[Guillain-Barre syndrome in pregnancy]. / Síndrome de Guillain-Barré en el embarazo.

TITLE: Síndrome de Guillain-Barré en el embarazo.

Pseudothrombocytopenia by ethylenediaminetetraacetic acid can jeopardize patient safety - report.

Pseudothrombocytopenia by ethylenediaminetetraacetic acid (EDTA) is an infrequent phenomenon of in vitro platelet agglutination due to the presence of antiplatelet autoantibodies. It has no clinical significance, but misdiagnosis may lead to clinical or therapeutic decision-making. In this study we report a case of an 8-year-old boy with no history of platelet disorder presenting a low platelet count and a peripheral blood smear showing clumping of platelets by EDTA. The initial diagnosis hypothesis was of an idiopathic thrombocytopenic purpura, and an unnecessary bone marrow aspirate was made even though he did not have personal or family history of bleeding. A second sample collected in sodium citrate confirmed the pseudothrombocytopenia by EDTA. In conclusion, the laboratory should enhance a strong relationship with clinicians trying to avoid misunderstandings as that reflected in this case report. It should be reminded that, in those cases where a pseudothrombocytopenia by EDTA is suspected, a blood smear is mandatory to confirm platelet clumps and blood must be tested anticoagulated with another anticoagulant (i.e., sodium citrate or heparin).

The Time-resolved and Extreme-conditions XAS (TEXAS) facility at the European Synchrotron Radiation Facility: the energy-dispersive X-ray absorption spectroscopy beamline ID24.

The European Synchrotron Radiation Facility has recently made available to the user community a facility totally dedicated to Time-resolved and Extreme-conditions X-ray Absorption Spectroscopy--TEXAS. Based on an upgrade of the former energy-dispersive XAS beamline ID24, it provides a unique experimental tool combining unprecedented brilliance (up to 10(14) photons s(-1) on a 4 µm × 4 µm FWHM spot) and detection speed for a full EXAFS spectrum (100 ps per spectrum). The science mission includes studies of processes down to the nanosecond timescale, and investigations of matter at extreme pressure (500 GPa), temperature (10000 K) and magnetic field (30 T). The core activities of the beamline are centered on new experiments dedicated to the investigation of extreme states of matter that can be maintained only for very short periods of time. Here the infrastructure, optical scheme, detection systems and sample environments used to enable the mission-critical performance are described, and examples of first results on the investigation of the electronic and local structure in melts at pressure and temperature conditions relevant to the Earth's interior and in laser-shocked matter are given.

Lessons from an outbreak of metallo-ß-lactamase-producing Klebsiella oxytoca in an intensive care unit: the importance of time at risk and combination therapy.

BACKGROUND: Outbreaks of nosocomial infection due to carbapenem-resistant Enterobacteriaceae (CRE), mostly Klebsiella spp., have become a worldwide phenomenon. AIM: To investigate the risk factors for the acquisition of clonal multidrug-resistant Klebsiella oxytoca (MDRKO) producing the metallo-ß-lactamase IMP-8 and hyperproducing chromosomal OXY-2 ß-lactamase during a well-characterized outbreak, and to describe the clinical features of infections due to MDRKO. METHODS: A four-wave outbreak due to MDRKO occurred in the intensive care unit of a Spanish hospital between 2009 and 2011. The risk factors for acquisition of MDRKO during waves 1 and 2 (in which colonized patients served as the main reservoir for the epidemic strain) were analysed using a case-control study by Cox regression and logistic regression analysis. Clinical data and treatments of patients infected with MDRKO were also analysed. FINDINGS: For the study of risk factors, 26 cases and 45 controls were studied. None of the variables studied in the Cox regression analysis showed an association with MDRKO acquisition; time at risk was the only associated variable by logistic regression analysis. Colonization pressure was not associated with earlier acquisition. Overall, 14 patients were infected with MDRKO; ventilator-associated pneumonia (seven patients) was the most frequent type of infection. Monotherapy tended to be associated with higher mortality than combination therapy [60% (3/5) vs 16.6% (1/6); P = 0.07]. CONCLUSIONS: Time at risk was the most significant risk determinant for the acquisition of carbapenem-resistant Enterobacteriaceae (CRE) in this epidemiological context and should be included in any study of risk factors for the acquisition of multidrug-resistant bacteria. Combination therapy may be superior to monotherapy for the treatment of CRE infections.

Enhanced production of a lutein-rich acidic environment microalga.

