Reliable resolution of ambiguous hepatitis C virus genotype 1 results with the Abbott HCV Genotype Plus RUO assay.

Accurate subtyping of hepatitis C virus genotype 1 (HCV-1) remains clinically and epidemiologically relevant. The Abbott HCV Genotype Plus RUO (GT Plus) assay, targeting the core region, was evaluated as a reflex test to resolve ambiguous HCV-1 results in a challenging sample collection. 198 HCV-1 specimens were analysed with GT Plus (38 specimens with and 160 without subtype assigned by the Abbott RealTime Genotype II (GT II) assay targeting the 5'NC and NS5B regions). Sanger sequencing of the core and/or NS5B regions were performed in 127 specimens without subtype assignment by GT II, with "not detected" results by GT Plus, or with mixed genotypes/subtypes. The remaining GT Plus results were compared to LiPA 2.0 (n = 45) or just to GT II results if concordant (n = 26). GT Plus successfully assigned the subtype in 142/160 (88.8%) samples. "Not detected" results indicated other HCV-1 subtypes/genotypes or mismatches in the core region in subtype 1b. The subtyping concordance between GT Plus and either sequencing or LiPA was 98.6% (140/142). Therefore, combined use of GT II and GT Plus assays represents a reliable and simple approach which considerably reduced the number of ambiguous HCV-1 results and enabled a successful subtyping of 98.9% of all HCV-1 samples.

A novel MYH7 founder mutation causing Laing distal myopathy in Southern Spain.

MYH7 gene mutations are associated with wide clinical and genetic heterogeneity. We report a novel founder mutation in MYH7 in Southern Spain (Andalucía). We studied two index patients and 24 family members from two apparently independent families by physical examination, serum creatine-kinase, muscle MRI, sequencing studies and genetic linkage analysis. Sixteen individuals were heterozygous for a (p.R1560P) variant in the MYH7 gene. Haplotype was consistent with a common ancestor for the two families. The patients displayed the classic Laing distal myopathy phenotype, with hanging first toe as the initial presentation, even in mildly affected patients who declared themselves asymptomatic, although neck flexor weakness was revealed as an early sign in some cases. MRI showed that the sartorius was the first muscle involved, even in two out of three asymptomatic carriers. Our findings support the novel variant p.R1560P in MYH7 as a founder mutation in Andalucía. The early involvement of the sartorius muscle in MRI may be useful as an indicator of affection status.

Clinical, virological and phylogenetic characterization of a multiresistant HIV-1 strain outbreak in naive patients in southern Spain.

BACKGROUND: We describe the characteristics of an HIV-1 strain with six viral reverse transcriptase mutations (D67N, T69N/D, V118I, V179D, T215S and K219Q), which we have called the Malaga strain. This strain was detected in treatment-naive patients from southern Spain. METHODS: The study was undertaken at the Virgen de la Victoria Hospital, Malaga, a reference centre for the study of HIV-1 genotype resistance in Andalusia (the 'Costa del Sol'), Spain. Genotypic resistance testing was done in an automated sequencer. Phylogenetic analysis was performed using a 630 bp region of the reverse transcriptase with the mutations mentioned. RESULTS: Between 2007 and 2014, we detected the Malaga strain in 30 treatment-naive patients. All were MSM, seen at five hospitals on the Costa del Sol. In all cases, the HIV-1 was subtype B with viral tropism R5. Phylogenetic analysis based on the reverse transcriptase sequence showed consistent grouping (with a bootstrap value of the common node of 100%) of the isolates that shared the mutation pattern mentioned. This strain has not been detected elsewhere or in previously treated patients. All of the patients treated with first-line combination ART responded. CONCLUSIONS: We report a cluster of an HIV-1 strain with multiple resistance mutations that was transmitted over a period of >8 years, affecting 30 naive patients from the same geographical area. The strain was susceptible to first-line combination ART.

Surface enhanced Raman scattering study of the antioxidant alkaloid boldine using prismatic silver nanoparticles.

Prismatic silver nanoparticles (PNps) were used in the surface enhanced Raman scattering (SERS) study of the antioxidant alkaloid boldine (5,6,6a,7-tetrahydro-1,10-dimethoxy-6-methyl-4H-dibenzo[de,g]quinoline-2,9-diol). Prismatic and quasi-spherical (QsNps) silver nanoparticles were synthesized and characterized by UV-Vis spectra, topographic profile (AFM) and zeta potential measurements. Raman and infrared (IR) spectra of the boldine were registered. Theoretical model calculations of the boldine onto the Ag surface predict a nearly coplanar orientation of the benzo[de]quinoline moiety and non-bonded interactions (electrostatic).

Surface-enhanced Raman scattering and theoretical studies of the C-terminal peptide of the ß-subunit human chorionic gonadotropin without linked carbohydrates.

