Probiotics reduce risk-taking behavior in the Elevated Plus Maze in the Flinders Sensitive Line rat model of depression.
Several studies have reported beneficial effects of probiotics on behavior and stress in humans and animals. The multi-strain probiotic product Ecologic® Barrier has shown antidepressant-like effects in rats when given in combination with high-fat diet. To circumvent the need for high-fat diet for future clinical use, the present study aimed to potentiate the behavioral effects of Ecologic® Barrier by developing a customized product that includes four additional bacterial species known to be associated with depression. Flinders Sensitive Line (FSL) rats, an animal model of depression, were randomized to receive vehicle, Ecologic® Barrier, or the customized product Ecologic® Barrier + 4. The control strain, Flinders Resistant Line rats, received vehicle to ascertain the depressive-like phenotype of FSL rats. Behavior was tested in the Elevated Plus Maze, Y-Maze, Novel Object Recognition Test, Open Field, and Forced Swim Test; HPA-axis reactivity was assessed by the Dexamethasone Suppression Test. Our results indicate that both probiotic products reduced open arm exploration in the Elevated Plus Maze, suggesting a reduction in risk-taking behavior. There were no intervention effects on cognition, locomotion, or depressive-like behavior. Moreover, all intervention groups had similar plasma corticosterone concentrations following the Dexamethasone Suppression Test. There was no difference between the two probiotic products, suggesting that the addition of four bacterial strains to Ecologic® Barrier did not potentiate its effect. Future studies should further investigate the effects of probiotics on risk-taking behavior and gain more mechanistic insights by studying individual strains and their cumulative contributions to the overall effect of a multi-strain product.


Detection of actin-binding proteins in human platelets by 125I-actin overlay of polyacrylamide gels.
Actin-binding proteins have been identified in human platelets with a gel-overlay technique that uses 125I-G-actin. Platelet proteins were separated on SDS polyacrylamide gels using the buffer system of Laemmli (1970, Nature [Lond.] 227:680-685). The proteins were fixed in the gels with methanol-acetic acid, the SDS was washed out, and the proteins were renatured. The gels were incubated with 125I-G-actin from rabbit skeletal muscle that was radiolabeled with 125I according to the method of Bolton and Hunter (1973, Biochem. J. 133:529-538) and has been shown to retain biological activity. After nonspecifically bound radioactivity was washed out, gels were dried and processed for autoradiography. The 125I-G-actin binds to several proteins in human platelets, platelet extracts, and the particulate fraction. Control experiments demonstrate that the 125I-G-actin can be displaced by use of increasing amounts of unlabeled actin, that the binding is stable to 0.6 M NaCl, and that preheating the 125I-G-actin to 90 degrees C for 3 min eliminates all binding. Prominent 125I-G-actin-binding activities were present at Mr 90,000 and 40,000. The binding to the 90,000 Mr protein appears to be at least partially Ca++ sensitive, whereas the binding to the 40,000 Mr protein does not. 125I-G-actin bound to proteins in the SDS gels can be fixed in situ and compared directly with the stained gel. This technique should prove generally useful in identification and purification of some actin-binding proteins from cells and tissues.


HLA-B44 allele frequencies and haplotypic associations in three European populations.
HLA-B44 is among the most frequent class I antigens in many populations studied so far. It has been subdivided into seven allelic forms that can only be discriminated by DNA typing. Using a simple PCR/sequence-specific oligonucleotide hybridization procedure, we have analysed the frequency distribution of B44 subtypes in three European populations from Slovenia, the Netherlands, and Switzerland. B*4402 and B*4403 were by far the predominant alleles, B*4404 and 4405 were rare, while B*4406 and B*4407 were not observed. Interestingly, B*4402 and 4403 occurred with different frequencies in the three populations, with B*4402 being most frequent in the Swiss (65% vs. 57% in the Dutch, and 46.5% in the Slovenes). Of the 139 individuals studied, 60 HLA-B44 ABDR haplotypes could be determined by family studies. In the respective populations, the linkage disequilibria between B44 and other HLA antigens occurred with different frequencies. A2-B*4402 haplotypes were very frequent in the Swiss sample, mostly associated with DRB1*0101, 0401 and 1301. B*4402 was more often linked with non-A2 antigens in the Slovenes (predominantly A24, A28) than in the Swiss and the Dutch. The predominant association of B*4403 was with DR7: this haplotype was very frequent in the Swiss (82% of the B*4403 haplotypes), while lower frequencies were found in the Dutch (72%) and Slovenian (59%) populations. In the Swiss population, more than half of the B44-DR7 haplotypes were A23-B*4403-DR7 (53% of all B*4403 haplotypes). This haplotype was significantly less frequent in the Slovenian (6%) and in the Dutch (14%) populations. The second most frequent B*4403 haplotype in both the Swiss and Slovenes is the A29-B*4403-CW*1601-DR7 haplotype (17.6 and 29.4%, respectively). Concomitant with the increased frequency of B*4403 in the Slovenes, a higher diversity of non-DR7 B44 haplotypes was observed in this population (41% of all B*4403 haplotypes). HLA-B44 oligotyping analysis allowed us to detect B44-subtype incompatibilities in several AB-sero, DRB1/B3/B5-oligo matched unrelated bone marrow donor/patient combinations. The different frequency distributions of HLA-B44 haplotypes in the three populations analysed in this study argue in favour of local volunteer bone marrow donor recruitment. This might significantly improve the chance of finding a highly matched donor for patients with less frequent A-B-DRB1 haplotypes.


Identification of novel putative rheumatoid arthritis susceptibility genes via analysis of rare variants.
Established loci for rheumatoid arthritis (RA), including HLA-DRB1 and PTPN22, do not fully account for the genetic component of susceptibility to the disease. One possible source of as yet undiscovered susceptibility genes are those mediated through effects of rare variants. We present a novel method for gene-based genome-wide scans of whole-genome association (WGA) data to identify accumulations of rare variants associated with disease. We apply our method to WGA SNP genotype data obtained from 868 RA cases and 1194 controls. Our results highlight novel putative RA susceptibility genes that have not previously been identified in large-scale WGA studies.


Biology of feline leukemia virus in the natural environment.
The feline leukemia virus (FeLV) was discovered in 1964 in a cluster of cats with lymphosarcoma. The observed clustering of cases of feline lymphosarcoma suggested that FeLV was an infectious agent for cats. The development of a simple immunofluorescent test for FeLV permitted a seroepidemiological study to be undertaken on the distribution of the virus in cats living in their natural environment. Over 2000 cats were tested, and the results showed conclusively that FeLV is an infectious agent for cats. This finding has now been independently confirmed using three different test procedures. After the infectious nature of FeLV was discovered, a simple FeLV test and removal program was devised to control the spread of the virus in the natural environment. The spread of FeLV was controlled in 45 households by removing the FeLV-infected cats, while in 25 households, where the infected cats were left in contact with the uninfected cats, 12% of the uninfected cats became infected. The ultimate control of FeLV awaits the development of an effective FeLV vaccine, which now seems feasible since we have already experimentally immunized some cats with attenuated FeLV. Although FeLV is infectious for cats there is no evidence that FeLV can infect humans.


An egg-hatch assay for resistance to levamisole in trichostrongyloid nematode parasites.
An in vitro technique is described for detecting resistance of nematodes to the anthelmintic levamisole hydrochloride. Samples of eggs are developed under controlled temperature conditions until just prior to the commencement of hatching. They are then exposed to different concentrations of the drug and, when hatching is almost complete, the test samples are killed and preserved. The proportion of unhatched eggs at each drug concentration can then be counted at leisure. Provided a suitable range of drug concentrations is chosen for each test isolate, this assay provides results which may be satisfactorily fitted to a log-concentration-probit regression model. Comparisons with in vivo anthelmintic assays have shown that the technique provides an accurate reflection of the resistance status of parasite populations.


The critical cytoplasmic regions of the alphaL/beta2 integrin in Rap1-induced adhesion and migration.
Rap1 is a potent inside-out signal that increases LFA-1 adhesive activity. In this study, we have defined the cytoplasmic region of the alphaL and beta2 integrin that are required for Rap1-stimulated adhesion and subsequent migration on ICAM-1. Human LFA-1 bearing truncated and point-mutated alphaL and beta2 cytoplasmic regions were reconstituted in mouse IL-3-dependent proB cells, BAF/3. Truncation of the alphaL, but not beta2 subunit cytoplasmic region, abolished Rap1V12-dependent adhesion to ICAM-1. The alanine substitution of two lysine residues (K1097/K1099) in the alphaL subunit was found to be critical in adhesion induced by Rap1V12, but not PMA. This mutation suppressed Rap1V12-induced LFA-1 conformation changes and ligand-binding affinity. The K1097/K1099 mutation also impaired binding to ICAM-1 induced by TCR cross-linking or SDF-1. In contrast, the alanine substitution for tyrosine in the beta2 subunit endocytosis motif inhibited internalization of LFA-1, and severely impaired detachment at the cell rear, which resulted in long-elongated cell shapes. This result demonstrates that internalization of LFA-1 is a critical step in the deadhesion process. Our study revealed novel requirements of amino acid residues of the LFA-1 cytoplasmic region in the response to the inside-out signaling and the subsequent deadhesion process.


Repeat coronary revascularization after coronary artery bypass surgery in older adults: the Society of Thoracic Surgeons' national experience, 1991-2007.
A major advantage of coronary artery bypass graft surgery (CABG) relative to percutaneous coronary intervention is its durability, yet there is a paucity of information on rates and predictors of repeat coronary revascularization after CABG in the modern era. We included patients ≥65 years from the Society of Thoracic Surgeons' National Adult Cardiac Surgery Database who were undergoing first-time isolated CABG from 1991 to 2007 (n=723 134, median age 73 years). After linking to Medicare claims data, long-term outcomes of CABG (up to 18 years after surgery) were examined by use of cumulative incidence curves. Multivariable Cox proportional hazard analysis was used to identify factors associated with 1- and 5-year repeat revascularization trends and variability. We found that the overall 18-year survival rate was 20%. Cumulative incidences of any repeat revascularization (percutaneous coronary intervention or CABG, yet most often percutaneous coronary intervention) were 2%, 7%, 13%, and 16% at 1, 5, 10, and 18 years after surgery, respectively. The rates of repeat CABG procedures were quite low for all time points (0.1%, 0.6%, 1.3%, and 1.7%, respectively). Female sex, disease severity represented by a history of percutaneous coronary intervention, preoperative dialysis, and partial revascularization were strongly associated with a higher revascularization rate, whereas advanced age, left main disease, and smoking were associated with a lower rate. There was approximately a 2-fold variation in repeat revascularization rates across centers at 1 year (interquartile range 1.7-3.6%) and 5 years (interquartile range 6.7-12.0%). Repeat revascularization is performed infrequently among older patients who undergo CABG; however, these rates vary substantially by patient subgroups and among providers.