AIMS: This study was aimed at increasing productivity of a novel lutein-rich acidic environment microalga, Coccomyxa onubensis, based on efficient inorganic carbon use. METHODS AND RESULTS: Productivity was determined based on dry weight data; inorganic carbon concentration mechanisms were determined by means of carbonic anhydrase activity; carotenoids were extracted with methanol and measured by HPLC techniques. The existence of carbon concentration mechanisms and conditions that might lead to use them for addressing increased productivity of C. onubensis was studied. Best growth and carbon uptake capacity occurred at acidic pH, proving acid-tolerant behaviour of C. onubensis. Incubation in air followed by shift to high carbon conditions enhanced carbon-use efficiency in terms of growth rate and biomass productivity, based on the action of both carbonic anhydrase activities. Lutein accumulated in the microalga at high concentrations above 5-6 g kg(-1) dry weight and did not depend on inorganic carbon conditions. CONCLUSIONS: Consequently, repeated cycles of air incubation and high CO2 incubation of C. onubensis might become a suitable tool to perform production processes of lutein-enriched biomass. SIGNIFICANT AND IMPACT OF THE STUDY: This study intends to show that acidic environment microalgae can be produced at similar productivities of nonextreme microalgae, with the added advantage of their growth in highly selective culture medium. Particularly, it is applied to C. onubensis which accumulates lutein at commercially relevant concentrations.

Wastewater drainage system as an occult reservoir in a protracted clonal outbreak due to metallo-ß-lactamase-producing Klebsiella oxytoca.

We describe the epidemiology of a protracted nosocomial clonal outbreak due to multidrug-resistant IMP-8 producing Klebsiella oxytoca (MDRKO) that was finally eradicated by removing an environmental reservoir. The outbreak occurred in the ICU of a Spanish hospital from March 2009 to November 2011 and evolved over four waves. Forty-two patients were affected. First basic (active surveillance, contact precautions and reinforcement of surface cleaning) and later additional control measures (nurse cohorting and establishment of a minimum patient/nurse ratio) were implemented. Screening of ICU staff was repeatedly negative. Initial environmental cultures, including dry surfaces, were also negative. The above measures temporarily controlled cross-transmission but failed to eradicate the epidemic MDRKO strain that reappeared two weeks after the last colonized patients in waves 2 and 3 had been discharged. Therefore, an occult environmental reservoir was suspected. Samples from the drainpipes and traps of a sink were positive; removal of the sink reduced the rate number but did not stop new cases that clustered in a cubicle whose horizontal drainage system was connected with the eliminated sink. The elimination of the horizontal drainage system finally eradicated the outbreak. In conclusion, damp environmental reservoirs (mainly sink drains, traps and the horizontal drainage system) could explain why standard cross-transmission control measures failed to control the outbreak; such reservoirs should be considered even when environmental cultures of surfaces are negative.

Therapeutic effect of an altered peptide ligand derived from heat-shock protein 60 by suppressing of inflammatory cytokines secretion in two animal models of rheumatoid arthritis.

Rheumatoid arthritis is a systemic autoimmune disease mediated by T cells. Productive engagement of T cell receptors by major histocompatibility complex-peptide leads to proliferation, differentiation and the definition of effector functions. Altered peptide ligands (APL) generated by amino acid substitutions in the antigenic peptide have diverse effects on T cell response. We predicted a novel T cell epitope from human heat-shock protein 60, an autoantigen involved in the pathogenesis of rheumatoid arthritis. Three APLs were designed from this epitope and it was demonstrated that these peptides induce the activation of T cells through their ability to modify cell cycle phase's distribution of CD4+T cells from RA patients. Also, IL-17, TNF-α and IL-10 levels were determined in PBMC from these patients. Unlike the wild-type peptide and the other two APLs, APL2 increased the IL-10 level and suppressed IL-17 secretion in these assays. Therapeutic effect of this APL in adjuvant arthritis (AA) and collagen-induced arthritis (CIA) models was also evaluated. Clinical score, histopathology, inflammatory and regulatory cytokine concentration were monitored in the animals. APL2 efficiently inhibited the progression of AA and CIA with a significant reduction of the clinical and histopathologic score. Therapeutic effect of APL2 on CIA was similar to that obtained with MTX; the standard treatment for RA. This effect was associated with a decrease of TNF-α and IL-17 levels. These results suggest that the therapeutic effect of APL2 is mediated in part by down-regulation of inflammatory cytokines and support the potential use of APL2 as a therapeutic drug in RA patients.

Geochemical behavior of heavy metals in a Zn-Pb-Cu mining area in the State of Mexico (central Mexico).

