Intracellular ATP concentrations of CD4 cells in kidney transplant patients with and without infection.

In the field of organ transplantation, overimmunosuppression is associated with severe side effects, such as infection, drug toxicity, and cancer, whereas underimmunosuppression is associated with acute rejection. Intracellular adenosine triphosphate (iATP) concentration following CD4 cell activation provides an assessment of cellular immune function to help monitor the immune status of immunosuppressed patients. This assay has shown to be the first post-transplant test related not only to the risk of acute rejection but also with the appearance of infection. The aim of our study was to compare the iATP concentrations of CD4 cells between healthy adults and kidney transplant recipients from a European population, analyzing the differences according to transplant clinical status. Samples from 81 kidney transplant patients who were admitted to our hospital over a nine-month period were drawn. T-cell activation was measured by determining the increase of iATP from CD4 cells. Results were compared with patient clinical status (rejection, infection, and stability). Three patients suffered an acute rejection episode and they were not included in the analysis (mean iATP concentration 247 +/- 87 ng/mL). iATP concentrations differed significantly between stable and infected patients (313 +/- 193 vs. 197 +/- 114 ng/mL; p = 0.008). iATP concentration values were not related to the length of admission, age, peak and current panel reactive antibodies, mismatches, leukocytes, weight, creatinine, days after transplantation and blood levels of cyclosporin, tacrolimus, and sirolimus. This assay measures global immune responses of CD4 T cells from a whole-blood sample, allowing for the assessment of the impact of immuno- suppressive drugs and of the patient's underlying clinical conditions. This assay identifies transplant patients at risk for infection or rejection, providing information which can guide immunosuppressive therapy.

Interleukin-12 plasma levels in drug-naïve patients with a first episode of psychosis: effects of antipsychotic drugs.

An overactivation of the Th1 activity in schizophrenia had been described. Interleukin-12 (IL-12), a proinflammatory cytokine, plays a key role in the regulation of the Th1 response. The aims of this study were to investigate the effect of first and second generation antipsychotic drugs on IL-12 production during the acute phase of the illness and its association with clinical features. Participants comprised 56 drug-naïve first episode psychotic patients and 28 healthy volunteers. Patients were initially randomly assigned to risperidone (n=16), olanzapine (n=20) or haloperidol (n=20); subject were maintained on the same medication throughout the study. Clinical assessments were conducted at baseline and at 6 weeks. IL-12 plasma levels were assessed at baseline and after 6 weeks of antipsychotic treatment. IL-12 haplotypes were also analysed. Patients showed higher IL-12 plasma levels at baseline compared with controls, and had a significant increase in IL-12 plasma level after 6 weeks of antipsychotic treatment. No significant differences in IL-12 level increase were found among the three antipsychotic treatments. IL-12 plasma levels at week 6 were not significantly associated with the severity of psychopathology at week 6. Thus, patients with a first episode of psychosis have inflammatory-like immunological function during early phases of the illness that it is independent of the antipsychotic treatment used.

Interleukin-1 receptor antagonist genotype and brain morphometry in first-episode non-affective psychosis.

Studies of schizophrenia that combine imaging and genetic approaches attempt to map structural brain anomalies associated with genetic risk variants. The aim of the present study was to investigate whether variations in the interleukin-1 receptor antagonist (IL-1RN) were associated with structural brain characteristics of 73 minimally medicated first-episode non-affective psychotic patients. We did not find evidence for association between genetic variation in the IL-1RN gene and brain morphometry at early phases of the illness.

Catechol-O-methyltransferase Val158Met polymorphism and clinical characteristics in first episode non-affective psychosis.

