Resilience to bullying victimization: the role of individual, family and peer characteristics.
Little research attention has been paid to bullied students who function better than expected and are therefore defined as "resilient". The present longitudinal study aimed to identify individual, family and peer factors that predict fewer than expected levels of depression and delinquency following experiences of bullying victimization. The sample consisted 3,136 adolescents. Self-report data were used to measure bullying victimization at age 13 and 14 and depression and delinquency at age 14. We examined the effects of gender, self-esteem, social alienation, parental conflict, sibling victimization and number of close friends on levels of emotional and behavioral resilience following bullying victimization. The resilience measures were derived by regressing depression and delinquency scores at age 14 on levels of bullying victimization at age 13 and 14, respectively. The adolescents who reported low depression despite frequently experiencing bullying tended to be male, had higher self-esteem, were feeling less socially alienated, were experiencing low levels of conflict with parents and were not victimized by siblings. On the other hand, the adolescents who reported low delinquency despite frequently experiencing bullying tended to be female, had higher self-esteem, were experiencing low levels of conflict with parents, were not victimized by siblings and had less close friends. Relationships with parents and siblings continue to play some role in promoting emotional and behavioral adjustment among victims of bullying and, therefore, interventions are more likely to be successful if they target both the psychosocial skills of adolescents and their relationships with their family.


Multiploid inheritance of HIV-1 during cell-to-cell infection.
During cell-to-cell transmission of human immunodeficiency virus type 1 (HIV-1), many viral particles can be simultaneously transferred from infected to uninfected CD4 T cells through structures called virological synapses (VS). Here we directly examine how cell-free and cell-to-cell infections differ from infections initiated with cell-free virus in the number of genetic copies that are transmitted from one generation to the next, i.e., the genetic inheritance. Following exposure to HIV-1-expressing cells, we show that target cells with high viral uptake are much more likely to become infected. Using T cells that coexpress distinct fluorescent HIV-1 variants, we show that multiple copies of HIV-1 can be cotransmitted across a single VS. In contrast to cell-free HIV-1 infection, which titrates with Poisson statistics, the titration of cell-associated HIV-1 to low rates of overall infection generates a constant fraction of the newly infected cells that are cofluorescent. Triple infection was also readily detected when cells expressing three fluorescent viruses were used as donor cells. A computational model and a statistical model are presented to estimate the degree to which cofluorescence underestimates coinfection frequency. Lastly, direct detection of HIV-1 proviruses using fluorescence in situ hybridization confirmed that significantly more HIV-1 DNA copies are found in primary T cells infected with cell-associated virus than in those infected with cell-free virus. Together, the data suggest that multiploid inheritance is common during cell-to-cell HIV-1 infection. From this study, we suggest that cell-to-cell infection may explain the high copy numbers of proviruses found in infected cells in vivo and may provide a mechanism through which HIV preserves sequence heterogeneity in viral quasispecies through genetic complementation.


Endorphin content of white blood cells and peritoneal cells in neonatally benzpyrene treated adult rats.
White blood cells of rats (lymphocytes, monocytes, macrophages, granulocytes and mast cells) contain beta-endorphin. Two months after a single neonatal benzpyrene treatment (imprinting) there is an elevated level of immunoreactive endorphin in the blood and peritoneal cells of female animals and blood cells of males. The endorphin content decreased in the peritoneal cells of males. In the blood, the granulocytes of female, and the lymphocytes of male rats contained the highest amount of endorphin. In the peritoneal fluid also the granulocytes of females contained the highest amount of endorphin, in contrast to males, where the endorphin content of cells decreased and the lowest level of it was present in the lymphocytes. The experiments justify that benzpyrene treatment can durably influence endorphin levels of white blood cells and gives new data to the already known lifelong health destroying effects of perinatal benzpyrene exposition (alterations of hormone receptor binding capacity and sexual behavior).


Analysis of T cell activation requirements with the use of alloantigens or an anti-clonotypic monoclonal antibody.
H-2Kb-specific alloreactive cytotoxic T lymphocyte (CTL) clones, which secrete macrophage-activating factor (MAF) in response to antigen, were used to study the antigenic and physiologic parameters for T cell-antigen receptor (Ti)-mediated activation. When H-2Kb was presented on untreated, formaldehyde (FOR)-fixed cells, or liposomes, optimal stimulation in the absence of co-factors was obtained only with the untreated cells. FOR cells, which were efficient inhibitors of CTL-target cell interactions for the CTL clones studied, had a greatly reduced activating capacity. Phorbol myristic acetate (PMA), which alone had no activating effect on the CTL clones, acted in synergy with FOR cells, restoring an unimpaired H-2Kb-dependent activation for some CTL clones. H-2Kb-liposomes were not stimulatory even in the presence of PMA and/or splenic feeder cells. For one H-2Kb-specific CTL clone (KB5-C20), activation was studied with the use of an anti-clonotypic (anti-Ti) monoclonal antibody (mAb), Désiré-1. By using this immunoglobulin (Ig) G2a mAb or its F(ab')2 or Fab fragments, it was observed that the association constant of the IgG2a (4.7 X 10(9) M-1) for KB5-C20 was 30-fold higher than that of the Fab fragment, whereas the molar concentration of mAb required to inhibit 50% of the H-2Kb-specific CTL activity of clone KB5-C20 was 70-fold to 90-fold higher for the Fab fragment than for the IgG2a (1 to 5 X 10(-10) M). It was also observed that activation as measured by MAF secretion of clone KB5-C20 was obtained by using the divalent IgG2a mAb or its F(ab')2 fragment at 10(-9) M to 10(-8) M in the absence of any added co-factor or feeder cells, whereas a 500-fold higher molar concentration of the Fab fragment only induced very low levels of MAF secretion. Also, in the presence of PMA, the mAb-mediated activation of MAF secretion by clone KB5-C20 was increased, but Fab-induced activation never resulted in high titers of MAF. We conclude that the intrinsic "affinity" of the H-2Kb-Ti interaction is probably so low for the Ti of clone KB5-C20 that efficient activation by H-2Kb is obtained only when multivalency is achieved on H-2Kb-expressing cells, and possibly when molecules on the T cells such as Lyt-2 and LFA-1 stabilize the interactions.(ABSTRACT TRUNCATED AT 400 WORDS)


Two new species of Fusarium section Liseola associated with mango malformation.
Mango malformation is an economically important disease of Mangifera indica globally. A recent DNA-based study indicated that two distinct, phylogenetic lineages previously identified as Fusarium subglutinans are associated with this disease in South Africa. The objective of this study was to characterize Fusarium isolates associated with mango malformation, including the two different F. subglutinans groups, based on morphological characteristics. For this purpose we examined Fusarium strains isolated from diseased mango inflorescences from diverse geographical origins. We also used sexual compatibility tests to determine whether sexual reproduction among the strains was possible. The morphological characters considered were shape of the conidia, presence of mono- and/or polyphialides, origin of the conidiophores from the substrate, presence of chlamydospores and the presence of sterile coiled hyphae. Three unique Fusarium spp. were identified. In this paper, we provide formal descriptions for the two new taxa in the section Liseola that we have named F. mangiferae and F. sterilihyphosum. Fusarium mangiferae is conspecific with strains that were previously identified as F. subglutinans and reported to be the causal agent of malformation in mango growing areas throughout the world. Fusarium sterilihyphosum, on the other hand, has been isolated only from malformed mango tissue in South Africa.


Predictors of acute kidney injury after lung resection surgery: a retrospective case-control study.
Patients undergoing lung resection are at increased risk for acute kidney injury (AKI) in the immediate postoperative period, with important consequences for longer term morbidity and mortality. Lung resection surgery has unique considerations that could increase the risk of AKI, including lung resection volume, duration of one-lung ventilation (OLV), and intraoperative fluid restriction. Yet, specific risk factor data are lacking. The objective of this study was to identify independent risk factors for early AKI after lung resection surgery. We conducted a retrospective case-control study of all patients presenting for elective lung resection surgery at an academic medical centre over a four-year period. Cases were patients who experienced an AKI and control patients were those who did not experience an AKI, based on KDIGO criteria. Baseline demographics and comorbidities along with duration of OLV and amount of lung resected were collected by retrospective chart review. The data were analyzed using multivariable logistic regression to identify independent predictors of AKI. Acute kidney injury occurred within 48 hr in 57/1,045 (5.5%; 95% confidence interval, 4.2 to 7.0) of patients. On multivariable analysis, our model of best fit included preoperative serum creatinine, male sex, use of angiotensin II receptor blockers, and duration of OLV. The rate of complications, intensive care unit admission, and risk of death were all higher in the group of patients who experienced AKI. Acute kidney injury occurs frequently after lung resection surgery and is associated with increased risk of postoperative complications. Increased duration of OLV may be a risk factor for AKI in this population.


Characterization of the canine type C enterotoxin produced by Staphylococcus intermedius pyoderma isolates.
The type C staphylococcal enterotoxins (SECs) are a group of highly conserved proteins with substantial antigenic cross-reactivity. Although Staphylococcus intermedius and coagulase-positive species of staphylococci are reported to produce SEC and other SEs, toxins produced by species other than Staphylococcus aureus have not been previously characterized. In this study we report the molecular, biological, and immunological properties of the canine SEC (SECcanine) expressed by pathogenic isolates of S. intermedius. The mature form of SECcanine has 239 amino acid residues and a pI of 7.0. Typical of the SEs, purified SECcanine induces an emetic response in monkeys and the proliferation of T cells in a Vbeta-dependent manner. Although SECcanine has >95% sequence identity to previously described SEC variants, its sequence is most related to SEC2 and SEC3. In contrast to the sequence similarity, the Vbeta profile induced by SECcanine is typical of that induced by SEC1. This result is likely explained by the conservation of a cysteine residue at position 26 in SECcanine; residues at this position have been previously shown to determine subtype-dependent differences in T-cell receptor interactions of other SEs. Overall, these results show that superantigen toxins produced by the multiple members of the genus Staphylococcus are highly conserved in respect to biological and structural properties. Further, the frequent association of SECcanine with pyoderma in dogs supports the notion that the toxins are important for staphylococcal survival and pathogenesis.