The geochemical behavior of zinc, lead and copper from sulfidic tailings in a mine site with potential to generate acidic drainage (pyrite (55%) and sphalerite (2%)) is reported in this paper. The mining area is divided in two zones, considering the topographic location of sampling points with respect to the tailings pile: (a) outer zone, out of the probable influence of acid mine drainage (AMD) pollution, and (b) inner zone, probably influenced by AMD pollution. Maximum total ions concentrations (mg/L) measured in superficial waters found were, in the outer zone: As (0.2), Cd (0.9), Fe (19), Mn (39), Pb (5.02), SO4(2-) (4650), Zn (107.67), and in the inner zone are As (0.1), Cd (0.2), Fe (88), Mn (13), Pb (6), SO4(2-) (4,880), Zn (46). The presence of these ions that exceeding the permissible maximum limits for human consume, could be associated to tailings mineralogy and acid leachates generated in tailings pile.

Caracterización de moléculas HLA tipo II y evaluación de citocinas en pacientes cubanos con artritis reumatoide

La Artritis Reumatoide es una enfermedad de etiología multifactorial, que involucra la presencia de factores genéticos, ambientales, inmunológicos y hormonales. En los últimos años se ha establecido una asociación entre la predisposición a padecer esta enfermedad y la existencia de determinados haplotipos del HLA clase II. El objetivo fundamental de este trabajo es la caracterización del polimorfismo de las moléculas HLA-DQB1 y HLA-DRB de un grupo de pacientes cubanos con Artritis Reumatoide, además analizar una posible correlación entre los niveles de varias citocinas proinflamatorias en un grupo de estos pacientes, con los alelos HLA tipo II genotipados, el sexo y el tiempo de diagnosticada la enfermedad. El estudio se llevo a cabo en 50 pacientes cubanos con el diagnóstico de Artritis Reumatoide y un grupo control, compuesto de 211 donantes sanos. Los haplotipos HLA-DQ y HLA-DRB1 fueron determinados a través de la reacción en cadena de la polimerasa. La cuantificación de las citocinas se realizó empleando inmuno-ensayos comerciales. Los resultados obtenidos en este análisis indican que los alelos con un radio de la razón de probabilidades superior a 2 fueron para el caso de la molécula HLA-DQB 1: HLA-DQB1*03 y *06, y para la molécula HLA-DRB 1 los alelos: *01, *04, *09 y *10. Encontramos además que los niveles de la citocina interferon ganma están significativamente aumentados en los pacientes con menos tiempo de diagnosticada la enfermedad. Este trabajo constituye el primer reporte de caracterización de las moléculas HLA tipo II, a través de técnicas moleculares, en pacientes cubanos con Artritis Reumatoide(AU)

Autoimmunity and molecular mimicry in tropical spastic paraparesis/human T-lymphotropic virus-associated myelopathy.

Viruses share antigenic sites with normal host cell components, a phenomenon known as molecular mimicry. It has long been suggested that viral infections might trigger an autoimmune response by several mechanisms including molecular mimicry. More than 600 antiviral monoclonal antibodies generated against 11 different viruses have been reported to react with 3.5% of cells specific for uninfected mouse organs. The main pathological feature of tropical spastic paraparesis/human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (TSP/HAM) is a chronic inflammation of the spinal cord characterized by perivascular cuffing of mononuclear cells accompanied by parenchymal lymphocytic infiltration. We detected the presence of autoantibodies against a 98- to 100-kDa protein of in vitro cultured human astrocytes and a 33- to 35-kDa protein from normal human brain in the serum of HTLV-I-seropositive individuals. The two cell proteins exhibited molecular mimicry with HTLV-I gag and tax proteins in TSP/HAM patients, respectively. Furthermore, the location of 33- to 35-kDa protein cross-reaction correlated with the anatomical spinal cord areas (in the rat model) in which axonal damage has been reported in several cases of TSP/HAM patients. Our experimental evidence strongly suggests that the demyelinating process occurring in TSP/HAM may be mediated by molecular mimicry between domains of some viral proteins and normal cellular targets of the spinal cord sections involved in the neurodegeneration.

