. Our findings highlight the urgent need to develop novel treatment approaches for this devastating disease.


[Atypical form of acute suppurative thyroiditis in pediatric patient: case report].
Acute thyroiditis is an unusual disease. In pediatric patients it is usually associated with congenital abnormalities towards the unilateral envelopment of the left thyroid lobe. In general, in the absence of preexisting thyroid disease, the thyroid function is most often normal. It is here described a case of a six-year-old girl who had presented a mass with flogistic signs in the anterior neck region for fifteen days, together with leukocytosis and increased levels of VSH and TSH. There was no evidence of previous thyroid disease and the cervical ultrasonography suggested thyroid abscess. Intravenous antibiotic and surgical drainage were done with a good response, however, the patient developed clinical hypothyroidism afterwards. Barium swallow did not demonstrate pyriform sinus fistula and the follow-up ultrasonography revealed heterogeneous internal texture suggesting Hashimoto's thyroiditis. The relevance of this case is due to its unusual findings: the absence of congenital abnormalities, bilateral envelopment of thyroid lobes, hypothyroidism and a probable Hashimoto's thyroiditis as a predisposing factor for acute thyroiditis.


Novel protein biomarkers associated with coronary artery disease in statin-treated patients with familial hypercholesterolemia.
Familial hypercholesterolemia (FH) is the most common and serious monogenic disorder of lipid metabolism. The incidence of coronary artery disease (CAD) varies among both treated and untreated FH patients. The aim of the study was to utilize proteomics to identify novel protein biomarkers that differentiate genetically confirmed heterozygous patients with FH at high CAD risk from those at low CAD risk. Sixty genetically confirmed FH patients were recruited and stratified into (1) asymptomatic FH with low atherosclerotic burden (FH, n = 20); (2) asymptomatic FH with high atherosclerotic burden (FH + Ca, n = 20); and (3) FH with previously confirmed symptomatic CAD (FH + CAD, n = 20). Six new potential proteins were identified; leucine-rich alpha-2-glycoprotein (LRG1), inter-alpha-trypsin inhibitor heavy chain H3, complement C4-B (C4B), complement C1q subcomponent subunit B (C1QB), monocyte differentiation antigen (CD14), and histidine-rich glycoprotein (HRG). There were significant associations between gender and C4B (Z = 2.31, P = .021), C1QB (Z = 2.49, P = .013), CD14 (Z = 2.17, P = .03), and HRG (Z = 2.14, P = .033). There were significant associations between smoking and LRG1 (χ22 = 6.59, P = .037), CB4 (χ22 = 7.85, P = .02), and HRG (χ22 = 6.11, P = .047). All the peptides were significantly associated with advanced CAD stages, independently of age and smoking. However, the absence of the proteins was the strongest marker. The most accurate association with CAD was HRG (area under the receiver operating characteristic curve = 0.922), whereas LRG1, C4B, and C1QB were also associated with CAD (area under the receiver operating characteristic curve >0.9). For either coronary atherosclerosis or CAD, LRG1, C4B, C1QB, and HRG were relatively well associated. The present study has identified 6 novel protein biomarkers that are associated with more advanced stages of atherosclerotic disease and subsequent coronary events in patients with heterozygous FH.


Removing noise and ambiguities from comparative maps in rearrangement analysis.
Comparison of genomic maps is hampered by errors and ambiguities introduced by mapping technology, incorrectly resolved paralogy, small samples of markers and extensive genome rearrangement. We design an analysis to remove or resolve most of these problems and to extract corrected data where markers occur in consecutive strips in both genomes. To do this we introduce the notion of pre-strip, an efficient way of generating these, and a compatibility analysis culminating in a Maximum Weighted Clique (MWC) search. The output can be directly analyzed with genome rearrangement algorithms, allowing the restoration of some of the data not incorporated into the clique solution. We investigate the trade-off between criteria for discarding excessive pre-strips to make MWC feasible, in terms of retaining as many markers as possible in the solution and producing an economical rearrangement analysis. We explore these questions through simulation and through comparison of the rice and sorghum genomes.


Osteopontin is down-regulated in hydatidiform mole.
Osteopontin (OPN) is a glycoprotein of the extracellular matrix that can bind to different types of receptors including integrins and CD44 receptors. Multiple binding affinity enables OPN to play a role in many physiological and pathological processes. OPN contributes to tumorigenesis in several types of cancers. OPN is also expressed by the endometrium and by trophoblast cells of the chorionic villus in human placenta, where OPN may regulate implantation and placentation in early pregnancies by promoting cell-cell interactions, adhesion, spreading, and migration of trophoblast. Our purpose was to determine the expression of OPN mRNA and protein in hydatidiform mole and in normal placenta of comparable gestational age. A total of 13 fresh tissues from complete hydatidiform moles, 2 from partial hydatidiform moles, and 9 from normal placentas were analyzed by performing quantitative real-time PCR on microdissected trophoblast cells and immunohistochemistry on frozen sections of tissue. Our results showed significantly lower expression of OPN mRNA and protein in hydatidiform mole, and in particular complete mole (P = 0.001 by real-time PCR and P < 0.001 by immunohistochemistry) as compared to nermal placenta. Although precise molecular mechanisms of gestational trophoblastic diseases have not yet been determined, down-regulation of osteopontin may play an important role in the pathogenesis of molar pregnancy.


Early-onset of sporadic basal-cell carcinoma: germline mutations in the TP53, PTCH, and XPD genes.
Basal cell carcinoma (BCC) is the most common non-melanoma skin cancer in the Western world. The objective of this study was to examine together germline mutations in the TP53, PTCH, and XPD genes as risk factors for developing BCC at a young age. We hypothesized that mutations in these genes significantly increase the risk of early-onset BCC (< or = 35 years). The PCR, DNA sequencing and Restriction Fragment Length Polymorphisms methods were utilized to study eight Puerto Rican patients with a confirmed diagnosis of BCC before age 35. A novel germline mutation (T:A transversion) was identified at the exon 4, codon 50 of the TP53 gene of one BCC patient. No other mutations were found at the TP53 or PTCH genes. The presence of the XPD mutant allele is associated with a seven-fold increase in risk (OR = 7.0, p = 0.03) for developing BCC prior to age 35. In addition, the DNA Repair Capacity (DRC) of these BCC patients showed a 47% reduction that was significant in relation to age-matched controls (p = 0.021). However, the XPD mutant allele was not associated with the decrease in DRC observed in BCC participants. The evaluated population presented BCC before age 35, a phenomenon that is so rare as to make very difficult the study of this subpopulation with a larger sample size. The results of this study, suggest that the XPD Lys751Gln polymorphism may have a significant role in the development of early-onset BCC in the Puerto Rican population.


Enzyme immunoassay for measurement of antibodies to herpes simplex virus infection: comparison with complement fixation, immunofluorescent-antibody, and neutralization techniques.
An enzyme immunoassay (EIA) was developed for detecting antibody to herpes simplex virus, and the results were compared with those of complement fixation, indirect immunofluorescent-antibody, and plaque reduction neutralization tests. Test sera showed very little nonspecific reactivity even at a starting dilution as low as 1:10. EIA results showed excellent correlation with results obtained by the neutralization test, with an average gain in sensitivity of 1.65. EIA proved very useful in detecting current herpes simplex virus infection, and antibody appeared in all cases soon after clinical onset. EIA appears to be a rapid, sensitive, and specific method for routine demonstration of herpes simplex virus antibody in a clinical setting.


How the associative strengths of stimuli combine in compound: summation and overshadowing.
When two conditioned stimuli (CSs) are presented in compound, the response is typically stronger than to the individual CSs, implying that their associative strengths combine. However, to identify exactly how associative strengths combine requires an accurate description of the relationship between associative strength and responding. The authors have used the delta rule (Rescorla & Wagner, 1972) to constrain the predicted growth of associative strength (V) to identify the relationship between V and responding across the course of Pavlovian conditioning of two CSs (one auditory, one visual). Responding to the compound was best predicted as 0.6xV(CS1)+0.6xV(CS2), suggesting that only 60% of the associative strength of each CS generalized to the compound. A second experiment confirmed this result and additionally showed that summation of responding between two same-modality CSs (both auditory or both visual) declined across training. A third experiment applied the procedure to compound conditioning, showing that responding to the compound was equal to the sum of the response rates to the individual CSs. The results are discussed in terms of configural and elemental models of Pavlovian conditioning.


Distribution of receptors for transferrin and low density lipoprotein on the surface of giant HeLa cells.
Giant HeLa cells, having a spread diameter of about 200 micrometers, were briefly surface-labeled at 0 degree C with 125I-labeled transferrin, low density lipoprotein, anti-HeLa cell antibody, or concanavalin A. The cells were washed at 0 degree C, fixed, and autoradiographed. The distribution of grains when either anti-HeLa cell antibodies or concanavalin A was used was roughly as expected: the cell surfaces appeared uniformly labeled. When either transferrin or low density lipoprotein was used, about half the labeled cells had a nonuniform distribution of grains. On round cells, the cell periphery was more densely labeled than the middle of the cell; on elongated cells, cell protrusions were often more highly labeled than the rest of the cell. The simplest interpretation of these results is that, during their endocytic cycles through these cells, the transferrin and low density lipoprotein receptors are returned to the cell surface at the cell's leading edge.