Obesity and poverty paradox in developed countries.
Obesity is a civilization disease and the proportion of people suffering from it continues to grow, especially in the developed countries. Number of obese people in Europe has increased threefold over the last 20 years. The paradox of obesity and poverty relationship is observed especially in the developed and developing countries. In developing countries, along with economic development and income growth, the number of people with overweight and obesity is increasing. This paradox has a relationship with both the easy availability and low cost of highly processed foods containing 'empty calories' and no nutritional value. To date, this paradox has been described in the United States and the United Kingdom, although many European countries are also experiencing high percentages of obese people. Among the reasons for the growing obesity in the population of poor people are: higher unemployment, lower education level, and irregular meals. Another cause of obesity is low physical activity, which among the poor is associated with a lack of money for sports equipment. Due to the large rate of deaths caused by diseases directly linked to obesity, the governments of many countries implement prevention programmes of overweight and obesity. These programmes are based primarily on educating the public about a healthy lifestyle based on healthy eating, daily physical activity and avoiding alcohol and cigarettes.


Competing-risks analysis for evaluating the prognosis of patients with microinvasive cutaneous squamous cell carcinoma based on the SEER database.
Utilizing the traditional Cox regression model to identify the factors affecting the risk of mortality due to microinvasive cutaneous squamous cell carcinoma (micSCC) may produce skewed results. Since cause-specific mortality can guide clinical decision-making, this study employed the Fine-Gray model based on the Surveillance, Epidemiology, and End Results (SEER) database to identify significant predictive variables for the risk of micSCC-related mortality. This study used the information of patients with micSCC who were listed in the SEER database during 2000-2015. Cox regression and Fine-Gray models were utilized for the multivariable analysis, and Gray's test and the cumulative incidence function were used for the univariable analyses. There were 100 patients who died from other reasons and 38 who died from micSCC among the 1259 qualified patients with micSCC. Most were female, white, married, had localized metastasis, etc. According to the univariable Gray's test (P < 0.05), the cumulative incidence rate for events of interest was strongly associated with age, sex, marital status, American Joint Committee on Cancer staging, radiation status, summary stage, chemotherapy status, surgery status, and tumor size. Multivariable Cox regression analysis and multivariable competing-risks analysis indicated that age, tumor size, and income were independent risk variables for the prognosis of patients with micSCC. In both age and tumor size variables, the competing-risks model showed a slight decrease in the hazard ratio and a slight narrowing of the 95% confidence interval compared with the Cox regression model. However, this pattern is not evident in the income variable. This study established a Fine-Gray model for identifying the independent risk factors that influence the risk of mortality among patients with micSCC. This study uncovers that, in the context of competing risks, age, tumor size, and income serve as independent risk factors influencing the risk of mortality due to micSCC among patients. Our findings have the potential to provide more accurate risk assessments for patient outcomes and contribute to the development of individualized treatment plans.


Electron-mediating Cu(A) centers in proteins: a comparative high field (1)H ENDOR study.
High field (W-band, 95 GHz) pulsed electron-nuclear double resonance (ENDOR) measurements were carried out on a number of proteins that contain the mixed-valence, binuclear electron-mediating Cu(A) center. These include nitrous oxide reductase (N(2)OR), the recombinant water-soluble fragment of subunit II of Thermus thermophilus cytochrome c oxidase (COX) ba(3) (M160T9), its M160QT0 mutant, where the weak axial methionine ligand has been replaced by a glutamine, and the engineered "purple" azurin (purpAz). The three-dimensional (3-D) structures of these proteins, apart from the mutant, are known. The EPR spectra of all samples showed the presence of a mononuclear Cu(II) impurity with EPR characteristics of a type II copper. At W-band, the g( perpendicular) features of this center and of Cu(A) are well resolved, thus allowing us to obtain a clean Cu(A) ENDOR spectrum. The latter consists of two types of ENDOR signals. The first includes the signals of the four strongly coupled cysteine beta-protons, with isotropic hyperfine couplings, A(iso), in the 7-15 MHz range. The second group consists of weakly coupled protons with a primarily anisotropic character with A(zz) < 3 MHz. Orientation selective ENDOR spectra were collected for N(2)OR, M160QT0, and purpAz, and simulations of the cysteine beta-protons signals provided their isotropic and anisotropic hyperfine interactions. A linear correlation with a negative slope was found between the maximum A(iso) value of the beta-protons and the copper hyperfine interaction. Comparison of the best-fit anisotropic hyperfine parameters with those calculated from dipolar interactions extracted from the available 3-D structures sets limit to the sulfur spin densities. Similarly, the small coupling spectral region was simulated on the basis of the 3-D structures and compared with the experimental spectra. It was found that the width of the powder patterns of the weakly coupled protons recorded at g(perpendicular) is mainly determined by the histidine H(epsilon)(1) protons. Furthermore, the splitting in the outer wings of these powder patterns indicates differences in the positions of the imidazole rings relative to the Cu(2)S(2) core. Comparison of the spectral features of the weakly coupled protons of M160QT0 with those of the other investigated proteins shows that they are very similar to those of purpAz, where the Cu(A) center is the most symmetric, but the copper spin density and the H(epsilon)(1)-Cu distances are somewhat smaller. All proteins show the presence of a proton with a significantly negative A(iso) value which is assigned to an amide proton of one of the cysteines. The simulations of both strongly and weakly coupled protons, along with the known copper hyperfine couplings, were used to estimate and compare the spin density distribution in the various Cu(A) centers. The largest sulfur spin density was found in M160T9, and the lowest was found in purpAz. In addition, using the relation between the A(iso) values of the four cysteine beta-protons and the H-C-S-S dihedral angles, the relative contribution of the hyperconjugation mechanism to A(iso) was determined. The largest contribution was found for M160T9, and the lowest was found for purpAz. Possible correlations between the spin density distribution, structural features, and electron-transfer functionality are finally suggested.


Baicalein Reduces Liver Injury Induced by Myocardial Ischemia and Reperfusion.
Baicalein is a component of the root of Scutellaria baicalensis Georgi, which has traditionally been used to treat liver disease in China. In the present study, we investigated baicalein' ability to reduce the liver injury induced by myocardial ischemia and reperfusion (I/R). Myocardial I/R was induced in this experiment by a 40[Formula: see text]min occlusion of the left anterior descending coronary artery and a 3[Formula: see text]h reperfusion in rats. The induced myocardial I/R significantly increased the serum levels of aspartate transaminase (AST) and alanine transaminase (ALT), indicating the presence of liver injury. Hepatic apoptosis was significantly increased. The serum levels of tumor necrosis factor-[Formula: see text] (TNF-[Formula: see text]), interleukin-1[Formula: see text] (IL-1[Formula: see text]), and interleukin-6 (IL-6) were significantly elevated, as was the TNF-[Formula: see text] level in the liver. Intravenous pretreatment with baicalein (3, 10, or 30[Formula: see text]mg/kg) 10[Formula: see text]min before myocardial I/R significantly reduced the serum level increase of AST and ALT, apoptosis in the liver, and the elevation of TNF-[Formula: see text], IL-1[Formula: see text], and IL-6 levels. Moreover, baicalein increased Bcl-2 and decreased Bax in the liver. Phosphorylation of the prosurvival kinases, including Akt and extracellular signal-regulated kinases 1 and 2 (ERK1/2), was also increased. In conclusion, we found that baicalein can reduce the liver injury induced by myocardial I/R. The underlying mechanisms are likely related to the inhibition of the extrinsic and intrinsic apoptotic pathways, possibly via the inhibition of TNF-[Formula: see text] production, the modulation of Bcl-2 and Bax, and the activation of Akt and ERK1/2. Our findings may provide a rationale for the application of baicalein or traditional Chinese medicine containing large amounts of baicalein to prevent liver injury in acute myocardial infarction and cardiac surgery.


Efficient upconverting carbon nitride nanotubes for near-infrared-driven photocatalytic hydrogen production.
We report a facile chemical technique for synthesizing nanotube-based hybrid materials for near-infrared-driven photocatalytic hydrogen (H2) production. Upconversion nanoparticles (UCNPs), NaYF4:Yb,Tm,Gd (NYFG) and NaYF4:Yb,Tm (NYF), were engineered on C3N4 nanotubes (C3N4 NTs) separately to construct heterojunction structures. With a UCNP loading content of 15 wt%, the NYFG/C3N4 NT heterojunction exhibits the highest H2 generation rate of 311.6 μmol g-1 with an apparent quantum efficiency of 0.80 ‰, about 1.4 times higher than that of the NYF/C3N4 NT nanocomposite under 980 nm laser irradiation. Comprehensive characterization reveals that the enhanced photocatalytic performance of the Gd doped nanostructure is attributed to the synergistic effect, stronger interaction, higher emission intensities, and faster charge transfer between the UCNPs and C3N4 NTs. Moreover, the steady-state and dynamic fluorescence spectra indicate that the energy from NYFG NPs was transferred to C3N4 NTs via a fluorescence-resonance energy-transfer process. Our work demonstrates the potential of developing near-infrared-responsive photocatalysts for energy and environmental applications.


Serially measured circulating microRNAs and adverse clinical outcomes in patients with acute heart failure.
Previous studies have identified candidate circulating microRNAs (circmiRs) as biomarkers for heart failure (HF) using relatively insensitive arrays, validated in small cohorts. The present study used RNA sequencing to identify novel candidate circmiRs and compared these with previously identified circmiRs in a large, prospective cohort of patients with acute HF (AHF). RNA sequencing of plasma from instrumented pigs was used to identify circmiRs produced by myocardium. Production of known myomiRs and microRNA (miR)-1306-5p was identified. The prognostic values of this and 11 other circmiRs were tested in a prospective cohort of 496 AHF patients, from whom blood samples were collected at up to seven time-points during the study's 1-year follow-up. The primary endpoint was the composite of all-cause mortality and HF rehospitalization. In the prospective AHF cohort, 188 patients reached the primary endpoint, and higher values of repeatedly measured miR-1306-5p were positively associated with risk for reaching the primary endpoint at the same time-point [hazard ratio (HR) 4.69, 95% confidence interval (CI) 2.18-10.06], independent of clinical characteristics and NT-proBNP. Baseline miR-1306-5p did not improve model discrimination/reclassification significantly compared with NT-proBNP. For miR-320a, miR-378a-3p, miR-423-5p and miR-1254, associations with the primary endpoint were present after adjustment for age and sex (HR 1.38, 95% CI 1.12-1.70; HR 1.35, 95% CI 1.04-1.74; HR 1.45, 95% CI 1.10-1.92; HR 1.22, 95% CI 1.00-1.50, respectively). Rates of detection of myomiRs miR-208a-3p and miR-499a-5p were very low. Repeatedly measured miR-1306-5p was positively associated with adverse clinical outcome in AHF, even after multivariable adjustment including NT-proBNP. However, baseline miR-1306-5p did not add significant discriminatory value to NT-proBNP. Low-abundance, heart-enriched myomiRs are often undetectable, which mandates the development of more sensitive assays.


