As part of a study on maternal anemia and offspring neurodevelopment, we analyzed blood lead level (BLL) with no prior knowledge of lead exposure in 225 mothers and 685 offspring 1 to 2 years old from Allada, a semi-rural area in Benin, sub-Saharan Africa, between May 2011 and May 2013. Blood samples were analyzed by inductively coupled plasma mass spectrometry. Environmental assessments in households and isotopic ratio measurements were performed for eight children with BLL > 100 µg/L. High lead levels (BLL > 50 µg/L) were found in 44% of mothers and 58% of children. The median BLL was 55.1 (interquartile range 39.2–85.0) and 46.6 (36.5–60.1) µg/L, respectively. Maternal BLL was associated with offspring’s consumption of piped water and animals killed by ammunition. Children’s BLL was associated with presence of paint chips in the house and consumption of animals killed by ammunition. In this population, with 98% of children still breastfed, children’s BLL was highly associated with maternal BLL on multivariate analyses. Environmental measures and isotopic ratios supported these findings. Offspring may be highly exposed to lead  Lead is well known to have detrimental effects on human health, including nervous, renal, cardiovascular, reproductive, immune and hematologic systems. In children, both postnatal and prenatal exposure to lead is associated with poor child neurodevelopment. Studies of postnatal exposure showed evidence of long-term effects on childhood intelligence quotient  as well A recent literature review of BLL in children <6 years old in sub-Saharan Africa (SSA) found only nine studies with data collection between 1991 and 2010 , but mosi.e., children with ID may be more susceptible to the detrimental effect of lead [Moreover, BLL may be increased with iron deficiency (ID) because lead and iron share common pathways for intestinal absorption . ID is h of lead ). TherefAs the investigation of elevated BLL was not initially planned but due to incidental discovery, the methods section contains successive steps.Malaria in Pregnancy Preventive Alternative Drugs” (MiPPAD) clinical trial (NCT00811421) comparing two intermittent preventive treatments of malaria in pregnancy [Tovi means child from the country) when the child was 12 months old [Our study sample included singletons born to pregnant women enrolled in the “al NCT008421 compaInformation on socioeconomic status and home environment were gathered during a home visit when the child was 12 months old. This questionnaire administered by a nurse or a psychologist included information on potential sources of lead including presence of paint and paint chips in the household, maternal and paternal occupation classified by risk of lead exposure , breastfWe collected 8 ml venous blood from each participant, 4 mL in a tube containing dipotassium EDTA and 4 mL in an iron-free dry tube. Blood samples were analyzed at the Centre de Toxicologie, Institut National de Santé Publique du Québec , by inductively coupled plasma mass spectrometry  before 20-fold dilution in amonia 0.5% v/v and 0.1% v/v surfactant Triton-X. The limit of detection for blood analysis was 0.2 µg/L. 3 and 2/3 HCl) by microwave  and dust by a graphite block digestion system . The LOQ was 0.1 mg/kg for food, 0.1 mg/kg for soil, 0.5 mg/L for gasoline and 2 µg/m2 for dust. Dishes were soaked in a solution of acetic acid (4%) for 24 h before analysis, and the LOQ was 2 µg/L. Furthermore, to investigate exposure pathways, we visited eight households for environmental and food assessments. Households were selected if the child’s BLL was > 100 µg/L (very high BLL) and a sufficient amount of blood was available to perform lead isotope ratio (LIR) measurements. The LIR is the ratio of abundance of lead with different atomic weight because of a different neutron number. LIR can provide information from lead contamination origins because the isotopic signature varies by the age of the original ore. The concentration of lead in soil, water, gasoline, dishes, food and paint, when present, was determined in each household. Ammunition for animal hunting was purchased from a local market. All samples were analyzed by ICP-MS (Agilent technologies 7500ce ICP-MS instrument) at a school of public health . For water, the limit of quantification (LOQ) was 1 µg/L. Before ICP-MS analysis, food, soil and gasoline were mineralized with a mixture of nitric and hydrochloric acid (1/3 HNOThe LIR in children’s blood and possible sources of lead was determined by the concentration of lead in each source for a given child. The LIR was determined in the most concentrated sample of each exposure media by using quadrupole ICP-MS (Agilent technologies 7500ce ICP-MS instrument) with environmental samples at EHESP. The mass bias was corrected with a certified reference material  with the standard bracketing technique . Lead stable isotope ratios in blood were established with low-resolution quadropole ICP-MS (Perkin Elmer NexION 300S instrument) after proper dilution to an obtained final lead concentration of 5 µg/L in a 0.5% (v/v) ammonia solution with 0.1% v/v surfactant Triton-X. Instrumental isotopic ratio responses were calibrated with certified reference materiel NIST SRM 981 and controlled with SRM NIST 982.Intercalibrated LIR was calculated by the two laboratories and involved two aqueous leaded samples. Each of the two laboratories used its own method for determining mass correction with the standard SRM 981 and correction of blanks. The results were comparable, except for LIR containing 204.vs. 207/206.Interpretation was based on the proximity of graphical isotopic ratios between blood and potential sources, accounting for measurement accuracy (a source was considered compatible with blood with recovery between the error bars of the source and blood)  and assep < 0.05; model 2 included socioeconomic factors in addition to sources of lead; model 3 included maternal BLL in addition to the former variables. The first two models were also used to analyze maternal BLL. In statistical analyses, BLL was log-transformed for normal distribution . We first described BLL in children and mothers, potential sources of lead, and sociodemographic characteristics of the study population. Second, we performed univariate analysis to assess the crude association between the potential sources of lead in both children and mothers, sociodemographic characteristics, and BLL. Third, we conducted multivariate analyses in three steps for children with logBLL and BLL >50 µg/L as dependent variables: model 1 included potential sources of lead identified on univariate analyses with t test and chi-square test were used to compare means and proportions, respectively. Multiple linear regression and logistic regression were used for continuous and binary outcomes, respectively. Statistical significance was defined as p < 0.05. Statistical analyses involved use of SAS 9.3.Student The study was approved by the institutional review boards of the University of Abomey-Calavi in Benin and New York University in the United States (IRB#09-1253). At recruitment, we obtained informed consent from all pregnant women and guardians of children who participated in this study. p < 0.001).Overall, 58% of children and 44% of mothers showed elevated BLL  . Child aMany children had eaten soil and most were drinking piped water; few children were living in households with painted walls or had activities that may involve use of lead . OverallOn multivariate analysis, paint in the household and eating animals killed by ammunition were significantly associated with children’s logBLL after adjustment for socioeconomic factors . In moden = 13) for children not breastfed compared with 55.7 µg/L (n = 668) in children breastfed.Because almost all children were still breastfed at 12 months (98%), breastfeeding was not included in the model. However, the median BLL was 47.5 µg/L , especially in children and pregnant women. These studies should include representative samples, larger numbers of households, comprehensive measurements of LIR and specific studies on infant hand-to-mouth behaviors. Indeed, in this hot and humid climate, children’s hands may be likely covered with dust and therefore their BLL may be higher than in children from high-income countries for a given concentration of lead in dust. Assessments of BLL and treatments with elevated BLL are not often available as is the case in Benin. There is a need for capacity building in environmental assessments, treatments and research in this field in SSA. BLL seems to have decreased in high-income countries but may be a growing concern in less developed countries. For example, the prevalence of elevated BLL in the United States in 2011 was 0.6% as compared with 57.8% in our study . Given t


Background: Radiation therapy (RT) is a treatment modality traditionally used in patients with multiple myeloma (MM), but little is known regarding the role and effectiveness of RT in the era of novel agents, i.e., immunomodulatory drugs and proteasome inhibitors.Methods: We retrospectively reviewed data from 449 consecutive MM patients seen at our institute in 2010–2012 to assess indications for RT as well as its effectiveness. Pain response was scored similarly to RTOG 0631 and used the Numerical Rating Pain Scale.Results: Among 442 evaluable patients, 149 (34%) patients and 262 sites received RT. The most common indication for RT was palliation of bone pain , followed by prevention/treatment of pathological fractures , spinal cord compression , and involvement of vital organs/extramedullary disease . Of the 55 patients evaluable for pain relief, complete and partial responses were obtained in 76.4 and 7.2%, respectively. Prior RT did not significantly decrease the median number of peripheral blood stem cells collected for autologous transplant, even when prior RT was given to both the spine and pelvis. Inadequacy of stem cell collection for autologous stem cell transplant (ASCT) was not significantly different and it occurred in 9 and 15% of patients receiving no RT and spine/pelvic RT, respectively. None of the three cases of therapy-induced acute myelogenous leukemia/MDS occurred in the RT group.Conclusion: Despite the introduction of novel effective agents in the treatment of MM, RT remains a major therapeutic component for the management in 34% of patients, and it effectively provides pain relief while not interfering with successful peripheral blood stem cell collection for ASCT. Multiple myeloma (MM) is a rare cancer, representing 1% of all malignancies, with an annual incidence of approximately 4-5/100,000 –4. The gIn the last few years, the introduction in clinical practice of several novel agents, i.e., the three immunomodulatory drugs (IMiDs) thalidomide, lenalidomide, and pomalidomide, and