Magnetic ionic plastic crystal: choline[FeCl4].

A novel organic ionic plastic crystal (OIPC) based on a quaternary ammonium cation and a tetrachloroferrate anion has been synthesized with the intention of combining the properties of the ionic plastic crystal and the magnetism originating from the iron incorporated in the anion. The thermal analysis of the obtained OIPC showed a solid-solid phase transition below room temperature and a high melting point above 220 °C, indicating their plastic crystalline behaviour over a wide temperature range, as well as thermal stability up to approximately 200 °C. The magnetization measurements show the presence of three-dimensional antiferromagnetic ordering below 4 K. The results from electrochemical characterization display a solid-state ionic conduction sufficiently high and stable (between 10(-2.7) and 10(-3.6) S cm(-1) from 20 to 180 °C) for electrochemical applications.

Analysis and management of HIV peptide microarray experiments.

OBJECTIVES: To develop a tool for then easy and user-friendly management of peptide microarray experiments and for the use of the results of these experiments for the study the immune response against HIV virus infection in clinical samples. METHODS: Applying bioinformatics and statistics for the analysis of data coming from microarray experiments as well as implementing a MIAME (Minimum Information About a Microarray Experiment) compliant database for managing and annotating experiments, results and samples. RESULTS: We present a new tool for managing not only nucleic acid microarray experiments but also protein microarray experiments. From the analysis of experimental data, we can detect different profiles in the reactivity of the sera with different genotypes. CONCLUSIONS: We have developed a new tool for managing microarray data including clinical annotations for the samples as well as the capability of annotating other microarray formats different to those based on nucleic acids. The use of peptide microarrays and bioinformatics analysis opens a new scope for the characterization of the immune response, and analyzing and identifying the humoral response of viruses with different genotypes.

Molecular epidemiology of HIV-1 in Madrid.

Thirteen HIV-1 isolates from patients of different risk groups in Madrid (Spain) have been analyzed at the genetic level. Two distinct lineages of subtype B have been detected among the HIV-1 circulating in this area: one was related to SF-2/RF strains, whereas the other consists of a more heterogeneous group related to reference strain III-B. Variants of each lineage appeared to circulate preferentially within a risk group: III-B among intravenous drug users, and RF/SF-2 among male homosexuals.

HIV type 1 non-B subtype prevalence in Spain, 1997-1998.

We have evaluated the prevalence of HIV-1 non-B subtypes in Spain by means of an enzyme immunoassay (EIA) for discrimination between B and non-B subtypes. Samples were obtained from newly diagnosed patients attended at internal medicine outpatient clinics between October 1997 and October 1998. Discrimination between HIV-1 B and non-B subtypes was carried out by means of the EIA, with V3 synthetic peptides specific to the different subtypes. Non-B-serotyped samples were genetically analyzed in the gp41 region from the original sera. During the study period, 909 samples were collected from 21 medical units located in various Spanish geographical regions. Serotyping was possible in 885 cases, of which 791 were assigned as B serotype (89.38%), 70 showed no reactivity to any of the peptides (7.91%), and the remaining samples displayed other reaction patterns (2.72%). Of the 94 non-B-assigned samples, 65 were genetically characterized in the gp41 region of the env gene: 55 were B subtype, 5 were A subtype (4 clustered with CRF02AG reference strains), 3 were C subtype, and 2 were G subtype. The prevalence rate for non-B subtypes in Spain was established at 1.13% (95% CI, 0.59-2.21). Although the B subtype is predominant in the Spanish population, other subtypes have been detected.

Molecular epidemiology of HIV type 1 subtypes in equatorial guinea.

Equatorial Guinea is endemic for HIV-1. This country borders to the north with Cameroon, where different subtypes belonging to group M, as well as group O strains, are circulating simultaneously. To assess the molecular epidemiology of HIV-1 in Equatorial Guinea we analyzed 76 plasma samples collected throughout 1999 from seropositive individuals. Phylogenetic analysis of the gp41 region revealed that 53 were of subtype A, with 64% of these sequences clustering with CRF02_AG reference strains; 11 were of subtype C; 4 were of subtype D; 2 (closely related to subtype F2) were of subtype F; 3 were of subtype G, two of them forming a separate cluster with the recombinant circulating forms CRF06_cpx; 1 was of subtype H; and 2 were unclassifiable. Although subtype A is predominant, the presence of 14% of subtype C is also noteworthy. This work represents the first HIV-1 subtype distribution study in Equatorial Guinea.

Molecular characterization of non-B HIV type 1 subtypes from Africa in Spain.

All of the known HIV-1 subtypes are present in sub-Saharan Africa. The B subtype is predominant in the United States and Europe, but previous studies have revealed that other subtypes are also in circulation. We report here on the genetic characterization of eight non-B subtype VIH-1 virus strains detected during 1999 in patients living in Spain and having epidemiological relationships with African countries. Five isolates clustering with recombinant form CRF02-AG came from West and Central Africa. One isolate was characterized as being of the D subtype in the gp41 region, and clustered with subtype A outside the CRF02-AG recombinant form, in regions C2V3 and p17. Another isolate was a G subtype, and the remaining isolate was an O subtype. In Spain, the B subtype is the most frequently detected HIV-1 subtype, although in more recent years non-B subtypes have been introduced through immigrant HIV-1-infected individuals coming from African countries, or through infected persons having relationships with endemically affected areas.