Catechol-O-methyltransferase (COMT) Val158Met polymorphism has been identified as a potential etiologic factor in schizophrenia. It has been proposed that this polymorphism could be associated with specific clinical markers. The aim of the study was to evaluate the influence of COMT Val158Met polymorphism genotype in the phenotypic expression of first episode psychosis at onset. Age of onset, DUP, SANS, and SAPS (positive, disorganized, and negative dimensions) were studied in 169 Caucasian drug-naïve patients with a first-episode of non-affective psychosis. The COMT Val158Met polymorphism was typed using PCR amplification of the relevant region followed by digestion with NlaIII and electrophoresis. A multivariate ANCOVA was performed with COMT and gender as independent variables. Patients with Val/Val genotype had significantly higher levels of SANS negative dimension scores (F: 3.539; P = 0.031) and had a younger age of onset (F: 4.649; P = 0.011) than Met carriers. Our findings suggest that the Val allele is associated with onset phenotypic features related to a poor prognosis of the illness. These data would indicate that COMT genotype may have a role in the etiological model for schizophrenia and other psychotic disorders.

Human herpesvirus-8 infection and Kaposi's sarcoma after liver and kidney transplantation in different geographical areas of Spain.

Since data on human herpesvirus 8 (HHV-8) infection in Spain is not available, our purpose was to determine the prevalence of HHV-8 infection and the risk of developing Kaposi's sarcoma (KS) among organ transplant recipients in different geographical areas of Spain. The study population consisted of 1019 liver and kidney transplant recipients recruited in four transplant units in Spain. Only post-transplant serum samples were available for all participants. IgG anti-HHV-8 latent and lytic antigens were detected by using an indirect immunofluorescence assay as well as enzyme-linked immunosorbant assays. In available samples, HHV-8 DNA genome was detected by using a nested polymerase chain reaction in sera, blood mononuclear cells and KS tissues. The prevalence of HHV-8 infection after transplantation was calculated. To determine risk factors for infection, odds ratios (OR) and 95% confidence intervals (CI) were also calculated. Of the 788 kidney transplants, 5 (0.6%) were HHV-8-positive shortly after transplantation. Of the 231 liver transplant individuals, 8 (3.4%) developed IgG anti-HHV-8 antibodies after transplantation. Thus, incidence of HHV-8 infection is significantly higher among liver transplant recipients in comparison with that in the control population (OR=6, 95% CI=1.2-28.5, p<0.05). In this series, HHV-8 prevalence in liver transplant recipients was higher in the northern (6.6-6.9%) than in the central (2.9%) or the southeastern (1.4%) areas of Spain. Four renal transplant recipients (0.5%) and five of the liver transplant recipients (2.16%) developed KS after transplantation. Time of KS diagnosis after transplant is significantly higher in kidney transplant patients (33.7 months) than in liver transplant recipients (10.4 months), indicating that the latter have a higher risk of developing KS.

Association of migraine and ESR1 G325C polymorphism.

Migraine is a genetically complex disorder in which sexual hormones influence the phenotype. ESR1 G594A polymorphism has been associated with migraine in Australians. We performed a case-control study with G594A and G325C polymorphisms to determine whether ESR1 is associated with migraine in our population. An association between G594A and migraine could not be demonstrated here. By contrast, we observed that the C325 allele conferred a 1.6 (95% confidence interval=1.1-2.4) higher risk for suffering from migraine in women than the G allele. Women carrying the C352C genotype were over 3 times more likely to suffer from migraine than those carrying the G325G genotype. Therefore, we conclude that ESR1 G325C polymorphism is associated with migraine in our population.

Sensitivity to bee venom antigen phospholipase A2: association with specific HLA class I and class II alleles and haplotypes in beekeepers and allergic patients.