the two proteasome inhibitors (PIs) bortezomib and carfilzomib, has produced significant disease responses and survival advantage in MM patients, revolutionizing the therapy of MM in all phases of treatment, i.e., induction therapy, maintenance, and in the setting of relapsed/refractory disease . The rolAfter permission from our Institutional Review Board, we performed a systematic retrospective review of medical charts of 449 consecutive MM patients seen in 2010–2012 followed at our institute, either directly or as consultants for community oncologists. Because a range of radiation doses were given, all dosing regimens were converted into biologic effective doses using an alpha beta ratio = 10 for myeloma cells  , both att-test for numerical variables, and χ2 test for categorical variables . Survival curves were obtained with the Kaplan-Meier method, and compared with the log-rank test. We used the Cox proportional hazard regression model to analyze the effect of several risk factors on survival. All p values were two-tailed, and values <0.05 were considered significant. Overall survival (OS) was calculated from diagnosis to last follow-up or death. The time of diagnosis was defined as the day of the initial bone marrow biopsy. Statistical analysis was performed using the program SAS® software, version 9.3 .Clinical characteristics of cases and controls were compared with the unpaired two-sample student’s n = 4), lung (3), skin (3), breast (1), uterus (1), esophagus (1), and pituitary adenoma (1). Seven of these 14 patients received RT only for reasons other than MM, and they were excluded from the analysis. Of the remaining 442 patients, 293 (66%) did not receive RT for MM , and 149 patients (34%) did . Table In our original dataset of 449 consecutive MM patients, 14 patients received RT for reasons different from MM, i.e., cancers of the prostate . Hazard ratios were 0.59 (95% CI 0.28–1.05) for stage I vs. stage III (p = 0.10), 3.17 (95% CI 2.13–4.96) for high-risk cytogenetics (p < 0.001), and 1.35 (95% CI 0.86–1.59) for presence of lytic lesions at skeletal survey (p = 0.39).In both groups, median follow-up was 28 months range, 0–184). Median OS was 108 months (95% CI 69–162) and 85 months (95% CI 49–98) in group A and B, respectively (84. Mediap = 0.76).Table The second most common indication for RT were pathologic fractures (28%), either for their prevention before an impending fracture (13%), or as “consolidation” RT after surgery for the fracture (15%). Another frequent reason for RT were neurological complications (16%), such as spinal cord compression (10%), or impingement of cranial nerves, spinal neural foramina, or cauda equina syndrome (6%). Other less common indications included involvement of bones adjacent to vital organs , extramedullary disease (4%), and esthetic reasons . The most common site of RT was the spine (36%), and spine and pelvis constituted almost half of the radiated sites. In 51% of cases, RT was administered at the time of diagnosis of MM, either before or soon after (within 2 months) the initiation of systemic therapy. In 42% of cases, RT was given later during the course of the disease, more than 1 year after the initial diagnosis of MM. Only nine patients (6%) required a second course of RT for relapsed disease in a previously radiated site.+ cells × 106/Kg, was 6.3 and 6.8 in group A (189 pts) and B (111 pts), respectively. The difference is not significant according to non-parametric Wilcoxon Rank-Sum t-test (p = 0.84). In group B, 48 pts underwent stem cell collection after RT to lumbar spine and pelvis, with a median dose of 45 Gy. Their median number of CD34+ cells × 106/Kg collected was 5.2, and only seven of them (15%) failed stem cell collection. Seventeen of 189 patients (9%) in Group A failed stem cell collection (p = 0.41).The median number of peripheral blood stem cells collected for autologous SCT, expressed as CD34p = 0.78 by Fisher’s exact test). There were three cases of secondary acute myelogenous leukemia (AML), but they were not related to RT: these patients belonged to group A, and their AML was presumably attributed to the previous exposure to high-dose melphalan used as conditioning regimen for SCT.Finally, we scrutinized our cohort of 442 patients for second malignancies, developed after the diagnosis of MM  or complete 76.4%) response of pain to RT was achieved in 84% of patients, and no progressive pain/failure of RT was observed. Interestingly, the pain response seems to be more effective in this era of IMiDs and PIs, because previously complete and partial responses were reported in 21.6–30 and 69.8–71% of MM patients, respectively , or later during the course of the disease, when MM progresses. Its use is rarely required when the disease is in remission, during the induction chemotherapy. In fact, according to our data, only 7% of MM patients required RT between 2 and 12 months of initiation of systemic therapy, presumably due to the effectiveness of modern chemotherapy regimens. Initial diagnosis and refractory disease at progression are critical moments in the course of MM, because they may require RT for rapid local tumor reduction and “debulking” of focal lesions. The excellent activity of RT against MM has been known for decades. In 1971, Bergsagel observed that a single RT dose of 10 Gy can produce a 3 log cell kill, whereas drugs such as melphalan and prednisone produce <1 log cell kill . In vitrpothesis , 25.We found that the two most common indications for RT were pain palliation and pathological fractures, either for their prevention, or as “consolidation” therapy after surgery. This was expected, as RT is known to be an effective treatment modality in reducing the incidence of new fractures and focal lesions in irradiated bones of MM patients with vertebral lesions . In our The baseline characteristics of our RT-treated and RT-naive groups were balanced for sex, race, and rates of high-risk cytogenetics. The statistical difference in age could theoretically be due to the fact that some very old patients have problems that prevent the administration of RT . ISS stage was significantly lower in the RT-treated group. This is probably due to a combination of factors: first, all MM patients who initially presented with a solitary plasmacytoma -clinically a more indolent plasma cell neoplasm than MM - received RT. Secondly, ISS stage III is defined by a serum level of beta-2 microglobulin >5.5 mg/dL, a finding that reflects not only high tumor load in the bones (which may need RT), but also renal insufficiency (which does not need RT). For reasons unknown to us, patients in the RT-treated group were more likely to have non-secretory disease. We can speculate that these patients come to medical attention for skeletal complications and require RT more frequently than patients with secretory MM, because the latter have useful tumor markers that may detect progressive disease at an earlier stage, or because they may present with MM manifestations that do not require RT, such as hyperviscosity syndrome due to IgG or IgA paraproteins, or cast nephropathy due to free light chain secretion . The assWe found that a median dose of 30 Gy was administered for successful palliation. Studies done in the prior era of treatment reported that even lower doses, e.g., 10–20 Gy over 1–2 weeks, can provide adequate pain relief . In the An important issue of RT in MM is its feasibility in transplant-eligible patients. Many MM experts avoid the use of RT - especially if directed to lumbar spine and pelvis – when they anticipate the need for collection of peripheral blood stem cells for autologous SCT, fearing toxicity of RT upon the hematopoietic stem cells. Published data on this subject remain controversial –30. In oFinally, regarding the development of secondary leukemias after RT, it is reassuring that we found no increased incidence of secondary malignancies among 149 RT-treated patients, albeit our median follow-up of 28 months is relatively short. Of note, all three cases of treatment-related AML observed in this time interval were found in patients who did not receive RT.In conclusion, despite the use of novel effective systemic agents and SCT, RT continues to play a prominent role in the palliative treatment of MM. In our series, approximately one-third of patients required the use of RT during the course of their disease, most commonly for pain control and prophylaxis/treatment of fractures and neurological complications. Radiation provided effective pain control in greater than 80% of patients. Exposure to RT did not compromise the feasibility of peripheral blood SCT, and it did not seem to increase the frequency of secondary malignancies.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Recent studies of genomic variation associated with autism have suggested the existence of extreme heterogeneity. Large-scale transcriptomics should complement these results to identify core molecular pathways underlying autism. Here we report results from a large-scale RNA sequencing effort, utilizing region-matched autism and control brains to identify neuronal and microglial genes robustly dysregulated in autism cortical brain. Remarkably, we note that a gene expression module corresponding to M2-activation states in microglia is negatively correlated with a differentially expressed neuronal module, implicating dysregulated microglial responses in concert with altered neuronal activity-dependent genes in autism brains. These observations provide pathways and candidate genes that highlight the interplay between innate immunity and neuronal activity in the aetiology of autism. Autism spectrum disorder (ASD) is a common, highly heritable neurodevelopmental condition characterized by marked genetic heterogeneity. In this study, the authors use RNA sequencing analyses to characterize differences in the transcriptome between autistic and typically developing brains. Autism is a neurodevelopmental disorder in which affected individuals show disruption of the normal course of social and communicative development, along with restrictive interests and repetitive behaviours. In the vast majority of cases, the cause of disease is not known. Despite a strong genetic component with heritability estimates between 50 and 90% refs , 2, 3, t4de novo variation contributing to autism. A major mode of action for genetic variation is through altered gene expression, so direct analysis of gene expression in a disease-relevant tissue is a complementary approach to genetic studies. Despite the extreme genetic heterogeneity observed in autism, it is possible that common downstream mechanisms may be alteredRecent studies to elucidate the molecular basis of autism have largely focused on genetic approaches, including both genome-wide association studies578et