[Placenta associated plasma protein-A (PAPP-A): clinical value in cardiovascular medicine].
PAPP-A is high molecular weight metalloproteinase originally identified in the serum of pregnant women. During pregnancy, the concentration of PAPP-A increases in maternal circulation with gestational age. Depressed levels, associated with an abnormal placental function, have formed the basis of the first trimester screening for Down syndrome. The role of PAPP-A in tissue other than placenta has only recently been explored. More recently elevated serum concentrations of PAPP-A have been identified in patients with unstable angina or acute myocardial infract. It has thus been proposed as a potential early indicator of acute coronary syndrome.


Reactive oxygen species amplify protein kinase C signaling in high glucose-induced fibronectin expression by human peritoneal mesothelial cells.
We previously demonstrated that high glucose up-regulates fibronectin mRNA and protein expression by human peritoneal mesothelial cells (HPMC) through activation of protein kinase C (PKC). PKC is known to induce cellular reactive oxygen species (ROS) and PKC-dependent activation of the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase has recently been shown to be responsible, in part, for increased oxidative stress in diabetes. On the other hand, high glucose-induced mitochondrial overproduction of superoxide anion was found to activate PKC. We, therefore, hypothesized that high glucose-induced activation of PKC in HPMC may increase cellular ROS and ROS, in turn, may activate PKC and thus provide signal amplification in high glucose-induced fibronectin up-regulation in HPMC. The role of ROS in high glucose- and PKC-induced fibronectin expression was examined by quantification of cellular ROS after stimulation with high glucose and phorbol 12-myristate 13-acetate (PMA), by the effect of hydrogen peroxide (H(2)O(2)) and PMA on fibronectin expression, and finally by inhibition of ROS and PKC. The source of cellular ROS was further examined by inhibition of NADPH oxidase and mitochondrial metabolism. D-glucose increased dichlorofluorescein (DCF)-sensitive cellular ROS in HPMC in a dose-dependent manner. l-glucose did not induce ROS generation and cytochalasin B completely blocked high glucose-induced ROS generation, suggesting that glucose uptake, but not media hyperosmolality, is required in ROS generation in HPMC. PMA increased cellular ROS and fibronectin secretion. A single dose of H(2)O(2) or H(2)O(2) continuously generated by glucose oxidase up-regulated fibronectin expression [corrected]. Antioxidants trolox and catalase inhibited high glucose- and PMA-induced fibronectin mRNA and protein expression. Inhibition of PKC inhibited high glucose-and H(2)O(2)-induced fibronectin secretion. NADPH oxidase inhibitors (diphenyleneiodinium and apocynin) and an inhibitor of mitochondrial electron transport chain subunit I (rotenone) all effectively inhibited high glucose-induced cellular ROS generation and fibronectin secretion. The present data demonstrate that high glucose increases cellular ROS in HPMC through activation of PKC, NADPH oxidase, and mitochondrial metabolism and that ROS, thus generated, up-regulate fibronectin expression by HPMC. ROS are not only downstream but also upstream signaling molecules to PKC and provide signal amplification in high glucose-induced fibronectin expression by HPMC. The present data imply that cellular ROS may be potential therapeutic targets in progressive accumulation of extracellular matrix in the peritoneal tissue of long-term peritoneal dialysis patients using high glucose-containing peritoneal dialysis solutions.


Arizona's tobacco control initiative illustrates the need for continuing oversight by tobacco control advocates.
In 1994, Arizona voters approved Proposition 200 which increased the tobacco tax and earmarked 23% of the new revenues for tobacco education programmes. To describe the campaign to pass Proposition 200, the legislative debate that followed the passage of the initiative, and the development and implementation of the tobacco control programme. This is a case study. Data were collected through semi-structured interviews with key players in the initiative campaign and in the tobacco education programme, and written records (campaign material, newspapers, memoranda, public records). Despite opposition from the tobacco industry, Arizonans approved an increase in the tobacco tax. At the legislature, health advocates in Arizona successfully fought the tobacco industry attempts to divert the health education funds and pass preemptive legislation. The executive branch limited the scope of the programme to adolescents and pregnant women. It also prevented the programme from attacking the tobacco industry or focusing on secondhand smoke. Health advocates did not put enough pressure at the executive branch to force it to develop a comprehensive tobacco education programme. It is not enough for health advocates to campaign for an increase in tobacco tax and to protect the funds at the legislature. Tobacco control advocates must closely monitor the development and implementation of tax-funded tobacco education programmes at the administrative level and be willing to press the executive to implement effective programmes.


Gelsolin regulates cisplatin sensitivity in human head-and-neck cancer.
Chemoresistance is a major challenge in cancer therapy. Cisplatin is commonly used for chemotherapy in patients with head-and-neck cancer (HNC), but it increases control of the disease by only 10-15%. Downregulation of proapoptotic pathways is a key determinant for chemoresistance in which gelsolin (GSN) is critically involved. We analyzed the association between GSN expression and cisplatin resistance in HNC cell lines, animals with HNC and cancer tissue samples from 58 cisplatin-treated patients with HNC. GSN expression levels were positively associated with chemoresistance in vitro and in vivo. Cisplatin-induced GSN downregulation was associated with the cleavage of GSN and the promotion of apoptosis. GSN silencing facilitated cisplatin-induced apoptosis in chemoresistant cells. In contrast, intact gelsolin was prosurvival in the presence of cisplatin by interacting with X-linked inhibitor of apoptosis protein (XIAP). In chemosensitive cells, cisplatin suppressed GSN-XIAP interaction, promoted translocation of XIAP from the perinuclear region to the nucleus and induced apoptosis. In chemoresistant cells, GSN was highly expressed, and cisplatin had no significant effect on GSN-XIAP interaction and apoptosis. We conclude that GSN is important for chemoresistance in HNC and may be an appropriate therapeutic target in chemoresistant cancers.


Analysis of machine learning algorithms as integrative tools for validation of next generation sequencing data.
While next generation sequencing (NGS) has become the technology of choice for clinical diagnostics, most genetic laboratories still use Sanger sequencing for orthogonal confirmation of NGS results. Previous studies have shown that when the quality of NGS data is high, most calls are indicated by Sanger sequencing, making confirmation redundant. We aimed at establishing a set of criteria that make it possible to distinguish NGS calls that need orthogonal confirmation from those that do not would significantly decrease the amount of work necessary to reach a diagnosis. A data set of 7976 NGS calls confirmed as true or false positive by Sanger sequencing was used to train and test different machine learning (ML) approaches. By varying the size and class balance of the training dataset, we measured the performance of the different algorithms to determine the conditions under which ML is a valid approach for confirming NGS calls in a diagnostic environment. Our results indicate that machine learning is a valid approach to find variant calls that need more investigation, but in order to reach the high accuracy required in a clinical environment, the training data set must include enough observations and these observations must be well-balanced between true/false positive NGS calls. Our results show that it is possible to integrate the diagnostic NGS validation workflow with a machine learning approach to reduce the number of Sanger confirmations of high- quality NGS calls, reducing the time and costs of diagnosis.


Effects of egg order on organic and inorganic element concentrations and egg characteristics in tree swallows, Tachycineta bicolor.
The laying order of tree swallow eggs was identified from the Housatonic River, Berkshire County, Massachusetts, USA, and eggs were chemically analyzed individually to document possible effects of laying order on organic contaminant and inorganic element concentrations. Effects of laying order on other parameters such as egg weight, size, and lipid and moisture content also were assessed. Some effects of egg order on total polychlorinated biphenyls (PCBs) were detected, but the effect was not uniform across individual females or between years. In 2004, clutches with higher total PCBs tended to have concentrations decline across egg order, whereas clutches with lower concentrations of PCBs tended to increase across egg order. In contrast, in 2005, there was a tendency for concentrations to increase across egg order. Polychlorinated biphenyl concentrations were highly variable within and among clutches in both years. The directionality of egg order associations (i.e., slopes) for trace elements was element dependent, was positive for Mn and Zn, was negative for B, and had no slope for Cr. Whole egg weight increased across egg order. Percentage lipid was variable within a clutch, with no pattern common across all females. Percentage lipid was also correlated with organic contaminant concentration. In highly contaminated environments, higher lipid content could have the unanticipated corollary of having higher concentrations of lipophilic contaminants such as PCBs. To reduce the effect of high variation within a clutch when assessing contamination exposure, it is recommended that two eggs per clutch be collected and pooled for chemical analysis. We further recommend that, as long as the two eggs are randomly collected, the additional effort needed to identify and collect specific eggs is not warranted.


