Fungal biodiversity in the air of Turin

The qualitative fungal composition of Turin 's atmospheric environment was surveyed, carrying out a twelve-month study and collecting with a single stage volumetric sieve sampler on Dermasel agar supplemented with 0.4 g l^–1 cycloheximide and 0.05 g l^–1 chloramphenicol. We isolated 165 species and 2 varieties of mesophilic fungi from 58 genera and 26 thermotolerant species from 12 genera. Penicillium, Aspergillus, Acremonium, Chrysosporium, Scopulariopsis, Malbranchea, Paecilomyces, Phialophora and Cladosporium were in sequence the genera most rich in mesophilic species; Aspergillus, Penicillium, Chrysosporium and Scopulariopsis the most rich in thermotolerant species. Many of the species isolated are rarely or never recorded in the atmospheric environment. Cycloheximide can thus be said to select among airborne fungi, giving a characteristic picture.Abbreviation CH Cycloheximide - cfu colony forming unit     

Enkephalin-binding systems in human plasma

Three amino acid-containing fractions present in human plasma are shown to bind both leu and met-enkephalin: serum albumin and two species of a much lower molecular weight, in all likelihood polypeptides. The amount of enkephalin associated with serum albumin seems comparatively smaller than that associated with the two low molecular weight systems. These systems jointly are apparently capable of binding a significant part of the circulating enkephalins. The possibility is suggested that the interactions described may play a role in maintaining the integrity of circulating enkephalins.     

8-Oxoguanine DNA-glycosylase repair activity and expression: a comparison between cryopreserved isolated lymphocytes and EBV-derived lymphoblastoid cell lines

Several lines of evidence suggest an association between oxidative DNA-damage repair capacity and cancer risk. In particular, a DNA-glycosylase assay for removal of 8-oxoguanine (8-oxoG) in peripheral blood mononuclear cells (PBMC) has been successfully applied to identify populations with increased risk for lung cancer and squamous cell carcinomas of head and neck. In order to verify whether EBV-transformed lymphoblastoid cell lines (LCL) are a suitable surrogate for PBMC in specific DNA-repair phenotypic assays, a validation trial was conducted. PBMC from 20 healthy subjects were collected and an aliquot was transformed with EBV to obtain LCL. The ability of cell-free extracts from both cell types to incise a 3'-fluorescently labelled duplex oligonucleotide containing a single 8-oxoG (OGG assay) was evaluated. Since this activity is mediated predominantly by OGG1, the OGG1 gene expression was also measured. 8-oxoG DNA-glycosylase activity and OGG1 expression were significantly higher (p<0.0001) in LCL than in PBMC. However, while this assay was shown to be robust and reproducible when used on PBMC (intra-assay CV=8%), a high intra-culture variability was observed with LCL (intra-culture CV=16.8%). Neither differences on OGG1 gene expression nor the cell-cycle distribution seemed to account for this variability. Inter-individual variability of OGG activity in PBMC and LCL was not associated with OGG1 gene expression. We have therefore established a non-radioactive cleavage assay that can be easily applied to measure OGG activity in human PBMC. The use of LCL for DNA-repair genotype-phenotype correlation studies seems to be inappropriate, at least with cell-free based functional assays.     

T Cell Activation and Cytokine Profile of Tuberculosis and HIV-Positive Individuals during Antituberculous Treatment and Efavirenz-Based Regimens

Introduction

The profile of immune activation markers in tuberculosis and HIV-infected patients is already known. The impact of simultaneous infections on the immune parameters is still not fully explored.

Methods

We conducted a prospective study to estimate trajectories of activated T cell subsets and the profile of anti- and pro-inflammatory cytokines in a group of HIV-TB individuals, previously naïve for HAART, recruited from a randomized clinical trial during TB treatment and first antiretroviral therapy with efavirenz. Patients were evaluated according to the immunosuppression levels at baseline as group 1 (CD4<200 cells/mm³) and group 2 (CD4>200 cells/mm³). These parameters were measured at the time of HAART initiation (started about 30 days after the onset of TB treatment) and at the follow-up visits after 30, 60, 90 and 180 days. Trajectories were estimated using least squares estimates of the coefficients of a restricted cubic spline function in time after adjusting for subject effects, bootstrapping it 500 times.

Results

Increase of CD4 T cell counts and suppression of HIV viral load were observed for all patients under HAART and TB treatment. Descendent trajectories were observed for the activated CD8⁺/CD38⁺ and CD3⁺/HLA-DR⁺ T cell subsets, and for plasma concentration of gamma- interferon (IFN-γ). Except for TNF-α and IL-2 discrete variations were observed for the other cytokines. Differences in the trajectories of these parameters were observed for groups 1 and 2. Higher values of IFN-γ, IL-2, IL-6 and IL-10 were observed for group 1 from the baseline to two months after treatment initiation, whereas reduced levels of TNF-α were observed for this group between 60 and 120 days of HAART.

Conclusion

Independent of the immunosuppression profile at baseline, HIV-TB patients under HAART were able to recover the CD4⁺ T cell counts, and control viral replication and immune activation parameters over time.     