al. recently utilized co-expressed gene networks from RNA-Sequencing (RNA-Seq) carried out in 19 autism brains and 17 controls to identify a set of co-expressed neuronal genes enriched for known autism susceptibility genes as well as a set of co-expressed genes enriched for both immune genes and glial markersGiven low tissue sample availability in autism research, efforts have focused on assessing gene expression in lymphoblastoid cell lines or whole blood11121314In the current study, we present results from the largest RNA sequencing of autism brains effort to date that allows for new insights into the aetiology of autism. We find clear differences in the transcriptome between control and ASD cortical brains. Using co-expression network analysis, we demonstrate that autism brains are specifically enriched for ‘activated’ M2 microglial and ‘immune response’ genes. Remarkably, the M2 microglial module is strongly negatively correlated with one of two differentially expressed neuronal modules, highlighting the interplay between innate immunity and neuronal activity in the aetiology of ASD.Transcriptomes from 104 human brain cortical tissue samples were resolved using next-generation RNA sequencing technology at single-gene resolution and through co-expressing gene clusters or modules. Multiple cortical tissues corresponding to Brodmann Area 19 (BA19), Brodmann Area 10 (BA10) and Brodmann Area 44 (BA44) were sequenced in 62, 14 and 28 samples, respectively, resulting in a total of 57  control and 47  autism samples . DiffereMyelin And Lymphocyte Protein  annotations and MSigDB curated gene sets. No gene-set enrichment with family-wise error rate (FWER) ≤0.05 was observed .de novo variation (RDNV) and genes involved in intellectual disability (ID) compiled from four published whole-exome sequencing studies578For common genetic variation, altered gene expression is a major mode of action20P<0.005), with mod5 (P=9.64 × 10−4) exceeding the multitest correction threshold  and ‘cytokine-mediated signalling pathway’  and mod7 (Phypergeometric=1.45 × 10−89)—were enriched for asdM16 signal. However, mod7 is not differentially expressed with respect to autism . By substantially increasing the sample size and number of genes evaluated, we are able to accurately pinpoint the relevant signal from the previously reported asdM16 module as coming from M2-state microglial cells and immunogenic responses  and rare classes of autism genetic variants (Phypergeometric=4.29 × 10−4) but comparably expressed between cases and controls , and the non-differentially co-expressed neuronal module, mod2, which showed a substantially stronger enrichment of 39% of FMRP targets  in autism has been provided by Darnell et al. organized the FMRP target genes into distinct temporally expressed subpopulations affected by different classes of genetic variation associated with ASDPhypergeometric=4.66 × 10−4), whereas mod2 was enriched for the FMRP targets in the modules that were expressed during fetal development  and mod1 , with samples originating from two different sites: the Harvard Brain Tissue Resource Center and the NICHD Brain and Tissue Bank at the University of Maryland. Tissue was obtained from post-mortem and written informed consent was obtained from next-of-kin or a legal guardian. This work was approved by the Institutional Review Board of the Johns Hopkins Hospital and University of Alabama at Birmingham and was conducted in accordance with institutional guidelines. The brain samples were dissected to obtain the cerebral cortex Brodmann area (BA) 19, anterior prefrontal cortex (BA10) and a part of the frontal cortex (BA44). Multiple cortical tissues corresponding to BA19, BA10 and BA44 were sequenced in 62, 14 and 28 samples, respectively, resulting in a total of 57  control and 47  autism samples. The average age at time of death of the 40 control and 32 autism individuals was similar , and there was no significant difference in cause of death between the two groups. RNA-Seq libraries were prepared from 50 μg of total RNA from post-mortem brain tissue extracted with Trizol reagent according to the manufacturer’s protocol (Invitrogen). The TruSeq RNAseq kit (Illumina) was used with minor modifications as follows. Total RNA pools were subjected to two rounds of hybridization and elution with oligo(dT) dynabeads (Invitrogen) to obtain purified polyadenylated (polyA) RNA. After mRNA selection, samples were randomly fragmented to minimize bias at the 3′ end of the transcript. First-strand cDNA synthesis was performed using random primers (Illumina) and SuperScriptII Reverse-Transcriptase (Invitrogen) followed by second strand cDNA synthesis using RNaseH and DNA polymerase I (Illumina). Illumina supplied adaptors (TruSeq kit) were ligated to the purified, end-repaired and 3′ adenylated cDNA, and we performed manual 200 bp size-selection of the final product by gel-excision. The 200-bp cDNA template molecules were then amplified by PCR to create the final library. Quality control measures during library amplification included PCR from reactions with no template, from libraries made with no ligase (hence no adaptors) and finally from libraries with no adaptor oligonucleotides included in the ligase reaction. In these cases, the library failed to amplify, thereby ensuring specificity of the expected product for each run. Each library was evaluated for uniformity on a 2100 Bioanalyzer (Agilent) before sequencing on a single lane of Illumina’s HiSeq 2000 to produce 100 base pair (bp) single-end reads. Each sequencing run included samples randomized by sex, collection site and case–control status.N=3–12) and contaminating adaptor sequences using a Python script, `cutadapt' (v1.2.1)et al.http://www-huber.embl.de/users/anders/HTSeq/)The sequenced reads for each sample were obtained as fastq files for 110 samples. To improve mapping, reads were trimmed to remove stretches of terminal A or Ts  and sequencing depth by methods implemented in Conditional Quantile NormalizationWe assessed summarized values on a per-gene basis, removing gene estimates for samples whose gene expression values were more than three standard deviations (s.d.) from the mean expression of each gene (per-gene outlier), as these outliers are artefactual in originhttp://picard.sourceforge.net, v1.87) command-line tools ‘CollectRnaSeqMetrics’ and ‘CollectGcBiasMetrics’ were used to provide RNA-Seq summary statistics (`prcomp' function in the `stats' package in R (http://www.R-project.org/). All 104 samples were within three s.d. of the mean of the first six principal components, which together explained ~55% of the variancePicard , site of sample collection, age, sex, brain region and ISVs were included as fixed effects. In addition, the model included a random intercept term to account for the correlation of gene expression estimated from multiple brain regions obtained from the same individual.P=4.76 × 10−7). We reiterate that the samples were obtained from two collection sites, and to estimate the threshold for transcriptome-wide significant differential expression, we permutated the case–control status within each site, maintaining the same phenotype for multiple samples  derived from a single individual.Permutation testing was used to estimate the threshold for transcriptome-wide significant differential expression   .Exon-level estimates were obtained using the ‘count.py’ script from DEXSeqftp://ftp.ncbi.nlm.nih.gov/gene/DATA/gene2go.gz). The number of genes differentially expressed at P<0.05, P<0.01, P<0.001 and P<0.0001 were 1,964, 749, 185 and 50, respectively. For each P value cutoff, we generated 2,000 random gene sets of equal size  and performed the same enrichment analysis as on the original data set. Minimum P values for each enrichment analysis were stored. The 0.05 FWER was then calculated to estimate false positives by setting the 100th  best P value as the threshold for a true discovery.To determine a common functional relationship among the top differentially expressed genes, we tested for the enrichment of biological processes using GO annotations (http://www.broadinstitute.org/gsea/msigdb/collections.jsp#C1). The P value of each gene was determined from a linear mixed model. We then mapped these P values to non-negative z-scores(z_i), whereWe also used an alternate method for pathway enrichment analysis for the identification of common functional categories represented by GO and curated gene sets. The pathways include all the pathways in GO and curated gene sets, which can be downloaded from MsigDB  among genetically associated genes  as described above.We also tested whether the genes associated with the risk of autism and ID were differentially expressed. Gene lists are in ed genes . We obtaS), age (A) and brain region (BR), yielding an artefact corrected (AC) data set. The sequencing artefacts used in the model were ‘percent coding bases’, ‘percent utr bases’, ‘percent intronic bases’, ‘percent intergenic bases’, ‘median CV coverage’, ‘median 5′ to 3′ bias’, ‘aligned reads’ and ‘AT dropout’. The correction formula was  was chosen in this study. This power function weights the network by transforming the pairwise correlation values and computing pairwise topological overlap (TO) between genesWe investigated the entire AC data set to obtain gene sets or modules that were differentially co-expressed between autism cases and control brains using WGCNA−3. The permutation threshold, P<2.0 × 10−3, was determined by permuting case–control labels (n=100 permutations) for the eigengene values and re-running the regression analysis. The fifth lowest P value was deemed as the study-wide empirical threshold for P<0.05.Once the co-expression modules were created, they were numerically labelled by module size, with mod1 denoting the largest module. The co-expression analysis on 13,443 genes from the AC data identified 12 modules, with each module being represented by its first principal component or eigengene  for each sample. We tested the association of each eigengene with case–control status using a univariate linear mixed regression model, with a random intercept term to account for the correlation among multiple samples derived from the same individual. No additional variables are included in the analysis, as the modules were constructed using the AC data (described above). The multiple test correction threshold using the Bonferroni method was 4.0 × 10Most of the 13,443 genes were clustered into mutually exclusive co-expressing modules. However, 5,075 genes were assigned into the predefined mod0, which is reserved for non-module genes. All 13,443 genes and their module membership along with the correlation of the genes to each module (kME) are tabulated in We investigated the global similarity in transcriptome organization in the autism case