Functional expression of costimulatory molecule CD86 on epithelial cells in the inflamed colonic mucosa.
Costimulatory signals are essential for T-cell activation. The aim of this study was to demonstrate expression of costimulatory molecules CD86 and CD80 in human colonic epithelial cells and assess their functional roles in the activation of T cells in inflamed colonic mucosa. CD86 and CD80 expression was assessed by immunohistochemistry of colonic mucosa, reverse-transcription polymerase chain reaction, and flow cytometric analysis of isolated colonic epithelial cells and cell lines. Costimulatory effect of CD86-expressing colonic epithelial cells on purified CD4(+) T-cell proliferation was also assessed at suboptimal phytohemagglutinin stimulation. CD86 and CD80 messenger RNA was detected in isolated colonic epithelial cells from normal and inflamed mucosa of patients with ulcerative colitis. Immunohistochemistry and flow cytometric analysis of colonic epithelial cells confirmed cell surface expression of CD86 protein in inflamed colonic mucosa. Cell surface expression of CD86 protein was increased in the colonic epithelial cell line HT29-18-N2 after interferon gamma stimulation. Purified CD4(+) T cells proliferated in response to suboptimal phytohemagglutinin costimulated with interferon gamma-stimulated HT29-18-N2, and these proliferative responses were efficiently inhibited by the addition of anti-CD86 monoclonal antibody. Costimulatory effect of CD86-expressing colonic epithelial cells isolated from inflamed mucosa of ulcerative colitis was also demonstrated at suboptimal phytohemagglutinin stimulation in CD4(+) T cells. Colonic epithelial cells may act as antigen-presenting cells, and contribute to the activation of T cells through costimulatory molecule CD86 expression in inflamed colonic mucosa.


Synergistic antibacterial effects of exopolysaccharides/nickel-nanoparticles composites against multidrug-resistant bacteria.
The need for an alternative treatment to fight infectious diseases caused by antibiotic-resistant bacteria is increasing. A possible way to overcome bacterial resistance to antibiotics is by reintroducing commonly used antibiotics with a sensitizer capable of enhancing their antimicrobial effect in resistant bacteria. Here, we use a composite composed of exopolysaccharide capped-NiO NPs, with antimicrobial effects against antibiotic-resistant Gram-positive and Gram-negative bacteria. It potentiated the antimicrobial effects of four different antibiotics (ampicillin, kanamycin, chloramphenicol, and ciprofloxacin) at lower concentrations than their minimal inhibitory concentrations. We observed that the Ni-composite synergistically enhanced, fourfold, the antibacterial effect of kanamycin and chloramphenicol against multidrug-resistant Staphylococcus aureus and Pseudomonas aeruginosa, as well as ampicillin against multidrug-resistant Staphylococcus aureus, and ciprofloxacin against multidrug-resistant Pseudomonas aeruginosa by eightfold. We also found that Ni-composite could not inhibit biofilm synthesis on the tested bacterial strains. Our results demonstrated the possibility of using metal nanoparticles, like NiO, as a sensitizer to overcome bacterial antibiotic resistance.


Partial inhibition of two genes that encode spliceosomal proteins in Giardia intestinalis.
Giardia intestinalis is an early divergent organism that was recently shown to have introns. The machinery responsible for the removal of introns in higher eukaryotes is the spliceosome, which consists of five ribonucleoproteins. Each of these ribonucleoproteins has a small nuclear RNA, a set of seven Sm proteins (B, D1, D2, D3, E, F and G) and several specific proteins. Some genes that encode spliceosome proteins have been bioinformatically identified in the parasite genome. Although it is assumed that the spliceosome is responsible for splicing in this parasite, biochemical characterization is lacking. Objective. To inhibit two G. intestinalis spliceosome protein genes in order to determine whether this inhibition affects parasite growth or encystation. Materials and methods. Antisense sequences of the genes encoding the spliceosomal parasite proteins SmB and SmD3 were cloned into a specific G. intestinalis vector. G. intestinalis individuals were subsequently transfected with the recombinant vectors and those parasites that incorporated the vector were selected. A decrease in mRNA levels by real-time PCR was confirmed and the growth and encystation in wild and transfected parasites was assessed. Results. A decrease of 40% and 70% of SmB and SmD3 mRNA levels, respectively, was observed. Growth and encystation in these parasites were not affected. Conclusion. Decrease of SmB and SmD3 mRNA levels does not affect the parasite, indicating that the spliceosome remains functional or that splicing is not essential for parasite viability.


Anury in two Cairn terriers.
Congenital anury (taillessness) was observed in 2 Cairn Terriers. Fecal staining of the hind limbs was evident in both dogs. A mating of the affected dogs produced a litter of 2 apparently normal pups. The small litter size may have been attributable to lethal gene combinations similar to those described in mice. The radiographic appearance of the caudal and sacral vertebrae was atypical in both tailless dogs. Results of myelography and electrodiagnostic testing of anal sphincter function were normal in one of the dogs. One of the tailless dogs was euthanatized and was found to lack specific muscles and muscle parts responsible for normal defecation, which resulted in fecal staining of the hind limbs. Pedigree analysis revealed the tailless dogs to be closely related and slightly inbred, suggesting a hereditary etiologic factor for anury.


Cranial irradiation alters dendritic spine density and morphology in the hippocampus.
Therapeutic irradiation of the brain is a common treatment modality for brain tumors, but can lead to impairment of cognitive function. Dendritic spines are sites of excitatory synaptic transmission and changes in spine structure and number are thought to represent a morphological correlate of altered brain functions associated with hippocampal dependent learning and memory. To gain some insight into the temporal and sub region specific cellular changes in the hippocampus following brain irradiation, we investigated the effects of 10 Gy cranial irradiation on dendritic spines in young adult mice. One week or 1 month post irradiation, changes in spine density and morphology in dentate gyrus (DG) granule and CA1 pyramidal neurons were quantified using Golgi staining. Our results showed that in the DG, there were significant reductions in spine density at both 1 week (11.9%) and 1 month (26.9%) after irradiation. In contrast, in the basal dendrites of CA1 pyramidal neurons, irradiation resulted in a significant reduction (18.7%) in spine density only at 1 week post irradiation. Analysis of spine morphology showed that irradiation led to significant decreases in the proportion of mushroom spines at both time points in the DG as well as CA1 basal dendrites. The proportions of stubby spines were significantly increased in both the areas at 1 month post irradiation. Irradiation did not alter spine density in the CA1 apical dendrites, but there were significant changes in the proportion of thin and mushroom spines at both time points post irradiation. Although the mechanisms involved are not clear, these findings are the first to show that brain irradiation of young adult animals leads to alterations in dendritic spine density and morphology in the hippocampus in a time dependent and region specific manner.


Treatment and Spiritual Care in Mental Health: Recovery As a Journey, Not a Destination.
Recovery in mental illness is a person-oriented process that involves self-determination/choice, and growth potential. This article discusses eight components of recovery-oriented treatment, and how nurses in any setting can facilitate recovery and encourage the healthy role of spirituality in clients' lives during the recovery process. Biblical references relating to each component of the recovery-oriented process are offered.


Pseudomonas canavaninivorans sp. nov., isolated from bean rhizosphere.
A novel canavanine-degrading bacterium, strain HB002T, was isolated from rhizosphere soil of a catch crop field collected from the island of Reichenau in Konstanz, Germany, and characterized by using polyphasic taxonomy. The facultative aerobe, rod-shaped, Gram-stain-negative bacterium was oxidase- and catalase-positive. The isolate was able to grow on canavanine as a sole carbon and nitrogen source. Results of phylogenetic analysis based on 16S rRNA gene sequences revealed highest similarities to Pseudomonas bijieensis (L22-9T, 99.93 %), Pseudomonas brassicacearum subsp. neoaurantiaca (ATCC 49054T, 99.76 %), Pseudomonas brassicacearum subsp. brassicacearum (DBK 11T, 99.63 %), Pseudomonas thivervalensis (DSM 13194T, 99.51 %), Pseudomonas kilonensis (DSM 13647T, 99.39 %) and Pseudomonas corrugata (ATCC29736T, 99.39 %). Marker gene analysis placed the strain in the intrageneric group of Pseudomonas fluorescens, subgroup P. corrugata. In silico DNA-DNA hybridization and average nucleotide identity values were both under the recommended thresholds for species delineation. The predominant fatty acids of strain HB002T were C16 : 0, C17 : 0 cyclo ω7c and C18 : 1 ω7c. The major respiratory quinone was Q9, followed by Q8 and minor components of Q7 and Q10. Results from the phenotypic characterization showd the strain's inability to hydrolyse gelatin and to assimilate N-acetyl glucosamide and a positive enzymatic activity of acid phosphatase and naphthol-AS-BI phosphohydrolase that distinguish this strain from closely related type strains. Taken together, these results show that strain HB002T represents a novel species in the genus Pseudomonas, for which the name Pseudomonas canavaninivorans sp. nov. is proposed. The type strain is HB002T (=DSM 112525T=LMG 32336T).