Flora Palinologica Italiana,Sezione Aeropalinologica — Specie esotiche — S 231:Sequoia sempervirens (Lamb.) Endl. (Taxodiaceae)

Riassunto La scheda morfopalinologica diSequoia sempervirens (Lamb.) Endl. continua lo studio sulle Pinohyta ornamentali più frequenti in Torino. È esaminato polline fresco ed acetolizzato proveniente da due stazioni e la provenienza non determina variazioni significative dei parametri morfometrici.

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Summary The morphopalynological card ofSequoia sempervirens (Lamb.) Endl. is a new contribution to the comprehensive study on pollen of ornamental Pinophyta in Turin. Fresh and acetolysed grains from two stations are examined, and for all parameters no significative differences were observed in relation to the collection site.

Parole chiave: Sequoia sempervirens(Lamb.) Endl., Flora Palinologica Italina, Sezione Aeropalinologica.     

Association between the allele compositions of major plant developmental genes and frost tolerance in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) germplasm of different origin

Changing climatic conditions with warming winters and shifts in the frequencies of drought, intense rainfall and cold spells together with associated changes in the geographical distribution of arable crops increase the challenges for selecting new varieties. In this context, we aim to contribute to a better understanding of the determinants of barley (Hordeum vulgare) frost tolerance (FRT) and consequent improvements to marker-assisted selection (MAS). Freezing injury in a diversity panel of 121 barley genotypes with different growth habits and origins was assessed using phenotyping based on chlorophyll fluorescence (F_v/F_m) measurements to screen genetic diversity in plants at an early growth stage. The haplotypes of vernalisation and photoperiod genes were determined with PCR, and correlation analyses were done using data from 12 laboratory and field-laboratory FRT tests. Previous results of allelic combinations of VRN-H1/VRN-H2 for FRT were confirmed with these experiments using a larger set of genotypes. The predictive power of polymorphisms in VRN-H1 intron 1 region for FRT was significantly higher than that of the VRN-H1 promoter polymorphism. The vrn-H1/vrn-H2 facultative genotypes had similar or higher FRT than vrn-H1/Vrn-H2 winter genotypes under suboptimal hardening conditions. Genes regulating long-day and short-day photoperiodic responses were significantly correlated with FRT. The most parsimonious model for prediction of FRT was based on polymorphisms in the VRN-H1 intron 1 region, VRN-H2 and PPD-H2 and explained 69% of the variation in FRT.     

Affective response to a loved one's pain: insula activity as a function of individual differences

Individual variability in emotion processing may be associated with genetic variation as well as with psychological predispositions such as dispositional affect styles. Our previous fMRI study demonstrated that amygdala reactivity was independently predicted by affective-cognitive styles (phobic prone or eating disorders prone) and genotype of the serotonin transporter in a discrimination task of fearful facial expressions. Since the insula is associated with the subjective evaluation of bodily states and is involved in human feelings, we explored whether its activity could also vary in function of individual differences. In the present fMRI study, the association between dispositional affects and insula reactivity has been examined in two groups of healthy participants categorized according to affective-cognitive styles (phobic prone or eating disorders prone). Images of the faces of partners and strangers, in both painful and neutral situations, were used as visual stimuli. Interaction analyses indicate significantly different activations in the two groups in reaction to a loved one's pain: the phobic prone group exhibited greater activation in the left posterior insula. These results demonstrate that affective-cognitive style is associated with insula activity in pain empathy processing, suggesting a greater involvement of the insula in feelings for a certain cohort of people. In the mapping of individual differences, these results shed new light on variability in neural networks of emotion.     

Association between the ACCN1 gene and multiple sclerosis in Central East Sardinia

Multiple genome screens have been performed to identify regions in linkage or association with Multiple Sclerosis (MS, OMIM 126200), but little overlap has been found among them. This may be, in part, due to a low statistical power to detect small genetic effects and to genetic heterogeneity within and among the studied populations. Motivated by these considerations, we studied a very special population, namely that of Nuoro, Sardinia, Italy. This is an isolated, old, and genetically homogeneous population with high prevalence of MS. Our study sample includes both nuclear families and unrelated cases and controls. A multi-stage study design was adopted. In the first stage, microsatellites were typed in the 17q11.2 region, previously independently found to be in linkage with MS. One significant association was found at microsatellite D17S798. Next, a bioinformatic screening of the region surrounding this marker highlighted an interesting candidate MS susceptibility gene: the Amiloride-sensitive Cation Channel Neuronal 1 (ACCN1) gene. In the second stage of the study, we resequenced the exons and the 3' untranslated (UTR) region of ACCN1, and investigated the MS association of Single Nucleotide Polymorphisms (SNPs) identified in that region. For this purpose, we developed a method of analysis where complete, phase-solved, posterior-weighted haplotype assignments are imputed for each study individual from incomplete, multi-locus, genotyping data. The imputed assignments provide an input to a number of proposed procedures for testing association at a microsatellite level or of a sequence of SNPs. These include a Mantel-Haenszel type test based on expected frequencies of pseudocase/pseudocontrol haplotypes, as well as permutation based tests, including a combination of permutation and weighted logistic regression analysis. Application of these methods allowed us to find a significant association between MS and the SNP rs28936 located in the 3' UTR segment of ACCN1 with p = 0.0004 (p = 0.002, after adjusting for multiple testing). This result is in tune with several recent experimental findings which suggest that ACCN1 may play an important role in the pathogenesis of MS.