and control brains by constructing signed networks for the autism case and control brains separately (power function threshold of 9). The construction of the signed networks separately for cases and controls identified 18 modules for each. We utilized the module preservation statistic ZsummaryWe functionally annotated the 12 modules with GO terms. The enrichment of the GO terms in each of the 12 modules was evaluated based on the hypergeometric test. To account for false positives, 12 random modules of the same size were generated 2,000 times, the hypergeometic test was carried out, and the 0.05 FWER was calculated. We have tabulated the GO term enrichment for each module at FWER=0.05 in www.arkinglab.org/resources). We present the enrichment of each module, with significance calculated based on the hypergeometric model and implemented in the GeneMerge software packageP value corresponding to FWER=0.05 is tabulated in To help provide insight into the interpretation of the gene expression data, we compiled gene sets that have been either implicated in ASD5152et. al. and are a manually curated set of candidate genes implicated by common variant association, candidate gene studies, genes within ASD-associated CNV, and, to a lesser extent, syndromic forms of ASD. This list from the SFARI AutDB was restricted to genes with strong genetic evidence by also filtering by the category S (syndromic) and evidence levels 1–4 . The ASD SFARI 2012 list excludes any exome sequencing-implicated RDNV genes; (ii) 235 genes (ASD SFARI 2014) from the SFARI AutDB databasede novo variant from the four whole-exome sequencing publications91011et al.et al. from four whole-exome sequencing publications578et al.de novo nonsense, frameshift or splice-site point mutations in autism probands that were obtained from Iossifov et al.et al.et al.et al.et al.As previously discussed with the single-gene analysis, the genes associated with autism and ID were taken from numerous sources presented in et al. identified two co-expression gene modules that were dysregulated in post-mortem ASD brains, asdM12  and asdM16 Voineagu FMRP interacting) implicated with translational regulation of synaptic proteinset al.FMRP 1 and FMRP 2), exhibit differential temporal expression: genes in FMRP 1 tend to be specifically upregulated during fetal development, whereas genes in FMRP 2 were generally upregulated in adolescence and adulthood. Numerous cell type markers were tested, as presented in et al. identified 3,955 exons mapping to 1,744 genes with high expression in the brain and a low burden of rare mutations, and designate these as ‘brain-critical exons’FMRP and its interacting partners .Accession codes: The RNA-Seq data have been deposited in the National Database for Autism Research (NDAR) under the accession code NDARCOL0002034.Supplementary Figures 1-3 and Supplementary ReferencesList of sample characteristics and phenotypesSamples with sequencing statistics as obtained from Picard tools3a: Correlation  of the ISV with sequencing artifacts.3b: Correlation  between the sequencing artifacts metrics.Differentially expressed genes.Differentially expressed exons.6a: Enrichment analysis of 749 genes differentially expressed at P < 0.01.6b: Enrichment analysis of 1964 genes differentially expressed at P < 0.05. 6c: Enrichment analysis of 185 genes differentially expressed at P < 0.001. 6d: Enrichment analysis of 50 genes differentially expressed at P < 0.0001Gene lists8a: Enrichment analysis of 1964, 749, 494 , 185 genes differentially expressed at P < 0.05, P < 0.01, P < 0.005, P < 0.001 , respectively 8b: Gene Set enrichment analysis to evaluate expression at the level of genetically associated genes.Module conservation between co-expressed modules constructed separately in cases and controlsGene and kME in different modulesAssociation of co-expression modules with disease statusModules and the enrichment of gene sets13a. mod1 and enrichment of Gene Ontology categories with FWER < 0.05. 13b. mod2 and enrichment of Gene Ontology categories with FWER < 0.05. 13c. mod3 and enrichment of Gene Ontology categories with FWER < 0.05. 13d. mod4 and enrichment of Gene Ontology categories with FWER < 0.05. 13e. mod5 and enrichment of Gene Ontology categories with FWER < 0.05. 13f. mod6 and enrichment of Gene Ontology categories with FWER < 0.05. 13g. mod7 and enrichment of Gene Ontology categories with FWER < 0.05. 13h. mod8 and enrichment of Gene Ontology categories with FWER < 0.05. 13i. mod9 and enrichment of Gene Ontology categories with FWER < 0.05. 13j. mod10 and enrichment of Gene Ontology categories with FWER < 0.05. 13k. mod11 and enrichment of Gene Ontology categories with FWER < 0.05.Correlation between the 12 module eigengenes (ME) in the network


Clostridium difficile infection (CDI) yet the immune deficiency predisposing to CDI is not well understood, despite an increasing incidence of CDI among such individuals. We aimed to estimate the incidence and to evaluate the risk factors of CDI among an HIV cohort in Italy.HIV infection is a risk factor for We conducted a retrospective case-control (1:2) study. Clinical records of HIV inpatients admitted to the National Institute for Infectious Disease “L. Spallanzani”, Rome, were reviewed (2002-2013). Cases: HIV inpatients with HO-HCFA CDI, and controls: HIV inpatients without CDI, were matched by gender and age. Logistic regression was used to identify risk factors associated with CDI.We found 79 CDI episodes . The mean age of cases was 46 years. At univariate analysis factors associated with CDI included: antimycobacterial drug exposure, treatment for Pneumocystis pneumonia, acid suppressant exposure, previous hospitalization, antibiotic exposure, low CD4 cell count, high Charlson score, low creatinine, low albumin and low gammaglobulin level. Using multivariate analysis, lower gammaglobulin level and low serum albumin at admission were independently associated with CDI among HIV-infected patients.Low gammaglobulin and low albumin levels at admission are associated with an increased risk of developing CDI. A deficiency in humoral immunity appears to play a major role in the development of CDI. The potential protective role of albumin warrants further investigation. Clostridium difficile infection (CDI) is increasing worldwide in both the general population and immunocompromised individuals [The incidence of ividuals -4.Several factors have been associated with the risk of CDI development, including: older age, antibiotic exposure, acid suppressant exposure, inflammatory bowel disease and immunosuppression -9. ImmunOur aim was to estimate the incidence of CDI among HIV hospitalized patients, and to evaluate the associated risk factors.We collected data from 2002 to 2013 on CDI among HIV-infected inpatients in our hospital, National Institute for Infectious Diseases “L. Spallanzani”, that is a a referral center for HIV infected individuals in our region of approximately 5.5 million inhabitants. CDI cumulative incidence was expressed as CDI episodes per 1000 hospital admissions of HIV-infected patients. CDI incidence rate was expressed as the number of CDI episodes per 10000 patient-days among HIV-infected inpatients. Also data on CDI among non HIV-infected inpatients were collected in the same period, in order to compare the CDI trend in both the groups, HIV-infected and not infected. In this case CDI cumulative incidence was expressed as CDI episodes per 1000 hospital admissions of non-HIV infected patients and CDI incidence rate was expressed as the number of CDI episodes per 10000 patient-days among non HIV-infected inpatients.C. difficile. Controls were randomly selected and matched for gender, age (±5 years) and year of admission with cases.Moreover, we conducted a retrospective case-control (1:2) study on adult HIV-infected patients. Cases were defined as HIV inpatients with CDI; controls were HIV inpatients without CDI that were hospitalized for at least 48 hours. Controls were patients without diarrhea or with diarrhea but with negative toxin test for Only healthcare facility (HCF)-onset, HCF-associated (HO-HCFA) CDI were included in the study. HO-HCFA CDI is defined as an episode of CDI occurring between 48 hours after admission and hospital discharge .C. difficile toxin assay in a stool sample from a patient with diarrhea. Diarrhea was defined as ≥ 3 unformed stools in a 24-hour period. The presence of toxin A and B was tested through enzyme immunoassays (EIA) for A/B toxins . CDI relapses were excluded from the analysis.A CDI episode was considered as a positive Demographic data , fever (i.e. temperature > 38.3°C) and biochemical parameters  measured at admission were recorded. Gammaglobulin levels were calculated through seroprotein electrophoresis from total proteins. Normal range for serum albumin was 3.5-5.5 g/dl and for serum gammaglobulin 800-1600 mg/dl according to our laboratory. In addition the following characteristics were compared between cases and controls: Charlson’s score index, length of hospital stay, antiretroviral treatment, antimycobacterial drug exposure, prophylaxis with trimethoprim-sulfamethoxazole (TMP-SMX) and/or azithromycin, chronic hepatitis C virus (HCV) infection, treatment for Pneumocystis pneumonia, antineoplastic chemotherapy, acid suppressants, prior hospitalization (in the previous 8 weeks), prior antibiotic exposure (in the previous 8 weeks), antibiotic exposure during hospitalization and methadone treatment. The recorded data were available for all cases and controls.The study involved the analysis of existing clinical and laboratory data that were anonymised before being included in the study database. The study was approved by the Ethics Committee of the National Institute for Infectious Diseases “L. Spallanzani”.Continuous variables are expressed as mean ± SD and categorical variables as percentages.Univariate analysis was performed for each recorded variable, with the exception of mortality and length of stay since such variables could be both a risk factor and/or a consequence of CDI. Odds ratio (OR), with 95% confidence interval, was calculated for qualitative variables; quantitative variables with normal distribution were compared using the Student’s t test.Multivariate regression analysis included all statistically significant variables in univariate analysis and all clinically relevant variables, whether statistically significant or not . The finThe Hosmer and Lemeshow test was applied to estimate the goodness of fit for the model.The statistical significance was set at P < 0,05.The analysis was performed using STATA 11 MP .From January 2002 to December 2013, 15 537 HIV-infected patients were admitted to our Institute. The distribution of HIV admission during this time is shown in Figure Seventy-nine cases of HO-HCFA CDI