[Significance of postoperative face-down positioning after surgery for idiopathic macular holes: consecutive case-control study].
Postoperative face-down positioning is considered to be a critical part in macular hole surgery with short-acting gas. However, the significance of this posturing remains uncertain using long-acting gas tamponade. Fifty consecutive eyes with idiopathic full-thickness macular holes underwent macular hole surgery with a follow up time of 6-12 months. The results of a cohort (25 eyes) with complete C3F8 (16%) vitreous fill without posturing was compared to a cohort (25 eyes) with SF6 (20%) with strict postoperative face-down positioning for two weeks. With SF6 tamponade the anatomic success rate was 64% in the C3F8 cohort a complete closure was achieved in 88%. In the SF6 cohort, visual recovery of three lines or better was attained in 44%, resulting in a visual acuity of 20/50 or better in 12%. The mean postoperative visual acuity was 20/125 with a mean rise of 1.4 lines. In the C3F8 cohort, 72% improved for at least three lines with a mean postoperative visual acuity of 20/63 and a mean visual recovery of 3.3 lines. Visual acuity of 20/50 or better was noted in 48% of eyes. The significantly higher success rate using C3F8 tamponade without positioning compared to SF6 gas tamponade with positioning confirms the cumulative tamponade duration being a crucial factor in macular hole surgery. Using a tamponade with short-acting gas only strict postoperative positioning ensures the contact of the gas bubble with foveal tissue and gains significance with reducing bubble size. While a potential for significant visual recovery in certain cases is evident, all in all a broad variability of the postoperative success rate can be stated. In contrast, even in supine position C3F8 attains a continuous tamponade enforcing a sufficient glial wound healing without any positioning.


Regular Physical Exercise Adherence Scale (REPEAS): a new instrument to measure environmental and personal barriers to adherence to regular physical exercise.
To create, develop, and validate a scale that identifies the environmental and personal barriers that make it difficult to adhere to the practice of physical exercise on a regular basis in a population of Brazilian adults. We include adult individuals, aged 18-59 years, practitioners or former practitioners of physical exercise, with Brazilian Portuguese as their mother tongue. In the development and validation phases of the process, 6 specialists in the field of the health assessed the content validity: firstly, the specialists were asked to freely list the questions they would ask to investigate the barriers to adherence to regulating physical activity. Secondly, after compiling all the suggestions listed and eliminating suggestions with similar content, the items suggested in the first round were sent to the specialists so that an evaluation of all questions using a 5-point Likert scale and the content validity coefficient was calculated. We then evaluated the structural validity, construct validity, reliability, internal consistency, and ceiling and floor effects of the Regular Physical Exercise Adherence Scale (REPEAS). Sixteen items were proposed to measure the factors that make it difficult to adhere to the regular practice of physical exercise. The internal structure of the REPEAS initially tested was based on the theoretical proposal of creating the instrument with two domains. After the structural analysis, we used the modification indices to identify the redundant items of the instrument. Consequently, the final version of the REPEAS after factor analysis had 12 items. Thus, the structure with 2 domains and 12 items presented adequate fit indices. With regard to construct validity, the REPEAS scores were compared in two distinct groups: irregular practitioners/ex-practitioners versus regular practitioners of physical exercise, in which a significant difference could be observed between groups (p < 0.001) for both the domains. Acceptable reliability was observed for the environment and personal domains, with ICC values of 0.86 and 0.94, in the same order. For internal consistency, Cronbach's alpha value was 0.908 (environmental domain) and 0.915 (personal domain), these values being adequate for the REPEAS. The REPEAS is a scale with a valid two-dimensional internal structure, consisting of 12 items, reliable and with a valid construct, which supports its use in the clinical, epidemiological, and research contexts in Brazil.


The potential application of European market research data in dietary exposure modelling of food additives.
Consumer exposure assessments for food additives are incomplete without information about the proportions of foods in each authorised category that contain the additive. Such information has been difficult to obtain but the Mintel Global New Products Database (GNPD) provides information about product launches across Europe over the past 20 years. These data can be searched to identify products with specific additives listed on product labels and the numbers compared with total product launches for food and drink categories in the same database to determine the frequency of occurrence. There are uncertainties associated with the data but these can be managed by adopting a cautious and conservative approach. GNPD data can be mapped with authorised food categories and with food descriptions used in the EFSA Comprehensive European Food Consumption Surveys Database for exposure modelling. The data, when presented as percent occurrence, could be incorporated into the EFSA ANS Panel's 'brand-loyal/non-brand loyal exposure model in a quantitative way. Case studies of preservative, antioxidant, colour and sweetener additives showed that the impact of including occurrence data is greatest in the non-brand loyal scenario. Recommendations for future research include identifying occurrence data for alcoholic beverages, linking regulatory food codes, FoodEx and GNPD product descriptions, developing the use of occurrence data for carry-over foods and improving understanding of brand loyalty in consumer exposure models.


A Phantom Study on Fetal Dose Reducing Factors in Pregnant Patients with Breast Cancer during Radiotherapy Treatment.
This study aims to investigate the factors that reduce fetal dose in pregnant patients with breast cancer throughout their radiation treatment. Two main factors in a standard radiation oncology center are considered as the treatment planning systems (TPSs) and simple shielding for intensity modulated radiation therapy technique. TPS factor was evaluated with two different planning algorithms: Anisotropic analytical algorithm and Acuros XB (external beam). To evaluate the shielding factor, a standard radiological purpose lead apron was chosen. For both studies, thermoluminescence dosimeters were used to measure the point dose, and an Alderson RANDO-phantom was used to simulate a female pregnant patient in this study. Thirteen measurement points were chosen in the 32nd slice of the phantom to cover all possible locations of a fetus up to 8th week of gestation. The results show that both of the TPS algorithms are incapable of calculating the fetal doses, therefore, unable to reduce them at the planning stage. Shielding with a standard lead apron, however, showed a slight radiation protection (about 4.7%) to the fetus decreasing the mean fetal dose from 84.8 mGy to 80.8 mGy, which cannot be disregarded in case of fetal irradiation. Using a lead apron for shielding the abdominal region of a pregnant patient during breast irradiation showed a minor advantage; however, its possible side effects (i.e., increased scattered radiation and skin dose) should also be investigated further to solidify its benefits.


Gastric Damage and Cancer-Associated Biomarkers in Helicobacter pylori-Infected Children.
Helicobacter pylori (H. pylori) is well-known to be involved in gastric carcinogenesis, associated with deregulation of cell proliferation and epigenetic changes in cancer-related genes. H. pylori infection is largely acquired during childhood, persisting long-term in about half of infected individuals, a subset of whom will go on to develop peptic ulcer disease and eventually gastric cancer, however, the sequence of events leading to disease is not completely understood. Knowledge on carcinogenesis and gastric damage-related biomarkers is abundant in adult populations, but scarce in children. We performed an extensive literature review focusing on gastric cancer related biomarkers identified in adult populations, which have been detected in children infected with H. pylori. Biomarkers were related to expression levels (RNA or protein) and/or methylation levels (DNA) in gastric tissue or blood of infected children as compared to non-infected controls. In this review, we identified 37 biomarkers of which 24 are over expressed, three are under expressed, and ten genes are significantly hypermethylated in H. pylori-infected children compared to healthy controls in at least 1 study. Only four of these biomarkers (pepsinogen I, pepsinogen II, gastrin, and SLC5A8) have been studied in asymptomatically infected children. Importantly, 13 of these biomarkers (β-catenin, C-MYC, GATA-4, DAPK1, CXCL13, DC-SIGN, TIMP3, EGFR, GRIN2B, PIM2, SLC5A8, CDH1, and VCAM-1.) are consistently deregulated in infected children and in adults with gastric cancer. Future studies should be designed to determine the clinical significance of these changes in infection-associated biomarkers in children and their persistence over time. The effect of eradication therapy over these biomarkers in children if proven significant, could lead to modifications in treatment guidelines for younger populations, and eventually promote the development of preventive strategies, such as vaccination, in the near future.


Interleukin-24 as a target cytokine of environmental aryl hydrocarbon receptor agonist exposure in the lung.
Exposure to environmental aryl hydrocarbon receptor (AhR) agonists, such as halogenated aromatic hydrocarbons and polycyclic aromatic hydrocarbons (PAHs), has great impacts on the development of various lung diseases. As emerging molecular targets for AhR agonists, cytokines may contribute to the inflammatory or immunotoxic effects of environmental AhR agonists. However, general cytokine expression may not specifically indicate environmental AhR agonist exposure. By comparing cytokine and chemokine expression profiles in human lung adenocarcinoma cell line CL5 treated with AhR agonists and the non-AhR agonist polychlorinated biphenyl (PCB) 39, we identified a target cytokine of environmental AhR agonist exposure of in the lungs. Thirteen cytokine and chemokine genes were altered in the AhR agonists-treated cells, but none were altered in the PCB39-treated cells. Interleukin (IL)-24 was the most highly induced gene among AhR-modulated cytokines. Cotreatment with AhR antagonist completely prevented IL-24 induction by AhR agonists in the CL5 cells. Knockdown AhR expression with short-hairpin RNA (shRNA) significantly reduced benzo[a]pyrene (BaP)-induced IL-24 mRNA levels. We further confirmed that gene transcription, but not mRNA stability, was involved in IL-24 upregulation by BaP. Particulate matter (PM) in the ambient air contains some PAHs and is reported to activate AhR. Oropharyngeal aspiration of PM significantly increased IL-24 levels in lung epithelia and in bronchoalveolar lavage fluid of mice 4weeks after treatment. Thus, our data suggests that IL-24 is a pulmonary exposure target cytokine of environmental AhR agonists.