Autoimmunity and molecular mimicry in tropical spastic paraparesis/human T-lymphotropic virus-associated myelopathy

Viruses share antigenic sites with normal host cell components, a phenomenon known as molecular mimicry. It has long been suggested that viral infections might trigger an autoimmune response by several mechanisms including molecular mimicry. More than 600 antiviral monoclonal antibodies generated against 11 different viruses have been reported to react with 3.5 percent of cells specific for uninfected mouse organs. The main pathological feature of tropical spastic paraparesis/human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (TSP/HAM) is a chronic inflammation of the spinal cord characterized by perivascular cuffing of mononuclear cells accompanied by parenchymal lymphocytic infiltration. We detected the presence of autoantibodies against a 98- to 100-kDa protein of in vitro cultured human astrocytes and a 33- to 35-kDa protein from normal human brain in the serum of HTLV-I-seropositive individuals. The two cell proteins exhibited molecular mimicry with HTLV-I gag and tax proteins in TSP/HAM patients, respectively. Furthermore, the location of 33- to 35-kDa protein cross-reaction correlated with the anatomical spinal cord areas (in the rat model) in which axonal damage has been reported in several cases of TSP/HAM patients. Our experimental evidence strongly suggests that the demyelinating process occurring in TSP/HAM may be mediated by molecular mimicry between domains of some viral proteins and normal cellular targets of the spinal cord sections involved in the neurodegeneration.

IL8 release, tight junction and cytoskeleton dynamic reorganization conducive to permeability increase are induced by dengue virus infection of microvascular endothelial monolayers.

Permeability alterations of microvascular endothelia may be a factor in the plasma leakage produced by dengue virus infection. Confluent monolayers of the human dermal microvascular endothelial cell line HMEC-1 were utilized as an experimental model to study the cellular responses induced by the virus. Infected monolayers showed increased permeability for [(3)H]mannitol, but no changes were observed for 4-70 kDa dextrans at 48 h post-infection (p.i.), a time at which viral titres reached maximal values and 40 % of the cells expressed viral proteins. A further increase in permeability occurred at 72 h, still without evident cytopathic effects on the monolayer. Coinciding with this, actin was reorganized in the infected cells and the tight junction protein occludin was displaced to the cytoplasm. Increments in the thickness of stress fibres and focal adhesions were observed in uninfected cells neighbouring infected cells. Culture medium from infected monolayers induced permeability changes and thickening of actin-containing structures in control cultures that resembled those observed 48 h p.i. Interleukin (IL) 8 was found in culture medium at concentrations ranging from 20 to 100 pg ml(-1). Neutralizing antibodies against IL8 partially inhibited the changes produced by the culture medium as well as those induced by addition of IL8. Genistein inhibited the effect of the culture medium and the phosphorylation of proteins associated with focal adhesions and indicated the participation of tyrosine kinases. These findings suggest that IL8 production by infected monolayers contributes to the virus-induced effect on the cytoskeleton and tight junctions and thereby modifies transendothelial permeability.

Application of a spectrophotometric method for the determination of post-antibiotic effect and comparison with viable counts in agar.

The post-antibiotic effects of gentamicin and ciprofloxacin at 1x, 2x and 4x MIC on Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213 were studied using a spectrophotometric method and the classic method of viable counts on agar as a reference. Monitoring of the growth kinetics was carried out by viability counting on the plate every hour and by means of the optical density of the cultures measured by spectrophotometry at a wavelength of 450 nm. No statistically significant differences were found between the results obtained with the spectrophotometric method and the reference method. The former method was much quicker, much easier to use and to replicate.

The effect of paprika, garlic and salt on rancidity in dry sausages.

Eight batches of ground fat and eight batches of a mixture of meat and fat (70 and 30%, respectively), were prepared by adding salt, paprika and garlic, in the proportions used for making chorizo - a dry fermented sausage - stuffed into natural casings and then ripened. The evolution of rancidity in these products was evaluated by means of total free fatty acid content, peroxide value and TBARS during the ripening period. Spanish paprika and salt showed antioxidant and prooxidant properties, respectively. Paprika was even able to inhibit the prooxidant effect of salt. Also, four batches of chorizo were made to compare the antioxidant effect of the spices (garlic and paprika) with a mixture of nitrate, nitrite and ascorbic acid. In this respect, paprika and garlic were as effective as the mixture of additives in inhibiting lipid oxidation.

[Role of oxidative stress in the maintenance of inflamation in patients with juvenile rheumatoid arthritis] / Papel do estresse oxidativo na manutenção da inflamação em pacientes com artrite reumatóide juvenil.