were identified among HIV-infected inpatients during the study period. Cumulative incidence was 5.1 per 1000 HIV hospital admissions, with an incidence rate of 3.4 CDI cases per 10,000 HIV patient-days. Figure Among CDI cases, 56 were male (71%), 23 were female (29%) and the mean age was 46 years. Forty-three percent of CDI cases had fever at diagnosis of CDI and the mean white cell count at diagnosis was 7061 per cubic millimeter (SD ±5167).Regarding antibiotic exposure, carbapenem, fluoroquinolone and rifamycin exposures were significantly more prevalent among cases compared to controls Table .Table 1Regimens used to treat CDI were metronidazole alone for 42 patients (53%), oral vancomycin alone for 11 patients (14%). Fourteen patients received both vancomycin and metronidazole (18%) and 12 patients (15% of cases) received no CDI treatment during the hospital stay because of death (1 patient) or voluntary discharge (11 patients).The mean duration of diarrhea among CDI cases was 13 days (SD ±12).CDI occurred, on average, 12.7 days after admission , with a median of 8 days after the admission (IQR 4-16). The time to onset of diarrhea during the hospital stay (from 48 hours after the admission to the discharge) is illustrated in Figure The risk factors analyzed for the 79 CDI cases and 158 matched control subjects are shown in Table On multivariable analysis Table  low gammEleven patients (13.9%) among cases and 10 patients (6.3%) among controls died during the hospital stay (p = 0.05). In 3 patients, in-hospital death was attributed to CDI.C. difficile among hospitalized HIV-infected patients [HIV infection is an established risk factor for CDI and the introduction of HAART has not effectively reduced the risk of patients ,16. Althpatients , in thispatients -20.et al. found that CD4 count was significantly associated with CDI among HIV-infected patients, but gammaglobulin levels were not evaluated among risk factors [However, in a study published in 1997, Barbut  factors . SimilarC. difficile toxins, is now believed to be a major determinant of the consequences of C. difficile acquisition [Host immunity, especially humoral responses to uisition -24. Indeuisition .C. difficile infections with advanced age and immunodeficiency-associated conditions, including HIV infection [per se is not significantly associated with development of CDI [C. difficile disease.Variation in antibody response may be a principal explanation for the association of symptomatic nfection . HIV infnfection ,27. The nfection ,29. In st of CDI ,21. TherC. difficile strains were often sensitive to rifamycins  [C. difficile strains, mainly ribotype 027, are resistant to rifamycins [In the 1990s, fabutin) , yet morfamycins . Therefofamycins  we thinket al. in non-HIV patients [Low serum albumin is a well known risk factor for CDI ,34. In opatients .C. difficile strain BI/NAP1/027. In support of this, the first case reports of C. difficile 027 in Italy were published in January 2010 [In this study, we observed a reduction in CDI incidence from 2002 to 2008 and a progressive increase from 2008 to 2013 that was similar to that observed among non-HIV patients hospitalized between 2008 and 2013 , therefore CDI episodes since this time could be potentially over-represented. In addition we only used the EIA toxins detection as diagnostic test for CDI; this test has a low sensitivity therefore it is possible that patients with true CDI but with false negative toxin EIA may have been inappropriately included as control. Moreover we did not perform culture for In our study, lower gammaglobulin and albumin levels are associated with the developing of CDI. It remains vital that the immune constitutents that play a role in susceptibility to CDI are further elucidated. Strategies to improve humoral immunity and albumin status in patients with immune dysfunction should be further explored in an effort to reduce the incidence of CDI.


Objective. This study aimed to evaluate the association between serum uric acid (SUA) levels within a normal to high range and the risk of metabolic syndrome (MetS) among community elderly and explore the sex difference. Design and Methods. A cross-sectional study was conducted in a representative urban area of Beijing between 2009 and 2010. A two-stage stratified clustering sampling method was used and 2102 elderly participants were included. Results. The prevalence of hyperuricemia and MetS was 16.7% and 59.1%, respectively. There was a strong association between hyperuricemia and four components of MetS in women and three components in men. Multiple logistic regression analysis showed ORs of hyperuricemia for MetS were 1.67 (95% CI: 1.11–2.50) in men and 2.73 (95% CI: 1.81–4.11) in women. Even in the normal range, the ORs for MetS increased gradually according to SUA levels. MetS component number also showed an increasing trend across SUA quartile in both sexes (P for trend < 0.01). Conclusion. This study suggests that higher SUA levels, even in the normal range, are positively associated with MetS among Chinese community elderly, and the association is stronger in women than men. Physicians should recognize MetS as a frequent comorbidity of hyperuricemia and take early action to prevent subsequent disease burden. Metabolic syndrome (MetS) is a complex of interrelated risk factors including obesity , hyperglycemia, elevated blood pressure, hypertriglyceridemia, and decreased high density-lipoprotein cholesterol (HDL-C) . CurrentSerum uric acid (SUA) is the end product of urine metabolism in human, and excess accumulation can lead to various diseases . The relMoreover, a rapidly increasing trend of MetS prevalence has been reported in the Chinese elderly in the past decade, with an absolute change of 7.7% . TherefoThis was a population-based cross-sectional survey conducted in Wanshoulu Community of Haidian district, a metropolitan area representative of the geographic and economic characteristics in Beijing, China. The eligible subjects were individuals aged ≥60 years who had lived in the local district for at least one year. Sampling and research methods were reported elsewhere . In brieEach participant was interviewed and completed a standardized questionnaire including demographic factors, medical history, family history of CVD, and lifestyles. The height, weight, waist, and blood pressure were measured according to standardized protocol. Physical examinations and face-to-face interviews were carried out by specially trained nurses and physicians. Height was measured in meters (without shoes), and weight was measured in kilograms (with heavy clothing removed and 1 kg deducted for remaining garments). Body mass index (BMI) was calculated as weight in kilograms divided by the square of height in meters. Waist circumference on standing participants was measured midway between the lower rib margin and iliac crest. Two blood pressure recordings were obtained from the right arm of participants in a sitting position after 30 minutes of rest. If the difference between the first and second measurement was more than 5 mmHg, then repeated measurements were performed. The average of last two measurements was used. Overnight fasting blood specimens were obtained for tests of serum lipids, glucose, and SUA level. Samples were sent to the Central Certified Laboratory of Chinese PLA General Hospital in less than 30 minutes.25, P50, and P75) as cut-off values. In this study, sex-specific SUA quartiles for participants without hyperuricemia were as follows: Q1: ≤4.71 mg/dL (n = 176), Q2: 4.72–5.43 mg/dL (n = 176), Q3: 5.44–6.48 mg/dL (n = 175), Q4: ≥6.49 mg/dL (n = 173) in men; Q1: ≤3.90 mg/dL (n = 267), Q2: 3.91 mg/dL–4.54 mg/dL (n = 263), Q3: 4.55 mg/dL–5.10 mg/dL (n = 260), Q4: ≥5.11 mg/dL (n = 260) in women. All the four groups were under normal range of SUA level.Participants were diagnosed with hyperuricemia if their SUA level was ≥7.0 mg/dL (417 mmol/L) in men or ≥6.0 mg/dL (357 mmol/L) in women. SUA levels of participants without hyperuricemia were categorized into 4 levels using the quartiles  central obesity: waist circumference (WC) ≥ 90 cm in Asian men and ≥80 cm in Asian women; (2) hypertriglyceridemia: fasting plasma triglycerides (TG) ≥1.7 mmol/L; drug treatment for elevated triglycerides is an alternate indicator; (3) decreased HDL-C: fasting HDL-C <1.0 mmol/L in men and <1.3 mmol/L in women; drug treatment for reduced HDL-C is an alternate indicator; (4) elevated blood pressure: systolic blood pressure (SBP) ≥ 130 mmHg and/or diastolic blood pressure (DBP) ≥ 85 mmHg; antihypertensive drug treatment in a patient with a history of hypertension is an alternate indicator; (5) hyperglycemia: fasting glucose level of ≥5.6 mmol/L (≥100 mg/dL); drug treatment of elevated glucose is an alternate indicator  central .Data were double entered using Epidata 3.1. All analyses were conducted using SPSS for windows . Reported probabilities were two-sided; all tests were set at the 0.05 level of statistical significance.n(%) for categorical variables unless otherwise specified. t-test and Chi-square test were used to examine differences in continuous and categorical variables. The relationship between SUA levels and other variables was assessed using the partial correlation coefficients. The trend test was used to determine MetS prevalence and its component number according to SUA quartiles. Finally, multivariable logistic regression was used to estimate the association between sex-specific SUA level and MetS prevalence. We calculated the odds ratio (OR) and 95% confidence intervals (CIs) of SUA for MetS and its five components.Descriptive data were expressed as Ethical approval was obtained from the Ethics Committees of Chinese PLA General Hospital (EC0411-2001). All eligible participants had given their written informed consent.A total of 2,102 participants completed the survey, with 848 (40.3%) men and 1254 (59.7%) women. The mean age was 71.2 ± 6.6 (60~95 yrs); the older elderly (aged ≥ 80 yrs) accounted for 9.2%. The baseline characteristics of participants according to sex and hyperuricemia are presented in P = 0.097).The prevalence of MetS and hyperuricemia in this community's elderly was 59.1%, and 16.7%, respectively. The percentage of participants who had both MetS and hyperuricemia was 12.4%; men were lower than women but with no statistical difference . Men had a higher prevalence in the younger elderly, but in the participants aged more than 70s, women had a higher prevalence.P < 0.05). The number of MetS components also showed the same trend , and, in women, four individual components (excluding elevated blood pressure) showed significant difference in participants with and without hyperuricemia (me trend .P for trend <0.05 in both sexes) for those with SUA level under normal range. The number of MetS