Electrical and dielectric properties of bovine trabecular bone--relationships with mechanical properties and mineral density.
Interrelationships of trabecular bone electrical and dielectric properties with mechanical characteristics and density are poorly known. While electrical stimulation is used for healing fractures, better understanding of these relations has clinical importance. Furthermore, earlier studies have suggested that bone electrical and dielectric properties depend on the bone density and could, therefore, be used to predict bone strength. To clarify these issues, volumetric bone mineral density (BMDvol), electrical and dielectric as well as mechanical properties were determined from 40 cylindrical plugs of bovine trabecular bone. Phase angle, relative permittivity, loss factor and conductivity of wet bovine trabecular bone were correlated with Young's modulus, yield stress, ultimate strength, resilience and BMDvol. The reproducibility of in vitro electrical and dielectric measurements was excellent (standardized coefficient of variation less than 1%, for all parameters), especially at frequencies higher than 1 kHz. Correlations of electrical and dielectric parameters with the bone mechanical properties or density were frequency-dependent. The relative permittivity showed the strongest linear correlations with mechanical parameters (r > 0.547, p < 0.01, n = 40, at 50 kHz) and with BMDvol (r = 0.866, p < 0.01, n = 40, at 50 kHz). In general, linear correlations between relative permittivity and mechanical properties or BMDvol were highest at frequencies over 6 kHz. In addition, a significant site-dependent variation of electrical and dielectric characteristics, mechanical properties and BMDvol was revealed in bovine femur (p < 0.05, Kruskall-Wallis H-test). Based on the present results, we conclude that the measurement of electrical and dielectric properties provides quantitative information that is related to bone quantity and quality.


Regions on adenylyl cyclase VII required for selective regulation by the G13 pathway.
Regulation of multiple adenylyl cyclases (AC) provides unique inputs to mediate the synthesis of cAMP, a ubiquitous second messenger that controls many aspects of cellular function. On stimulation by G(s), the activities of ACs can be further selectively modulated by other pathways to ensure precise control of intracellular cAMP responses to specific stimuli. Recently, we reported that one of the AC isoforms, AC7, is uniquely regulated by the G(13) pathway. To understand more fully the molecular mechanism of this regulation, we compared the regulation of AC7 with that of AC2 in bone marrow-derived macrophages devoid of AC7. Although both enzymes could fully restore regulation of cAMP by Gβγ, activation of the G(13) pathway preferentially synergized with AC7. Exchange of domains between the two isoforms indicates that the C1b domain and the N-terminus of the C1a domain are important for directing selective regulation of AC7 by the G(13) pathway. A mutagenesis screen identified more specific regions of AC7 that differentially mediate its regulation by distinct pathways.


A framework towards computational discovery of disease sub-types and associated (sub-)biomarkers.
Biomarker related patient data is generally assessed in order to determine relevant but generalized subset of the biomarkers. However, it fails to identify specific sub-groups of the patients or their corresponding (subset of) the biomarkers. This paper therefore proposes a novel framework that is capable of discovering disease sub-groups (types) and associated subset of biomarkers, which is expected to lead to enable the discovery of personalized bio-marker set. The framework is based on the utilization of a histogram obtained by using the Euclidean distances between the samples in a given data set. The t-test method is used for the selection of sub-set(s) of the biomarkers whereas the classification is performed by means of k-nearest neighbor, support vector machines and naive Bayes (NBayes) classifiers. For the assessment of the methods, leave-out-out cross validation is employed. As a case study, the method is applied in the analysis of male hypertension microarray data that consists of 159 patients and 22184 gene expressions. The method has helped identify specific sub-groups of the patients and their corresponding bio-marker sub-sets. The results therefore suggest that the generalized bio-marker sub-sets are not representative of the disease and therefore more focus should be on the sub-groups of the patients and their biomarker subsets identified through the proposed approach. It is particularly observed that the threshold values over the histogram are crucial to discover both sub-sets of the samples and biomarkers, and therefore can be used to determine complexity level of the study.


Standing centre of pressure alters the vibration transmissibility response of the foot.
Vibration-white foot as an occupational disease has underscored the need to better understand the vibration response of the foot. While vibration transmissibility data exist for a natural standing position, it is anticipated that weight distribution will affect the response. The purpose of this study was to determine the effects of changes in centre of pressure (COP) on the foot's biomechanical response. Twenty-one participants were exposed to vertical vibration of 30 mm/s, with a sine sweep from 10-200 Hz. Z-axis (vertical) vibration was measured at 24 locations on the right foot, with the COP shifted forward or toward the heel. A mixed model analysis at each location revealed significant differences (p < .001) in the transmissibility response when the COP was altered to the forefoot and rearfoot. In general, the peak frequency of the average vibration response increased for a region of the foot when the COP was shifted toward that region. Practitioner Summary: Altering the centre of pressure location resulted in changes in the transmission of vibration through the foot. The forward lean position was associated with the greatest amplitude of vibration transmissibility at the toes. This information is relevant for clinicians studying vibration-induced white-foot and engineers designing protective equipment.


Targeting the inflammatory response in healing myocardial infarcts.
Healing of myocardial infarcts depends on an inflammatory cascade that ultimately results in clearance of dead cells and matrix debris and formation of a scar. Myocardial necrosis activates complement, Nuclear Factor (NF)-kappaB and Toll-like Receptor (TLR)-dependent pathways, and generates free radicals, triggering an inflammatory response. Chemokines and cytokines are markedly induced in the infarct and mediate recruitment and activation of neutrophils and mononuclear cells. Extravasation of platelets and plasma proteins, such as fibrinogen and fibronectin, results in formation of a clot, consisting of platelets embedded in a mesh of crosslinked fibrin. This provisional matrix provides a scaffold for migration of cells into the infarct. Monocytes differentiate into macrophages and secrete fibrogenic and angiogenic growth factors inducing formation of granulation tissue, containing myofibroblasts and neovessels. Repression of proinflammatory cytokine and chemokine synthesis, mediated in part through Transforming Growth Factor (TGF)-beta and Interleukin (IL)-10, is critical for resolution of the inflammatory infiltrate and transition to fibrous tissue deposition. Infarct myofibroblasts deposit extracellular matrix proteins and a collagen-based scar is formed. As the wound matures, fibroblasts undergo apoptosis and neovessels regress, resulting in formation of a scar with a low cellular content containing dense, cross-linked collagen. The pathologic and structural changes associated with infarct healing directly influence ventricular remodeling and affect prognosis in patients with myocardial infarction. Understanding the mechanisms involved in the regulation of the post-infarction inflammatory response, and the spatial and temporal parameters of wound healing is necessary in order to identify specific molecular targets for therapeutic intervention.


Penetrating atherosclerotic ulcer of the aorta: treatment by endovascular stent-graft placement.
To summarize the current clinical experience with endovascular stent-graft repair in patients presenting with penetrating atherosclerotic ulcer of the descending thoracic aorta. Penetrating atherosclerotic ulcer is increasingly acknowledged as a pathologic variant of classic false lumen aortic dissection with a high incidence of bleeding complications and rupture in up to 40% of patients. So far, no generally accepted therapeutic regimen has been established, as the natural history of penetrating atherosclerotic ulcer is not yet fully understood. Recently, however, penetrating atherosclerotic ulcer is increasingly considered to be treated more aggressively (preferentially surgically). Given the high morbidity and mortality of aortic surgery, endovascular stent-graft repair may be an attractive, less invasive alternative in selected patients with penetrating atherosclerotic ulcer. Sealing of a penetrating ulcer by the stent-graft reduces wall stress and thus provides stabilization of the diseased aortic segment. To date, there is limited experience with endovascular repair in penetrating atherosclerotic ulcer, suggesting that endovascular stent-graft repair is safe and effective. Long-term results are, however, required to fully establish the efficacy of endovascular repair in penetrating atherosclerotic ulcer.


Seasonality, annual trends, and characteristics of dengue among ill returned travelers, 1997-2006.
We examined seasonality and annual trends for dengue cases among 522 returned travelers reported to the international GeoSentinel Surveillance Network. Dengue cases showed region-specific peaks for Southeast Asia (June, September), South Central Asia (October), South America (March), and the Caribbean (August, October). Travel-related dengue exhibited annual oscillations with several epidemics occurring during the study period. In Southeast Asia, annual proportionate morbidity increased from 50 dengue cases per 1,000 ill returned travelers in nonepidemic years to an average of 159 cases per 1,000 travelers during epidemic years. Dengue can thus be added to the list of diseases for which pretravel advice should include information on relative risk according to season. Also, dengue cases detected at atypical times in sentinel travelers may inform the international community of the onset of epidemic activity in specific areas.


Blood pressure and sodium balance in hypertensive patients in hospital.
Ninety-one subjects with uncomplicated essential hypertension were admitted to hospital. They received a diet with a fixed amount of sodium (55 mmol/day), without calorie restriction. Blood pressure was measured at 2-hr intervals and averaged for each day. In patients who apparently adhered to a low salt diet before admission (group I), BP dropped 5 mmHg during the first 24 hr and then remained stable. In patients who previously ingested more salt (group II) a similar drop in pressure occurred during the first day, but pressure declined another 6 mmHg during the second day, to stabilize thereafter. Both neurovegetative and sodium-dependent factors appear to contribute to the decrease in BP while the patient is in hospital.