Raman and surface-enhanced Raman scattering (SERS) spectra of the synthetic carboxy terminal peptide of human chorionic gonadatropin ß-subunit free of carbohydrate moieties(P37) are reported. The spectral analysis is performed on the basis of our reported Raman spectrum and SERS data of oligopeptides displaying selected amino acids sequences MRKDV, ADEDRDA, and LGRGISL. SERS samples of P37 were prepared by coating the solid peptide with metal colloids on a quartz slide. This treatment makes possible to obtain high spectral batch to batch reproducibility. Amino acids components of P37 display net charges and hydrophobic characteristics, which are related to particular structural aspects of the adsorbate-substrate interaction. The spectroscopic results are supported by quantum chemical calculations performed by using extended Hückel theory method for a model of P37 interacting with an Ag surface. The P37-metal interaction is drove by positively charged fragments of selected amino acids,mainly threonine 109, lysine 122, and arginine in positions 114 and 133. Data here reported intend to contribute to the knowledge about the antigen-antibody interaction and to the drugs delivery research area

SERS and theoretical studies of arginine.

Arginine amino acid (Arg) has been vibrationally studied through its infrared, Raman and surface-enhanced Raman scattering (SERS) spectra, and theoretical calculations. Net charge is used to predict the possibility to obtain the SERS spectrum of Arg in colloidal solution. The interpretation of the SERS spectral data suggests that the Arg-Ag nanoparticles interaction in a colloidal solution and in the case of the Arg coated by Ag is mainly verified through the guanidinium moiety. Theoretical calculations performed by using extended Hückel theory method for a model of Arg interacting with an Ag cluster support the observed SERS experimental result.

Role of within-individual variation in capitulum size and achene mass in the adaptation of the annual Centaurea eriophora to varying water supply in a Mediterranean environment.

To clarify the adaptive value of variation in capitulum size and achene mass, plants of Centaurea eriophora were studied in a glasshouse and in a natural population. C. eriophora plants consist of a basal leaf rosette from which an erect stem grows, with lateral branches of various orders ending in capitula of different orders. Primary, secondary and tertiary capitula are comparable in size and they produce similar numbers of achenes, which are similar in weight (large achenes). These capitula are formed during April, May and early June, and constitute the normal or primary flowering. Following ripening of tertiary capitula, leaves senesce, but, later during June and the first half of July, a secondary flowering of a variable number of smaller capitula may occur if wet conditions persist for longer than usual. Plants that have almost senesced develop small lateral branches 1-2 cm long bearing a few small leaves and ending in a capitulum about half the diameter of capitula from the primary flowering period. The number of achenes produced in these capitula (small achenes) and their weight are 70 and 30% less, respectively, than those of capitula formed during primary flowering. These reductions appear to result from restricted availability of resources. Large and small achenes have similar dispersal characteristics and possess similar germination potential. However, large achenes produce seedlings that are capable of emerging from greater burial depths, providing the resulting plants with a potential advantage. The normal flowering period coincides with the optimum time of year for flowering and fruiting in the south of Spain, and only if rainfall lasts longer than usual does secondary flowering occur. Secondary flowering extends the normal flowering and fruiting periods, thereby providing a supplementary crop of smaller, yet viable, fruits. It can be considered to be an adaptive response to the unpredictable Mediterranean climate, optimizing the use of available resources.

Evaluation of three methods to measure anti-Brucella IgM antibodies and interference of IgA in the interpretation of mercaptan-based tests.

The results of a dipstick assay for the detection of immunoglobulin M (IgM) to Brucella smooth lipopolysaccharide (S-LPS) correlated with those of an enzyme-linked immunosorbent assay (ELISA) for IgM and of the serum agglutination test (SAT) performed with and without dithiothreitol. Two sera which were dithiothreitol-sensitive and were dipstick negative were shown to contain specific IgA. The dipstick assay is recommended as a simple method for detecting specific IgM antibodies in acute-phase brucellosis patients.

[Temporal evolution of tuberculosis and human immunodeficiency virus infection in a population cared for in a hospital in Málaga]. / Evolución temporal de la tuberculosis e infección por el virus de la inmunodeficiencia humana en la población atendida por un hospital de Málaga.

BACKGROUND: We have studied the incidence of tuberculosis++ disease, HIV infection and their association during a period of 6 years using samples analysed in a third level hospital laboratory. MATERIAL AND METHOD: 21,242 samples for mycobacteria and 63,425 for HIV antibodies were analysed between 1993 and 1998. The protocol used for mycobacteria consisted of Lowenstein-Jensen, hemoculture, biochemical tests for identification and DNA probe. The diagnosis of HIV was performed using screening with mix EIA HIV 1 + 2, confirmed with Western-blot. Spearman coefficient correlation was used for study of tendency. RESULTS: 1,613 samples (7.5%) positive for mycobacteria from 566 patients (98 females and 428 males) aging between 31 and 40 years (46.9%) were detected. The highest incidence for tuberculosis was observed in 1995 (49.2 x 10(5) and was followed by a decreasing linear tendency. Mycobacterium tuberculosis (96%) and M. bovis (2.7%) were most frequent agents. 2,295 samples (3.6%) showed anti-HIV (406 females and 1,889 males). 54.1% of the infected patients belonged to the age interval 21 to 40 years. The highest incidence was observed in 1994 and was followed by a significant decrease (p < 0.05). The percentage of patients co-infected with tuberculosis-HIV was 39.7% with a maximum in 1995 followed by a linear decrease. CONCLUSIONS: The incidence of both infections and co-infection was very high in the first triennium and was followed by a progressive decrease. The decrease of HIV preceded tuberculosis. The result suggest a possible epidemiological correlation between both infections.