components also increased gradually with increasing SUA quartiles. In men, three of the individual components (excluding elevated blood pressure and hyperglycemia) and, in women, all five components showed increasing trends according to SUA quartiles (P for trend <0.05).There were sex differences of the ORs for MetS. Women had higher ORs of SUA level for MetS than men. The ORs for elevated blood pressure and hyperglycemia were not statistically significant in men using all three forms of SUA, but in women the ORs for the two components were significant using SUA levels as continuous variable and quartiles.n = 105, 5.0%) were excluded, the adjusted ORs of hyperuricemia for MetS were 1.64 (95 CI: 1.09–2.47) in men and 2.68 (95 CI: 1.78–4.03) in women , the adjusted ORs of hyperuricemia for MetS were 1.96 (95% CI: 1.26–3.06) in men and 2.55 (95% CI: 1.63–3.99) in women.We also ascertained the association of SUA level and MetS in the sensitivity analysis and the results were similar Tables  and 7. Win women . Also, iIn this study, we evaluated the association of SUA level and MetS in a Chinese community elderly population. We found that higher SUA level, even within normal range, was associated with an increased prevalence of MetS even after adjusting for confounding factors. This association of MetS risk by SUA levels was more robust in women than in men.MetS and CVD are rapidly growing threats to public health worldwide, especially in economically developing countries such as China. Just in the past decade, prevalence of MetS in China has increased about 8% . ConsideA number of studies have researched the association between SUA levels and MetS or its individual components, primarily in adults or institutionalized people. There is little information about the situation in the community elderly, a population with higher prevalence of hyperuricemia and MetS and higher risk of developing CVD events. In this study, we investigated the main components of MetS in the aging population and their potential association with SUA levels. In this representative sample of the Chinese urban elderly, we found a graded positive association between hyperuricemia and the prevalence of MetS. Even in the normal range, SUA level still has higher remarkable increased risk of MetS. And MetS component number was found to rise along with the increase of SUA levels too. This is in accordance with previous studies. Several studies reported that there are significant associations of hyperuricemia with visceral or central obesity, high BMI, TGs, and vascular endothelial dysfunction . A cohorP = 0.475). After we calculate the age-specific prevalence, we can see men had a higher prevalence in the younger elderly, while in the participants aged more than 70s, the prevalence of hyperuricemia in women was higher than men. This is contrary to previous studies focused on adults [The prevalence of hyperuricemia was 16.2% in this population, with 17.5% in men and 16.3% in women, but with no significant difference (n adults , 20. A mn adults . Differen adults . Gouty an adults , 24.The present study also provides evidence that elevated SUA level was more strongly associated with MetS in women than in men. Previous studies that performed sex-specific analyses also reported similar results. Sex is clearly an important factor in the relationship between hyperuricemia and MetS. Women with a higher SUA concentration had a higher incidence of hypertension, hypertriglyceridemia, and low HDL-C, as well as increased cardiovascular morbidity and mortality compared to that in men . A cohorThis is a population-based cross-sectional study with strict training process and quality assurance programs. Wanshoulu district is a representative metropolitan area of the geographic and economic characteristics in Beijing, such as income levels, residential status, and lifestyle factors. The response rate was high and there was no statistically significant difference between participants who completed and those who did not complete the data. Consequently, the findings are more representative than the previous ones, which are mainly about institutionalized patients or health screeners, and potentially reduce the selection bias. Second, our study showed that higher SUA levels, even in the normal range, are strongly associated with the prevalence of MetS. Most studies have revealed the association between obvious hyperuricemia and MetS. The results of our study showed higher SUA levels, even not high enough to the criteria of hyperuricemia, are of elevated risk of MetS. Third, this study explored the sex difference between the association of SUA level and prevalence of MetS, and a stronger association was revealed in women.There are several limitations in this study which need to be considered. First, the nature of cross-sectional study did not allow us to assess the temporal relationship. Further cohort study with follow-up data is needed to verify the results. Second, the study only included participants who were aged ≥60 years and therefore may not be generalized to young people. But on the other hand, this aged population has a higher prevalence of developing MetS and CVD and would bring on bigger disease burden, so as to give us a critical alert to pay more attention to uric acid in the elderly with MetS and take early prevention actions to prevent serious CVD events. Third, the sex difference in the association of SUA level and MetS risk needs future investigations to explore the underlying mechanisms. Also, we did not have the information whether the women included in the study were taking hormone replacement therapy, which would have impact on both SUA level and MetS prevalence.In summary, the present study showed a strong association between SUA and prevalence of MetS in Chinese community elderly population living in urban Beijing, even in the normal range of SUA level. It also provides additional evidence that SUA is more closely associated with the risk of developing MetS in women than in men. Physicians should recognize MetS as a frequent comorbidity of hyperuricemia and take early action to prevent subsequent chronic disease and related potential socioeconomic burden in the Chinese elderly.


Cardiovascular disease (CVD) results from a combination of abnormalities in lipoprotein metabolism, oxidative stress, chronic inflammation, and susceptibility to thrombosis. Atherosclerosis is the major cause of CVD. CD36 has been shown to play a critical role in the development of atherosclerotic lesions by its capacity to bind and promote endocytosis of oxidized low-density lipoprotein (oxLDL) and is implicated in the formation of foam cells. The purpose of this research was to evaluate whether there is an association of sCD36 and oxLDL levels with cardiovascular risk factors in young subjects.A total of 188 subjects, 18 to 25 years old, 133 normal-weight and 55 obese subjects from the state of Guerrero, Mexico were recruited in the study. The lipid profile and glucose levels were measured by enzymatic colorimetric assays. Enzyme-linked immunosorbant assays (ELISA) for oxLDL and sCD36 were performed. Statistical analyses of data were performed with Wilcoxon- Mann Whitney and chi-square tests as well as with multinomial regression.P < 0.05). Obese subjects had 5.8 times higher risk of sCD36 in the third tertil (>97.8 ng/mL) than normal-weight controls (P = 0.014), and 7.4 times higher risk of oxLDL levels in third tertile (>48 U/L) than control group. The subjects with hypercholesterolemia, hypertriglyceridemia, fasting impaired LDL-C had a higher risk of oxLDL levels in the third tertile (>48 U/L) than the control group (P < 0.05).TC, LDL-C, TG, oxLDL and sCD36 levels were higher in obese subjects than in normal-weight controls, as well as, monocyte and platelet counts . Cardiovascular disease (CVD) risk factors such as advanced age, obesity, smoking, hyperlipidemia, diabetes, and hypertension, account for 30–40% of the worldwide prevalence of this disease . The patA key process in the pathogenesis of atherosclerosis is the deposition of cholesterol in the arterial wall. Lipoproteins are involved in this process, including cholesterol carried by very low-density (VLDL), remnant lipoproteins and low-density lipoproteins (LDL), particularly the small and dense forms; conversely, cholesterol is carried away from the arterial wall by high-density lipoprotein (HDL) .Oxidative modification of LDL (oxLDL) in the arterial wall is central to the pathogenesis of atherosclerosis . OxLDL iCD36, an 88 kDa glycoprotein, was originally described as platelet receptor glycoprotein which belongs to the class B scavenger receptor family ,12. CD36Monocyte/macrophage CD36 has been shown to play a critical role in the development of atherosclerotic lesions by its capacity to bind and promote endocytosis of oxLDL, and is also implicated in the formation of foam cells ,19. MoreA soluble form of CD36 (sCD36), a marker of altered tissue CD36 expression, was recently identified in human plasma, and elevated levels were found in obesity and type 2 diabetes. In this regard, sCD36 in plasma has been reported up to 4-fold higher in obese T2D-patients compared to lean healthy control subjects . The cirA total of 188 subjects were randomly selected, 18 to 25 years old, 133 normal-weight controls (BMI 18.5 to 24.9 kg/m2) and 55 obese subjects (BMI ≥30 kg/m2) from the state of Guerrero, Mexico. There were 117 women and 71 men; participants were not under any medication or had evidence of metabolic disease other than obesity. All subjects gave their written informed consent previous explanation of the purpose and nature of the study. The protocol was approved by the Research Ethics Committee of the University of Guerrero.Blood pressure was measured in the sitting position with the use of an automatic sphygmomanometer on the left arm after 10 min rest. The systolic blood pressure (SBP) and diastolic blood pressure (DBP) were calculated from two readings with a minimal interval of 10 min. Hypertension was defined as mean SBP ≥140 mmHg and/or DBP ≥90 mmHg .A venous blood sample of 5 mL was obtained from each subject after at least a 12 hours fasting. All serum lipid levels and glucose were determined by enzymatic methods with commercially available kits (Spinreact). Abnormal biochemical levels were identified when total-cholesterol (TC) ≥ 200 mg/dL, triglycerides (TG) ≥ 150 mg/dL, low-density lipoprotein cholesterol (LDL-C) > 100 mg/dL, high-density lipoprotein cholesterol (HDL-C) < 40 mg/dL and glucose >100 mg/dL, based on the criteria of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) .Enzyme-linked immunosorbant assays (ELISA) for oxLDL (Mercodia Oxidized LDL ELISA) and sCD36  were performed, according to the manufacturer’s instructions, with an intra-assay CV <6% and