FXXLF and WXXLF sequences mediate the NH2-terminal interaction with the ligand binding domain of the androgen receptor.
The nuclear receptor superfamily members of eukaryotic transcriptional regulators contain a highly conserved activation function 2 (AF2) in the hormone binding carboxyl-terminal domain and, for some, an additional activation function 1 in the NH(2)-terminal region which is not conserved. Recent biochemical and crystallographic studies revealed the molecular basis of AF2 is hormone-dependent recruitment of LXXLL motif-containing coactivators, including the p160 family, to a hydrophobic cleft in the ligand binding domain. Our previous studies demonstrated that AF2 in the androgen receptor (AR) binds only weakly to LXXLL motif-containing coactivators and instead mediates an androgen-dependent interaction with the AR NH(2)-terminal domain required for its physiological function. Here we demonstrate in a mammalian two-hybrid assay, glutathione S-transferase fusion protein binding studies, and functional assays that two predicted alpha-helical regions that are similar, but functionally distinct from the p160 coactivator interaction sequence, mediate the androgen-dependent, NH(2)- and carboxyl-terminal interaction. FXXLF in the AR NH(2)-terminal domain with the sequence (23)FQNLF(27) mediates interaction with AF2 and is the predominant androgen-dependent interaction site. This FXXLF sequence and a second NH(2)-terminal WXXLF sequence (433)WHTLF(437) interact with different regions of the ligand binding domain to stabilize the hormone-receptor complex and may compete with AF2 recruitment of LXXLL motif-containing coactivators. The results suggest a unique mechanism for AR-mediated transcriptional activation.


Cigarette Smoking Among US Adults With Selected Chronic Diseases Associated With Smoking, 2010-2019.
People who smoke cigarettes are at greater risk of developing chronic diseases and related complications. Our study provides recent estimates and trends in cigarette smoking among people with respiratory and cardiovascular diseases, cancers, and diabetes. Using data from the 2019 National Health Interview Survey, we calculated the prevalence of current and former cigarette smoking among adults aged 18 to 44 years, 45 to 64 years, and 65 years or older with chronic diseases. Those diseases were cancers associated with smoking, chronic obstructive pulmonary disease, diabetes, coronary heart disease, and/or stroke (N = 3,741). Using data from the 2010-2019 National Health Interview Surveys, we assessed trends in current cigarette smoking by chronic disease by using the National Cancer Institute's Joinpoint Regression Program. In 2019, current cigarette smoking prevalence among adults with chronic diseases associated with smoking ranged from 6.0% among adults aged 65 or older with diabetes to 51.9% among adults aged 18 to 44 years with 2 or more chronic diseases. During 2010 through 2019, a significant decrease occurred in current cigarette smoking among adults aged 45 to 64 years with diabetes. Overall, smoking prevalence remains high and relatively unchanged among people with chronic diseases associated with smoking, even as the overall prevalence of cigarette smoking in the US continues to decrease. The lack of progress in smoking cessation among adults with chronic diseases associated with smoking suggests that access, promotion, and integration of cessation treatment across the continuum of health care (ie, oncology, pulmonology, and cardiology settings) may be important in the success of smoking cessation in this population.


Direct and indirect retinal input into degenerated dorsal lateral geniculate nucleus after striate cortical removal in monkey: implications for residual vision.
We removed the striate cortex of one cerebral hemisphere in a macaque monkey, causing almost total retrograde degeneration of the corresponding dorsal lateral geniculate nucleus (dLGN) and extensive transneuronal degeneration of ganglion cells in the corresponding hemi-retina of each eye. The rare surviving geniculate projection neurons were retrogradely labelled by horseradish peroxidase (HRP) from extra-striate cortex and retinogeniculate terminals were labelled by an intraocular injection of HRP. Retinal terminals in the degenerated dLGN made synaptic contact exclusively with the dendrites of interneurons immunopositive for gamma-aminobutyric acid (GABA) in both parvocellular and magnocellular regions of dLGN. As well as being post-synaptic to retinal terminals these vesicle-containing dendrites were pre- and postsynaptic to other similar dendrites, and presynaptic to relay cells. Surviving labelled projection neurons received retinal input indirectly, via both the GABA-immunopositive interneurons and GABA-immunonegative terminals characteristic of those from the superior colliculus. In the degenerated, as opposed to the normal dLGN, about 20% of retinal terminals were GABA-immunopositive and GABA-immunoreactivity was prominently elevated in the ganglion and amacrine cell layers of the degenerated half of the retina. The optic nerve also contained numerous GABA-immunopositive axons but very few such axons were found in a normal optic nerve processed in identical manner. The surviving pathways from the retina must underlie the visual abilities that survive striate cortical removal in monkeys and human patients and may involve the degenerated dLGN as well as the mid-brain.


Multi-system disorder syndromes associated with cystinuria type I.
Cystinuria type I is an autosomal recessive disorder with an exclusively renal phenotype caused by inactivating mutations in SLC3A1. Recently 3 similar but distinct syndromes associated with cystinuria type I have been described: 2p21 deletion syndrome, Hypotonia-Cystinuria Syndrome (HCS) and atypical HCS. Genetic analysis indicated that these are recessive contiguous gene deletion syndromes which differ in the number of genes affected. Patients with HCS are missing both alleles of SLC3A1 and PREPL. In atypical HCS an additional gene (C2orf34) is deleted, and finally, in the 2p21 deletion syndrome the open reading frame of PPM1B is also disrupted. With the exception of SLC3A1, the gene products have not been fully characterized. The severity of the different syndromes reflects the number of genes which are deleted. HCS, a relatively mild syndrome, is characterised by cystinuria type I, generalised hypotonia at birth, growth retardation and minor facial dysmorphic features. On the other end of the spectrum is the 2p21 deletion syndrome, a severe syndrome with a number of additional features including a moderate to severe psychomotor retardation and a decrease in activity of the respiratory chain complexes I, III, IV and V. Finally, atypical HCS displays an intermediate phenotype comparable with classical HCS but associated with mild to moderate mental retardation and a decrease in activity of only the respiratory chain complex IV. This review will focus on the phenotypic similarities and differences observed in these syndromes. Furthermore, we speculate on the function of the gene products, based on the available data.


Effect of age on the pharmacokinetics of oral levodopa in patients with Parkinson's disease.
The influence of age on the kinetics of a standard oral dose of levodopa administered with an inhibitor of peripheral dopa decarboxylase enzymes (benserazide) has been evaluated in 40 patients with Parkinson's disease (age 34-78 y) on chronic therapy. They were divided into 2 groups, on the basis of age below (21 patients, Group A) or above (19 patients, Group B) 65 y. The area under the plasma concentration-time curve (AUC) of levodopa was significantly greater in the older group (547 versus 428 mumol.l-1.min in Group B), coupled with a reduced apparent oral clearance (8.1 versus 10.7 ml.min-1.kg-1) and a longer plasma elimination half-life (67.6 versus 54.6 min). The age of the patients was positively correlated with the AUC of levodopa (r = 0.474) and its plasma elimination half-life (r = 0.391), and was negatively correlated with clearance (r = -0.489). The findings confirm previous data on volunteers that showed a reduction in the systemic clearance of levodopa due to age, which would probably account for the finding of a greater AUC of levodopa in older patients. The observed, age-mediated differences in levodopa pharmacokinetics, albeit statistically significant, were moderate and were likely to be of only minor importance for the dosing schedule.


Reliability and Accuracy of a Standardized Shallow Water Running Test to Determine Cardiorespiratory Fitness.
A standardized fitness assessment is critical for the development of an individualized exercise prescription. Although the benefits of aquatic exercise have been well established, there remains the need for a standardized nonswimming protocol to accurately assess cardiorespiratory fitness (CRF) in shallow water. The present investigation was designed to assess (a) the reliability of a standardized shallow water run (SWR) test of CRF and (b) the accuracy of a standardized SWR compared with a land-based treadmill (LTM) test. Twenty-three healthy women (20 ± 3 years), with body mass index (23.5 ± 3 kg·m), performed 2 shallow water peak oxygen consumption (V[Combining Dot Above]O2peak) running tests (SWRa and SWRb), and 1 V[Combining Dot Above]O2max LTM. Intraclass correlation coefficients indicated moderately strong reliability for V[Combining Dot Above]O2peak (ml·kg·min) (r = 0.73, p < 0.01), HRpeak (b·min) (r = 0.82; p < 0.01), and O2pulse (V[Combining Dot Above]O2 [ml·kg·min]·HR [b·min]) (r = 0.77, p < 0.01). Using paired t-tests and Pearson's correlations, SWR V[Combining Dot Above]O2peak and HRpeak were significantly lower than during LTM (p ≤ 0.05) and showed moderate correlations of 0.60 and 0.58 (p < 0.001) to LTM. O2pulse was similar (p > 0.05) for the SWR and LTM tests with a moderate correlation of 0.63. A standardized SWR test as a measure of CRF is a reliable, and to some degree, valid alternative to conventional protocols and may be used by strength and conditioning professionals to measure program outcomes and monitor training progress. Furthermore, this protocol provides a water-based option for CRF assessment among healthy women and offers insight toward the development of an effective protocol that can accommodate individuals with limited mobility, or those seeking less musculoskeletal impact from traditional land-based types of training.


[A complete atrioventricular block during exertion].
Exercise-induced atrioventricular (AV) block in patients with normal electrocardiogram at rest is uncommon. We report the clinical features of two patients with AV block during treadmill test. The first patient was a woman of 54 years of age with presyncope on exercise. She developed complete AV block during exercise testing without evidence of ischemic myocardial disease. Electrophysiologic study documented distal AV block. The second patient was a man 31 years old who developed angina and third-degree AV block with depression of ST segment during treadmill test. Myocardial perfusion study suggested ischemic heart disease. He received anti-ischemic drugs with improvement of symptoms. A control exercise testing demonstrated normal AV conduction and electrophysiologic study was normal. In the first case, exercise AV block was probably due to abnormal His Purkinje conduction system refractoriness to autonomic modulation, while in second case AV block was secondary to ischemic heart disease. Third-degree AV block at exercise can be present in patients without conduction system abnormalities at rest. Exercise-induced infra-Hisian AV block must be treated with pacing until ischemic heart disease has been ruled out.