OBJECTIVE: To determine the level of cellular oxidative stress blood markers and the enzymatic system of antioxidant defense establishing the oxidative profile in patients with Juvenile Rheumatoid Arthritis. METHODS: Case-control study that included 64 patients (46 of female sex) with Juvenile Rheumatoid Arthritis (JRA) following clinical control in the Pediatric Rheumatology Service of the Vall dacute;Hebron Hospital, Barcelona, Spain. The patients were separated in three subtypes based on the pattern of onset within the first six months of disease: polyarticular, pauciarticular and systemic. The control group included 60 patients (38 of female sex) following clinical control to diseases of non inflammatory nature, in the same hospital. The plasmatic levels of malondialdehyde (MDA), lipoperoxide (LPO), hydroperoxide (HPX), carbonile groups (CG) of proteins and gluthathione and the enzymatic activities of Superoxide dismutase (SOD), gluthathione peroxidase (GSH-Px) and gluthathione reductase were determined. RESULTS: The group of patients with JRA presented high concentrations of lipid peroxidation products, evaluated by determining the plasmatic levels of MDA, LPO, and HPX; oxidative damage of the circulate protein, determined by CG contents of plasma proteins; elevation of enzymatic activity of SOD and GSH-Red; decrease of GSH-Px activity and GSH levels. CONCLUSIONS: Our results show the presence of molecular damage determined by oxygen free radicals in the JRA patients. The SOD activity and the changes of gluthathione redox enzymatic cycle confirm the decrease of capacity of cellular defense system against the induced toxicity of oxidative stress in these patients.

Recurrence of the D409H mutation in Spanish Gaucher disease patients: description of a new homozygous patient and haplotype analysis.

Gaucher disease results, in most patients, from mutations in the gene encoding glucocerebrosidase. Mutation D409H is the third most frequent in Spanish patients, accounting for 5.7% of all mutated alleles. This allele is associated mainly with the neurological forms of the disease. Recently, homozygosity for the D409H mutation has been associated with a particular phenotype, including specific cardiovascular symptoms. Here we report a second Spanish patient bearing the D409H/D409H genotype with a very early manifestation of the disease. The patient started enzyme replacement therapy at 3 months of age. A common origin for the Spanish D409H alleles was ruled out by haplotype analysis using an internal polymorphism of the glucocerebrosidase gene and two external microsatellite markers.

Broadly reactive antibodies against a gp120 V3 loop multi-epitope polypeptide neutralize different isolates of human immunodeficiency virus type 1 (HIV-1).

A gene encoding for a novel multi-epitope polypeptide (TAB4) was synthesized and expressed in Escherichia coli. The protein was composed of 15 amino acid fragments derived from the V3 loop of HIV-1 isolates MN, IIIB, RF, JY1, BRVA and LR150, joined by five-amino-acid linkers. Immunogenicity of TAB4 in rabbits was studied, and the antibody response against individual peptides investigated. TAB4 was shown to be immunogenic in Complete Freund's Adjuvant in a dose-dependent manner, and was able to elicit a humoral response against all V3 epitopes included on the protein. Sera from some of the animals were able to neutralize the replication of viral strain MN, and in one case IIIB, with moderate titers. Some sera also neutralized several Cuban clinical strains, isolated in peripheral blood mononuclear cells, after one round of amplification in MT4 cells.

[Bacteremias due to the Streptococcus milleri group. An analysis of 18 episodes]. / Bacteriemias por el grupo Streptococcus milleri. Análisis de 18 episodios.

OBJECTIVE: To study the epidemiological and clinical characteristics of bacteremia caused by Streptococcus milleri group streptococci (SMG). METHODS: Prospective evaluation of all bacteremic episodes with clinical significance from 1990 to 1995 in two general hospitals. In this study all episodes caused by SMG were analyzed. RESULTS: A total of 905 bacteremic episodes with clinical significance were detected; 18 (1.98%) were caused by SMG (0.16/1,000 admissions). The mean age of patients were 43 years and the male/female ratio 1.6. Seventeen patients (94.4%) had some underlying disease; nine patients had diabetes, four were parenteral drug abusers, and two had neoplasms. The most common sources of bacteremia were intraabdominal in four episodes (two liver abscesses, one subphrenic abscess and one pancreatic pseudocyst), cutaneous and/or soft tissues in four, surgical wound in two and respiratory in two; no source was identified in five episodes. Four episodes had a polymicrobial origin. In 13 isolates the identification was at species level (Streptococcus anginosus eight, Streptococcus intermedius four and Streptococcus constellatus one). All strains were susceptible to penicillin. Six patients (33.3%) required surgery. In ten episodes a favorable outcome was recorded, although four patients required surgery. The infection associated mortality rate was 31.2%. The mean age of deceased patients was higher than for cured patients (62.2 +/- 20.2 versus 35.3 +/- 20.3; p < 0.05). CONCLUSIONS: SMB bacteremia is uncommon. It involved mainly diabetic patients or parenteral drug abusers, commonly with an intraabdominal suppurative source or in skin or soft tissues. The mortality rate was high despite surgery in one third of patients. Patients with advanced age had a poorer prognosis. All isolates investigated were susceptible to penicillin.