Bee venom hypersensitivity is a clinical entity of outstanding importance because bee stings are a leading cause of mortality worldwide. Individuals with immediate-type bee venom hypersensitivity, beekeepers, and healthy controls were examined for HLA-DRB1, DQB1, and DQA1 alleles by sequence-specific oligonucleotide probe typing. Defined hypersensitivity to bee venom antigen phospholipase A2 (vbPLA2) is significantly associated with the presence of susceptible HLA class II alleles: DRB1*0101 (RR = 2.7, p < 3 x 10(-3)), DRB1*0103 (RR = 21.2, p < 7.5 x 10(-11)), DQA1*0101 (RR = 1.2, p < 38.52 x 10(-10)), and DQB1*0501 (RR = 4, p < 2.18 x 10(-10)). Some HLA class I alleles were also associated with risk to bee venom allergy: A*01 (RR = 2.4, p < 7.5 x 10(-4)), B*57 (RR = 35.1, p < 3.5 x 10(-7)), and B*5901 (p < 3.5 x 10(-5)), but they are probably of secondary significance. Three- (DRB1*0103-DQA1*0101-DQB1*0501) (RR = 21.24, p < 7.5 x 10(-11)) and five-loci (A*01-B*59-DRB1*0103-DQA1*0101-DQB1*0501) (p < 2.3 x 10(-6)) extended haplotypes are also significantly carried by vbPLA2 allergic patients. When HLA allele frequencies from patients are compared with those from beekeepers, only HLA-DRB1*0103 (RR = 11.7, p < 8.5 x 10(-5)) and HLA-DQA1*0101 (p < 0.02) were significantly increased in the former. These observations emphasize the importance of the DRB1*0103-DQA1*0101-DQB1*0501 haplotype as a strong candidate for susceptibility to vbPLA2 hypersensitivity, at least in our region.

Thymidylate synthase promoter tandem repeat and MTHFD1 R653Q polymorphisms modulate the risk for migraine conferred by the MTHFR T677 allele.

There is growing evidence that folate metabolism is involved in migraine pathophysiology, mainly in migraine with aura. Even though folate metabolism is regulated by a number of enzymes, only two functional polymorphisms have been tested in association studies with migraine. Here, we have explored the possible role in migraine of other folate-metabolizing enzymes which are in close interdependency with 5',10'-methylenetetrahydrofolate reductase analyzing functional polymorphisms of these enzymes in a case-control study. Individually, thymidylate synthase (TS), methenyltetrahydrofolate cyclohydrolase formyltetrahydrofolate synthase (MTHFD1), or methionine synthase (MS) polymorphisms did not modify the general risk for suffering migraine. Nevertheless, we observed a strong interaction between TS and MTHFR mutated genotypes, which increased over 8-fold the risk for experiencing aura among migraineurs; MTHFD1 and MTHFR mutated genotypes also increased together the risk for migraine in general (OR = 3.08; 95% CI = 1.3-7.4). We conclude that the pathogenetic role of the MTHFR T677 allele in migraine is modulated by functional polymorphisms of TS and MTHFD1.

Mannose-binding lectin promoter polymorphisms and gene variants in pulmonary tuberculosis patients from cantabria (northern Spain).

Mannose-binding lectin is a central molecule of the innate immune system. Mannose-binding lectin 2 promoter polymorphisms and structural variants have been associated with susceptibility to tuberculosis. However, contradictory results among different populations have been reported, resulting in no convincing evidence of association between mannose-binding lectin 2 and susceptibility to tuberculosis. For this reason, we conducted a study in a well genetically conserved Spanish population in order to shed light on this controversial association. We analysed the six promoter and structural mannose-binding lectin 2 gene variants in 107 patients with pulmonary tuberculosis and 441 healthy controls. Only D variant and HYPD haplotype were significantly more frequents in controls which would indicate that this allele could confer protection against pulmonary tuberculosis, but this difference disappeared after statistical correction. Neither the rest of alleles nor the haplotypes were significantly associated with the disease. These results would indicate that mannose-binding lectin promoter polymorphisms and gene variants would not be associated with an increased risk to pulmonary tuberculosis. Despite the slight trend of the D allele and HYPD haplotype in conferring protection against pulmonary tuberculosis, susceptibility to this disease would probably be due to other genetic factors, at least in our population.

Characterization of a Listeria monocytogenes protein interfering with Rab5a.