[Paraneoplastic syndromes in bronchogenic carcinomas].
During 5 years the authors followed up 35 patients with bronchogenic carcinomas. Of these, 34.3% had diverse paraneoplastic manifestations. The paraneoplastic reactions determined by ectopic production of different hormones and other substances as well as so-called "masks" of the systemic diseases are reviewed. An attempt is made to specify the basic mechanisms of the pathogenesis of each paraneoplastic sign.


Assessing the likely effectiveness of screening older people for impaired vision in primary care.
Screening for visual impairment is frequently included in multiphasic screening assessments for older people, although evidence for the effectiveness of screening from randomized trials is lacking. This paper uses previously developed criteria for assessing the likely effectiveness of community screening programmes to review the non-trial evidence around visual screening. Unreported or undiagnosed visual impairment is common among older people and is associated with considerable morbidity. Testing for visual acuity is easy and quick, but may not accurately reflect the level of functional disability caused by the visual problem in everyday living. Effective therapeutic interventions exist for most symptomatic patients, but the effects of treating unreported visual impairment detected by screening have not been evaluated. Existing barriers to effective treatment for older people with symptomatic visual problems include financial costs to the patient, and an inability of ophthalmic services to meet demand. These same factors may be barriers to the uptake of treatment following screening. Further work is needed to assess the needs of older people with unreported visual problems, and to clarify barriers to effective screening.


Visualizing Movement Control Optimization Landscapes.
A large body of animation research focuses on optimization of movement control, either as action sequences or policy parameters. However, as closed-form expressions of the objective functions are often not available, our understanding of the optimization problems is limited. Building on recent work on analyzing neural network training, we contribute novel visualizations of high-dimensional control optimization landscapes; this yields insights into why control optimization is hard and why common practices like early termination and spline-based action parameterizations make optimization easier. For example, our experiments show how trajectory optimization can become increasingly ill-conditioned with longer trajectories, but parameterizing control as partial target states-e.g., target angles converted to torques using a PD-controller-can act as an efficient preconditioner. Both our visualizations and quantitative empirical data also indicate that neural network policy optimization scales better than trajectory optimization for long planning horizons. Our work advances the understanding of movement optimization and our visualizations should also provide value in educational use.


Charged stacks of dithiin, diselenin, thianthrene and selenanthrene radical cations: long range multicenter bonds.
The bonding mechanism of charged stacks of thianthrene and selenanthrene radical cations is studied using quantum-chemical methods. The investigation of the nature of the electronic ground state and the electronic structure via gas-phase multireference calculations brings insight into the interactions of such dimers and trimers as found in molecular crystals. Since thianthrene and selenanthrene are the dibenzo-homologues of dithiin and diselenin, the latter are taken as model systems to study the influence of the ring systems. For all investigated systems, the singlet state is the ground state. Multicenter bonds are formed between the singular occupied orbitals of the radicals. All dimers are found to be metastable. The thianthrene trimers, which are experimentally found in molecular crystals are also stable in the gas phase, while the analogue selenanthrene trimers are not.


The mechanisms and physiological relevance of glycocalyx degradation in hepatic ischemia/reperfusion injury.
Hepatic ischemia/reperfusion (I/R) injury is an inevitable side effect of major liver surgery that can culminate in liver failure. The bulk of I/R-induced liver injury results from an overproduction of reactive oxygen and nitrogen species (ROS/RNS), which inflict both parenchymal and microcirculatory damage. A structure that is particularly prone to oxidative attack and modification is the glycocalyx (GCX), a meshwork of proteoglycans and glycosaminoglycans (GAGs) that covers the lumenal endothelial surface and safeguards microvascular homeostasis. ROS/RNS-mediated degradation of the GCX may exacerbate I/R injury by, for example, inducing vasoconstriction, facilitating leukocyte adherence, and directly activating innate immune cells. Preliminary experiments revealed that hepatic sinusoids contain a functional GCX that is damaged during murine hepatic I/R and major liver surgery in patients. There are three ROS that mediate GCX degradation: hydroxyl radicals, carbonate radical anions, and hypochlorous acid (HOCl). HOCl converts GAGs in the GCX to GAG chloramides that become site-specific targets for oxidizing and reducing species and are more efficiently fragmented than the parent molecules. In addition to ROS/RNS, the GAG-degrading enzyme heparanase acts at the endothelial surface to shed the GCX. The GCX seems to be degraded during major liver surgery, but the underlying cause remains ill-defined. The relative contribution of the different ROS and RNS intermediates to GCX degradation in vivo, the immunogenic potential of the shed GCX fragments, and the role of heparanase in liver I/R injury all warrant further investigation.


Integrated genome-wide methylation and expression analyses reveal functional predictors of response to antidepressants.
Major depressive disorder (MDD) is primarily treated with antidepressants, yet many patients fail to respond adequately, and identifying antidepressant response biomarkers is thus of clinical significance. Some hypothesis-driven investigations of epigenetic markers for treatment response have been previously made, but genome-wide approaches remain unexplored. Healthy participants (n = 112) and MDD patients (n = 211) between 18-60 years old were recruited for an 8-week trial of escitalopram treatment. Responders and non-responders were identified using differential Montgomery-Åsberg Depression Rating Scale scores before and after treatment. Genome-wide DNA methylation and gene expression analyses were assessed using the Infinium MethylationEPIC Beadchip and HumanHT-12 v4 Expression Beadchip, respectively, on pre-treatment peripheral blood DNA and RNA samples. Differentially methylated positions (DMPs) located in regions of differentially expressed genes between responders (n = 82) and non-responders (n = 95) were identified, and technically validated using a targeted sequencing approach. Three DMPs located in the genes CHN2 (cg23687322, p = 0.00043 and cg06926818, p = 0.0014) and JAK2 (cg08339825, p = 0.00021) were the most significantly associated with mRNA expression changes and subsequently validated. Replication was then conducted with non-responders (n = 76) and responders (n = 71) in an external cohort that underwent a similar antidepressant trial. One CHN2 site (cg06926818; p = 0.03) was successfully replicated. Our findings indicate that differential methylation at CpG sites upstream of the CHN2 and JAK2 TSS regions are possible peripheral predictors of antidepressant treatment response. Future studies can provide further insight on robustness of our candidate biomarkers, and greater characterization of functional components.


Antenatal diagnosis of congenital diaphragmatic hernia.
The mortality rate in cases of antenatally diagnosed congenital diaphragmatic hernia (CDH) remains high: 80% to 100% mortality was reported in early series and 73% in a more recent series. In addition, the early reports described 100% mortality for cases diagnosed before 24 weeks gestational age. To determine the mortality rate in patients at their institution (a specialized tertiary-care hospital), the authors reviewed 11 cases of CDH diagnosed from 1987 to 1990, for all of which antenatal ultrasonography (US) had been performed at the hospital. Overall, 7 of the 11 patients survived. In seven of the cases the condition had been diagnosed antenatally. Of these patients, four survived, including two of the three in whom the diagnosis had been made before 21 weeks gestational age. All four false-negative cases involved a right-side hernia, and in three of these the prenatal US examinations had been performed before 16 weeks gestational age. Three of these four patients survived. This study suggests that the prognosis in antenatally diagnosed CDH is improving and confirms that this condition is not uniformly fatal in cases diagnosed before 24 weeks gestational age.


Phase 1/dose expansion trial of brentuximab vedotin and lenalidomide in relapsed or refractory diffuse large B-cell lymphoma.
New therapies are needed for patients with relapsed/refractory (rel/ref) diffuse large B-cell lymphoma (DLBCL) who do not benefit from or are ineligible for stem cell transplant and chimeric antigen receptor therapy. The CD30-targeted, antibody-drug conjugate brentuximab vedotin (BV) and the immunomodulator lenalidomide (Len) have demonstrated promising activity as single agents in this population. We report the results of a phase 1/dose expansion trial evaluating the combination of BV/Len in rel/ref DLBCL. Thirty-seven patients received BV every 21 days, with Len administered continuously for a maximum of 16 cycles. The maximum tolerated dose of the combination was 1.2 mg/kg BV with 20 mg/d Len. BV/Len was well tolerated with a toxicity profile consistent with their use as single agents. Most patients required granulocyte colony-stimulating factor support because of neutropenia. The overall response rate was 57% (95% CI, 39.6-72.5), complete response rate, 35% (95% CI, 20.7-52.6); median duration of response, 13.1 months; median progression-free survival, 10.2 months (95% CI, 5.5-13.7); and median overall survival, 14.3 months (95% CI, 10.2-35.6). Response rates were highest in patients with CD30+ DLBCL (73%), but they did not differ according to cell of origin (P = .96). NK cell expansion and phenotypic changes in CD8+ T-cell subsets in nonresponders were identified by mass cytometry. BV/Len represents a potential treatment option for patients with rel/ref DLBCL. This combination is being further explored in a phase 3 study (registered on https://clinicaltrials.org as NCT04404283). This trial was registered on https://clinicaltrials.gov as NCT02086604.


[Dermoid cyst of the parotid gland].
Dermoid cyst of the parotid gland is a rare clinical entity. Definitive clinical diagnosis is often difficult to determine preoperatively because of the lack of pathognomonic features. The most frequent location of a parotid dermoid cyst is a triangular area lying above the pinna. Imaging studies do not definitely diagnose a parotid dermoid cyst. Although parotid dermoid cyst is (generally) well-encapsulated, complete removal of the cyst wall is not sufficient to cure it, so it is mandatory to perform careful excision of the cyst by parotidectomy, in terms of preserving facial nerve integrity. Histopathology of the parotid gland tumor removed by parotidectomy makes a diagnosis of certainty, by revealing a cyst wall with keratinization of the squamous epithelium and the presence of skin annexes (hair follicles, sweat glands, sebaceous glands). We present a rare case of parotid dermoid cyst in a 21-year-old male patient with symptoms and imaging rather suggestive of arch I branchial cyst and a brief review of data in the medical literature of the last 20 years.