Protease-activated receptor 2 sensitizes the capsaicin receptor transient receptor potential vanilloid receptor 1 to induce hyperalgesia.
Inflammatory proteases (mast cell tryptase and trypsins) cleave protease-activated receptor 2 (PAR2) on spinal afferent neurons and cause persistent inflammation and hyperalgesia by unknown mechanisms. We determined whether transient receptor potential vanilloid receptor 1 (TRPV1), a cation channel activated by capsaicin, protons, and noxious heat, mediates PAR2-induced hyperalgesia. PAR2 was coexpressed with TRPV1 in small- to medium-diameter neurons of the dorsal root ganglia (DRG), as determined by immunofluorescence. PAR2 agonists increased intracellular [Ca2+] ([Ca2+]i) in these neurons in culture, and PAR2-responsive neurons also responded to the TRPV1 agonist capsaicin, confirming coexpression of PAR2 and TRPV1. PAR2 agonists potentiated capsaicin-induced increases in [Ca2+]i in TRPV1-transfected human embryonic kidney (HEK) cells and DRG neurons and potentiated capsaicin-induced currents in DRG neurons. Inhibitors of phospholipase C and protein kinase C (PKC) suppressed PAR2-induced sensitization of TRPV1-mediated changes in [Ca2+]i and TRPV1 currents. Activation of PAR2 or PKC induced phosphorylation of TRPV1 in HEK cells, suggesting a direct regulation of the channel. Intraplantar injection of a PAR2 agonist caused persistent thermal hyperalgesia that was prevented by antagonism or deletion of TRPV1. Coinjection of nonhyperalgesic doses of PAR2 agonist and capsaicin induced hyperalgesia that was inhibited by deletion of TRPV1 or antagonism of PKC. PAR2 activation also potentiated capsaicin-induced release of substance P and calcitonin gene-related peptide from superfused segments of the dorsal horn of the spinal cord, where they mediate hyperalgesia. We have identified a novel mechanism by which proteases that activate PAR2 sensitize TRPV1 through PKC. Antagonism of PAR2, TRPV1, or PKC may abrogate protease-induced thermal hyperalgesia.


A new value of proximal femur geometry to evaluate hip fracture risk: true moment arm.
This study was undertaken to determine the influence of proximal femur geometry on hip fracture risk independent of bone mineral density. We examined 34 hip fracture subjects (17 men, 17 women) and 36 control subjects (18 men, 18 women). The control subjects were matched with the hip fracture patients by femoral neck bone mineral density (+ or - 0.100 g/cm(2)). Hip axis length (HAL), femoral axis length (FAL), femoral neck-shaft angle (Theta angle), lateral and medial femoral cortical thickness were measured on standart pelvic radiographs. In the literature, there are conflicting views of the relationship between femur geometry and hip fracture risk which may be explained by different definitions of some parameters. We investigated the effect of a new parameter called true moment arm (TMA) on hip fracture risk. Longer TMA may be correlated to higher transmission of impact energy to the femoral neck. Thus it may be useful to define fracture prone individuals. The results of this study showed that HAL, FAL and TMA were significantly longer in the hip fracture subjects compared to the control group (p<0.001). Hip fracture patients had thinner lateral and medial femoral cortical thickness (p<0.001). Theta angle was wider in the hip fracture group than in the control group (p<0.001). In conclusion, our study showed that evaluation of TMA in addition to HAL, FAL, Theta angle, MCT and LCT can be used to determine of the fracture risk independently of BMD.


Chikungunya virus non-structural protein 2-mediated host shut-off disables the unfolded protein response.
The unfolded protein response (UPR) is a cellular defence mechanism against high concentrations of misfolded protein in the endoplasmic reticulum (ER). In the presence of misfolded proteins, ER-transmembrane proteins PERK and IRE1α become activated. PERK phosphorylates eIF2α leading to a general inhibition of cellular translation, whilst the expression of transcription factor ATF4 is upregulated. Active IRE1α splices out an intron from XBP1 mRNA, to produce a potent transcription factor. Activation of the UPR increases the production of several proteins involved in protein folding, degradation and apoptosis. Here, we demonstrated that transient expression of chikungunya virus (CHIKV) (family Togaviridae, genus Alphavirus) envelope glycoproteins induced the UPR and that CHIKV infection resulted in the phosphorylation of eIF2α and partial splicing of XBP1 mRNA. However, infection with CHIKV did not increase the expression of ATF4 and known UPR target genes (GRP78/BiP, GRP94 and CHOP). Moreover, nuclear XBP1 was not observed during CHIKV infection. Even upon stimulation with tunicamycin, the UPR was efficiently inhibited in CHIKV-infected cells. Individual expression of CHIKV non-structural proteins (nsPs) revealed that nsP2 alone was sufficient to inhibit the UPR. Mutations that rendered nsP2 unable to cause host-cell shut-off prevented nsP2-mediated inhibition of the UPR. This indicates that initial UPR induction takes place in the ER but that expression of functional UPR transcription factors and target genes is efficiently inhibited by CHIKV nsP2.


S-acylation-dependent membrane microdomain localization of the regulatory Kvβ2.1 subunit.
The voltage-dependent potassium (Kv) channel Kvβ family was the first identified group of modulators of Kv channels. Kvβ regulation of the α-subunits, in addition to their aldoketoreductase activity, has been under extensive study. However, scarce information about their specific α-subunit-independent biology is available. The expression of Kvβs is ubiquitous and, similar to Kv channels, is tightly regulated in leukocytes. Although Kvβ subunits exhibit cytosolic distribution, spatial localization, in close contact with plasma membrane Kv channels, is crucial for a proper immune response. Therefore, Kvβ2.1 is located near cell surface Kv1.3 channels within the immunological synapse during lymphocyte activation. The objective of this study was to analyze the structural elements that participate in the cellular distribution of Kvβs. It was demonstrated that Kvβ peptides, in addition to the cytoplasmic pattern, targeted the cell surface in the absence of Kv channels. Furthermore, Kvβ2.1, but not Kvβ1.1, targeted lipid raft microdomains in an S-acylation-dependent manner, which was concomitant with peptide localization within the immunological synapse. A pair of C-terminal cysteines (C301/C311) was mostly responsible for the specific palmitoylation of Kvβ2.1. Several insults altered Kvβ2.1 membrane localization. Therefore, growth factor-dependent proliferation enhanced surface targeting, whereas PKC activation impaired lipid raft expression. However, PSD95 stabilized Kvβ2.1 in these domains. This data shed light on the molecular mechanism by which Kvβ2.1 clusters into immunological synapses during leukocyte activation.


Variance Based Measure for Optimization of Parametric Realignment Algorithms.
Neuronal responses to sensory stimuli or neuronal responses related to behaviour are often extracted by averaging neuronal activity over large number of experimental trials. Such trial-averaging is carried out to reduce noise and to diminish the influence of other signals unrelated to the corresponding stimulus or behaviour. However, if the recorded neuronal responses are jittered in time with respect to the corresponding stimulus or behaviour, averaging over trials may distort the estimation of the underlying neuronal response. Temporal jitter between single trial neural responses can be partially or completely removed using realignment algorithms. Here, we present a measure, named difference of time-averaged variance (dTAV), which can be used to evaluate the performance of a realignment algorithm without knowing the internal triggers of neural responses. Using simulated data, we show that using dTAV to optimize the parameter values for an established parametric realignment algorithm improved its efficacy and, therefore, reduced the jitter of neuronal responses. By removing the jitter more effectively and, therefore, enabling more accurate estimation of neuronal responses, dTAV can improve analysis and interpretation of the neural responses.


Proteins associated with emergence of the resistance to lethal glucocorticoid actions in P1798 mouse lymphosarcoma cells.
Two-dimensional gel electrophoresis was used to detect [35S]methionine-labeled newly synthesized proteins from corticosteroid-sensitive and corticosteroid-resistant P1798 mouse lymphosarcoma cells. When the protein patterns from corticosteroid-sensitive cells are compared with those from highly resistant cells, they are very similar except for seven changes. One of the most obvious changes is the appearance of a new protein with a molecular weight of about 36,000 in highly resistant cells. There are also four other proteins that increase in the resistant state, as well as two that decrease. The control experiments indicate that protein differences are present to approximately the same degree when the sensitive tumor is carried in adrenalectomized animals, suggesting that the protein differences are not induced by endogenous glucocorticoids. Other experiments show that the protein differences are neither components of the small amounts of fibrous tissue that are associated with the cells nor components of blood. These results suggest that resistance of the tumor cells to glucocorticoid killing in vivo could be related to the relative rates of synthesis of a few proteins.


The effects of prenatal and postnatal malnutrition on the morphology, differentiation, and metabolism of skeletal striated muscle tissue in rats.
To study the contractile properties, metabolism and morphological characteristics of muscles submitted to prenatal and postnatal protein malnutrition. Animals were distributed into two groups: Control, normoprotein diet (CG; n = 15; 5/5/5), and Malnourished, hypoprotein diet (MG; n = 15; 5/5/5), and examined on the 7th, 14th, and 28th days of the experiment. Total body mass, weight, and the contractile properties and morphology of the anterior tibial muscle were assessed. Several 8 microm-thick tissue samples were taken from 7, 14, and 28 day old rats and stained with HE or subjected to NADH-TR or m-ATPase (pH = 4.4) techniques. Body and muscle weight were lower in the malnourished group. On the 7th day of malnutrition, muscle samples exhibited fibers with smaller diameter, higher polymorphism and higher endomysial conjunctive tissue content. Histochemical methods were unable to precisely determine the types of fiber present. On the 14th day, there were smaller muscle fibers, more polymorphism, many of them with central nuclei and moderate endomysial conjunctive tissue content. With reference to contractile properties, the m-ATPase reaction identified both slow and fast fibers. The NADH-TR reaction revealed the following types of fiber: slow oxidative (SO), fast oxidative glycolytic (FOG) and fast glycolytic (FG). On the 28th day smaller, bunched muscle fibers varying shapes. All three types of fiber exhibited unclear recognition limits with respect to contraction and metabolism. Our experimental results suggest that, in addition to the reduction in numbers of fibers, malnutrition retards the differentiation of the morphological, metabolic, and contractile characteristics of skeletal muscle fibers in growing rats.