Listeria monocytogenes (LM) phagocytic strategy implies recruitment and inhibition of Rab5a. Here, we identify a Listeria protein that binds to Rab5a and is responsible for Rab5a recruitment to phagosomes and impairment of the GDP/GTP exchange activity. This protein was identified as a glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Listeria (p40 protein, Lmo 2459). The p40 protein was found within the phagosomal membrane. Analysis of the sequence of LM p40 protein revealed two enzymatic domains: the nicotinamide adenine dinucleotide (NAD)-binding domain at the N-terminal and the C-terminal glycolytic domain. The putative ADP-ribosylating ability of this Listeria protein located in the N-terminal domain was examined and showed some similarities to the activity and Rab5a inhibition exerted by Pseudomonas aeruginosa ExoS onto endosome-endosome fusion. Listeria p40 caused Rab5a-specific ADP ribosylation and blocked Rab5a-exchange factor (Vps9) and GDI interaction and function, explaining the inhibition observed in Rab5a-mediated phagosome-endosome fusion. Meanwhile, ExoS impaired Rab5-early endosomal antigen 1 (EEA1) interaction and showed a wider Rab specificity. Listeria GAPDH might be the first intracellular gram-positive enzyme targeted to Rab proteins with ADP-ribosylating ability and a putative novel virulence factor.

Cutting edge: a novel nonoxidative phagosomal mechanism exerted by cathepsin-D controls Listeria monocytogenes intracellular growth.

Deciphering how Listeria monocytogenes exploits the host cell machinery to invade mammalian cells is a key issue in understanding the pathogenesis of this food-borne pathogen, which can cause diseases ranging from gastroenteritis to meningitis and abortion. In this study, we show that the lysosomal aspartyl-protease cathepsin-D (Ctsd) is of considerable importance for nonoxidative listericidal defense mechanisms. We observed enhanced susceptibility to L. monocytogenes infection of fibroblasts and bone-marrow macrophages and increased intraphagosomal viability of bacteria in fibroblasts isolated from Ctsd-deficient mice compared with wild type. These findings are further supported by prolonged survival of L. monocytogenes in Ctsd-deficient mice after infection. Transient transfection of Ctsd in wild-type cells was sufficient to revert these wild-type phagosomes back to microbicidal compartments. Based on infection experiments with mutant bacteria, in vitro degradation, and immunoprecipitation experiments, we suggest that a major target of cathepsin D is the main virulence factor listeriolysin O.

Association between the interleukin-1 receptor antagonist gene and negative symptom improvement during antipsychotic treatment.

The contribution of immune system to schizophrenia has been an important area of focus in schizophrenia research. Several genetic variants in the cytokine system have been associated with the pathogenesis of schizophrenia. The purpose of this study was to determine whether a pharmacogenetic relationship exists between a variable number of tandem repeats (VNTR) polymorphism in the interleukin-1 receptor antagonist gene (IL-1RN) and clinical improvement during antipsychotic treatment in patients with a first non-affective psychotic episode. One hundred and fifty-four subjects presenting with a first non-affective psychotic episode were randomly assigned to treatment with haloperidol, risperidone, or olanzapine and rated with the Brief Psychiatric Rating Scale (BPRS), the Scale for the Assessment of Negative Symptoms (SANS) and the Scale for the Assessment of Positive Symptoms (SAPS) both at baseline and after 6 weeks of treatment. A control sample of 336 blood bank donors was also included. No differences in genotype or allele distributions were found between patients and controls. However, after controlling for baseline SANS scores, the genotype in the VNTR polymorphism in the IL-1RN gene significantly predicted negative symptom improvement, accounting for approximately 7% of the variance (F = 5.23, df = 2, P = 0.006). The mean decrease in SANS scores was 58% for the IL-1RN* 2/2, 44% for the IL-1RN* 1/2, and 14% for the IL-1RN* 1/1 subjects, respectively. These results suggest that the VNTR polymorphism in the IL-1RN gene may be a useful predictor of negative symptom improvement in schizophrenic patients treated with antipsychotic drugs.

Inhibition of Rab5a exchange activity is a key step for Listeria monocytogenes survival.