How are multifactorial beliefs about the role of genetics and behavior in cancer causation associated with cancer risk cognitions and emotions in the US population?
People who believe that cancer has both genetic and behavioral risk factors have more accurate mental models of cancer causation and may be more likely to engage in cancer screening behaviors than people who do not hold such multifactorial causal beliefs. This research explored possible health cognitions and emotions that might produce such differences. Using nationally representative cross-sectional data from the US Health Information National Trends Survey (N = 2719), we examined whether endorsing a multifactorial model of cancer causation was associated with perceptions of risk and other cancer-related cognitions and affect. Data were analyzed using linear regression with jackknife variance estimation and procedures to account for the complex survey design and weightings. Bivariate and multivariable analyses indicated that people who endorsed multifactorial beliefs about cancer had higher absolute risk perceptions, lower pessimism about cancer prevention, and higher worry about harm from environmental toxins that could be ingested or that emanate from consumer products (Ps < .05). Bivariate analyses indicated that multifactorial beliefs were also associated with higher feelings of risk, but multivariable analyses suggested that this effect was accounted for by the negative affect associated with reporting a family history of cancer. Multifactorial beliefs were not associated with believing that everything causes cancer or that there are too many cancer recommendations to follow (Ps > .05). Holding multifactorial causal beliefs about cancer are associated with a constellation of risk perceptions, health cognitions, and affect that may motivate cancer prevention and detection behavior.


Metformin-attenuated sepsis-induced oxidative damages: a novel role for metformin.
Sepsis can result in severe organ injury by provoking inflammatory cascades and oxidative stress. Several studies are currently underway to find a drug with anti-inflammatory effects to prevent mortality and morbidity during sepsis. The present study was undertaken to assess the effects of metformin on oxidative stress and antioxidant status in sepsis induced by the Cecal Ligation and Puncture (CLP) method. Male Wistar rats were divided into 4 groups (n=10): sham, CLP, and 50 and 100 mg/kg metformin-treated CLP groups. After 12 hr, blood samples were collected and lung tissue was removed for histopathological study to detect tissue damage and degree of inflammation based on neutrophil infiltration and assay of the oxidative stress biomarkers superoxide dismutase (SOD), total antioxidant capacity (TAC), malondialdehyde (MDA), myeloperoxidase (MPO), glutathione peroxidase (GPx), and plasminogen activator inhibitor-1 (PAI-1). The MPO activity and MDA level were decreased in the metformin-treated groups (P<0.05). Moreover, the groups receiving metformin showed lower inflammation scores than the CLP group (P<0.05). No significant differences in SOD, GPx, or PAI in the different groups were observed. The TAC level was reduced in the CLP group compared to the sham group (P<0.05), and interestingly, this value was reduced even further in the metformin-treated groups (P<0.05 compared with the CLP group). It was concluded that metformin protects lung tissue against sepsis-induced oxidative damage, and this protective effect may be more related to its anti-inflammatory and reduced neutrophil accumulation and less to its anti-oxidative properties.


The international cooperative Gaucher group (ICCG) Gaucher registry.
Gaucher disease GD), is a rare lysosomal storage disorder caused by deficient acid β-glucosylceramidase activity and accumulation of glucosylceramide in tissue macrophages. With the 1991 advent of alglucerase enzyme replenishment therapy (ERT), the manufacturer (Genzyme Corporation) created the ICGG Gaucher Registry to collect longitudinal observational "real word" information about GD world-wide in heterogeneous patient populations, to annotate phenotypes and genotypes that define the natural history of GD in untreated patients, and to document and analyze treatment outcomes for alglucerase and any other future treatments. For 32 years, the ICGG Gaucher Registry has functioned as an educational tool for patients, clinicians, and other stakeholders to increase scientific knowledge of GD, to provide practical management guidance, and to positively impact patient care. This paper illustrates how an industry sponsored registry guided by a company independent scientific advisory board has successfully addressed its mission and evolved in step with technologic and scientific advances.


Abilities of the mCP Agar method and CRENAME alpha toxin-specific real-time PCR assay to detect Clostridium perfringens spores in drinking water.
We first determined the analytical specificity and ubiquity (i.e., the ability to detect all or most strains) of a Clostridium perfringens-specific real-time PCR (rtPCR) assay based on the cpa gene (cpa rtPCR) by using a bacterial strain panel composed of C. perfringens and non-C. perfringens Clostridium strains. All non-C. perfringens Clostridium strains tested negative, whereas all C. perfringens strains tested positive with the cpa rtPCR, for an analytical specificity and ubiquity of 100%. The cpa rtPCR assay was then used to confirm the identity of 116 putative C. perfringens isolates recovered after filtration of water samples and culture on mCP agar. Colonies presenting discordant results between the phenotype on mCP agar and cpa rtPCR were identified by sequencing the 16S rRNA and cpa genes. Four mCP(-)/rtPCR(+) colonies were identified as C. perfringens, whereas 3 mCP(+)/rtPCR(-) colonies were identified as non-C. perfringens. The cpa rtPCR was negative with all 51 non-C. perfringens strains and positive with 64 of 65 C. perfringens strains. Finally, we compared mCP agar and a CRENAME (concentration and recovery of microbial particles, extraction of nucleic acids, and molecular enrichment) procedure plus cpa rtPCR (CRENAME + cpa rtPCR) for their abilities to detect C. perfringens spores in drinking water. CRENAME + cpa rtPCR detected as few as one C. perfringens CFU per 100 ml of drinking water sample in less than 5 h, whereas mCP agar took at least 25 h to deliver results. CRENAME + cpa rtPCR also allows the simultaneous and sensitive detection of Escherichia coli and C. perfringens from the same potable water sample. In itself, it could be used to assess the public health risk posed by drinking water potentially contaminated with pathogens more resistant to disinfection.


CXCL16 induces angiogenesis in autocrine signaling pathway involving hypoxia-inducible factor 1α in human umbilical vein endothelial cells.
Chemokine (C-X-C motif) ligand 16 (CXCL16) is a new angiogenic factor inducing angiogenesis via extracellular signal-regulated kinases pathway. To further understand the molecular mechanism underlying CXCL16‑induced angiogenesis, we explored involvement of other relevant pathways in CXCL16-induced angiogenesis. In the present study, we investigated the mechanisms underlying CXCL16-induced angiogenesis in human umbilical vein endothelial cells (HUVECs). CXCL16 promoted HUVEC proliferation, tube formation and migration. Enzyme-linked immunosorbent assay revealed that CXCL16 induced vascular endothelial growth factor secretion from HUVECs. Western blot analysis showed that CXCL16 increased the level of hypoxia‑inducible factor 1α, p-extracellular signal-regulated kinases (ERK), p-p38 and p-Akt dose- and time-dependently. ERK-, p38- and Akt-selective inhibitors significantly suppressed HUVEC proliferation, migration, tube formation and hypoxia-inducible factor 1α (HIF-1α) expression induced by CXCL16. Furthermore, CXCL16 peptides induced CXCL16 secretion via ERK, p38 and Akt pathways, which was suppressed by HIF-1α-selective inhibitor PX12. Our data suggest that CXCL16 induces angiogenesis in autocrine manner via ERK, Akt, p38 pathways and HIF-1α modulation.


Topical anesthesia for laser hair removal: comparison of spot sizes and 755 nm versus 800 nm wavelengths.
The Candela alexandrite and the Coherent diode laser systems come equipped with built-in skin cooling systems that are designed to both protect the epidermis, allowing higher fluences, and to alleviate discomfort. Nevertheless, pain can be a significant problem especially with treatment of larger areas. Scatter of the laser beam is reportedly affected by the spot size. It is claimed that larger spot sizes are more effective at identical fluences. This study evaluated the effectiveness of a topical 5% lidocaine cream (ELA-Max) to control pain and compared pain levels at identical fluences between the 8 mm and 12 mm spot size of the alexandrite laser and between the alexandrite and diode laser with its 9 mm spot size. The study was conducted in the axillae of 12 patients. Each axilla was divided in half for side-by-side comparison. Half of the right axilla was treated with the alexandrite 8 mm and the other half with the 12 mm spot size at identical fluences. The left axilla was treated at the maximum tolerated fluences with the alexandrite 12 mm spot size and the diode 9 mm spot size. At identical fluences and other parameters, there was significantly more pain with the alexandrite 12 mm spot size than with the alexandrite 8 mm spot size, both with or without topical anesthesia. The alexandrite laser was significantly less painful than the diode laser both with and without topical anesthetic. The difference was most noticeable between the alexandrite 8 mm spot size and the diode 9 mm spot size. Topical 5% lidocaine anesthesia was effective in reducing pain, though not completely, for both the alexandrite and the diode lasers. Topical 5% lidocaine cream is a simple and effective method for reducing patient discomfort during laser hair removal procedures, even when a skin cooling device is being used. A larger spot size causes more pain than a smaller spot size at identical fluences. The 800 nm diode laser causes greater discomfort than the 755 nm alexandrite laser.


Laparoscopic division of a portosystemic shunt to treat chronic hepatic encephalopathy.
In the event of liver cirrhosis with severe portal hypertension, voluminous portosystemic shunt may lead to refractory encephalopathy. Obliteration of the shunt has been described as a satisfactory therapeutic solution, and reported procedures are mainly endovascular embolization and surgical shunt ligation through laparotomy. The former procedure is less invasive and seems to be as efficient. Laparoscopy, which is widely recognized to minimize mortality and morbidity in cirrhotic patients undergoing surgery, has never been used for such a procedure. Shunt division can therefore be considered using this modern approach to good effect and reduced morbidity. In support of this view, we report a case of severe chronic encephalopathy cured by laparoscopic surgical division of a large shunt after failure of the percutaneous technique.


The Challenges of Participant Photography: A Critical Reflection on Methodology and Ethics in Two Cultural Contexts.
Photovoice and photo-elicitation are two common methods of participant photography used in health research. Although participatory photography has many benefits, this critical reflection provides fellow researchers with insights into the methodological and ethical challenges faced when using such methods. In this article, we critically reflect on two studies that used participatory photography in different cultural contexts. The first study used photo-elicitation to investigate mothers' experiences of infant settling in central Vietnam. The second study used photovoice to explore pregnant embodiment in Australia. Following a discussion of the literature and a detailed overview of the two studies, we examine the methodological challenges in using participant photography before, during and after each study. This is followed by a discussion of ethical concerns that arose in relation to the burden of participation, confidentiality, consent, and the photographing of families and children. To conclude, we highlight implications for using participatory photography in other settings.