Fine-root respiration in a loblolly pine (Pinus taeda L.) forest exposed to elevated CO2 and N fertilization.
Forest ecosystems release large amounts of carbon to the atmosphere from fine-root respiration (R(r)), but the control of this flux and its temperature sensitivity (Q(10)) are poorly understood. We attempted to: (1) identify the factors limiting this flux using additions of glucose and an electron transport uncoupler (carbonyl cyanide m-chlorophenylhydrazone); and (2) improve yearly estimates of R(r) by directly measuring its Q(10)in situ using temperature-controlled cuvettes buried around intact, attached roots. The proximal limits of R(r) of loblolly pine (Pinus taeda L.) trees exposed to free-air CO(2) enrichment (FACE) and N fertilization were seasonally variable; enzyme capacity limited R(r) in the winter, and a combination of substrate supply and adenylate availability limited R(r) in summer months. The limiting factors of R(r) were not affected by elevated CO(2) or N fertilization. Elevated CO(2 )increased annual stand-level R(r) by 34% whereas the combination of elevated CO(2) and N fertilization reduced R(r) by 40%. Measurements of in situ R(r) with high temporal resolution detected diel patterns that were correlated with canopy photosynthesis with a lag of 1 d or less as measured by eddy covariance, indicating a dynamic link between canopy photosynthesis and root respiration. These results suggest that R(r) is coupled to daily canopy photosynthesis and increases with carbon allocation below ground.


Optomechanical design and characterization of a printed-circuit-board-based free-space optical interconnect package.
We present a proof of concept and a feasibility demonstration of a practical packaging approach in which free-space optical interconnects (FSOI's) can be integrated simply on electronic multichip modules (MCM's) for intra-MCM-board interconnects. Our system-level packaging architecture is based on a modified folded 4f imaging system that has been implemented with only off-the-shelf optics, conventional electronic packaging, and passive-assembly techniques to yield a potentially low-cost and manufacturable packaging solution. The prototypical system as built supports 48 independent FSOI channels with 8 separate laser and detector chips, for which each chip consists of a one-dimensional array of 12 devices. All the chips are assembled on a single substrate that consists of a printed circuit board or a ceramic MCM. Optical link channel efficiencies of greater than 90% and interchannel cross talk of less than -20 dB at low frequency have been measured. The system is compact at only 10 in.3 (25.4 cm3) and is scalable, as it can easily accommodate additional chips as well as two-dimensional optoelectronic device arrays for increased interconnection density.


Ultrafast bond softening in bismuth: mapping a solid's interatomic potential with X-rays.
Intense femtosecond laser excitation can produce transient states of matter that would otherwise be inaccessible to laboratory investigation. At high excitation densities, the interatomic forces that bind solids and determine many of their properties can be substantially altered. Here, we present the detailed mapping of the carrier density-dependent interatomic potential of bismuth approaching a solid-solid phase transition. Our experiments combine stroboscopic techniques that use a high-brightness linear electron accelerator-based x-ray source with pulse-by-pulse timing reconstruction for femtosecond resolution, allowing quantitative characterization of the interatomic potential energy surface of the highly excited solid.


The Impact of a Social Marketing Campaign on HIV and Sexually Transmissible Infection Testing Among Men Who Have Sex With Men in Australia.
In response to increasing HIV and other sexually transmissible infection (HIV/STI) notifications in Australia, a social marketing campaign Drama Downunder (DDU) was launched in 2008 to promote HIV/STI testing among men who have sex with men (MSM). We analyzed prospective data from (1) an online cohort of MSM and (2) clinic-level HIV/STI testing to evaluate the impact of DDU on HIV, syphilis, gonorrhea, and chlamydia testing. (1) Cohort participants who completed 3 surveys (2010-2014) contributed to a Poisson regression model examining predictors of recent HIV testing.(2) HIV, syphilis, gonorrhea, and chlamydia tests among MSM attending high caseload primary care clinics (2007-2013) were included in an interrupted time series analysis. (1) Although campaign awareness was high among 242 MSM completing 726 prospective surveys, campaign recall was not associated with self-reported HIV testing. Reporting previous regular HIV testing (adjusted incidence rate ratio, 2.4; 95% confidence interval, 1.3-4.4) and more than 10 partners in the previous 6 months (adjusted incidence rate ratio, 1.2; 95% confidence interval, 1.1-1.4) was associated with recent HIV testing. (2) Analysis of 257,023 tests showed increasing monthly HIV, syphilis, gonorrhea, and chlamydia tests pre-DDU. Post-DDU, gonorrhea test rates increased significantly among HIV-negative MSM, with modest and nonsignificant increasing rates of HIV, syphilis, and chlamydia testing. Among HIV-positive MSM, no change in gonorrhea or chlamydia testing occurred and syphilis testing declined significantly. Increasing HIV/STI testing trends among MSM occurred pre- and post-DDU, coinciding with other plausible drivers of testing. Modest changes in HIV testing post-DDU suggest that structural changes to improve testing access may need to occur alongside health promotion to increase testing frequency.


Rapid B cell receptor-induced unfolded protein response in nonsecretory B cells correlates with pro- versus antiapoptotic cell fate.
The adaptive unfolded protein response (UPR) is essential for the development of antibody-secreting plasma cells. B cells induced by lipopolysaccharide (LPS) to differentiate into plasma cells exhibit a nonclassical UPR reported to anticipate endoplasmic reticulum stress prior to immunoglobulin production. Here we demonstrate that activation of a physiologic UPR is not limited to cells undergoing secretory cell differentiation. We identify B cell receptor (BCR) signaling as an unexpected physiologic UPR trigger and demonstrate that in mature B cells, BCR stimulation induces a short lived UPR similar to the LPS-triggered nonclassical UPR. However, unlike LPS, BCR stimulation does not induce plasma cell differentiation. Furthermore, the BCR-induced UPR is not limited to cells in which BCR induces activation, since a UPR is also induced in transitional immature B cells that respond to BCR stimulation with a rapid apoptotic fate. This response involves sustained up-regulation of Chop mRNA indicative of a terminal UPR. Whereas sustained Chop expression correlates with the ultimate fate of the BCR-triggered B cell and not its developmental stage, Chop-/- B cells undergo apoptosis, indicating that CHOP is not required for this process. These studies establish a system whereby a terminal or adaptive UPR can be alternatively triggered by physiologic stimuli.


Obscured by administrative data? Racial disparities in occupational injury.
Objectives Underreporting of occupational injuries is well documented, but underreporting patterns may vary by worker characteristics, obscuring disparities. We tested for racial and ethnic differences in injury reporting patterns by comparing injuries reported via research survey and administrative injury database in the same group of healthcare workers in the US. Methods We used data from a cohort of 1568 hospital patient-care workers who were asked via survey whether they had been injured at work during the year prior (self-reported injury; N=244). Using the hospital's injury database, we determined whether the same workers had reported injuries to the hospital's occupational health service during that year (administratively reported injury; N=126). We compared data sources to test for racial and ethnic differences in injury reporting practices. Results In logistic regression models adjusted for demographic and occupational characteristics, black workers' odds of injury as measured by self-report data were 1.91 [95% confidence interval (95% CI) 1.04-3.49] compared with white workers. The same black workers' odds of injury as measured by administrative data were 1.22 (95% CI 0.54-2.77) compared with white workers. Conclusions The undercount of occupational injuries in administrative versus self-report data may be greater among black compared to white workers, leading to underestimates of racial disparities in workplace injury.


The consequences of not accounting for background selection in demographic inference.
Recently, there has been increased awareness of the role of background selection (BGS) in both data analysis and modelling advances. However, BGS is still difficult to take into account because of tractability issues with simulations and difficulty with nonequilibrium demographic models. Often, simple rescaling adjustments of effective population size are used. However, there has been neither a proper characterization of how BGS could bias or shift inference when not properly taken into account, nor a thorough analysis of whether rescaling is a sufficient solution. Here, we carry out extensive simulations with BGS to determine biases and behaviour of demographic inference using an approximate Bayesian approach. We find that results can be positively misleading with significant bias, and describe the parameter space in which BGS models replicate observed neutral nonequilibrium expectations.


Delayed atrophy of the retinal pigment epithelium after submacular surgery.
We report a case of delayed atrophy of the retinal pigment epithelium (RPE) eighteen months after apparently successful excision of submacular choroidal new vessels (CNV) in a patient with age-related macular degeneration (AMD). Submacular surgery for CNV was achieved without visible disturbance of the underlying RPE in an 83 year old man diagnosed with AMD. At the time of surgery the CNV displayed clinical features consistent with lying internal to Bruchs membrane (Type 2 configuration). There was no visible RPE defect at the fovea and vision improved during the subsequent 12 months follow-up. Eighteen months later, however, an atrophic central RPE defect appeared, with a similar shape to the CNV originally excised. This case demonstrates that submacular CNV with Type 2 configuration can occur in AMD and lead to an initially favourable outcome following submacular surgery. Atrophy of the RPE nevertheless did eventually occur and in a pattern consistent with damage during the original operation. It is important to consider results of longer term follow-up when interpreting success rates for surgery in AMD. (Eur J Ophthalmol 2005; 15: 170-2).


Shared decision making in the UK: Moving towards wider uptake.
Shared decision making (SDM) is firmly on the policy agenda in the UK and a recent legal ruling has confirmed its importance. Policymakers, ethicists, professional regulators and societies, patient organisations and now the courts are committed to ensuring that SDM becomes the norm throughout the NHS, but an unfavourable economic climate makes this especially challenging. Considerable progress has been made over the last few years, with new learning from demonstration sites, various initiatives in capacity building and training, wider availability of patient decision aids, and important leadership initiatives. Enthusiasm for this way of working is growing among clinicians, patients and managers, but it could be undermined if SDM comes to be seen primarily as a means of cost control.