[Factors affecting the clinical course of bacteremia. Prospective study at a university hospital]. / Factores que influyen en la evolución de la bacteriemia. Estudio prospectivo en un hospital universitario.

BACKGROUND: The aim of this study was to identify the risk factors mainly influencing the mortality of a series of bacteremic patients. METHODS: A prospective study of the clinically significant bacteremias detected in the Hospital Clínico Universitario in Malaga (Spain) over the period from June 1994 to May 1995 was performed. Univariate analysis of the results was carried out with the chi 2 test and multivariate analysis was undertaken by logistic regression (Stepwise backward). RESULTS: The incidence of bacteremia was of 19.5 cases/1,000 admissions and the mortality was of 21.9%. According to the univariate analysis, 11 variables were significantly associated with greater risk death: age > 60 years, stay in the intensive care unit, respiratory diseases as the main diagnosis, etiology by Pseudomonas aeruginosa, absence of fever, septic shock, presence of chronic renal insufficiency, cirrhosis or heart disease with underlying diseases, performance of invasive procedures prior to and hospital stay of less than 10 days. Logistic regression analysis determined the factors which mainly influenced in the prognosis of bacteremia: septic shock (p < 0.0001, odds ratio [OR]: 17.97), cardiovascular diseases (p = 0.004, OR: 9.86), AIDS (p = 0.03; OR: 1.02). CONCLUSIONS: Although the prognosis of bacteremia is strongly influenced by determined conditions of the patient, it may be improved, overall through the control of possible hemodynamic complications of the patients and to a lesser extent by antibiotic treatment.

Analysis of risk factors associated with nosocomial bacteraemias.

A prospective study of 2676 blood cultures was performed to identify the factors associated with clinically, significant nosocomial bacteraemia that occurred during a one year period in the Malaga University Clinical Hospital. Three hundred and fifty-five episodes of bacteraemia were considered clinically significant. The overall incidence of bacteraemia was 19.5/1000 admissions, of which 46% were hospital-acquired. A multivariate model showed that only six factors were significantly, and independently, responsible for nosocomial bacteraemias: intravascular catheterization (P < 0.0001, OR = 18.37), invasive procedures (P < 0.0001, OR = 10.38), malignancy (P = 0.035, OR = 3.11), indwelling devices (P = 0.005, OR = 3.05), stay in intensive care or surgical departments (P = 0.05, OR = 2.63) and length of hospital stay (P = 0.051, OR = 1.02). These results show that the factors which had most influence on the development of nosocomial bacteraemias were those factors associated with the treatment received by patients during their hospital stay.

Valvuloplastia mitral percutánea con cateter balon de inoue: resultados de 74 pacientes / Percutanea mitral Valvuloplastia with cateter ball of inoue: results of 74 patients

Expone que el uso del balón de INOE, facilitó la realización de la valvuloplastia mitral, con disminución en el tiempo del procedimiento, con incremento del éxito y redujo el número de insuficiencia mitral secundaria a valvuloplastia. Desde 1990 año en que se inició en nuestro servicio, el uso del balón de Inoue, hasta mar. 1996, se han realizado 74 valvuloplastias mitrales percutáneas, de estos el 87xcto fueron pacientes de sexo femenino y 13xcto del masculino, las edades variarion entre los 58 y 16 años (media 36+-9). En todos los pacientes se requirió un Score ecocardiográfico previo, realizándose la valvuloplastia hasta con un puntaje de 11, (media 6,95+-1,62). El área mitral se evaluó pre y post tratamiento por eco y por cateterismo cardiaco se usó la fórmula de Gorling. El área se incrementó de 1.01+-0.23 cm a 1.89+-0.4 cm (P menor que 0.05). El gradiente diastólico transvalvular mitral disminuyó de 20.16...

[Conjunctival ophthalmomyiasis caused by Oestrus ovis]. / Oftalmomiasis conjuntival causada por Oestrus ovis.

BACKGROUND: Microbiologic diagnosis of conjunctival ophthalmomyiasis. MATERIAL AND METHODS: Material extracted from three patients' conjunctiva who complained of sudden presence of foreign body and were diagnosed as conjunctival ophthalmomyiasis has been studied. Patients were recruited over two months from the Emergency Department. Diagnosis was performed through observation using the light microscope. RESULTS: Oestrus ovis first-instance larvas were observed in all the samples studied. CONCLUSIONS: This observation of similar cases in a short period of time and the low frequency of diseased people who lived in a urban environment makes necessary to think in this agent as cause of ophthalmomyiasis in our area and therefore perform an adequate differential diagnosis.