The effect on haemoglobin of the use of iron cooking pots in rural Malawian households in an area with high malaria prevalence: a randomized trial.
Innovative low-cost sustainable strategies are required to reduce the high prevalence of iron-deficiency anaemia in developing countries. We undertook a community-based randomized controlled intervention trial to assess the effects of cooking in iron or aluminium cooking pots in Malawian households in an area with high malaria prevalence. Analysis was by intention to treat and consistency of use. The primary outcomes were change in haemoglobin and iron status. The study population comprised 164 participants eating from aluminium cooking pots and 158 from iron cooking pots. The mean haemoglobin change was significantly increased after 6 weeks in adults who consistently ate from an iron cooking pot (+3.6 g/l compared to -3.2 g/l, mean difference between groups 6.8 g/l, 95% CI +0.86, +12.74). In children, no significant haemoglobin change was observed in consistent pot users, although they showed a significant reduction in iron deficiency (iron 8.6 ZP/g and aluminium 10.8 ZP/g, mean difference 2.2 ZP/g, 95% CI +1.08, +3.32). Rural Malawian adults in a high malaria transmission area who consistently consume food prepared in iron cooking pots show a significant rise in haemoglobin after 6 weeks use. Children showed a reduction in iron deficiency, but no significant improvement in haemoglobin, possibly because of their high malaria parasite prevalence. Using iron cooking pots in developing countries could provide an innovative way to prevent iron deficiency and anaemia in malarious areas where regular iron supplementation is problematic.


Population groups: indexing, coverage, and retrieval effectiveness of ethnically related health care issues in health sciences databases.
This study examined methods of accessing (for indexing and retrieval purposes) medical research on population groups in the major abstracting and indexing services of the health sciences literature. The study of diseases in specific population groups is facilitated by the indexing of both diseases and populations in a database. The MEDLINE, PsycINFO, and Embase databases were selected for the study. The published thesauri for these databases were examined to establish the vocabulary in use. Indexing terms were identified and examined as to their representation in the current literature. Terms were clustered further into groups thought to reflect an end user's perspective and to facilitate subsequent analysis. The medical literature contained in the three online databases was searched with both controlled vocabulary and natural language terms. The three thesauri revealed shallow pre-coordinated hierarchical structures, rather difficult-to-use terms for post-coordination, and a blurring of cultural, genetic, and racial facets of populations. Post-coordination is difficult because of the system-oriented terminology, which is intended mostly for information professionals. The terminology unintentionally restricts access by the end users who lack the knowledge needed to use the thesauri effectively for information retrieval. Population groups are not represented adequately in the index languages of health sciences databases. Users of these databases need to be alerted to the difficulties that may be encountered in searching for information on population groups. Information and health professionals may not be able to access the literature if they are not familiar with the indexing policies on population groups. Consequently, the study points to a problem that needs to be addressed, through either the redesign of existing systems or the design of new ones to meet the goals of Healthy People 2000 and beyond.


Dyadic Model of Adaptation to Life-Limiting Illness.
Context: We previously developed the reintegration model to describe the adjustment process for individuals at the end of life. However, caregivers and loved ones also require significant support and must work to reimagine their relationship with one another. Objectives: We sought to develop a dyadic version of the reintegration model that delineates key parts of the adjustment process that occur between the patient and another significant person rather than as two separate individuals. Methods: We refined an initial conceptual model of this dyadic process with findings from a narrative literature review on spousal dyadic mutuality. We assessed emergent themes regarding dyadic adjustment from the literature for their fit with our original reintegration model and through consensus discussion, applied the findings to a final proposed conceptual model of dyadic reintegration at the end of life. Results: Examples of dyadic adjustment in the literature relate to the comprehension, creative adaptation, and reintegration processes described in the original reintegration model. Evidence also supported three substantive additions in the new dyadic model: (1) shared understanding that the harmony of the dyad is interrupted; (2) consideration of the "we" (the dyad) and the "I" (the individual) in mutual reflection to create a shared narrative; and (3) emphasis on relationship as a factor impacting adjustment processes. Conclusions: Available evidence supports interdependent relationships between members of dyads for the three adaptation processes of comprehension, creative adaptation, and reintegration in the model. This dyadic reintegration model can be useful in clinical practice to support dyads facing life-limiting illness.


Thresholding implemented in the frequency domain.
Image processing procedures are usually carried out in the spatial domain where the images are acquired and presented/utilized. The linear nature of the Fourier transform allows only those operations that are linear to be mapped into the frequency domain. In contrast, nonlinear operations and manipulations cannot be realized directly in the frequency domain. One of these nonlinear operations is thresholding. When operating in the spatial domain to segment image contents into object and background, thresholding is simple and efficient. However, it has no obvious representation in the frequency domain and cannot be carried out there in a straightforward fashion. In this paper, a means to relax the rigid linear limitation of the Fourier transform was investigated. A novel approach was established to achieve spatial thresholding using only frequency domain operations. The spatial grayscale or scalar data set (two-dimensional (2-D) image or three-dimensional (3-D) volume) was expanded into a binary volume in hyperspace having one more dimension than the original data set. The extended dimension is the gray level of the original data. Manipulating only on that dimension produces the effect of thresholding.


[The Zurich Specialist Clinic for Adolescent with Gender Dysphoria - Preliminary Follow-up Results].
The Zurich Specialist Clinic for Adolescent with Gender Dysphoria - Preliminary Follow-up Results The specialist clinic for children and adolescents with gender dysphoria (GD) of the Psychiatric University Hospital of Zurich shows an increasing number of referrals since its foundation in 2009. Since 2014 we started an observational study including adolescents aged 13 years and older. At the time of the first appointment (T0) N = 77 participants completed a battery of questionnaires assessing demographic factors, general psychopathology, quality of life as well as gender identity, social transitioning and GD treatment modalities. Few of the adolescents were socially transitioned and had hormone therapy but 77.9 % wished to get hormone therapy. Follow up assessment T1 was performed after at least one year of treatment in our specialist clinic. 51 adolescents completed an online follow-up examination including the same questionnaires and baseline parameters as well as a scale measuring treatment satisfaction. At T0, 77.3 % of the adolescents scored in the clinical range of the Youth Self Report (YSR) total score, which did not decrease significantly until T1 in our preliminary follow up sample. Puberty blocking before T0 correlated negatively with the YSR score, indicating less psychopathology in treated patients. Preliminary longitudinal analysis suggests that social transitioning influences quality of life (Kidscreen subscale autonomy and parental relationship). At T1, 52 % of the adolescents were socially transitioned in all contexts and 70 % received gender affirming hormonal treatment. Gender identity changed between T0 and T1 in about 18 % of the cases. Treatment satisfaction in most cases was high.


Role of Foxp3-positive tumor-infiltrating lymphocytes in the histologic features and clinical outcomes of hepatocellular carcinoma.
The role of Foxp3-positive regulatory T cells (Foxp3 Tregs) in suppression of antitumoral immune response is well documented in patients with cancer. However, it is not known whether Foxp3 Tregs are associated with specific clinicopathologic characteristics of hepatocellular carcinoma (HCC). The aims of the present study were: (1) to investigate the relationship between Foxp3 Tregs and histologic differentiation, Edmondson-Steiner (ES) nuclear grade, vascular invasion, and pathologic stage of HCC in patients undergoing surgery for their disease; and (2) to evaluate any Foxp3 Treg-defined difference in the risk for tumor recurrence or death. The study sample included 131 histologic sections of HCC. The number of tumor-infiltrating CD3, CD8, and Foxp3 lymphocytes was assessed by immunohistochemistry. An increased Foxp3:CD3 ratio was associated with more poorly differentiated HCC (P=0.0016) and higher ES nuclear grade (P=0.0407). An increased Foxp3:CD8 ratio was also associated with poorer differentiation (P=0.0044), higher ES nuclear grade (P=0.0179), recurrence (P=0.0183), decreased overall survival (hazard ratio=1.153; 95% confidence interval, 1.019-1.304; P=0.0235), and decreased disease-free survival (hazard ratio=1.138; 95% confidence interval, 1.016-1.273; P=0.0249). Tumor size and type of surgery (surgical resection) were associated with decreased disease-free survival on univariate analysis but not on multivariate analysis. In conclusion, a higher concentration of tumor-infiltrating Foxp3 Tregs in HCC is associated with higher grade and poorly differentiated tumors and signifies an unfavorable prognosis.


[Treatment of experimental corneal injuries by dioxidine and urotropin solutions].
The efficiency of 1% dioxidine and 5% urotropin solutions in suppurative penetrating corneal wounds was studied in 2 experimental series on 54 rabbits. In series A toxic effects of these drugs on eyeball tissues were studied on 6 rats, in series B drug effects were studied on a model of suppurative penetrating corneal wounds. A 21-day course of therapy with these drugs (instillations and subconjunctival parabulbar injections) caused no toxic changes in ocular tissues. Therapy with 1% dioxidine and 5% urotropin accelerated clinical cure by 4-6 days in comparison with other methods of treatment. Clinical cure was observed 3-5 days sooner after combined use of both drugs in comparison with their individual application, during during combined use their antibacterial effect was synergistic. Hence, a combination of 1% dioxidine and 5% urotropin solutions is an effective method for treating suppurative penetrating wounds of the cornea.