Primary structure of IgE monoclonal antibodies expressing an intrastrain crossreactive idiotype.
We have obtained amino acid sequences (by mRNA and amino acid sequencing) for two IgE kappa mAb that have specificity for the Ars hapten group and are related to the major idiotypic family, CRIA (crossreactive idiotype A), in the A strain of mouse. One mAb, SE20.2, fully expresses CRIA; the other, SE1.3, possesses some but not all of the characteristic idiotopes. Both IgE proteins contain VH and V kappa segments that are closely related to those associated with CRIA. The D segment of SE20.2 is also typical of CRIA+ mAb, but that of SE1.3 is one amino acid residue longer. Chain recombination experiments indicated that the L chain of SE1.3 is fully capable of supporting CRIA expression. Its deficiency with respect to idiotopes of CRIA was attributed to the extra amino acid in the D region and/or substitutions in the VH segment. A major objective was to ascertain the frequency of somatic mutations in IgE. For the VH segment (amino acids 1-98) of SE20.2, there are only three nucleotide differences and one uncertainty with respect to the nucleotide sequence of the germline gene associated with CRIA. A somewhat higher frequency of substitutions is present in the VH segment of SE1.3. The VK amino acid sequences of the IgE proteins are nearly identical to those of a prototype of the CRIA family, mAb R16.7. The results are discussed with reference to the mechanism of the IgM to IgE switch.


Patterns and trends of alcohol consumption in rural and urban areas of China: findings from the China Kadoorie Biobank.
In China, alcohol consumption has increased significantly in recent decades. Little evidence exists, however, about temporal trends in levels and patterns of alcohol consumption and associated factors in adult populations. In 2004-08, the China Kadoorie Biobank recruited ~ 512,000 adults (41% men, mean age 52 years [SD 10.7]) from 10 (5 urban, 5 rural) geographically diverse regions across China, with ~ 25,000 randomly selected participants resurveyed in 2013-14. The self-reported prevalence and patterns (e.g., amount, beverage type, heavy drinking episodes) of alcohol drinking at baseline and resurvey were compared and related to socio-demographic, health and other factors. At baseline, 33% of men drank alcohol at least weekly (i.e., current regular), compared to only 2% of women. In men, current regular drinking was more common in urban (38%) than in rural (29%) areas at baseline. Among men, the proportion of current regular drinkers slightly decreased at resurvey (33% baseline vs. 29% resurvey), while the proportion of ex-regular drinkers slightly increased (4% vs. 6%), particularly among older men, with more than half of ex-regular drinkers stopping for health reasons. Among current regular drinkers, the proportion engaging in heavy episodic drinking (i.e., > 60 g/session) increased (30% baseline vs. 35% resurvey) in both rural (29% vs. 33%) and urban (31% vs. 36%) areas, particularly among younger men born in the 1970s (41% vs. 47%). Alcohol intake involved primarily spirits, at both baseline and resurvey. Those engaging in heavy drinking episodes tended to have multiple other health-related risk factors (e.g., regular smoking, low fruit intake, low physical activity and hypertension). Among Chinese men, the proportion of drinkers engaging in harmful drinking behaviours increased in the past decade, particularly among younger men. Harmful drinking patterns tended to cluster with other unhealthy lifestyles and health-related risk factors.


Safety and efficacy of vagus nerve stimulation in fibromyalgia: a phase I/II proof of concept trial.
We performed an open-label Phase I/II trial to evaluate the safety and tolerability of vagus nerve stimulation (VNS) in patients with treatment-resistant fibromyalgia (FM) as well as to determine preliminary measures of efficacy in these patients. Of 14 patients implanted with the VNS stimulator, 12 patients completed the initial 3-month study of VNS; 11 patients returned for follow-up visits 5, 8, and 11 months after start of stimulation. Therapeutic efficacy was assessed with a composite measure requiring improvement in pain, overall wellness, and physical function. Loss of both pain and tenderness criteria for the diagnosis of FM was added as a secondary outcome measure because of results found at the end of 3 months of stimulation. Side effects were similar to those reported in patients treated with VNS for epilepsy or depression and, in addition, dry mouth and fatigue were reported. Two patients did not tolerate stimulation. At 3 months, five patients had attained efficacy criteria; of these, two patients no longer met widespread pain or tenderness criteria for the diagnosis of FM. The therapeutic effect seemed to increase over time in that additional participants attained both criteria at 11 months. Side effects and tolerability were similar to those found in disorders currently treated with VNS. Preliminary outcome measures suggested that VNS may be a useful adjunct treatment for FM patients resistant to conventional therapeutic management, but further research is required to better understand its actual role in the treatment of FM.


Establishing the natural history and growth rate of ameloblastoma with implications for management: systematic review and meta-analysis.
Ameloblastoma is the second most common odontogenic tumor, known to be slow-growing, persistent, and locally aggressive. Recent data suggests that ameloblastoma is best treated with wide resection and adequate margins. Following primary excision, bony reconstruction is often necessary for a functional and aesthetically satisfactory outcome, making early diagnosis paramount. Despite earlier diagnosis potentially limiting the extent of resection and reconstruction, an understanding of the growth rate and natural history of ameloblastoma has been notably lacking from the literature. A systematic review of the literature was conducted by reviewing relevant articles from PubMed and Web of Science databases. Each article's level of evidence was formally appraised according to the Centre of Evidence Based Medicine (CEBM), with data from each utilized in a meta-analysis of growth rates for ameloblastoma. Literature regarding the natural history of ameloblastoma is limited since the tumor is immediately acted upon at its initial detection, unless the patient voluntarily refuses a surgical intervention. From the limited data, it is derived that the highest estimated growth rate is associated with solid, multicystic type and the lowest rate with peripheral ameloblastomas. After meta-analysis, the calculated mean specific grow rate is 87.84% per year. The growth rate of ameloblastoma has been demonstrated, offering prognostic and management information, particularly in cases where a delay in management is envisaged.


Valuation, categories and attributes.
Existing research on categories has only examined indirectly the value associated with being a member of a category relative to the value of the set of attributes that determine membership in that category. This study uses survey data to analyze consumers' preferences for the "organic" label versus for the attributes underlying that label. We found that consumers generally preferred products with the category label to those with the attributes required for the organic label but without the label. We also found that the value accorded to the organic label increased with the number of attributes that an individual associated with the category. Category membership nevertheless still had greater value than even that of the sum of the attributes associated with it.


Erythrocyte dyscrasia, anemia, and hypothyroidism in chronically underweight llamas.
A syndrome characterized by anemia, erythrocyte dyscrasia, low body weight, and hypothyroidism was observed in 8 llamas (Lama glama). At initial examination (1 to 23 months of age; median, 7.5 months), llamas (3 males, 5 females) were markedly underweight (29 to 55 kg; median, 36 kg) and anemic (PCV, 12.9 to 25.5% [median, 19%]). Five of the llamas became progressively more anemic over time; in 2 of them, PCV decreased to less than 10%. Erythrocyte changes included severe poikilocytosis, anisocytosis, asymmetric distribution of hemoglobin within the cytoplasm, and cytoplasmic extensions from one or both poles. Six llamas had moderate to severe valgus deformities of the carpus. All llamas had low baseline serum thyroxine concentration and diminished response to thyrotropin administration. Baseline and post-thyrotropin triiodothyronine concentrations did not have consistent patterns. Five llamas were hypophosphatemic and 7 had low serum iron concentration (iron concentration was not determined in 1 llama). Orally administered iron supplementation did not induce clinical improvement. Because 3 of the affected llamas were full sisters, a genetic basis for the problem has to be considered. It was not possible to evaluate the familial relationship of the other 5 affected llamas. Although the underlying cause of the problem was not established, the prognosis for affected llamas is guarded to poor.


The postnatal maturation of the immunoglobulin heavy chain IgG repertoire in human preterm neonates is slower than in term neonates.
During the perinatal period the development of the IgH chain CDR3 (CDR-H3) repertoire of IgM transcripts is maturity-dependent and not influenced by premature exposure to Ag. To study whether maturity-dependent restrictions also predominate in the perinatal IgG repertoire we compared 1000 IgG transcripts from cord blood and venous blood of extremely preterm neonates (24-28 wk of gestation) and of term neonates from birth until early infancy with those of adults. We found the following. First, premature contact with the extrauterine environment induced the premature development of an IgG repertoire. However after preterm birth the diversification of the IgG repertoire was slower than that after term birth. Second, the IgG repertoire of preterm neonates retained immature characteristics such as short CDR-H3 regions and overrepresentation of D(H)7-27. Third, despite premature exposure to the extrauterine environment, somatic mutation frequency in IgG transcripts of preterm infants remained low until they reached a postconceptional age corresponding to the end of term gestation. Thereafter, somatic mutations accumulated with age at similar rates in preterm and term neonates and reached 30% of the adult level after 6 mo. In conclusion, class switch was inducible already at the beginning of the third trimester of gestation, but the developing IgG repertoire was characterized by similar restrictions as those of the developing IgM repertoire. Those B cells expressing more "mature" H chain sequences were not preferentially selected into the IgG repertoire. Therefore, the postnatal IgG repertoire of preterm infants until the expected date of delivery differs from the postnatal repertoire of term neonates.


Dual binding mode of a novel series of DHODH inhibitors.
Human dihydroorotate dehydrogenase (DHODH) represents an important target for the treatment of hyperproliferative and inflammatory diseases. In the cell DHODH catalyzes the rate-limiting step of the de novo pyrimidine biosynthesis. DHODH inhibition results in beneficial immunosuppressant and antiproliferative effects in diseases such as rheumatoid arthritis. Here, we present high-resolution X-ray structures of human DHODH in complex with a novel class of low molecular weight compounds that inhibit the enzyme in the nanomolar range. Some compounds showed an interesting dual binding mode within the same cocrystal strongly depending on the nature of chemical substitution. Measured in vitro activity data correlated with the prevailing mode of binding and explained the observed structure-activity relationship. Additionally, the X-ray data confirmed the competitive nature of the inhibitors toward the putative ubiquinone binding site and will guide structure-based design and synthesis of molecules with higher activity.


Bystander activation of iNKT cells occurs during conventional T-cell alloresponses.
It is well established that iNKT cells can be activated by both exogenous and a limited number of endogenous glycolipids. However, although iNKT cells have been implicated in the immune response to transplanted organs, the mechanisms by which iNKT cells are activated in this context remain unknown. Here we demonstrate that iNKT cells are not activated by allogeneic cells per se, but expand, both in vitro and in vivo, in the presence of a concomitant conventional T-cell response to alloantigen. This form of iNKT activation was found to occur independently of TCR-glycolipid/CD1d interactions but rather was a result of sequestration of IL-2 produced by conventional alloreactive T cells. These results show for the first time that IL-2, produced by activated conventional T cells, can activate iNKT cells independently of glycolipid/CD1d recognition. Therefore, we propose that the well-documented involvement of iNKT cells in autoimmunity, the control of cancer as well as following transplantation need not involve recognition of endogenous or exogenous glycolipids but alternatively may be a consequence of specific adaptive immune responses.