Listeria monocytogenes (LM) modifies the phagocytic compartment by targeting Rab5a function through an unknown mechanism. Inhibition of Rab5a exchange by LM can be considered the main virulence mechanism as it favours viability of the parasite within the phagosome as well as the exclusion of putative listericidal lysosomal proteases such as cathepsin-D. The significance of this survival mechanism is evidenced by the overexpression of Rab5a mutants in CHO cells that promoted GDP exchange on Rab5a and eliminated pathogenic LM. The following mutants showed listericidal effects: Rab5a:Q79L, a constitutively active mutant with accelerated GDP exchange and Rab5a GEF, Vps9, which overactivates the endogenous protein. Clearance of LM from these phagosomes was controlled by the hydrolytic action of cathepsin-D as suggested by the lysosomal protease inhibitor chloroquine, or the cathepsin-D inhibitor, pepstatin A, which caused a reversion of listericidal activity. Moreover, the effects of LM on Rab5a phagocytic function mimics those reported for the GDP locked dominant negative Rab5a mutant, S34N. Transfection of these mutants into CHO cells increased pathogen survival as they showed higher numbers of viable bacteria, complete inhibition of GDP exchange on Rab5a and impairment of the listericidal action probably exerted by cathepsin-D. We cotransfected functional Rab5a GEF into this dominant negative mutant and restored normal LM intraphagosomal viability, Rab5a exchange and listericidal action of cathepsin-D.

Significance of micro-geographical population structure in forensic cases: a bayesian exploration.

We studied the influence of population structure at the microgeographical level on the analysis of forensic cases. A total of nine autosomal STRs and seven Y-STRs were analyzed in the general mixed population and in two relatively isolated valleys of Cantabria, a region in Northern Spain. Statistically significant differences existed in the frequency distribution of four autosomal STRs, with an overall Fst value of 0.3%. A simulation of virtual trio cases revealed that it did not have a practical influence on the analysis of paternity disputes. Significant differences also existed in most Y-STRs, with an overall Fst value of 3%. Thus, using the general database instead of the specific valley database resulted in 5-fold or higher overestimation of the likelihood ratio of matching in up to 30% of cases. A bayesian analysis revealed that this had a significant impact on the estimation of the probability of identity in scenarios of low "a priori" odds of suspicion.

Correlation between transcranial interleukin-6 gradient and outcome in patients with acute brain injury.

OBJECTIVE: This study was performed to examine both brain and systemic interleukin-6 (IL-6) release in patients with an acute brain injury (ABI), to study whether a correlation exists between the transcranial IL-6 gradient during the first days after injury and prognosis, and finally, to investigate the relationship between a nucleotide polymorphism at position -174 in the promoter of the gene encoding IL-6, IL-6 responsiveness, and clinical evolution. DESIGN: Prospective clinical investigation. SETTING: A 19-bed intensive care unit in a university hospital. PATIENTS AND METHODS: A total of 62 patients were followed up for 3 days after acute brain injury, and both their arterial and jugular IL-6 levels were measured serially and at the moment of brain death diagnosis. Genetic polymorphism of IL-6 was also determined in all patients. Data were correlated with those from score procedures for clinical severity. Neurologic outcome was graded according to the Glasgow Outcome Scale 6 months after injury. IL-6 levels and IL-6 genotyping was performed in control healthy individuals. MAIN RESULTS: There is a significant transcranial IL-6 gradient at admission and at the moment of brain death. The gradient is higher in those patients who evolved toward a fatal outcome during the first 6 months after injury (p <.001). There is significant correlation between the transcranial IL-6 gradient and the acute brain injury severity. CONCLUSIONS: IL-6 is elevated in patients with acute brain injury, and a significant relationship exits between the severity of acute brain injury and the transcranial IL-6 gradient at admission. It can be considered to be a prognosis marker at admission. When data at the moment of brain death are considered, venous IL-6 (p <.01) and the transcranial IL-6 gradient (p <.005) are significantly higher than at the time of admission. Although the IL-6 C allele is associated with significantly lower concentrations of IL-6, there was no correlation between low or high IL-6 responders and patient outcome.