interassay CV <7% for oxLDL ELISA assay, and an intra-assay CV <5% and interassay CV <9% for CD36 ELISA assay.P value <0.05 was considered statistically significant.The statistical analyses were performed with the statistical software package SPSS 15.0 and STATA software 9.0. Quantitative variables were expressed as medians and 25th to 75th percentiles and significant differences between groups were determined using the Wilcoxon-Mann Whitney test. Qualitative variables were expressed as percentages and the differences between groups were determined using the chi-square test. The association analysis between serum levels of oxLDL and sCD36 with lipid phenotypes was carried out with multinomial regression, where a P < 0.05). There were no significant differences by gender (P = 0.162).General and biochemical characteristics of the study subjects are shown in Table P = 0.028)   Table .P = 0.014) , since there are no established reference values. Obese subjects had 5.8 times higher risk of sCD36 in the third tertile than normal-weight controls, adjusted for age and gender  than in normal weight subjects (6.1%), with a prevalence of 13.6% in the total study subjects. These findings are similar to those reported in another study in U.S. teenagers; where hypertension showed a prevalence of 30% in obese subjects . It is kIn this study, as in previous studies the lipid profile  was higher in obese subjects than normal-weight controls. This may be due to increased adiposity since adipose tissue undergoes morphological and physiological changes which include the release of proinflammatory cytokines such as tumour necrosis factor alpha (TNF-α), and in turn, decrease insulin sensitivity and increase lipolysis. These morphological changes contribute to insulin resistance and dyslipidemia .An interesting finding in this study was that obese subjects have a higher number of monocytes and platelets than the normal-weight subjects. The leukocyte count is considered as an indicator of inflammatory status in obesity . In addiP = 0.002), these results are congruent with previous studies [In this study, we observed that sCD36 was higher in obese subjects than in normal weight subjects , similar results have been reported in previous studies [We observed a high correlation of sCD36 with BMI in obese subjects  were associated with obesity, although there is a lack of studies that support this association, sCD36 have been correlated with BMI ,50. HighWe also found that oxLDL levels in third tertile >48.0 U/L) were associated with hypercholesterolemia, impaired fasting LDL-C, hypertriglyceridemia and obesity. The association between dyslipidemia and oxidation of LDL has been demonstrated in individuals in the pre-diabetic state .0 U/L we. The assOur research has some limitations. We could not study the associations between sCD36 and oxLDL levels with early atherosclerosis. As our study is comprised of young subjects without clinical atherosclerotic diseases, we were only able to study associations between traditional cardiovascular risk factors with sCD36 and oxLDL levels. Whether the increase of these markers in young subjects is associated with the silent phase of atherosclerosis remains to be elucidated. The authors believe that a greater obese group is desirable to improve the power of the study.In this research, sCD36 and oxLDL levels are associated with cardiovascular risk factors, particularly with obesity, hypercholesterolemia, impaired fasting LDL-C and HDL-C and hypertriglyceridemia. Therefore, sCD36 and oxLDL levels may be potential early markers for CVD. However, these associations should be investigated in further studies.The authors declare that they have no competing interests.LERA performed ELISA assays, statistical analysis and writing the manuscript. UDCM carried out the immunoassays. ABSB performed biochemical measurements and quality control. JFMV and NCA participated in the critical revision of the manuscript. IPR conceived the study and participated in manuscript preparation. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2261/14/54/prepub


Patients undergoing SFPG treatment showed significant (P < 0.05), durable improvement of three major clinical indices of disease severity: reduced bleeding (day 7), plaque and gingival conditions (day 14), and consistent gingival pocket depth reduction (day 45). Proinflammatory nitric oxide metabolites reached normal values in plasma (day 14) and gingival crevicular fluid (GCF) at day 45 with SFPG applications compared to controls that did not reach normalisation. Levels of highly increased proinflammatory  and suppressed anti-inflammatory (IL-10) cytokines normalised in the SFPG group by days 14 (plasma) and 45 (GCF), but never in the control group. Although not acting directly as antibiotic, SFPG acted in synergy with human granulocytes blocking adaptive catalase induction in S. aureus in response to granulocyte-derived oxidative stress, thus enhancing intracellular bacterial killing.The clinical efficacy of topical administration of standardised fermented papaya gel (SFPG), known to have antioxidant and anti-inflammatory properties, Periodo Porphyromonas gingivalis [The chronic inflammation of periodontal disease is induced by polymicrobial infection , 11, witngivalis . The sevngivalis , which mngivalis . The micngivalis –18. For ngivalis , 19–21.α (TNF-α) and interleukin-6 (IL-6), in the gingival fluid and tissue, that define the severity of destructive processes in both the gingival epithelium and bone tissue [κB) ligand, were closely associated with total oxidant balance in serum and gingival crevicular fluid (GCF) [The presence of distinct microbes in the periodontal environment has been associated to increased levels of host-produced proinflammatory cytokines, such as tumour necrosis factor-e tissue . Interlee tissue . It has e tissue  and/or le tissue  may affee tissue –28. PeriAlthough many efforts have been made to develop clinically effective and cost-effective therapies, none has proven to have sufficient efficacy, and the urgent need for better treatments and preventive strategies remains. in vitro and in vivo experiments [In the folk medicine of South East Asia, papaya preparations have been traditionally used as antimicrobial, anti-inflammatory, and wound healing remedies. Standardised fermented papaya preparations have shown remarkable free radical scavenging, antioxidant, anti-inflammatory, phagocyte-stimulating, and wound healing activities in theeriments , 30, as eriments . Of parteriments , 33. In The study enrolled a group of 84 patients of both sexes (age range 38–62 years) suffering from chronic moderate-to-severe periodontitis and visiting dentists at Medical Department of the Kabardino-Balkar Berbekov's State University . The study protocol was scrutinised and approved by the local Ethical Committee (Protocol MD-023-2013). The patients were randomly assigned to experimental or control groups. The demographic distribution of periodontitis patients in the groups is shown in Traditional treatment protocols included education to oral hygiene, plaque removal, teeth enamel polishing, and surgical elimination of tartar. In addition, local instillations of 0.06% chlorohexidine gluconate into periodontal pockets were regularly performed as antimicrobial and anti-inflammatory procedures.The patients of the experimental group additionally received intragingival pocket applications (7 g of SFPG per patient per day) for 10 consecutive days. The SFPG was kept inside the pocket for 15 min and then washed out by physiological solution. Since SFPG consists of a complex mixture of fermented papaya fruit ingredients each of which possesses a biological activity, and water as a solvent of these organic active agents of plant origin, we omitted the use of pure water as a placebo in the controls group.n = 25) were recruited from the Medical Department staff and trainees, who donated blood and GCF after signing the informed consent form.Healthy donors matched by sex and age . PMN from the interface were resuspended in phosphate buffer saline, centrifuged at 1650 rpm for 10 min, and then aliquoted at 5 × 106/vial. Freshly isolated PMN were used in phagocytosis assays and measurements of reactive oxygen species production. Samples of whole blood (5 mL) were allowed to sediment for 40 min at room temperature and then plasma supernatant was collected, aliquoted, and stored at −80°C until quantitative analyses of nitrites/nitrates and cytokines.Peripheral venous blood (20 mL) was drawn after overnight fasting into vacutainers with ethylene diamine tetraacetic disodium salt (EDTA) as anticoagulant. Both patient and donor samples were processed and analysed in parallel. Circulating polymorphonuclear leukocytes (PNM) were obtained by double density gradient centrifugation of 15 mL of total blood ; kits for enzyme activities and nitrite/nitrate assays were from Cayman Chem. Co. ; monoclonal antibodies for ELISA interleukin kits were from R&D Systems ; kits for protein determination were from Bio-Rad Laboratories .The majority of chemical reagents and solvents, H Staphylococcus aureus strains 241 and 242 , 548 , and 556 . S. aureus were grown in tryptic soy broth at 37°C under continuous shaking as described previously [9 cells/mL) was mixed with the same volume of 0.9% SFPG and coincubated for 1 hour at 37°C under shaking conditions at 180 rpm. Finally, S. aureus cultures were thoroughly washed trice by big volumes of fresh physiological solution.Bacterial strains used in this study were as follows:eviously . To asse6 cells/mL) was mixed with 1 mL of bacterial suspension (107 cells/mL). The mixture was incubated under continuous shaking at 37°C for 30 min. Then, smears were prepared on microscopic slides, fixed, and stained by Romanowsky-Giemsa dye. The smears were examined under microscope, and percent of phagocytozing PMN was determined. The remaining mixture was used to assess intracellular bacterial killing. After centrifugation at 1500 ×g for 10 min, bacterial sediments were collected and diluted to an OD600 of 0.1 with fresh medium, spread onto Petri dishes with appropriate medium with agar, and were allowed to grow at 37°C for 24 hours. Bacteria survival rates were calculated as colony-forming-units (CFU) of cells coincubated with granulocytes, divided for that of untreated bacteria. The results were expressed in percent.Intensity of phagocytosis was assessed by routine clinical bacteriology methods. Briefly, 1 mL of PMN suspension . One unit of catalase activity was defined as the decomposition of 1 μmol H2O2 per min at pH 7.0 at 25°C. The results were expressed in Units/g protein.Bacterial cells pretreated and nonpretreated with SFPG were grown in appropriate mediums to an ODs method , using aμL or cps/106 cells, resp.) response to different