Is the association between irritable bowel syndrome and abuse explained by neuroticism? A population based study.
In outpatients and the community, an association between abuse (particularly sexual abuse) and irritable bowel syndrome (IBS) has been observed, but whether there is a causal link continues to be disputed. To test the hypothesis that psychological factors explain the apparent association between abuse and IBS. A sample of residents of Penrith (a Sydney suburb sociodemographically similar to the Australian population) selected randomly from the electoral rolls (that by law include the entire population of age 18 years and above) was mailed a validated self report questionnaire. Measured were gastrointestinal (GI) symptoms including the Rome criteria for IBS, abuse (including the standardised Drossman questions), neuroticism (Eysenck Personality Questionnaire), and psychological morbidity (General Health Questionnaire). The response rate was 64% (n = 730); 12% fulfilled the Rome criteria for IBS. Overall abuse in childhood (odds ratio (OR) = 2.02, 95% confidence interval (CI) 1.29 to 3.15) but not adulthood (OR = 1.39, 95% CI 0.88 to 2.19) was associated with IBS univariately. Neuroticism and psychological morbidity were also univariately associated with abuse in childhood, abuse in adulthood and IBS, respectively. However, by logistic regression, abuse in childhood was not associated with IBS after controlling for age, gender, and psychological factors (OR = 1.34, 95% CI 0.83 to 2.17). The results were not altered by restricting the analyses to more severe forms of abuse, and were not explained by interactions between abuse and psychological variables. There is an association between abuse and IBS in the community, but this may be explained in part by other psychological factors. Based on a path analysis, we postulate that abuse may induce the expression of neuroticism that in turn leads to IBS.


Site-selected incorporation of 5-carboxymethylaminomethyl(-2-thio)uridine into RNA sequences by phosphoramidite chemistry.
5-Carboxymethylaminomethyluridine (cmnm(5)U) and 5-carboxymethylaminomethyl-2-thiouridine (cmnm(5)s(2)U) are located at the wobble position in several cytosolic and mitochondrial tRNA sequences. In this paper, we report the first site-selected incorporation of cmnm(5)U and cmnm(5)s(2)U into RNA sequences by phosphoramidite chemistry on a CPG solid support. Trifluoroacetyl and 2-(trimethylsilyl)ethyl were selected for the protection of the amine and carboxyl functions, respectively.


Investigating the healing mechanisms of an angiogenesis-promoting topical treatment for diabetic wounds using multimodal microscopy.
Impaired skin wound healing is a significant comorbid condition of diabetes that is caused by poor microcirculation, among other factors. Studies have shown that angiogenesis, a critical step in the wound healing process in diabetic wounds, can be promoted under hypoxia. In this study, an angiogenesis-promoting topical treatment for diabetic wounds, which promotes angiogenesis by mimicking a hypoxic environment via inhibition of prolyl hydroxylase resulting in elevation or maintenance of hypoxia-inducible factor, was investigated utilizing a custom-built multimodal microscopy system equipped with phase-variance optical coherence tomography (PV-OCT) and fluorescence lifetime imaging microscopy (FLIM). PV-OCT was used to track the regeneration of the microvasculature network, and FLIM was used to assess the in vivo metabolic response of mouse epidermal keratinocytes to the treatment during healing. Results show a significant decrease in the fluorescence lifetime of intracellular reduced nicotinamide adenine dinucleotide, suggesting a hypoxic-like environment in the wounded skin, followed by a quantitative increase in blood vessel density assessed by PV-OCT. Insights gained in these studies could lead to new endpoints for evaluation of the efficacy and healing mechanisms of wound-healing drugs in a setting where delayed healing does not permit available methods for evaluation to take place.


Early physical rehabilitation in intensive care patients with sepsis syndromes: a pilot randomised controlled trial.
Survivors of sepsis syndromes have poor outcomes for physical and cognitive function. No investigations of early physical rehabilitation in the intensive care unit have specifically targeted patients with sepsis syndromes. To determine whether early physical rehabilitation improves physical function and associated outcomes in patients with sepsis syndromes. Fifty critically ill adults admitted to a general intensive care unit with sepsis syndromes were recruited into a prospective double-blinded randomised controlled trial investigating early physical rehabilitation. Primary outcomes of physical function (acute care index of function) and self-reported health-related quality of life were recorded at ICU discharge and 6 months post-hospital discharge, respectively. Secondary measures included inflammatory biomarkers; Interleukin-6, Interleukin-10 and tumour necrosis factor-α, blood lactate, fat-free muscle mass, exercise capacity, muscle strength and anxiety. A significant increase in patient self-reported physical function (81.8 ± 22.2 vs. 60.0 ± 29.4), p = 0.04) and physical role (61.4 ± 43.8 vs. 17.1 ± 34.4, p = 0.005) for the SF-36 at 6 months was found in the exercise group. Physical function scores were not significantly different between groups. Muscle strength scores were (51.9 ± 10.5 vs. 47.3 ± 13.6, p = 0.24) with the standard care mean Medical Research Council Muscle Score (MRC) <48/60. The mean change of Interleukin-10 increased and was significantly higher in the exercise group (1.8 pg/ml, 180 % vs. 0.9 pg/ml, 90 %, p = 0.04). There was no significant difference between groups for lactate, Interleukin-6, tumour necrosis factor-α, muscle strength, exercise capacity, fat-free mass or hospital anxiety. Implementation of early physical rehabilitation can improve self-reported physical function and induce systemic anti-inflammatory effects.


Transition radiation from graphene plasmons by a bunch beam in the terahertz regime.
The terahertz band is an increasingly important spectrum in a wide range of applications from bioimaging and medical diagnostics to security and wireless communications. We propose a tunable terahertz coherent radiation source based on graphene plasmon-induced transition radiation. The transition radiation in terahertz regime arises from the graphene plasmons, which are excited by a normally incident bunched electron beam. We analyze the field-intensities and spectral-angular distributions of the transition radiation with respect to Fermi energy, substrate dielectric permittivity, and electron bunch energy for both the coherent and incoherent radiation. The effect of electron bunching on the radiation pattern is discussed. The mechanism of plasmon frequency-selective transition radiation is discovered.


Histopathological and biochemical changes in lung tissues of rats following administration of fluoride over several generations.
The possible effects of multigenerational administration of sodium fluoride (NaF) via drinking water on lung tissue morphology and biochemistry and body and lung weight were investigated in second-generation adult male rats. For this purpose we selected 45 Albino adult Wistar rats in nine cages, each of which consisted of four females and one male. Twenty-eight pregnant rats were selected for the experiment, divided into four groups of seven rats given 1 (control group), 10, 50 and 100 mg l(-1) NaF in drinking water during the gestation period. After gestation the rats had 165 pups in total. The mothers received fluoridated water during the lactation period and the offspring of the first generation had access to fluoridated water during the suckling period (21 days) and after the weaning period (30 days) until they became mature and at the start of the second part of the experiment. During this time 23 pups died and 79 female and 63 male first-generation rats survived. These first-generation rats were then used to obtain the second-generation offspring in the same manner as before, which were subjected to the same treatments. At the end of 6 months the rats were sacrificed and autopsied. Serum fluoride levels and the activities of principal antioxidant enzymes were determined in lung tissue samples taken from all groups. In addition, the lung tissues were submitted for histopathological examination. Histological findings showed alveolar congestion, alveolar cell hyperplasia and necrosis, prominent alveolar septal vessels, epithelial desquamation and macrophages in the alveolar spaces in the experimental groups. Additionally, there were inflammatory infiltrations in peribronchial, perivascular, intraparenchymal and respiratory tract lumen; intraparenchymal hyperaemic vessels; respiratory epithelial desquamation and proliferation; intraparenchymal thick walled vessels; parenchymal fibrosis; bronchiolitis; pneumonic and focal emphysematous areas. Furthermore, the lung parenchyma was observed to have a distorted appearance with loss of alveolar architecture. These histopathological findings were more pronounced for the rat groups of 50 and 100 mg l(-1) fluoride. No significant histopathological changes were observed in the rats of the control group. The increased activities of superoxide dismutase (SOD) and reduced glutathione peroxidase (GSH-Px) and the decreased activity of catalase (CAT) in the lung tissues with 10 mg l(-1) fluoride might indicate activation of the antioxidant defence mechanism. The decrease in SOD, GSH-Px and CAT activities with 50 and 100 mg l(-1) fluoride and the increase in thiobarbituric acid-reactive substance levels might be related to oxidative damage that occurred in the lung. This multigenerational evaluation of the long-term effect of different doses of fluoride intake through drinking water on lung damage shows that the lung tissues were damaged, there was emphysema and inflammation of lung parenchyma associated with loss of alveolar architecture and the degree of lung damage seemed to correlate with the increased dosage of fluoride. A similar relationship was observed between the degree of lung damage, body and lung weight and serum fluoride levels according to the fluoride dose. Therefore, these results contribute to a better understanding of chronic fluoride toxicity in lung tissue of second-generation rats, especially via drinking water, and the biochemical findings were in agreement with histological observations. In addition, increased fluoride concentration did not affect reproduction or the number of pups dying but the body weight and lung weight ratios were affected by the high dose of fluoride in a dose-related pattern.


Induction of donor-type chimerism and transplantation tolerance across major histocompatibility barriers in sublethally irradiated mice by Sca-1(+)Lin(-) bone marrow progenitor cells: synergism with non-alloreactive (host x donor)F(1) T cells.
Induction of transplantation tolerance by means of bone marrow (BM) transplantation could become a reality if it was possible to achieve engraftment of hematopoietic stem cells under nonlethal preparatory cytoreduction of the recipient. To that end, BM facilitating cells, veto cells, or other tolerance-inducing cells, have been extensively studied. In the present study, we show that BM cells within the Sca-1(+)Lin(-) cell fraction, previously shown to be enriched for early hematopoietic progenitors, are capable of reducing specifically antidonor CTL-p frequency in vitro and in vivo, and of inducing split chimerism in sublethally 7-Gy-irradiated recipient mice across major histocompatibility complex barriers. The immune tolerance induced by the Sca-1(+)Lin(-) cells was also associated with specific tolerance toward donor-type skin grafts. The minimal number of cells required to overcome the host immunity remaining after 7 Gy total body irradiation is very large and, therefore, it may be very difficult to harvest sufficient cells for patients. This challenge was further addressed in our study by demonstrating that non-alloreactive (host x donor)F(1) T cells, previously shown to enhance T-cell-depleted BM allografts in lethally irradiated mice, synergize with Sca-1(+)Lin(-) cells in their capacity to overcome the major transplantation barrier presented by the sublethal mouse model.


Histone H1 structural changes and its interaction with DNA in the presence of high glucose concentration in vivo and in vitro.
Histone H1 has an important role in the packing of chromatin and controlling gene expression. The effect of nonenzymatic glycation on the structure of histone H1 (a basic protein), the inhibitory role of spermine on this phenomenon and interaction of H1 with DNA is investigated here. H1 was extracted from the liver of normal and diabetic rats with or without receiving spermine as a chemical chaperone. Rat liver H1 was also incubated with glucose (50 mM) and its structure was compared with the protein obtained from diabetic rat. The results indicated a lower fluorescence emission and alpha-helical content of glycated H1; i.e., alteration in its folding; and reduced DNA (high molecular weight or specific sequences) binding in comparison with normal protein. Spermine partially (not completely) returns the studied parameters on glycated H1 toward the normal values. The changes in the structure and function of histone H1 is suggested as one of the possible mechanisms involved in diabetic complications.