Modular TRAPP complexes regulate intracellular protein trafficking through multiple Ypt/Rab GTPases in Saccharomyces cerevisiae.
Ypt/Rab are key regulators of intracellular trafficking in all eukaryotic cells. In yeast, Ypt1 is essential for endoplasmic reticulum (ER)-to-Golgi transport, whereas Ypt31/32 regulate Golgi-to-plasma membrane and endosome-to-Golgi transport. TRAPP is a multisubunit complex that acts as an activator of Ypt/Rab GTPases. Trs85 and Trs130 are two subunits specific for TRAPP III and TRAPP II, respectively. Whereas TRAPP III was shown to acts as a Ypt1 activator, it is still controversial whether TRAPP II acts as a Ypt1 or Ypt31/32 activator. Here, we use GFP-Snc1 as a tool to study transport in Ypt and TRAPP mutant cells. First, we show that expression of GFP-Snc1 in trs85Δ mutant cells results in temperature sensitivity. Second, we suggest that in ypt1ts and trs85Δ, but not in ypt31Δ/32ts and trs130ts mutant cells, GFP-Snc1 accumulates in the ER. Third, we show that overexpression of Ypt1, but not Ypt31/32, can suppress both the growth and GFP-Snc1 accumulation phenotypes of trs85Δ mutant cells. In contrast, overexpression of Ypt31, but not Ypt1, suppresses the growth and GFP-Snc1 transport phenotypes of trs130ts mutant cells. These results provide genetic support for functional grouping of Ypt1 with Trs85-containing TRAPP III and Ypt31/32 with Trs130-containing TRAPP II.


Stable size distribution of amyloid plaques over the course of Alzheimer disease.
Amyloid β plaques are a key pathologic feature of Alzheimer disease (AD), but whether plaque sizes increase or stabilize over the course of AD is unknown. We measured the size distribution of total immunoreactive (10D5-positive) and dense-core (Thioflavin S-positive) plaques in the temporal neocortex of a large group of subjects with AD and age-matched plaque-bearing subjects without dementia to test the hypothesis that amyloid plaques continue to grow along with the progression of the disease. The size of amyloid β (10D5)-positive plaques did not differ between groups, whereas dense-core plaques from the group with AD were slightly larger than those from the group without dementia (∼25%-30%, p = 0.01). Within the group with AD, dense-core plaque size did not independently correlate with duration of clinical disease (from 4 to 21 years, p = 0.68), whereas 10D5-positive plaque size correlated negatively with disease duration (p = 0.01). By contrast, an earlier age of symptom onset strongly predicted a larger postmortem plaque size; this effect was independent of disease duration and the presence of the APOE[Latin Small Letter Open E]4 allele (p = 0.0001). We conclude that plaques vary in size among patients, with larger size distributions correlating with an earlier age of onset, but plaques do not substantially increase in size over the clinical course of the disease.


The Effects of Angelica Sinensis Polysaccharide on Tumor Growth and Iron Metabolism by Regulating Hepcidin in Tumor-Bearing Mice.
Iron plays a fundamental role in cell biology and its concentration must be precisely regulated. It is well documented that excess iron burden contributes to the occurrence and progression of cancer. Hepcidin secreted by liver plays an essential role in orchestrating iron metabolism. In the present study, we aimed to investigate the ability of angelica sinensis polysaccharide (ASP) to decrease iron burden in tumor-bearing mice and the mechanism of ASP regulation hepcidin expression. Western blot, RT-PCR, immunohistochemistry (IHC), and enzyme-linked immunosorbent assay (ELISA) were used to detect the regulation of hepcidin and related cytokines by ASP. The role of ASP in tumor proliferation was investigated using in vivo assays. Iron depositions and iron concentrations in organs were determined by hematoxylin-eosin (H&E) staining and atomic absorption spectrophotometer. We found that ASP could inhibit tumor growth in mice xenografted with 4T1 and H22 cancer cells. In vivo experiments also showed that ASP could potently regulate hepcidin expression in liver and serum and decrease iron burden in liver, spleen and grafted tumors in mouse model. Treatment with ASP in hepatic cell lines reproduced comparable results in decreasing hepcidin as in mouse liver. Furthermore, we found that ASP markedly suppressed the expression of interleukin-6 (IL-6), JAK2, p-STAT3, and p-SMAD1/5/8 in liver, suggesting that JAK/STAT and BMP-SMAD pathways were involved in the regulation of hepcidin expression by ASP. We also found down-regulation of iron-related cytokines in ASP treated mice. The present study provides new evidence that ASP decreases hepcidin expression, which can reduce iron burden and inhibit tumor proliferation. These findings might aid ASP developed as a potential candidate for cancer treatment in patients with iron overload.


Discrete particle modeling and micromechanical characterization of bilayer tablet compaction.
A mechanistic particle scale model is proposed for bilayer tablet compaction. Making bilayer tablets involves the application of first layer compaction pressure on the first layer powder and a second layer compaction pressure on entire powder bed. The bonding formed between the first layer and the second layer particles is crucial for the mechanical strength of the bilayer tablet. The bonding and the contact forces between particles of the first layer and second layer are affected by the deformation and rearrangement of particles due to the compaction pressures. Our model takes into consideration the elastic and plastic deformations of the first layer particles due to the first layer compaction pressure, in addition to the mechanical and physical properties of the particles. Using this model, bilayer tablets with layers of the same material and different materials, which are commonly used pharmaceutical powders, are tested. The simulations show that the strength of the layer interface becomes weaker than the strength of the two layers as the first layer compaction pressure is increased. The reduction of strength at the layer interface is related to reduction of the first layer surface roughness. The reduced roughness decreases the available bonding area and hence reduces the mechanical strength at the interface. In addition, the simulations show that at higher first layer compaction pressure the bonding area is significantly less than the total contact area at the layer interface. At the interface itself, there is a non-monotonic relationship between the bonding area and first layer force. The bonding area at the interface first increases and then decreases as the first layer pressure is increased. These results are in agreement with findings of previous experimental studies.


Test-retest reliability and task order effects of emotional cognitive tests in healthy subjects.
Little is known of the retest reliability of emotional cognitive tasks or the impact of using different tasks employing similar emotional stimuli within a battery. We investigated this in healthy subjects. We found improved overall performance in an emotional attentional blink task (EABT) with repeat testing at one hour and one week compared to baseline, but the impact of an emotional stimulus on performance was unchanged. Similarly, performance on a facial expression recognition task (FERT) was better one week after a baseline test, though the relative effect of specific emotions was unaltered. There was no effect of repeat testing on an emotional word categorising, recall and recognition task. We found no difference in performance in the FERT and EABT irrespective of task order. We concluded that it is possible to use emotional cognitive tasks in longitudinal studies and combine tasks using emotional facial stimuli in a single battery.


[Clinical significance of sentinel node navigation surgery in the treatment of early gastric cancer].
Radical lymphadenectomy is the standard surgical approach even for early-stage gastric cancer with a relatively low incidence of lymph node metastasis because of the limited sensitivity of diagnostic imaging to detect micrometastasis in regional lymph nodes. The sentinel node (SN) concept is one topic among novel diagnostic procedures for micrometastasis. The SN is defined as the first draining node from the primary lesion and it would be the first site of micrometastasis. SN biopsy has been clinically validated and applied for the surgical treatment of malignant melanoma and breast cancer. Although the feasibility of this technique in other solid tumors including gastric cancer is still controversial, there are several reports demonstrating the diagnostic significance of SN mapping in early gastric cancer. We have established radio-guided SN mapping for early gastric cancer, and the diagnostic accuracy using this procedure in cT1N0 cases reached 98%. The radio-guided method allows us to detect the SN in endoscopic surgery quantitatively and reproducibly. Validation of the SN concept in gastric cancer in a multi-centric clinical trial is essential for clinical application of this procedure, including the establishment of a novel, minimally invasive approach for early-stage gastric cancer.


Organotypic tissue culture of adult rodent retina followed by particle-mediated acute gene transfer in vitro.
Organotypic tissue culture of adult rodent retina with an acute gene transfer that enables the efficient introduction of variable transgenes would greatly facilitate studies into retinas of adult rodents as animal models. However, it has been a difficult challenge to culture adult rodent retina. The purpose of this present study was to develop organotypic tissue culture of adult rodent retina followed by particle-mediated acute gene transfer in vitro. We established an interphase organotypic tissue culture for adult rat retinas (>P35 of age) which was optimized from that used for adult rabbit retinas. We implemented three optimizations: a greater volume of Ames' medium (>26 mL) per retina, a higher speed (constant 55 rpm) of agitation by rotary shaker, and a greater concentration (10%) of horse serum in the medium. We also successfully applied this method to adult mouse retina (>P35 of age). The organotypic tissue culture allowed us to keep adult rodent retina morphologically and structurally intact for at least 4 days. However, mouse retinas showed less viability after 4-day culture. Electrophysiologically, ganglion cells in cultured rat retina were able to generate action potentials, but exhibited less reliable light responses. After transfection of EGFP plasmids by particle-mediated acute gene transfer, we observed EGFP-expressing retinal ganglion cells as early as 1 day of culture. We also introduced polarized-targeting fusion proteins such as PSD95-GFP and melanopsin-EYFP (hOPN4-EYFP) into rat retinal ganglion cells. These fusion proteins were successfully transferred into appropriate locations on individual retinal neurons. This organotypic culture method is largely applicable to rat retinas, but it can be also applied to mouse retinas with a caveat regarding cell viability. This method is quite flexible for use in acute gene transfection in adult rodent retina, replacing molecular biological bioassays that used to be conducted in isolated cultured cells.