Typing human T-cell lymphotropic virus (HTLV-I and HTLV-II) by nested polymerase chain reaction: application to clinical specimens.

Human T-cell lymphotropic virus type I and II provirus DNA was detected by polymerase chain reaction (PCR). MT-2 (HTLV-I infected), C3/44 Mo (HTLV-II infected) cell lines and peripheral blood mononuclear cells (PBMNC) from HTLV seropositive samples were used. The procedure consists of first amplification which detects both HTLV-I and HTLV-II, and a second amplification (nested-PCR) to discriminate between the two viruses and to improve sensitivity. Optimal conditions of MgCl2 concentration and annealing temperature were found for maximal amplification and specificity. This method was used for the amplification of conserved regions of pol and env genes. 1.5 pg of MT-2 and 5 pg of C3/44 Mo cell line DNAs were detected using nested-PCR and liquid hybridization in the pol system. The env system could detect 1.5 pg of MT-2 and 1.5 pg of C3/44 Mo cell lines DNAs using nested-PCR and liquid hybridization. The pol system can type both HTLV-I and HTLV-II in only two steps without the use of type-specific radiolabeled probes. Furthermore, this method can detect and discriminate the two viruses in one step PCR using the primers used in the nested-PCR. Nevertheless, there is a decrease in sensitivity of 100-fold. The results of five seropositive samples confirmed by Western blot are compared with PCR. PCR typed one of these samples as HTLV-I and the rest as HTLV-II. This technique is useful in cases such as window period, perinatal studies and when serologic results are not satisfactory.

[Trend of HIV seroprevalence among mothers of new-born infants from 1996 to 1999]. / Evolución de la seroprevalencia del virus de la inmunodeficiencia humana en madres de recién nacidos entre 1996 y 1999.

BACKGROUND: To analyze the evolution of HIV prevalence in mothers of Spanish new-borns. SUBJECTS AND METHOD: Unlinked anonymous testing of HIV in blood spots for detection of metabolic diseases of all new-borns in 1996-1999 in seven regions: Baleares, Canarias, Castilla-La Mancha, Castilla y León, Galicia, Melilla and Murcia. HIV antibody detection was done with ELISA and confirmation with a immunoblot. RESULTS: The prevalence of HIV antibodies was 0.99 per 1,000 in 1996, 1.29 in 1997, 1.42 in 1998 and 1.54 in 1999. There was an upward trend both in the global sample (p = 0.0015) and in those from Canarias (p < 0.0001) and Castilla y León (p = 0.0389). The prevalence of HIV-1 for the whole period was 1.31 per 1.000 and of 1.13 per 100.000 for HIV-2. CONCLUSIONS: There is a need to offer systematic counselling and HIV testing to all pregnant women.

[Isolation, sequencing and ultrastructure of HTLV-I and HTLV-II retroviruses. Presence of the HTLV-II subtype b among Spanish intravenous drug addicts]. / Aislamiento, secuenciación y ultraestructura de los retrovirus HTLV-I y HTLV-II. Presencia del HTLV-II subtipo b entre adictos a drogas por vía parenteral españoles.

BACKGROUND: Six cases of HTLV-I/II infection were selected for isolation and characterization of these retrovirus. METHODS: Detection of anti-HTLV antibodies was carried out by enzyme immunoassay (EIA), immunofluorescence (IFI), and Western blot (WB). Analysis of proviral DNA was performed by PCR. Viral culture and partial sequencing of the pol and pX genes were carried out. Electron microscopy morphologically characterized the viral particles. RESULTS: Serologic study demonstrated four cases of HTLV-II, one of HTLV-I, and one non-typeable HTLV infections. This last case was confirmed as positive for HTLV-II by PCR. Five new HTLV-II and one HTLV-I infected cell lines have been established by co-culture. Electron microscopy allowed morphologic characterization of the viral particles found in the infected cells. The sequence of the five strains of HTLV-II was identical demonstrating a divergence of 0.49% in the pX region and of 4.5% in the pol region compared with the HTLV-II Mo prototype. Comparison of these sequences with those corresponding to different strains of HTLV-II isolates from American Indians (subtypes b) suggest that these Spanish strains are more closely related with the subtype b than with the subtype a (HTLV-II Mo). Genetic variability study did not reveal any change in the sequence of these stains suggesting that the variability of these retroviruses in very infrequent in the regions studied. The analysis of the pol region of the HTLV-I strain demonstrated a divergence of 3.4% with respect to the sequence of the ATK-1 prototype (Japan) and of 1.7% of the strain HS-35 (Caribbean) showing a greater relation with the Caribbean strains than with those from Japan. CONCLUSIONS: The presence of HTLV-II subtype has been confirmed among intravenous drug addicts in Spain. Isolation and characterization of the HTLV-I strain demonstrated that this also circulating around Spain despite its South American origin.