The Study of Postmortem Human Synaptosomes for Understanding Alzheimer's Disease and Other Neurological Disorders: A Review.
Synaptic dysfunction is thought to play important roles in the pathophysiology of many neurological diseases, including Alzheimer's disease, Parkinson's disease, and schizophrenia. Over the past few decades, there have been systematic efforts to collect postmortem brain tissues via autopsies, leading to the establishment of dozens of human brain banks around the world. From cryopreserved human brain tissues, it is possible to isolate detached-and-resealed synaptic terminals termed synaptosomes, which remain metabolically and enzymatically active. Synaptosomes have become important model systems for studying human synaptic functions, being much more accessible than ex vivo brain slices or primary neuronal cultures. Here we review recent advances in the establishment of human brain banks, the isolation of synaptosomes, their biological activities, and various analytical techniques for investigating their biochemical and ultrastructural properties. There are unique insights to be gained by directly examining human synaptosomes, which cannot be substituted by animal models. We will also discuss how human synaptosome research has contributed to better understanding of neurological disorders, especially Alzheimer's disease.


Identification of an Increased Alveolar Macrophage Subpopulation in Old Mice That Displays Unique Inflammatory Characteristics and Is Permissive to Mycobacterium tuberculosis Infection.
The elderly population is more susceptible to pulmonary infections, including tuberculosis. In this article, we characterize the impact of aging on the phenotype of mouse alveolar macrophages (AMs) and their response to Mycobacterium tuberculosis. Uninfected AMs were isolated from bronchoalveolar lavage of young (3 mo) and old (18 mo) C57BL/6 mice. AMs from old mice expressed higher mRNA levels of CCL2, IFN-β, IL-10, IL-12p40, TNF-α, and MIF than young mice, and old mice contained higher levels of CCL2, IL-1β, IFN-β, and MIF in their alveolar lining fluid. We identified two distinct AM subpopulations, a major CD11c+ CD11b- population and a minor CD11c+ CD11b+ population; the latter was significantly increased in old mice (4-fold). Expression of CD206, TLR2, CD16/CD32, MHC class II, and CD86 was higher in CD11c+ CD11b+ AMs, and these cells expressed monocytic markers Ly6C, CX3CR1, and CD115, suggesting monocytic origin. Sorted CD11c+ CD11b+ AMs from old mice expressed higher mRNA levels of CCL2, IL-1β, and IL-6, whereas CD11c+ CD11b- AMs expressed higher mRNA levels of immune-regulatory cytokines IFN-β and IL-10. CD11c+ CD11b+ AMs phagocytosed significantly more M. tuberculosis, which expressed higher RNA levels of genes required for M. tuberculosis survival. Our studies identify two distinct AM populations in old mice: a resident population and an increased CD11c+ CD11b+ AM subpopulation expressing monocytic markers, a unique inflammatory signature, and enhanced M. tuberculosis phagocytosis and survival when compared with resident CD11c+ CD11b- AMs, which are more immune regulatory in nature.


Spin photonics in 3D whispering gallery mode resonators.
Whispering gallery modes are known for possessing orbital angular momentum, however the interplay of local spin density, orbital angular momentum, and the near-field interaction with quantum emitters is far less explored. Here, we study the spin-orbit interaction of a circularly polarized dipole with the whispering gallery modes (WGMs) of a spherical resonator. Using an exact dyadic Green's function approach, we show that the near-field interaction between the photonic spin of a circularly polarized dipole and the local electromagnetic spin density of whispering gallery modes gives rise to unidirectional behaviour where modes with either positive or negative orbital angular momentum are excited. We show that this is a manifestation of spin-momentum locking with the whispering gallery modes of the spherical resonator. We also discuss requirements for possible experimental demonstrations using Zeeman transitions in cold atoms or quantum dots, and outline potential applications of these previously overlooked properties. Our work firmly establishes local spin density, momentum and decay as a universal right-handed electromagnetic triplet for near-field light-matter interaction.


Salute to the sun: a new dawn in yoga therapy for breast cancer.
Interest in the application of yoga for health benefits in western medicine is growing rapidly, with a significant rise in publications. The purpose of this systematic review is to determine whether the inclusion of yoga therapy to the treatment of breast cancer can improve the patient's physical and psychosocial quality of life (QoL). A search of peer reviewed journal articles published between January 2009 and July 2014 was conducted. Studies were included if they had more than 15 study participants, included interventions such as mindfulness-based stress reduction (MBSR) or yoga therapy with or without comparison groups and had stated physical or psychological outcomes. Screening identified 38 appropriate articles. The most reported psychosocial benefits of yoga therapy were anxiety, emotional and social functioning, stress, depression and global QoL. The most reported physical benefits of yoga therapy were improved salivary cortisol readings, sleep quality and lymphocyte apoptosis. Benefits in these areas were linked strongly with the yoga interventions, in addition to significant improvement in overall QoL. The evidence supports the use of yoga therapy to improve the physical and psychosocial QoL for breast cancer patients with a range of benefits relevant to radiation therapy. Future studies are recommended to confirm these benefits. Evidence-based recommendations for implementation of a yoga therapy programme have been derived and included within this review. Long-term follow-up is necessary with these programmes to assess the efficacy of the yoga intervention in terms of sustainability and patient outcomes.


Relationship of drusen and abnormalities of the retinal pigment epithelium to the prognosis of neovascular macular degeneration. The Macular Photocoagulation Study Group.
We graded macular features of 127 fellow eyes of participants in the Macular Photocoagulation Study who had an extrafoveal choroidal neovascular membrane secondary to age-related macular degeneration in the first eye and no initial evidence of the neovascular form of age-related macular degeneration in the fellow eye. Our aims were to determine the relationship of drusen characteristics and retinal pigment epithelial abnormalities to the risk of subsequent development of neovascularization in the fellow eye and the risk of subsequent development of recurrent neovascular membranes after photocoagulation in the first eye. Regression analysis demonstrated that the presence of large drusen and focal hyperpigmentation of the retinal pigment epithelium were independent risk factors for the subsequent development of neovascularization in the fellow eye (relative risk, 2.4 and 2.5, respectively). Only 10% of eyes with no large drusen or any retinal pigment epithelial hyperpigmentation compared with 58% of eyes with both large drusen and retinal pigment epithelial hyperpigmentation developed neovascularization in the fellow eye within 5 years. Using multivariate Cox regression analysis, we noted that the risk of developing recurrent neovascular membranes in the first eye was significantly increased when large drusen (relative risk, 2.8) were noted in the fellow eye at the time of laser treatment in the first eye. Fundus features in the fellow eye appear to help identify patients at high risk of developing visual loss from recurrent neovascular membranes following laser treatment in the first eye and from development of a neovascular membrane in the fellow eye.


Human pancreatic ductal cells: large-scale isolation and expansion.
The in vitro differentiation of pancreatic stem cells has recently been shown to represent a new source of beta cells for cell therapy in diabetes. Human ductal cell differentiation, in vitro, has been documented in three-dimensional (3D) culture and recently substantiated. Although encouraging, the optimization of the ductal cell source, expansion and differentiation ex vivo are mandatory for clinical relevance. We compared three sources of human ductal cells (hDC) (method A1-2, B, and C). The classical main duct isolation of hDC by explant (A1), or enzymatic digestion (A2), was compared with two indirect methods: from 3D cultured human islet/duct-enriched fractions (B) and dedifferentiated exocrine fractions (C). Method A: few viable hDC were obtained from the main duct. Method B: embedding islet/duct rich fraction in 3D collagen gels expands the cytokeratin 19 (CK19)-positive ductal component in the form of ductal cysts, as we described previously; monolayers derived from digested cysts were 80% ductal (CK19). Method C: initially adherent amylase-positive exocrine clusters contained 12% (CK19) to 22% (CK7) ductal cells. One-week exocrine cultures were amylase negative and 46% (CK19) to 63% (CK7) ductal. Cell viability varied: <20% (A1), 81+/-12% (B), 91+/-2% (C). Extrapolating total yields we obtained (+/-SEM): 10.5+/-4.6 x 10(3) (A1), 36+/-18 x 10(3) (A2), 292+/-50 x 10(6) (B), 1696+/-526 x 10(6) (C) viable hDC per pancreas. A secondary monolayer expansion of cyst-derived hDC (method B) was achieved with NuSerum (4.2-fold on plastic, 2.6-fold on 804G matrix; p < 0.05 vs. control cells on plastic). First passage exocrine-derived ductal cells also responded to matrix and to growth factors, albeit not significantly. In conclusion, this study demonstrated that an abundant hDC supply can be obtained from islet/duct or exocrine fractions followed by monolayer expansion with NuSerum. If their differentiation capacity is confirmed, in particular exocrine-derived ductal cells may represent a promising abundant source of islets for allogenic and autologous diabetes cell therapy.