Relationship between 25-OH-D serum level and relapse rate in multiple sclerosis patients before and after vitamin D supplementation.
Vitamin D could play a protective role in multiple sclerosis. In an observational, uncontrolled study, vitamin D3 supplementation (3010 IU/day on average) was given to 156 consecutive patients with relapsing-remitting multiple sclerosis, under first-line immunomodulatory therapy and with initial 25-OH-D serum level lower than 100 nmol/l (40 ng/ml). Relapses were determined for 29.1 ± 8.4 months during vitamin D and 29.8 ± 10.1 months before supplementation. The 25-OH-D level was measured before supplementation and several times during supplementation. The incidence rate of relapses before and during supplementation was estimated using negative binomial regression models with follow-up durations as offset terms. The incidence rate and incidence rate ratio of relapses at various 25-OH-D levels were also calculated using negative binomial regression models. In 76 patients, immunomodulatory therapy preceded vitamin D supplementation (by 4.2 ± 2.7 years) and in 80 patients both treatments were started simultaneously. Under supplementation, the 25-OH-D level increased from 49 ± 22 nmol/l to 110 ± 26 nmol/l on average. Pooling data collected before and during supplementation, we found a significant strong inverse relationship between the relapse incidence rate and the 25-OH-D level (p < 0.0001), suggesting that vitamin D did indeed influence the relapse rate. Results of univariate, bivariate and multivariate analyses were analogous: in the multivariate model adjusted for age, disease duration and previous use of immunomodulatory therapy, every 10 nmol increase in 25-OH-D level was associated with a reduction in the relapse incidence rate of 13.7%. Dividing iteratively the population made up of pooled periods into two subgroups according to the 25-OH-D levels, the relapse incidence rate ratio decreased as the 25-OH-D level increased up to 110 nmol/l, but a plateau effect was observed beyond this limit. Further studies are warranted for accurate quantification of the vitamin D effect.


Mental Health during COVID-19 Pandemic among Caregivers of Young Children in Kenya's Urban Informal Settlements. A Cross-Sectional Telephone Survey.
The emergence of COVID-19 has profoundly affected mental health, especially among highly vulnerable populations. This study describes mental health issues among caregivers of young children and pregnant women in three urban informal settlements in Kenya during the first pandemic year, and factors associated with poor mental health. A cross-sectional telephone survey was administered to 845 participants. Survey instruments included the Patient Health Questionnaire-9, General Anxiety Disorder-7 scale, COVID-19 Anxiety Scale, and questions on the perceived COVID-19 effects on caregiver wellbeing and livelihood. Data were analyzed using descriptive statistics, and univariate and multivariate analysis. Caregivers perceived COVID-19 as a threatening condition (94.54%), affecting employment and income activities (>80%). Caregivers experienced discrimination (15.27%) and violence (12.6%) during the pandemic. Levels of depression (34%), general anxiety (20%), and COVID-19 related anxiety (14%) were highly prevalent. There were significant associations between mental health outcomes and economic and socio-demographic factors, violence and discrimination experiences, residency, and perceptions of COVID-19 as a threatening condition. Caregivers high burden of mental health problems highlights the urgent need to provide accessible mental health support. Innovative and multi-sectoral approaches will be required to maximize reach to underserved communities in informal settlements and tackle the root causes of mental health problems in this population.


Fascial suspension mastopexy.
The objective of this study was to assess a new procedure for breast mastopexy, fascial suspension mastopexy. Between December of 2001 and July of 2004, 52 patients (102 breasts) underwent fascial suspension mastopexy. Sixteen women had grade 1 ptosis (30 percent), 31 had grade 2 ptosis (60 percent), two had grade 3 ptosis (4 percent), two (4 percent) had breast asymmetry and underwent unilateral mastopexy, and one had pseudoptosis (2 percent). Of the 52 patients, 43 (83 percent) underwent fascial suspension mastopexy only, whereas nine patients (17 percent) underwent fascial suspension mastopexy combined with breast parenchymal excision of less than 120 g in each breast. Three patients (6 percent) underwent combined procedures. The complication rate was 7.7 percent (four patients). Two patients underwent conversion of the vertical scar into a horizontal scar. One patient underwent drainage of a hematoma 1 day after her operation, and one patient developed a hypertrophic scar. The average follow-up was 1.4 years, with the longest follow-up being 3.5 years. On a scale of 1 (very poor) to 10 (excellent), postoperative ptosis correction was rated as 8.8, breast symmetry as 8.4, postoperative upper pole fullness as 9.1, postoperative medial breast fullness as 7.8, and overall breast shape and contour as 8.6. Fascial suspension mastopexy is safe, simple, and versatile; does not require pectoralis major flap harvesting; and achieves an excellent suspension of breast parenchyma, creating a projected and rejuvenated breast shape with upper pole fullness and long-standing breast contour.


Differences in cortical serotonergic innervation among humans, chimpanzees, and macaque monkeys: a comparative study.
In this study, we assess the possibility that the evolution of human intellectual capacities was supported by changes in the supply of serotonin to the frontal cortex. To this end, quantitative comparative analyses were performed among humans, chimpanzees, and macaques. Immunohistochemical methods were used to visualize serotonin transporter-immunoreactive (SERT-ir) axons within the cerebral cortex. Areas 9 and 32 were chosen for evaluation due to their roles in working memory and theory of mind, respectively. Primary motor cortex was also evaluated because it is not associated with higher cognitive functions. The findings revealed that humans do not display a quantitative increase in serotonin innervation. However, the results indicated region- and layer-specific differences among species in serotonergic innervation pattern. Compared with macaques, humans and chimpanzees together displayed a greater density of SERT-ir axons relative to neuron density in layers V/VI. This change was detected in cortical areas 9 and 32, but not in primary motor cortex. Further, morphological specializations, coils of axons, were observed in humans and chimpanzees that were absent in macaques. These features may represent a greater capacity for cortical plasticity exclusive to hominoids. Taken together, these results indicate a significant reorganization of cortical serotonergic transmission in humans and chimpanzees.


Social, contextual, and individual factors affecting the occurrence and acoustic structure of drumming bouts in wild chimpanzees (Pan troglodytes).
The production of structured and repetitive sounds by striking objects is a behavior found not only in humans, but also in a variety of animal species, including chimpanzees (Pan troglodytes). In this study we examined individual and social factors that may influence the frequency with which individuals engage in drumming behavior when producing long distance pant hoot vocalizations, and analyzed the temporal structure of those drumming bouts. Male chimpanzees from Budongo Forest, Uganda, drummed significantly more frequently during travel than feeding or resting and older individuals were significantly more likely to produce drumming bouts than younger ones. In contrast, we found no evidence that the presence of estrus females, high ranking males and preferred social partners in the caller's vicinty had an effect on the frequency with which an individual accompanied their pant hoot vocalization with drumming. Through acoustic analyses, we demonstrated that drumming sequences produced with pant hoots may have contained information on individual identity and that qualitatively, there was individual variation in the complexity of the temporal patterns produced. We conclude that drumming patterns may act as individually distinctive long-distance signals that, together with pant hoot vocalizations, function to coordinate the movement and spacing of dispersed individuals within a community, rather than as signals to group members in the immediate audience.


Perceptual Visual Distortions in Adult Amblyopia and Their Relationship to Clinical Features.
Develop a paradigm to map binocular perceptual visual distortions in adult amblyopes and visually normal controls, measure their stability over time, and determine the relationship between strength of binocular single vision and distortion magnitude. Perceptual visual distortions were measured in 24 strabismic, anisometropic, or microtropic amblyopes (interocular acuity difference ≥ 0.200 logMAR or history of amblyopia treatment) and 10 controls (mean age 27.13 ± 10.20 years). The task was mouse-based target alignment on a stereoscopic liquid crystal display monitor, measured binocularly five times during viewing dichoptically through active shutter glasses, amblyopic eye viewing cross-hairs, fellow eye viewing single target dots (16 locations within central 5°), and five times nondichoptically, with all stimuli visible to either eye. Measurements were repeated over time (1 week, 1 month) in eight amblyopic subjects, evaluating test-retest reliability. Measurements were also correlated against logMAR visual acuity, horizontal prism motor fusion range, Frisby/Preschool Randot stereoacuity, and heterophoria/heterotropia prism cover test measurement. Sixty-seven percent (16/24) of amblyopes had significant perceptual visual distortions under dichoptic viewing conditions compared to nondichoptic viewing conditions and dichoptic control group performance. Distortions correlated with the strength of motor fusion (r = -0.417, P = 0.043) and log stereoacuity (r = 0.492, P = 0.015), as well as near angle of heterotropic/heterophoric deviation (r = 0.740, P < 0.001), and, marginally, amblyopia depth (r = 0.405, P = 0.049). Global distortion index (GDI, mean displacement) remained, overall, consistent over time (median change in GDI between baseline and 1 week = -0.03°, 1 month = -0.08°; x-axis Z = 4.4256, P < 0.001; y-axis Z = 5.0547, P < 0.001). Perceptual visual distortions are stable over time and associated with poorer binocular function, greater amblyopia depth, and larger angles of ocular deviation. Assessment of distortions may be relevant for recent perceptual learning paradigms specifically targeting binocular vision.


Parental decisions, child health and valuation of avoiding arsenic in drinking water in rural Bangladesh.
Arsenic contamination of groundwater in Bangladesh is a widespread public health hazard. Water sources without high arsenic levels are scarce, affecting people's availability for work and other activities when they have to seek safe water to drink. While children are particularly susceptible to chronic arsenic exposure, limited information and heavy constraints on resources may preclude people in developing countries from taking protective actions. Since parents are primary decision-makers for children, a model of stochastic decision-making analytically linking parent health and child health is used to frame the valuation of avoiding arsenic exposure using an averting behavior model. The results show that safe drinking water programs do work and that people do take protective actions. The results can help guide public health mitigation policies, and examine whether factors such as child health and time required for remediation have an effect on mitigation measures.


[Translated article] Scabies in Spain: Trends in Public Interest and Ectoparasiticide Use.
Scabies is an ectoparasitic dermatosis caused by the Sarcoptes scabiei var. hominis mite, which lives and reproduces in humans. Its incidence in Spain has increased in recent years. The aim of this study was to complement existing evidence on the increasing number of scabies cases in our country by analyzing changes in ectoparasiticide prescriptions and Internet searches for scabies infestations measured by Google Trends. We also examined correlations between these two variables. Our results show that public interest in scabies has increased in recent years and is positively and significantly correlated with an increasing use of ectoparasiticides. We believe that Google Trends should be considered as a complementary tool for monitoring real-time trends in scabies infestations in Spain.


Intravascular optical coherence tomography imaging at 3200 frames per second.
We demonstrate intravascular optical coherence tomography (OCT) imaging with frame rate up to 3.2 kHz (192,000 rpm scanning). This was achieved by using a custom-built catheter in which the circumferential scanning was actuated by a 1.0 mm diameter synchronous motor. The OCT system, with an imaging depth of 3.7 mm (in air), is based on a Fourier domain mode locked laser operating at an A-line rate of 1.6 MHz. The diameter of the catheter is 1.1 mm at the tip. Ex vivo images of human coronary artery (78.4 mm length) were acquired at a pullback speed of 100 mm/s. True 3D volumetric imaging of the entire artery, with dense and isotropic sampling in all dimensions, was performed in <1 second acquisition time.