[HTLV-I myelopathy: presentation of a new case]. / Mielopatía por HTLV-I: presentación de un nuevo caso.

INTRODUCTION: HTLV-I is a human retrovirus which has been implicated in the genesis of tropical spastic paraparesis (HTLV-I-associated myelopathy). So far five cases of this illness have been detected in Spain, five of them in immigrants. We present a new case in Spain, with a characteristic chronic clinical picture. CASE REPORT: A 36-year-old black woman native of Ecuatorial Guinea, developed along 10 years a progressive paraparesis of asymmetric onset with important back pain, that arrives to paraplegic spastic phase at the present time. She presents distal amyotrophies, ulcers of decubitus and loss of control of sphincters, with normal mental status. Laboratory tests: blood, biochemistry and microbiologic studies: normal, or negative. She presented positive Western Blot serology for HTLV-I, confirmed by means of PCR technique. Cranial MRI: small and hyperintense subcortical lesions on T2 weighted images; spinal MRI: local atrophy at high thoracic level. A lumbar puncture was performed, with no cells, and with presence of oligoclonal bands, and a high IgG index. Urodynamic study: neurogenic spastic bladder. EMG: mild axonal polyneuropathy with prevalence in legs. CONCLUSIONS: In the differential diagnosis of progressive paraperesis, and mainly with epidemic antecedents, it is necessary to include a determination of HTLV-I between the diagnostic tests.

Prevalence and determinants of high-risk human papillomavirus (HPV) infection and cervical cytological abnormalities in imprisoned women.

The aim of the study was to estimate the prevalence and risk factors associated with infection by high-risk human papillomavirus (HR-HPV) in cervix and squamous intra-epithelial lesions (SIL) in imprisoned women. This was done by a cross-sectional study of imprisoned women attending the gynaecological clinic in Foncalent prison in Alicante, Spain. The study period was from May 2003 to December 2005. HR-HPV infection was determined through Digene HPV Test, Hybrid Capture II (HC-II). HPV typing was determined by multiplex nested PCR assay combining degenerate E6/E7 consensus primers. Multiple logistic regression modelling was used for the analysis of associations between variables where some were considered possible confounders after checking for interactions. A total of 219 women were studied. HR-HPV prevalence was 27.4% and prevalence of SIL was 13.3%. HIV prevalence was 18%, higher in Spaniards than in migrant women (24.6% vs. 14.3%, P<0.05). In multivariate analyses, risk factors for HPV infection were younger age (P for trend=0.001) and tobacco use (OR 2.62, 95% CI 1.01-6.73). HPV infection (OR 4.8, 95% CI 1.7-13.8) and HIV infection were associated with SIL (OR 4.8, 95% CI 1.6-14.1). The commonest HPV types were HPV16 (29.4%), HPV18 (17.6%), HPV39 (17.6%) and HPV68 (17.6%). The prevalence of both HR-HPV infection and SIL in imprisoned women found in this study is high. Determinants for each of the outcomes studied were different. HPV infection is the most important determinant for SIL. A strong effect of HIV co-infection on the prevalence of SIL has been detected. Our findings reinforce the need to support gynaecological clinics in the prison setting.

Oncogenic human papillomavirus (HPV) type distribution and HPV type 16 E6 variants in two Spanish population groups with different levels of HPV infection risk.

The aim of this study is to determine oncogenic human papillomavirus (HPV) types and HPV type 16 (HPV16) variant distribution in two Spanish population groups, commercial sex workers and imprisoned women (CSW/IPW) and the general population. A multicenter cross-sectional study of 1,889 women from five clinical settings in two Spanish cities was conducted from May to November 2004. Oncogenic HPV infection was tested by an Hybrid Capture II (HC2) test, and positive samples were genotyped by direct sequencing using three different primer sets in L1 (MY09/11 and GP5+/GP6+) and E6/E7. HPV16 variants were identified by sequencing the E6, E2, and L1 regions. Four hundred twenty-five samples were positive for the HC2 test, 31.5% from CSW/IPW and 10.7% from the general population. HPV16 was the most frequent type. Distinct profiles of oncogenic HPV type prevalence were observed across the two populations. In order of decreasing frequency, HPV types 16, 31, 58, 66, 56, and 18 were most frequent in CSW/IPW women, and types 16, 31, 52, 68, 51, and 53 were most frequent in the general population. We analyzed HPV16 intratype variants, and a large majority (78.7%) belonged to the European lineage. AA variants were detected in 16.0% of cases. African variants belonging to classes Af1 (4.0%) and Af2 (1.3%) were detected. Different HPV types and HPV16 intratype variants are involved in oncogenic HPV infections in our population. These results suggest that HPV type distribution differs in CSW/IPW women and in the general population, although further analysis is necessary.