Effect of separate sampling on classification accuracy.
Measurements are commonly taken from two phenotypes to build a classifier, where the number of data points from each class is predetermined, not random. In this 'separate sampling' scenario, the data cannot be used to estimate the class prior probabilities. Moreover, predetermined class sizes can severely degrade classifier performance, even for large samples. We employ simulations using both synthetic and real data to show the detrimental effect of separate sampling on a variety of classification rules. We establish propositions related to the effect on the expected classifier error owing to a sampling ratio different from the population class ratio. From these we derive a sample-based minimax sampling ratio and provide an algorithm for approximating it from the data. We also extend to arbitrary distributions the classical population-based Anderson linear discriminant analysis minimax sampling ratio derived from the discriminant form of the Bayes classifier. All the codes for synthetic data and real data examples are written in MATLAB. A function called mmratio, whose output is an approximation of the minimax sampling ratio of a given dataset, is also written in MATLAB. All the codes are available at: http://gsp.tamu.edu/Publications/supplementary/shahrokh13b.


HDAC inhibition facilitates the switch between memory systems in young but not aged mice.
Chromatin modifications, especially histone acetylation, are critically involved in gene regulation required for long-term memory processes. Increasing histone acetylation via administration of histone deacetylase inhibitors before or after a learning experience enhances memory consolidation for hippocampus-dependent tasks and rescues age-related memory impairments. Whether acutely and locally enhancing histone acetylation during early consolidation processes can operate as a switch between multiple memory systems is less clear. This study examined the short- and long-term behavioral consequences of acute intra-CA1 administration of the histone deacetylase inhibitor Trichostatin A (TSA) on cue versus place learning strategy selection after a cue-guided water maze task and competition testing performed 1 or 24 h later in mice. Here, we show that intra-CA1 TSA infusion administrated immediately post-training biased young mice away from striatum-dependent cue strategy toward hippocampus-dependent place strategy under training condition that normally promotes cue strategy in vehicle controls. However, concomitant infusions of TSA with either PKA inhibitor, Rp-cAMPS, into CA1 or cAMP analog, 8Br-cAMP, into dorsal striatum failed to bias young mice to place strategy use. Behavioral and immunohistochemical analyses further indicated that post-training TSA infusion in aged mice rescued aging-associated deregulation of H4 acetylation in the CA1 but failed to reverse phosphorylated CREB deficits and to produce strategy bias on the 24 h probe test. These findings suggest that post-training intra-CA1 TSA infusion promotes dynamic shift from striatum toward the hippocampal system in young but not aged animals, and support the possibility of a role for CREB in the TSA-mediated switch between these two memory systems.


Exploring the impact of wheelchair design on user function in a rural South African setting.
Wheelchairs provide mobility that can enhance function and community integration. Function in a wheelchair is influenced by wheelchair design. To explore the impact of wheelchair design on user function and the variables that guided wheelchair prescription in the study setting. A mixed-method, descriptive design using convenience sampling was implemented. Quantitative data were collected from 30 wheelchair users using the functioning every day with a Wheelchair Scale and a Wheelchair Specification Checklist. Qualitative data were collected from ten therapists who prescribed wheelchairs to these users, through interviews. The Kruskal-Wallis test was used to identify relationships, and content analysis was undertaken to identify emerging themes in qualitative data. Wheelchairs with urban designs were issued to 25 (83%) participants. Wheelchair size, fit, support and functional features created challenges concerning transport, operating the wheelchair, performing personal tasks, and indoor and outdoor mobility. Users using wheelchairs designed for use in semi-rural environments achieved significantly better scores regarding the appropriateness of the prescribed wheelchair than those using wheelchairs designed for urban use (p = <0.01). Therapists prescribed the basic, four-wheel folding frame design most often because of a lack of funding, lack of assessment, lack of skills and user choice. Issuing urban type wheelchairs to users living in rural settings might have a negative effect on users' functional outcomes. Comprehensive assessments, further training and research, on long term cost and quality of life implications, regarding provision of a suitable wheelchair versus a cheaper less suitable option is recommended.


Small molecule-mediated reprogramming of Xenopus blastula stem cells to a neural crest state.
Neural crest cells are a stem cell population unique to vertebrates that give rise to a diverse array of derivatives, including much of the peripheral nervous system, pigment cells, cartilage, mesenchyme, and bone. Acquisition of these cells drove the evolution of vertebrates and defects in their development underlies a broad set of neurocristopathies. Moreover, studies of neural crest can inform differentiation protocols for pluripotent stem cells and regenerative medicine applications. Xenopus embryos are an important system for studies of the neural crest and have provided numerous insights into the signals and transcription factors that control the formation and later lineage diversification of these stem cells. Pluripotent animal pole explants are a particularly powerful tool in this system as they can be cultured in simple salt solution and instructed to give rise to any cell type including the neural crest. Here we report a protocol for small molecule-mediated induction of the neural crest state from blastula stem cells and validate it using transcriptome analysis and grafting experiments. This is an powerful new tool for generating this important cell type that will facilitate future studies of neural crest development and mutations and variants linked to neurocristopathies.


Effect of Electroacupuncture Intervention on Expression of CGRP, SP, COX-1, and PGE2 of Dorsal Portion of the Cervical Spinal Cord in Rats with Neck-Incision Pain.
The present study was aimed to determine if cervicospinal substance P (SP) and its neurokinin-1 receptor (NK-1R), calcitonin gene-related peptide (CGRP), cyclooxygenase-1 (COX-1), and prostaglandin E2 (PGE2) were involved in electroacupuncture (EA) analgesia in neck-incision pain rats. EA intervention was applied to bilateral Futu (LI18), Hegu (LI4)-Neiguan (PC6), and Zusanli (ST36)-Yanglingquan (GB34) for 30 min. Cervicospinal SP and CGRP immunoactivity was detected by immunofluorescence technique, NK-1R and COX-1 protein and mRNA expression levels were determined using Western blot and real-time PCR, respectively, and PGE2 content was measured using ELISA. Outcomes indicated that EA of EA-LI18 and LI4-PC6 (not ST36-GB34) significantly suppressed neck-incision induced decrease of thermal pain threshold (P < 0.05). EA stimulation of LI18 and LI4-PC6 markedly inhibited neck-incision induced upregulation of SP and CGRP immunoactivity, NK-1 R and COX-1 mRNA and protein expression levels, as well as the increase of PGE2 content in the dorsal cervicospinal cord (P < 0.05). These findings showed that LI18 and LI4-PC6 EA stimulation-induced downregulation of SP, CGRP, NK-1R, COX-1, and PGE2 levels in the dorsal cervicospinal cord may contribute to their effects in relieving neck-incision pain. This study highlights the targets of EA intervention for reducing post-thyroid-surgery pain for the first time.


Dravet syndrome: seizure control and gait in adults with different SCN1A mutations.
Dravet syndrome (DS) is an aggressive epileptic encephalopathy. Pharmacoresistant seizures of several types plague most patients with DS throughout their lives. Gait difficulties are a common, but inconsistent finding. The majority of cases are caused by mutations in the SCN1A gene, but little information is available about how particular mutations influence the adult phenotype. The purpose of this study is to correlate different types of SCN1A mutations and (1) seizure control, (2) occurrence of convulsive status epilepticus (cSE), and (3) the presence of crouch gait in adult patients. In a cohort of 10 adult patients with DS caused by SCN1A mutations, we investigated seizure frequency, history of cSE, and gait. All patients were identified in the epilepsy clinic between 2009 and 2011. SCN1A mutations were divided into four different groups based on location or effect of the mutation. Retrospective chart review and recent physical examination were completed in all cases. All patients had a pathogenic mutation in the SCN1A gene. Four SCN1A mutations have not been described previously. Greater than 90% seizure reduction was observed (compared to childhood frequency) in six of seven patients with missense mutations in the pore-forming region (PFR) of the Na(v) 1.1 protein (group A) and nonsense mutations (group B). One patient with a splice-site mutation (group C) and another with a mutation outside the PFR (group D) became free of all types of seizures. cSE after the age of 19 years was observed in only one patient. Crouch gait, without spasticity, is identified as an element of the adult DS phenotype. However, only one half of our adult DS cohort demonstrated crouch gait. This feature was observed in five of seven patients from groups A and B. This study shows that seizure control improves and cSE become less frequent in DS as patients age, independent of their SCN1A mutation type. Complete seizure freedom was seen in two patients (groups C and D). Finally, this study shows that in DS, crouch gait can be observed in up to 50% of adults with SCN1A mutation. Although no definite statistical correlations could be made due to the small number of patients, it is interesting to note that crouch gait was observed only in those patients with nonsense mutations or mutations in the PFR. Future studies with larger cohorts will be required to formally assess an association of gait abnormalities with particular SCN1A mutations.


Calcification of superficial scalp veins secondary to intravenous infusion of sodium bicarbonate and calcium chloride.
Soft tissue calcification has been reported following the intramuscular injection of calcium salts and the intravenous administration of calcium gluconate. The present case report describes a neonate in whom calcification of the superficial scalp veins developed following administration of calcium chloride and sodium bicarbonate through a scalp vein needle.


Remarkably high thermal-driven MoS2 grain boundary migration mobility and its implications on defect healing.
Two-dimensional (2D) transition-metal dichalcogenides (TMDs) hold great potential for many important device applications, such as field effect transistors and sensors, which require a robust control of defect type, density, and distribution. However, how to control the defect type, density, and distribution in these materials is still a challenge. In this study, we explore the kinetics and dynamics of four types of grain boundaries (GBs) in monolayer MoS2, which are composed of S-polar dislocation (S5|7), Mo-polar dislocation (Mo5|7), dislocation-double S vacancy complex (S4|6), and dislocation-double S interstitial complex (S6|8), respectively. Our study shows that these four GBs in monolayer MoS2 exhibit a great disparity in their migration behavior. More specifically, the S4|6 and S6|8 GBs possess a much higher migration mobility than the S5|7 and Mo5|7 GBs under the same thermal fluctuations or temperature gradient. Interestingly, the S4|6 and S6|8 GBs follow an abnormal relationship with temperature, due to the change in defect configurations with temperature. Our study further shows that the remarkably high mobilities of the S4|6 and Mo6|8 GBs may enable the reactions of GBs, leading to the annihilation and reduction of defect density. In addition, the movement of GBs in MoS2 under a temperature gradient field can cause defect redistribution, which in turn changes the thermal conductivity. The present study not only deepens our understanding of the dynamic evolution of GBs in TMDs, but also presents new opportunities to engineer GBs for novel electronic applications.