Planar silicon nitride waveguides for biosensing.
The principles of attenuation of the light intensity due to multiple reflections are realised in a planar silicon oxide (SiO(2))silicon nitride (Si(3)N(4)) waveguiding structure for the purpose of developing optical biosensors with improved sensitivity. The analysis of the experimental data shows that the large difference in refractive indices of core and cladding layers gives rise to an increase in sensitivity by a factor of 3 over previously reported structures. Composite polyelectrolyte self-assembled thin films containing cyclo-tetra-chromotropylene as an indicator and enzymes glucose oxidase or urease were employed in the superstrate as a sensing membrane. Individual enzyme reactions as well as their inhibition by pesticides were studied by monitoring the intensity of light output from the planar waveguide. The results were compatible with those obtained by conventional ultraviolet-visible absorption spectroscopy. The instrument detection limit for Imidacloprid pesticide was found to be as low as 10 ppb in concentration.


Ambulatory care provided by office-based specialists in the United States.
Increasing use of specialist services in the United States is leading to a perception of a specialist shortage. Little is known, however, about the nature of care provided by this secondary level of services. The aim of this study was to examine the content of care provided by specialists in community settings, including visits for which the patient had been referred by another physician. Nationally representative visit data were obtained from the National Ambulatory Medical Care Survey (NAMCS) for the years 2002 through 2004. To describe the nature of care, we developed a taxonomy of office-based visit types and constructed logistic regression models allowing for adjusted comparisons of specialty types. Overall, 46.3% of visits were for routine follow-up and preventive care of patients already known to the specialist. Referrals accounted for only 30.4% of all visits. Specialists were more likely to report sharing care with other physicians for referred, compared with not referred, patients (odds ratio [OR] = 2.99; 95% confidence interval [CI], 2.52-3.55). Overall, 73.6% of all visits resulted in a return appointment with the same physician, in more than one-half of all cases as a result of a routine or preventive care visit. Ambulatory office-based activity of specialists includes a large share of routine and preventive care for patients already known, not referred, to the physician. It is likely that many of these services could be managed in primary care settings, lessening demand for specialists and improving coordination of care.


Effect of Resistance Training and Various Sources of Protein Supplementation on Body Fat Mass and Metabolic Profile in Sarcopenic Overweight Older Adult Men: A Pilot Study.
The decrease in resting energy expenditure (REE) and fat oxidation with aging is associated with an increase in fat mass (FM), and both could be prevented by exercise such as resistance training. Dairy consumption has also been shown to promote FM loss in different subpopulations and to be positively associated with fat oxidation. Therefore, we sought to determine whether resistance exercise combined with dairy supplementation could have an additive impact on FM and energy metabolism, especially in individuals with a deficit in muscle mass. Twenty-six older overweight sarcopenic men (65 ± 5 years old) were recruited for the study. They participated in 4 months of resistance exercise and were randomized into three groups for postexercise shakes (control, dairy, and nondairy isocaloric and isoprotein supplement with 375 ml and ~280 calories per shake). Body composition was measured by dual X-ray absorptiometry and REE by indirect calorimetry. Fasting glucose, insulin, leptin, inflammatory profile, and blood lipid profile were also measured. Significant decreases were observed with FM only in the dairy supplement group; no changes were observed for any other variables. To conclude, FM may decrease without changes in metabolic parameters during resistance training and dairy supplementation with no caloric restriction without having any impact on metabolic properties. More studies are warranted to explain this significant decrease in FM.


Current practice of cord clamping in the Netherlands: a questionnaire study.
Recent meta-analyses recommend delayed cord clamping (DCC) after uncomplicated births as well as preterm births, but there is no clear definition of timing and uniform national guidelines are lacking. We aimed to investigate if guidelines for the timing of cord clamping (CC) are followed and what the national practice entails. A postal questionnaire concerning CC after uncomplicated vaginal, Caesarean term and preterm deliveries was sent to all midwifery practices (n = 526) and obstetrical departments (n = 94) in the Netherlands. The response rate was 81% (500/620). CC protocols were present in 16 and 38% of midwifery and obstetric practices, respectively. Early cord clamping (ECC) was recommended in 54%, DCC in 33%, 6% indicated a specific time point and 7% did not specify. In current practice, DCC was applied after uncomplicated vaginal term deliveries in 90% and ECC in 6%, and no timing was specified in 4%. Midwives used DCC more often than obstetricians (97 vs. 75%). Cessation of cord pulsations was often (54%) used as a time point, 40% used a fixed time point, 2% waited for placental expulsion and 4% did not specify. ECC was preferred in obstetric practices after Caesarean deliveries (in 81%). In preterm births, ECC was practised by 36%, DCC by 54 and 10% did not specify. In the Netherlands, although often not protocolized, DCC is widely used after uncomplicated vaginal term and preterm deliveries, but not after Caesareans. Cessation of cord pulsation is often used as the time point for CC.


How DMSO, a widely used solvent, affects spinal cord injury.
The aim of this experimental study was to investigate whether dimethylsulfoxide (DMSO) has protective effects on spinal cord ischemia-reperfusion (I/R) injury. New Zealand rabbits were enrolled in the study. In addition to the control group, the study group received 0.1 mL/kg DMSO prior to ischemia. Blood samples were taken to obtain nitrite-nitrate levels during the surgical procedure. After neurological evaluation at 24 hr of reperfusion, lumbar spinal cords were removed for electron microscopic evaluation and malondialdehyde and myeloperoxidase measurements. The mean Tarlov score of the DMSO group was higher than that of the control group. Electron microscopic examination was carried out with tissue samples at 24 hr of reperfusion. The DMSO group had better preservation with the electron microscopic scoring compared to the control group. Malondialdehyde and myeloperoxidase levels were decreased in the DMSO group compared to the control group. Nitrite-nitrate levels were also lower in the DMSO group compared to control at 5 and 30 min of reperfusion. This study demonstrates a considerable neuroprotective effect of DMSO on neurological, biochemical, and histopathological analyses during periods of spinal cord I/R injury in rabbits. Although there was a difference between the DMSO and control groups in all measured parameters in our study, this was not statistically significant. DMSO deserves further investigation related with spinal cord ischemia and reperfusion. We should also consider the effect of DMSO when we use it as a solvent or vehicle during experimental I/R models.


From sink to source: Regional variation in U.S. forest carbon futures.
The sequestration of atmospheric carbon (C) in forests has partially offset C emissions in the United States (US) and might reduce overall costs of achieving emission targets, especially while transportation and energy sectors are transitioning to lower-carbon technologies. Using detailed forest inventory data for the conterminous US, we estimate forests' current net sequestration of atmospheric C to be 173 Tg yr(-1), offsetting 9.7% of C emissions from transportation and energy sources. Accounting for multiple driving variables, we project a gradual decline in the forest C emission sink over the next 25 years (to 112 Tg yr(-1)) with regional differences. Sequestration in eastern regions declines gradually while sequestration in the Rocky Mountain region declines rapidly and could become a source of atmospheric C due to disturbances such as fire and insect epidemics. C sequestration in the Pacific Coast region stabilizes as forests harvested in previous decades regrow. Scenarios simulating climate-induced productivity enhancement and afforestation policies increase sequestration rates, but would not fully offset declines from aging and forest disturbances. Separating C transfers associated with land use changes from sequestration clarifies forests' role in reducing net emissions and demonstrates that retention of forest land is crucial for protecting or enhancing sink strength.


Late-term elective abortion and susceptibility to posttraumatic stress symptoms.
The primary aim of this study was to compare the experience of an early abortion (1st trimester) to a late abortion (2nd and 3rd trimester) relative to Posttraumatic Stress Disorder (PTSD) symptoms after controlling for socio-demographic and personal history variables. Online surveys were completed by 374 women who experienced either a 1st trimester abortion (up to 12 weeks gestation) or a 2nd or 3rd trimester abortion (13 weeks gestation or beyond). Most respondents (81%) were U.S. citizens. Later abortions were associated with higher Intrusion subscale scores and with a greater likelihood of reporting disturbing dreams, reliving of the abortion, and trouble falling asleep. Reporting the pregnancy was desired by one's partner, experiencing pressure to abort, having left the partner prior to the abortion, not disclosing the abortion to the partner, and physical health concerns were more common among women who received later abortions. Social reasons for the abortion were linked with significantly higher PTSD total and subscale scores for the full sample. Women who postpone their abortions may need more active professional intervention before securing an abortion based on the increased risks identified herein. More research with diverse samples employing additional measures of mental illness is needed.


Concordance between self-reported substance use and toxicology among HIV-infected and uninfected at risk youth.
Substance use by youth living with HIV (YLWH) is a concern, given potential interactions with virus-associated immune suppression and adverse effects on risk behaviors, neurocognition, and adherence. Self-report substance use measures provide efficient cost-effective assessments. Analyses describe self-reported substance use among YLWH and examine agreement with toxicology assays. Seventy-eight youth age 18-24 years (87% male, 71% African-American) with behaviorally acquired HIV-1 infection and 55 uninfected youth completed the Alcohol, Smoking, and Substance Involvement Screening Test to assess drug use frequency, including tobacco, marijuana, cocaine, and alcohol, over the prior three months. Elisa-based toxicology assays were used to detect 27 substances in plasma. Chi-square tests compared substance use between YLWH and uninfected youth; Kappa statistics compared agreement between self-report and toxicology. YLWH reported marijuana (49%), tobacco (56%), and alcohol (87%) use, with 20%, 28% and 3% reporting daily use of each substance, respectively; other substance use was uncommon. Uninfected youth reported less tobacco use but otherwise similar substance use. All youth who reported daily use of marijuana or tobacco had positive plasma toxicology results, while concordance decreased with less frequent self-reported use. Among youth reporting no substance use, few tested positive (4% YLWH, 2% uninfected youth for cannabis; 8%YLWH for tobacco). Youth report high rates of marijuana, tobacco, and alcohol use. Concordance between self-report and toxicology for marijuana and tobacco use, particularly for daily users, supports self-report as a valid indicator of substance use in research studies of youth with or without HIV-1 infection.