strains of S. aureus was quantified by a Victor2 1420 counter, equipped with Wallac 1420 Software . In brief, 105 cells or 10 μL of whole blood was added to 1 mL of preheated Hank's Balanced Salt Solution (pH 7.4) containing 0.2 mM luminol. Then, 107/mL of S. aureus was added and the chemiluminescence response was monitored for 5 min. Each measurement was repeated three–five times. Results were expressed in cps per 1 mL of whole blood or 106 PMN.Whole blood and PMN luminol-dependent chemiluminescence  dependent cytochrome c reduction method described previously  and expr2−/NO3−, expressed as μmoles/L) were measured spectrophotometrically by Griess reagent kit, following manufacturer's instructions.Plasma and GCF levels of nitrites/nitrates  IL-6, and interleukin-10 (IL-10) were measured by enzyme-linked immunosorbent assay (ELISA) purchased from R&D Systems , following manufacturer's instructions. Cytokine concentrations were expressed in pg/mL of plasma, and each protein factor was quantified in the linear range of its calibration curve.The plasma and GCF levels of interleukin-1U test for independent samples was employed for comparison between periodontitis groups and healthy controls. Significance was assumed at a P value < 0.05.All biochemical measurements were done in triplicate and data were statistically evaluated. Statistical analysis of clinical and laboratory data was performed using STATISTICA 6.0 program . Reported data were treated as continuous. Normality of data was checked using the Shapiro-Wilk test. Since the distribution of the data in the three groups was significantly different from normal, nonparametric statistics was used. Values were presented as mean ± standard error of the mean of triplicate analyses. The Mann-Whitney P < 0.05) difference of Muhlemann's index  between the group treated additionally with SFPG and the control one appeared by the 7th day, was observed for the entire period of SFPG administration (10 days), and persisted in the follow-up period at the 14th and 45th days  they were found later, by day 45 during the follow-up period  proved to be nontoxic and well tolerated by the patients, among which no adverse effect or withdrawal from trial was registered throughout the whole study. Recruited patients assigned to both control and experimental groups exhibited clinical features of moderate-to-severe periodontitis that was revealed by visual clinical observation and by assessment with conventional objective indexes of gingival and periodontal conditions, that is, PMA, Silness-Löe, and Muhlemann's indexes . The bas5th days . Similar5th days , while fp period .Decreased levels of periodontal bone resorption were evident for the experimental group patients by day 45, when second orthopanoramic images were registered, while no differences in periodontal bone conditions were observed in the control group of patients during the study course (data not shown).n = 25) matching in age and sex. The concentration of NO2−/NO3− in GCF was highly elevated in periodontitis versus normal values  between the GCF levels of NO2−/NO3− for two groups reached statistical significance by the 14th day of the trial.The dynamics of nitrite/nitrate levels in GCF and blood serum is shown in l values . Also, psubjects . At the β and IL-6) and of the anti-inflammatory cytokine IL-10 are shown in Figures β and IL-6 were highly and equally elevated . When human granulocytes were mixed with several strains of S. aureus (the ratio of human cells to microbes was 1 : 100) which differed by catalase expression, all these strains even without opsonisation induced ROS production by phagocytes, assessed both by luminol-dependent chemiluminescence  and by the spectrophotometric superoxide assay , superoxide release from phagocytes was constant and not connected with catalase activity. Of particular importance, although catalase activity in S. aureus did not influence the intensity of phagocytosis (data not shown), it strongly and directly correlated with bacterial survival after phagocytosis .The SFPG did not exhibit direct bacteriostatic or bactericidal action towardsde assay . While t S. aureus with elevated or absent catalase activities were pretreated with SFPG for 1 hour and then mixed with human granulocytes  and suppressed level of anti-inflammatory cytokine IL-10 involved in the development of periodontitis and its severity in both GCF and blood plasma  by granulocytes is a key host defence against Staphylococcus aureus, Escherichia coli, Porphyromonas gingivalis, and other pathogenic bacteria [ P. gingivalis. OxyR is required to induce oxidative stress-related genes, such as sod (encoding superoxide dismutases), kat , pdgX (encoding peroxiredoxin-glutaredoxin), and ahpC  [ P. gingivalis connected to a transcriptional regulation by PG2212 zinc finger protein has been described [2O2 and its toxic metabolites like hypochlorite (OCl−) produced by granulocytes, known as key cellular mediators of inflammation in periodontitis [ S. aureus, which correlated with lower bacterial survival after phagocytosis. On the basis of these results, we assume that SPFG could be regarded as exerting an indirect antibiotic action, and a possible adjuvant function with direct antibiotics, owing to its capacity to inhibit bacterial adaptive antioxidant response, such as catalase induction, to granulocyte-induced or antibiotic-induced oxidative stress  , 59. Recescribed . The antescribed . The hypescribed . SPGF didontitis . The cate stress .The present clinical laboratory study showed the remarkable and durable clinical efficacy of topical administration of standardised fermented papaya gel to patients suffering from moderate-to-severe chronic periodontitis. Positive clinical results were associated with a normalisation of the inflammatory/anti-inflammatory cytokine pattern and of the proinflammatory metabolites of nitric oxide in gingival crevicular fluid and blood plasma. Clinical and biochemical anti-inflammatory effects of fermented papaya gel preparation were partly attributed to its indirect antibacterial impact through the inhibition of adaptive antioxidant response in bacteria to intracellular oxidative stress in phagocytes.


We report the case of a 24-year-old nondiabetic, nonhypertensive lady with history of fatigue, dyspnoea and limb claudication. She has been diagnosed with Takayasu's arteritis. Subsequently she developed rash, alopecia, joint pain, and various other laboratory abnormalities which led to a diagnosis of SLE. Takayasu's arteritis (TA) rarely coexists with systemic lupus erythematosus (SLE). The absence of specific SLE markers in patients with TA who subsequently develop SLE suggests that the coexistence of these conditions may be coincidental. The antiphospholipid syndrome in patients with SLE may mimic the occlusive vasculitis of TA. Takayasu's arteritis involves narrowing or obliteration of the major arteries and branches of the aorta resulting in the terminology “pulseless disease” . TubercuA 24-year-old unmarried Indian woman consulted her doctor for dyspnoea, fatigue, and upper limb claudication 3 years ago. She was diagnosed as a case of Takayasu's arteritis depending on impalpable pulse and unrecordable blood pressure on left arm, carotid artery bruit on left side, low velocity biphasic flow of all arteries of both upper limbs in Doppler study, and high erythrocyte sedimentation rate (ESR) (60 mm/hr). Antinuclear antibody (ANA) report was negative. She was on daily oral prednisolone (10 mg/day) but she was not adherent to the treatment and follow-up. After 3 years she presented with small and large joints arthritis, malar rash, discoid rash, oral ulcer, nonscarring alopecia, and photosensitivity . PeripheLaboratory investigations showed hemoglobin 8.3 gm/dL, reticulocyte count 6%, and erythrocyte sedimentation rate (ESR) 60 mm/hr. Urine for 24 hr protein was 2.41 gm/day. Serum antinuclear antibody (ANA) which was negative 3 years back became positive in high titre  and serum anti-dsDNA (titre was >1 : 180) came out to be positive. Further serum analysis revealed Blood Urea Nitrogen (BUN) 67 mg/dL, creatinine (Cr) 2.3 mg/dL, CRP 6.0 mg/L, C3 and C4 decrease, and positive anti-Sm antibody. Prothrombin time and activated partial prothrombin time were normal. Direct Coombs test was positive and antiphospholipid antibody was negative. Chest X-ray and ultrasonography of abdomen were normal. Echocardiography showed chink of pericardial effusion. Echo-Doppler study revealed that (1) right subclavian artery cannot be visualized beyond proximal part with wall thickening and mild luminal narrowing of right common carotid artery. (2) There were proximal stenosis of left subclavian and left common carotid artery . On CT aAccording to SLEDAI (SLE Disease Activity Index), the patient had a score of 22, so the patient had a severe flare. Then we put the patient on steroid at a dose of 1 mg/kg body weight and intravenous monthly cyclophosphamide. Joint pain, dyspnea, and fatigue were subsided within 2 months of treatment.Serum reports showed BUN 22 mg/dL, Cr 1.4 mg/dL, and CRP 2.0 mg/L and urine for 24 hr protein was 300 mg/day after 2 months of treatment.Takayasu's arteritis is more prevalent in Japan and Afro-Asian countries. Autopsy incidence in Japan is quoted as 33% . Lupi-HeAll of the American College of Rheumatology criteria of Takayasu's arteritis  were metVascular disease is frequent in patients with systemic lupus erythematosus and represents the most frequent cause of death in established disease. In this context, vasculopathy can be directly etiologically implicated in the pathogenesis of the disease, presenting as an acute/subacute manifestation of lupus . Alternatively, it can develop as an important accompanying comorbidity (steroid-related atherosclerotic disease) or represent the synergistic pathogenetic outcome of augmented atherosclerosis within a proinflammatory environment .Takayasu's arteritis involves mononuclear infiltration, granulomatous change, and fibrosis in the media with intimal thickening and obliterative aortitis of the large vessels. Immune mechanisms (mainly cell-mediated immunity) probably play a major role . VasculiThe claudication and asymmetric pulse preceded the appearance of rash and arthritis by more than a year. Antinuclear antibody was negative also at that time. Usually, SLE involves small and peripheral vessels but here involvement spans the medium to large vessels. There is a temporal association between Takayasu's arteritis and systemic lupus erythematosus, and aortoarteritis was the initial manifestation of systemic lupus erythematosus in our case, which is very rare. The natural history of Takayasu's arteritis is variable. But it usually progresses slowly with the development of hypertension, retinopathy, and aneurysm formation . The the