Impact of HIV subtype on performance of the limiting antigen-avidity enzyme immunoassay, the bio-rad avidity assay, and the BED capture immunoassay in Rakai, Uganda.
Previous studies demonstrated that individuals with subtype D HIV infection who had been infected for 2 or more years were frequently misclassified as assay positive using cross-sectional incidence assays. Samples from 510 subjects (212 subtype A, 298 subtype D) who were infected for 2.2 to 14.5 years (median 5.4 years) and were not virally suppressed were tested using an LAg-Avidity enzyme immunoassay (LAg-Avidity EIA), Bio-Rad Avidity assay, and BED capture enzyme immunoassay (BED-CEIA). The performance of these three assays was evaluated using various assay cutoff values [LAg-Avidity EIA: <1.0 OD-n and <2.0 OD-n; Bio-Rad Avidity assay: <40% avidity index (AI) and <80% AI; BED-CEIA: <0.8 OD-n]. The mean LAg-Avidity EIA result was higher for subtype A than D (4.54±0.95 vs. 3.86±1.26, p<0.001); the mean Bio-Rad Avidity assay result was higher for subtype A than D (88.9%±12.5% vs. 75.1±30.5, p<0.001); and the mean BED-CEIA result was similar for the two subtypes (2.2±1.2 OD-n for subtype A, 2.2±1.3 OD-n for subtype D, p<0.9). The frequency of misclassification was higher for individuals with subtype D infection compared to those with subtype A infection, using either the LAg-Avidity EIA with a cutoff of <2.0 OD-n or the Bio-Rad Avidity assay with cutoffs of <40% or <80% AI. No subtype-specific differences in assay performance were observed using the BED-CEIA. Sex and age were not significantly associated with misclassification by any assay. The LAg-Avidity EIA with a cutoff <1.0 OD-n had the lowest frequency of misclassification in this Ugandan population.


The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene.
This paper presents the nucleotide sequence of the Herpes Simplex Virus thymidine kinase (tk) gene. The position on the DNA sequence corresponding to the 5' and 3' termini of tk messenger RNA have been mapped. The mRNA termini are separated by slightly more than 1,300 nucleotides. The same 2,300 nucleotide segment of tk coding strand DNA is fully protected from S1 nuclease digestion when hybridized to tk mRNA. The location and size of the mRNA-coding segment corresponds to a region of the viral DNA that is essential for tk gene expression in microinjected frog oocytes. The nucleotide sequence of the HSV tk gene exhibits an open translational reading frame of 376 codons that extends from the methionine codon most proximal to the 5' terminus of tk mRNA to a UGA stop codon approximately 70 nucleotides from the poly-A addition site. The results of these experiments indicate that the tk gene is not interrupted by intervening DNA sequences, and that certain oligonucleotide sequences adjacent to the termini of the tk gene are homologous to similarly positioned sequences common to structural genes of eukaryotic cells.


Posttransplant hyperglycemia is associated with increased risk of liver allograft rejection.
Intensive glycemic control has been shown to positively impact outcomes in an intensive care setting. Whether this practice is beneficial after liver transplantation (LT) is not known. A retrospective review of patients undergoing LT from February 2002 to July 2007 was conducted to analyze the association between perioperative hyperglycemia and outcomes after LT. Covariates included preexisting diabetes, mean glucose 3 months pre-LT, need for insulin drip post-LT, mean total glucose during the post-LT hospitalization, age, sex, type of transplant, and model for end-stage liver disease score. Outcomes within 1 year of LT included rejection, infection, rehospitalization, prolonged ventilation, and patient/graft survival. One hundred thirteen LT and 31 liver-kidney recipients were included. By multivariate logistic regression adjusting for covariates, the rejection rate was significantly lower for patients with postoperative glucose levels less than 200 mg/dL (n=114) vs. more than 200 mg/dL (n=30) (odds ratio: 0.055; 95% confidence interval: 0.0154-0.200; P<0.001). The need for prolonged ventilation was more common in patients with glucose less than 200 vs. more than 200 mg/dL (odds ratio: 4.30; 95% confidence interval: 1.284-14.388; P=0.018). Although other outcomes, infection, rehospitalization, patient/graft survival, were not different among the glucose control groups, rejection was associated with increased rehospitalizations and infections. Our data demonstrate an association between the immediate posttransplant glycemic control and the development of subsequent rejection. Prospective trials investigating the effects of perioperative glycemic control on outcomes and morbidity after LT are warranted.


Community patterns of the small riverine benthos within and between two contrasting glacier catchments.
Ongoing glacial retreat is expected to lead to numerous changes in glacier-fed rivers. This study documents the development of community composition of the hitherto widely neglected micro- and meiobenthos (MMB: bacteria, fungi, algae, protists, and meiofauna) in glacier rivers in response to the distinct habitat conditions driven by different stages of (de)glacierization. Our model is based on the glacier catchments of the Möll River (MC) and Kleinelendbach stream (KC), in the Austrian Alps, with 60% and 25% glacierization and glacier retreats of 403 and 26 m, respectively, since 1998. Analyses of overall catchment diversity and resemblance patterns showed that neither intense glacierization nor rapid deglacierization were predominant MMB determinants. This was ascribed to the specific environmental conditions at the MC, where the rapidly retreating Pasterze glacier has formed a harsh unstable proglacial, but also a benign floodplain area, with the former suppressing and the latter supporting the structural development of the MMB. Comparisons of similarly aged riverine habitats of the MC proglacial and the KC main channel further evidenced developmental suppression of the MMB (64 taxa) by the rapidly retreating MC glacier, unlike the moderate glacial retreat in the KC (130 taxa). Habitat conditions interacting with melt periods explained the differences in MMB resemblance patterns, which themselves differentially reflected the spatiotemporal habitat settings imposed by the different glacier activities. The varying glacial influences were represented by a glaciality index (GIm) based on water temperature, electrical conductivity, and stream bed stability. The taxonomic richness of nematodes, rotifers, algae, and diatoms was distinctly related to this index, as were most MMB abundances. However, the strongest relationships to the GIm were those of nematode abundances and maturity. Our observations highlight the intense response of the MMB to ongoing glacier retreat and the utility of a simple index to reveal such patterns.


Epidemiology of squamous cell carcinomas in rudd Scardinius erythrophthalmus from SE Ireland.
An epidemiological study was carried out to investigate the possible aetiology of squamous cell carcinomas which occur in a population of rudd Scardinius erythrophthalmus (L.) from Lough Aderry in south-east Ireland. A total of 1343 rudd were sampled from Lough Aderry and 2 nearby small lakes in spring, summer, autumn, and winter over 2 yr, 1986 to 1988. Fish were weighed, measured, sexed, aged, and examined for lesions. Water quality parameters and natural radioactivity were assessed as possible influencing factors in the disease. The prevalence of squamous cell carcinoma was 6.1% overall, with no significant difference between the lakes, seasons or years. Both male and female fish were affected, of ages from 1+ to 5+ yr. No fish of 6+ or 7+ yr was found with tumours, indicating that the neoplasm caused premature death. The female:male sex ratio of rudd with tumours was higher than that of healthy rudd overall, suggesting that males are more susceptible to the neoplasm. Of the parasites observed, Posthodiplostomum cuticola was common, but rarely found in rudd with tumours. Sphaerospora sp.was also common, but not in sufficient densities for statistical inference, and Argulus sp. was present on 7 fish. No viral particles were found. Natural radiation levels in the vicinity of the lakes were low. The lakes sampled are in an agricultural catchment, and the waters were eutrophic. While rudd are tolerant, it is likely that environmental conditions were stressful at least some of the time, with possible consequences for the immunocompetence of the rudd. The aetiology of the carcinoma is still unknown, but the possibilities of carcinogenic compounds resulting from the high nutrient levels, or of carcinogenic algal toxins produced during algal blooms, should be examined.


Favorable Effects of Astaxanthin on Brain Damage due to Ischemia- Reperfusion Injury.
Activated inflammation and oxidant stress during cerebral ischemia reperfusion injury (IRI) lead to brain damage. Astaxanthin (ASX) is a type of carotenoid with a strong antioxidant effect. The aim of this study was to investigate the role of ASX on brain IRI. A total of 42 adult male Sprague-Dawley rats were divided into 3 groups as control (n=14) group, IRI (n=14) group and IRI + ASX (n=14) group. Cerebral ischemia was instituted by occluding middle cerebral artery for 120 minutes and subsequently, reperfusion was performed for 48 hours. Oxidant parameter levels and protein degradation products were evaluated. Hippocampal and cortex cell apoptosis, neuronal cell count, neurological deficit score were evaluated. In the IRI group, oxidant parameter levels and protein degradation products in the tissue were increased compared to control group. However, these values were significantly decreased in the IRI + ASX group (p<0.05). There was a significant decrease in hippocampal and cortex cell apoptosis and a significant increase in the number of neuronal cells in the IRI + ASX group compared to the IRI group alone (p<0.05). The neurological deficit score which was significantly lower in the IRI group compared to the control group was found to be significantly improved in the IRI + ASX group (p<0.05). Astaxanthin protects the brain from oxidative damage and reduces neuronal deficits due to IRI injury.