Preschool and childcare center characteristics associated with children's physical activity during care hours: an observational study.
Preschools and childcare settings offer opportunities to promote adequate levels of physical activity. Research is needed to identify the key features of these settings to optimize young children's activity. The aims of this study were to determine if differences existed in preschool children's physical activity during care hours compared with outside care hours and to examine a comprehensive range of potential center-based correlates of physical activity for preschool boys and girls. Data are from the Healthy Active Preschool and Primary Years study: 71 childcare centers, 65 preschools and 1002 preschool children. Percent of time in total (light- to vigorous-intensity) physical activity was measured using Actigraph GT1M accelerometers. Center physical environment characteristics, policies and practices were assessed by trained research staff using comprehensive audit tools. Data were collected in 2008/9 and were analyzed separately for boys and girls in Stata using multilevel mixed effects models. Boys and girls were less active during care than outside care hours (51.1 % vs. 52.4 %, p = 0.01; 48.0 % vs. 51.5 %, p < 0.0001, respectively). In the final adjusted models, number of outdoor spaces with natural ground coverings was associated with boys' physical activity (coeff = 0.477, 95 % CI 0.089, 0.867) and the amount of time girls spent indoors before going outdoors was inversely associated with their physical activity (coeff = -0.035, 95 % CI -0.065, -0.004). The models explained 12 and 10 % of boys' and girls' physical activity during care hours, respectively. This study identified that children are significantly less active during than outside care hours. Few center-based correlates of preschool children's physical activity were identified. Future research should explore other aspects of centers, such as what children actually do while they are outside, and broader potential influences on children's behaviours including social, cultural and policy contexts within which centers operate.


Visible second-harmonic light generated from a self-organized centrosymmetric lattice of nanospheres.
We designed and fabricated a centrosymmetric material where one may be able to consider an efficient quadratic nonlinear interaction. We followed a solid phase-supported organic synthesis methodology to covalently bind a large number of highly nonlinear molecules to the surface of polystyrene nanospheres. Such chemically modified optically nonlinear latex spheres, when suspended in water, are seen to perfectly self-organize into a centrosymmetric lattice. Taking advantage of the nonlinear interaction located at the sphere-water interface and the photonic crystal properties of the fabricated material we were able to generate second-harmonic light visible to the naked eye.


Validation of mental health screening instruments in the Kashmir Valley, India.
The present study aimed to culturally adapt, translate, and validate the Hopkins Symptom Checklist-25 (HSCL-25) and the Harvard Trauma Questionnaire-Posttraumatic Stress Symptoms Checklist (HTQ-16) prior to use in a cross-sectional mental health population survey in the Kashmir Valley. Cultural adaptation and translation of the HSCL-25 and the HTQ-16 employed multiple forms of transcultural validity check. The HSCL-25 and HTQ-16 were compared against a "gold standard" structured psychiatric interview, the Mini International Neuropsychiatric Interview (MINI). Interviews were conducted with 290 respondents recruited using consecutive sampling from general medical outpatient departments in five districts of the Kashmir Valley. Receiver operating characteristics (ROC) analysis was used to estimate the cut point with optimal discriminatory power based on sensitivity and specificity. Internal reliability of the HSCL-25 was high, Cronbach's alpha (α) = .92, intraclass correlation coefficient (ICC) = 0.75, with an estimated optimal cut point of 1.50, lower than the conventional cut point of 1.75. Separation of the instruments into subscales demonstrated a difference in the estimated cut point for the anxiety subscale and the depression subscale, 1.75 and 1.57, respectively. Too few respondents were diagnosed with posttraumatic stress disorder (PTSD) during structured psychiatric interview, and therefore the HTQ-16 could not be validated despite the fact that high internal reliability was demonstrated (α = .90). This study verified the importance of culturally adapting and validating screening instruments in particular contexts. The use of the conventional cut point of 1.75 would likely have misclassified depression in our survey, leading to an underestimate of this condition.


MicroRNA-182-5p relieves murine allergic rhinitis via TLR4/NF-κB pathway.
Allergic rhinitis (AR) is one of the most common chronic diseases. This study examined whether microRNA (miR)-182-5p plays a role in AR by regulating toll-like receptor 4 (TLR4). First, data demonstrated that TLR4 was a target of miR-182-5p. Subsequently, AR mouse model was established to explore the role of miR-182-5p and TLR4 in AR in vivo. Initially, quantitative reverse transcription-PCR (qRT-PCR) analysis indicated that miR-182-5p was downregulated, while TLR4 expression was upregulated in AR mice. Then we found that miR-182-5p mimic reduced the frequency of sneezing and nose rubbing of the AR mice. In addition, miR-182-5p mimic significantly increased ovalbumin (OVA)-specific IgE and leukotriene C4 expression levels in nasal lavage fluid (NLF) and serum of AR mice. miR-182-5p mimic decreased the number of inflammatory cells in NLF of AR mice. It also reduced the levels of inflammatory factors in the serum of AR mice, such as interleukin (IL)-4, IL-5, IL-13, IL-17 and tumor necrosis factor (TNF)-α, while increasing the release of IFN-γ and IL-2. Finally, miR-182-5p mimic inhibited NF-κB signaling pathway activation in AR mice. However, all effects of miR-182-5p mimic on AR mice were reversed by TLR4-plasmid. In conclusion, miR-182-5p/TLR4 axis may represent a novel therapeutic target for AR.


The usefulness of CD26 in flow cytometric analysis of peripheral blood in Sézary syndrome.
The loss of CD26 expression was proposed to be a constant feature of circulating Sézary cells by flow cytometric immunophenotyping (FCIP), but the experience with CD26 is limited. To establish its usefulness, CD26 results were correlated with morphologic, molecular, and immunophenotypic findings. Based on FCIP of 179 samples of peripheral blood, CD26 negativity was found in 59.3% of cases with Sézary syndrome (SS), 33.3% of mycosis fungoides (MF), 14.2% of benign dermatosis (BD), and no control cases. In diagnostic subgroups of SS based on morphologic, molecular, and immunophenotypic criteria, the percentage of CD26- cases varied from 41.1% to 63.6%. The specificity of a CD26- result was inferior to that of T-cell antigen loss in differentiating SS from MF and BD. CD26 offers lower diagnostic performance than previously suggested; however, in addition to the findings of major T-cell antigen loss, it could improve sensitivity of FCIP in patients with SS.


Spatial and temporal structuring of arbuscular mycorrhizal communities is differentially influenced by abiotic factors and host crop in a semi-arid prairie agroecosystem.
Agroecosystems are dynamic systems that experience frequent chemical inputs and changes in plant cover. The objective of this study was to test whether abiotic (soil chemical properties and climate) and biotic (plant host identity) factors influence the spatial and temporal structuring of arbuscular mycorrhizal fungal (AMF) communities in a semi-arid prairie agroecosystem. 454 GS FLX+ high-throughput sequencing technology was successfully utilized to characterize the AMF communities based on long reads (mean length: 751.7 bp) and generated high-resolution data with excellent taxonomic coverage. The composition of the AMF community colonizing roots of the three crops (pea, lentil, and wheat) significantly differed, but plant host identity had a minimal effect on the composition of the AMF community in the soil. We observed a temporal shift in the composition of AMF communities in the roots and surrounding soil of the crops during the growing season. This temporal shift was particularly evident in the root-associated AMF community and was correlated with soil phosphate flux and climatic variables. In contrast, the spatial structuring of the AMF community in the site was correlated with soil pH and electrical conductivity. Individual AMF taxa were significantly correlated with pH, electrical conductivity, and phosphate flux, and these relationships were phylogenetically conserved at the genus level within the Glomeromycota.


Sixteen-year single-surgeon experience with coil embolization for ruptured intracranial aneurysms: recurrence rates and incidence of late rebleeding. Clinical article.
Over a 16-year period, 570 patients presenting with acute aneurysmal subarachnoid hemorrhage were successfully treated using endosaccular coil embolization within 30 days of hemorrhage by a single surgeon. Patients were followed to assess the stability of aneurysm occlusion and its longer-term efficacy in protecting against rebleeding. Patients were followed for 6 to 191 months (mean 73.7 months, median 67 months) by clinical review, angiography performed at 6 and 24 months posttreatment, and questionnaires sent via the postal service every 5 years. Late rebleeding was defined as > 30 days after treatment. Stable angiographic occlusion was evident in 74.5% of small, 72.2% of large, and 60% of giant aneurysms. Recurrent filling was found in 119 (26.3%) of 452 aneurysms. Rebleeding was diagnosed in 9 patients (6 treated aneurysms) and occurred between 2 and 114 months posttreatment. It was due to aneurysm recurrence in 6 patients, rupture of a coincidental untreated aneurysm in 2 patients, and rupture of a de novo aneurysm in 1 patient. Rebleeding occurred in 3 (2.5%) of 119 unstable aneurysms and in 3 (0.9%) of 333 stable aneurysms, as seen on initial follow-up angiography studies. Annual rebleeding rates ranged from 0.2% to 0.6% for all causes and from 0.2% to 0.4% for rebleeding of treated aneurysms. No rebleeding was recorded after the first decade, with 138 patients having more than 10 years of follow-up. Periodic follow-up with angiographic studies after coil embolization is recommended to identify aneurysm recurrence and patients at a high risk of late rebleeding in the medium term. More frequent follow-up is recommended for patients harboring coincidental unruptured aneurysms.


Clinical implications from a single-center study of colorectal adenocarcinoma in transplant recipients.
With the increasing burden of organ transplant recipients and improvements in allograft outcome, the incidence of neoplasms rising from these patients is an important issue. In this study, we investigated transplant recipients with colorectal cancer to determine its incidence, clinicopathological characteristics, and prognosis. The database of Severance Hospital was queried for all cases of colorectal adenocarcinoma among transplant recipients from August 2005 to January 2013. A total of 29 patients were diagnosed with colorectal adenocarcinoma after transplantation, and the median age at diagnosis was 58.6 years. As for primary tumor stage, 17 (58.6%) patients had stage ≥3, and distant metastasis was found in 10 (34.4%) patients. The mean time from transplantation to tumor detection was 13.7 years. The median disease-free survival was 11.0 months and the median overall survival (OS) was 18.1 months. In multivariate analysis of prognostic factors for OS, surgical resection was a positive prognostic factor (HR 1.357, p = 0.010) and the presence of distant metastasis at diagnosis was a negative prognostic factor (HR 1.047, p = 0.006). The behavior of colorectal cancer in posttransplant patients is more aggressive and refractory to treatment. A separate guideline for the colorectal screening program for the posttransplant patients needs to be established.