Identification of oda6 as a Chlamydomonas dynein mutant by rescue with the wild-type gene.
We find that two Chlamydomonas outer arm dynein assembly loci, oda6 and oda9, are located on the left arm of linkage group XII, in the vicinity of the previously mapped locus for a 70,000 Mr dynein intermediate chain protein. Restriction fragment length polymorphism mapping indicates that this dynein gene is very closely linked to the oda6 locus. A cDNA clone encoding the 70,000 Mr protein was isolated, sequenced, and used to select genomic clones spanning the corresponding locus from both wild-type and oda6 libraries. When wild-type clones were introduced into cells containing an oda6 allele, the mutant phenotype was rescued, while no rescue was observed after transformation with oda6 clones. Genetic analysis further revealed that newly introduced gene copies were responsible for the rescued phenotype and thus confirms that ODA6 encodes the 70,000 Mr dynein intermediate chain protein. The inability of oda6 mutants to assemble any major outer arm dynein subunits shows that this protein is essential for assembly of stable outer dynein arms. This is the first use of transformation with a wild-type gene to identify the product of a Chlamydomonas mutant.


Sharing organs with foreign nationals.
Organs for transplantation are an absolute scarcity throughout the world, and many countries do not offer transplantation. Developed countries with transplant programs receive requests to list foreign nationals for transplantation. Any national standard deserves justification by a thorough exploration of the issues. In this article, the issues regarding organ transplantation for foreign nationals in Canada are explored. Currently Canada has no policy on listing foreign nationals for transplantation. Three topics are reviewed: (1) arguments for and against the transplantation of organs from deceased donors to foreign nationals, (2) relevant legislation and position statements, and (3) relevant practices in other countries. Finally, practical policy options are suggested. This article's analysis of the issues will provide guidance for health care professionals and policy makers in Canada and developed countries exploring listing foreign nationals for transplantation.


Rosiglitazone induces interleukin-1 receptor antagonist in interleukin-1beta-stimulated rat synovial fibroblasts via a peroxisome proliferator-activated receptor beta/delta-dependent mechanism.
To study the potency of 2 peroxisome proliferator-activated receptor gamma (PPARgamma) agonists, 15-deoxy-Delta(12,14)-prostaglandin J(2) (15-deoxy-PGJ(2)) and rosiglitazone, to modulate the expression of interleukin-1 receptor antagonist (IL-1Ra) in rat synovial fibroblasts. Levels of messenger RNA for IL-1Ra and PPAR isotypes (alpha, beta/delta, gamma) were assessed by real-time polymerase chain reaction in rat synovial fibroblasts exposed to 10 ng/ml of IL-1beta. PPAR levels were assessed by Western blotting and secreted IL-1Ra levels by immunoassay. The potency of PPARgamma agonists and the PPARbeta/delta agonist GW-501516 on IL-1Ra levels was tested in the range of 1-10 microM and at 100 pM, respectively. The contribution of PPARgamma to the effects of rosiglitazone on IL-1Ra secretion was examined either by its overexpression or by inhibition using wild-type or dominant-negative constructs and the antagonist GW-9662 (10 microM), respectively. The dominant-negative strategy was also performed to investigate the possible contribution of PPARbeta/delta and NF-kappaB activation. IL-1beta-induced IL-1Ra production was increased by 10 microM rosiglitazone but was reduced dose-dependently by 15-deoxy-PGJ(2). Both agonists lowered IL-1beta secretion, but rosiglitazone alone reduced the imbalance of IL-1beta/IL-1Ra toward basal levels. Enhancement of IL-1beta-induced IL-1Ra production by rosiglitazone was not affected by PPARgamma overexpression or by its inhibition with dominant-negative PPARgamma or GW-9662. Inhibition of NF-kappaB was also ineffective against rosiglitazone but abolished the stimulating effect of IL-1beta on IL-1Ra. All PPAR isotypes were expressed constitutively in rat synoviocytes, but PPARgamma decreased dramatically upon IL-1beta exposure, whereas PPARbeta/delta remained stable. Dominant-negative PPARbeta/delta abolished the enhancement of IL-1Ra by rosiglitazone, whereas GW-501516 reproduced the effect of rosiglitazone on IL-1Ra secretion. Rosiglitazone stimulates IL-1Ra production by a PPARbeta/delta mechanism in activated rat synovial fibroblasts, further contributing to its potential antiarthritic properties and opening new perspectives for the modulation of inflammatory genes by specific PPAR agonists in articular cells.


Targeting immune privilege to prevent pathogenic neovascularization.
PURPOSE. Current studies suggest that the immune system plays a critical role in blinding eye disorders. The eye is an immune-privileged site, and FasL expression is a major part of that mechanism because Fas/FasL interactions regulate inflammation and neovascularization, preventing damage to delicate ocular structures. These studies were undertaken to test the idea that modulating immune privilege might be an effective therapeutic approach to pathogenic angiogenesis in the eye. METHODS. C57BL/6 mice or FasL-defective B6-gld mice were laser treated to induce choroidal neovascularization (CNV). Mice were injected with cytotoxic FasL in the vitreous cavity or were treated with oral doxycycline in the drinking water. They were evaluated for CNV 7 days later. In some experiments eye tissue was harvested and evaluated for FasL expression, macrophage influx by immunohistochemistry, and release of sFasL. RESULTS. Injection of cytotoxic FasL successfully prevented neovascularization in a mouse model of CNV. Oral doxycycline increased functional FasL in the eye and substantially inhibited neovascularization. Doxycycline treatment increased FasL expression on the RPE cells and reduced circulating and tissue-associated sFasL. Treatment was ineffective in B6-gld mice, demonstrating that CNV inhibition was mediated by FasL. CONCLUSIONS. Targeting immune privilege using cytotoxic molecules or by increasing expression of the proapoptotic protein FasL may be a viable approach to treating neovascular eye disease.


Association of the yellow leaf (y10) mutant to soybean chromosome 3.
At least 19 single recessive gene yellow leaf mutants and one duplicate recessive gene mutant have been described in soybean. This study was conducted to associate a yellow leaf mutant, y10, with a specific soybean chromosome by using primary trisomics (2n = 41). Seven soybean primary trisomics were hybridized as female parent with genetic stock strain, T161, carrying y10. F(1) disomic and primary trisomic plants were identified cytologically. One disomic (control) and all primary trisomic plants were allowed to self-pollinate and F(2) populations were classified for green versus yellow leaf mutant. The F(2) population of Triplo 3 segregated in a 17:1 ratio, while a disomic (3:1) ratio was observed with Triplo 8-, 17-, 18-, and 20-derived F(2) populations, suggesting that the y10 locus is on chromosome 3. The y10 locus was examined with four simple sequence repeat (SSR) markers (Satt584, Sat_033, Satt387, and Satt022) from molecular linkage group (MLG) N and y10 was found linked with Satt022. Therefore we confirmed the association of MLG N with chromosome 3. The possible association of y10 with Triplo 16 and Triplo 19 are discussed.


LC-ESI-TOF MS method for the evaluation of the immunostimulating activity of soybeans via the determination of the functional peptide soymetide.
Bioactive peptides content in foodstuffs can seriously vary with many factors such as crop variety, food processing, animal breeding, etc. Because of this variability, quantitative methodologies are required to evaluate the content of bioactive peptides in foodstuffs. Progress in liquid chromatography and mass spectrometry technologies offer a great opportunity for the quantitation of bioactive peptides. This study undertook the development of a liquid chromatography-electrospray ionization-time-of-flight mass spectrometry method using a fused-core technology column for the sensitive and unambiguous determination of the immunostimulating peptide soymetide in soybean varieties. Soymetide precursor protein (α' subunit of β-conglycinin) was extracted with a Tris-HCl buffer (pH 8.0) containing urea and digested with trypsin. Soymetide separation conditions by reversed phase liquid chromatography (ion-pairing reagent, organic modifier, separation column, and elution gradient) and detection by MS were optimized, and a study of soymetide stability was also conducted. The method selectivity having been demonstrated, the linearity, accuracy, precision, and limits of detection and quantitation were evaluated. The developed method enabled the detection and quantitation of soymetide concentrations in the ppb range (7.5 ng/mL and 25 ng/mL, respectively), and about 30 times lower than those detected and determined in a previous work by capillary liquid chromatography with UV detection. These values could allow the quantitation of only 17 μg of soymetide per gram of soybean. The developed methodology was applied to the quantitation of soymetide in different soybean varieties from Europe, Japan, and USA observing great differences in soymetide content that ranged from 40 to 600 μg per gram of soybean depending on the soybean variety.


Seasonal variation in plasma levels of lithium in the Indian population: is there a need to modify the dose?
Lithium still remains an important choice in the therapy of manic-depressive psychosis (MDP), and though there are reports of seasonal variation in lithium levels from a few countries, such studies have not been conducted in India. Variability in the lithium level can lead to lack of efficacy or toxicity, making seasonal variation clinically relevant. A retrospective case sheet audit was performed for 101 MDP patients for recording plasma lithium level, oral lithium dose, age and gender for one year. The overall average oral lithium dose and level were recorded; the monthly average to which it most closely matched was noted as the control month, and values of other months were compared with this control month by Friedman's test followed by Dunn's test. The mean age of patients was 38.22 (standard deviation 12.07) years, and 72 out of 101 patients were male. The mean lithium dose in November (938.61 +/- 243.40 mg/day), which was the closest to the overall mean dose (938.24 +/- 241.78 mg/day) was taken as the control month, which when compared with other monthly values, did not show any significant difference. The June (0.54 +/- 0.23 meq/L), July (0.55 +/- 0.24 meq/L) and August (0.55 +/- 0.24 meq/L) mean plasma lithium values were significantly high when compared to the October value (0.45 +/- 0.22 meq/L) as control. High-low variability between the plasma lithium values of different months was found to be 25 percent. The present study showed a significant high variability of lithium levels in different months of the year, therefore frequent plasma level monitoring and oral lithium dose adjustment to prevent situations of toxicity and lack of efficacy in MDP.