Molecular effect of fenofibrate on PBMC gene transcription related to lipid metabolism in patients with metabolic syndrome.

BACKGROUND: Both fasting and postprandial hypertriglyceridaemia are considered independent risk factors for atherosclerosis. Treatment of hypertriglyceridaemia is based on fibrates, which activate the peroxisome proliferator-activated receptor alpha (PPARα). However, the metabolic pathways that activate or inhibit fibrates, and how the postprandial triglyceride levels are modified, have not yet been fully described. Accordingly, the aim of this study was to determine the feasibility of peripheral blood mononuclear cells (PBMC) to study the effects of fenofibrate in patients with the metabolic syndrome. MATERIALS AND METHODS: A fat overload was given to 50 patients before and after treatment with fenofibrate for 3 months. Anthropometric and biochemical variables as well as gene expression in PBMC were analysed. RESULTS: After treatment with fenofibrate, we observed a decrease in both baseline and postprandial (3 h after the fat overload) levels of serum triglycerides, cholesterol and uric acid and an increase in HDL cholesterol and apolipoprotein AI levels. After treatment, there was also a rise in PPARα and RXRα expression and changes in genes regulated by PPARα, both baseline and postprandial. Furthermore, in vitro experiments showed that a PPARα agonist changed the expression of genes related with lipid metabolism. CONCLUSION: Treatment with fenofibrate reduced fasting and postprandial serum triglyceride levels, possibly through a mechanism related with an increase in the expression of RXRα and PPARα, by activating the pathways involved in the uptake and degradation of triglycerides and increasing the synthesis of apolipoprotein. These results suggest that PBMC may be useful for the easy study of fenofibrate actions.

Endotoxin increase after fat overload is related to postprandial hypertriglyceridemia in morbidly obese patients.

The low-grade inflammation observed in obesity has been associated with a high-fat diet, though this relation is not fully understood. Bacterial endotoxin, produced by gut microbiota, may be the linking factor. However, this has not been confirmed in obese patients. To study the relationship between a high-fat diet and bacterial endotoxin, we analyzed postprandial endotoxemia in morbidly obese patients after a fat overload. The endotoxin levels were determined in serum and the chylomicron fraction at baseline and 3 h after a fat overload in 40 morbidly obese patients and their levels related with the degree of insulin resistance and postprandial hypertriglyceridemia. The morbidly obese patients with the highest postprandial hypertriglyceridemia showed a significant increase in lipopolysaccharide (LPS) levels in serum and the chylomicron fraction after the fat overload. Postprandial chylomicron LPS levels correlated positively with the difference between postprandial triglycerides and baseline triglycerides. There were no significant correlations between C-reactive protein (CRP) and LPS levels. The main variables contributing to serum LPS levels after fat overload were baseline and postprandial triglyceride levels but not glucose or insulin resistance. Additionally, superoxide dismutase activity decreased significantly after the fat overload. Postprandial LPS increase after a fat overload is related to postprandial hypertriglyceridemia but not to degree of insulin resistance in morbidly obese patients.

Host cell-specific protein expression in vitro in Ehrlichia ruminantium.

Ehrlichia ruminantium, a tick-transmitted pathogen, is the causative agent of heartwater in ruminants. In this study, a proteomic approach was used to identify host cell-specific E. ruminantium proteins encoded by the map1 multigene family, expressed in vitro in bovine endothelial and tick cell cultures. Two-dimensional gel electrophoresis combined with mass spectrometry analysis was used to establish the identities of immunodominant proteins. Proteins extracted from E. ruminantium-infected endothelial cells were shown to be products of the map1 gene, whereas tick cell-derived E. ruminantium proteins were products of a different gene, map1-1. The expressed proteins were found to be glycosylated. Differential expression of MAP1 family proteins in vitro in mammalian and tick cell cultures indicates that the map1 multigene family might be involved in the adaptation of E. ruminantium to the mammalian host and vector tick.

Differential transcription of the major antigenic protein 1 multigene family of Ehrlichia ruminantium in Amblyomma variegatum ticks.

The rickettsial pathogen Ehrlichia ruminantium causes heartwater in ruminants and is transmitted by ticks of the genus Amblyomma. The map1 gene, encoding the major surface protein MAP1, is a member of a multigene family containing 16 paralogs. In order to investigate differential transcription of genes of the map1 multigene family in vivo in unfed and feeding ticks, RNA was extracted from midguts and salivary glands of E. ruminantium-infected adult female Amblyomma variegatum ticks and analysed by RT-PCR using MAP1 paralog-specific primers. In unfed ticks, only transcripts from the map1-1 gene were observed in midguts and no transcripts were detected in salivary glands. In feeding ticks, map1-1 transcripts were more abundant in midguts whereas high levels of map1 transcripts were observed in salivary glands. Our results show that differential transcription of genes of the E. ruminantium map1 cluster occurs in vivo in different tissues of infected ticks before and during transmission feeding, indicating that this multigene family may be involved in functions of biological relevance in different stages of the life cycle of E. ruminantium.

Lumbar segmental artery pseudoaneurysm after L5 pedicle screw placement. A rare vascular complication. / Pseudoaneurisma de arteria segmentaria lumbar tras introducción de tornillo transpedicular L5. Una rara complicación vascular.

Posterior lumbar screw fixation is a common surgical procedure nowadays. However, it can sometimes produce complications that can be devastating. One of the less common causes of major complication is the misplacement of a pedicle screw. This highlights the importance of being methodical when placing pedicle screws, and checking that the pathway has been created correctly and their placement. We present a case of a massive bleed after a pedicular screw placement during lumbar canal stenosis surgery. Screw malposition led to intraoperative haemodynamic instability after failed attempts to control bleeding in the surgical site. Contrast enhanced CT imaging revealed a lumbar intersegmentary artery injury that was eventually controlled by means of a coil embolisation.

Malaria diagnosis by the polymerase chain reaction: a field study in south-eastern Venezuela.

A polymerase chain reaction (PCR) method that amplifies genus- and species-specific sequences present within the small subunit of ribosomal ribonucleic acid (ssRNA) genes of the human malaria parasites was used for the diagnosis of malaria in south-eastern Venezuela. One hundred blood samples were submitted to deoxyribonucleic acid extraction, PCR amplification and electrophoretic analysis of the PCR products, and the results were compared to those of routine microscopical diagnosis. The sensitivity of PCR for detection of Plasmodium vivax and P. falciparum malaria was 99% and 100%, respectively. However, 6 patients (6%) harboured parasites undetected by microscopy. The PCR assay detected a high proportion of mixed infections: 29% (17/59) of the infections microscopically diagnosed as P. vivax were shown to be mixed infections of P. vivax and P. falciparum. Forty per cent (7/17) of the individuals with a missed P. falciparum infection had received chloroquine in the previous 30 d. These results suggest that, in places where transmission of both P. vivax and P. falciparum occurs, PCR detection of malaria parasites can be a very useful complement to microscopical diagnosis in order to ascertain the true incidence of each species and for the follow-up of patients after specific treatment.

Deep subsurface sulfate reduction and methanogenesis in the Iberian Pyrite Belt revealed through geochemistry and molecular biomarkers.

The Iberian Pyrite Belt (IPB, southwest of Spain), the largest known massive sulfide deposit, fuels a rich chemolithotrophic microbial community in the Río Tinto area. However, the geomicrobiology of its deep subsurface is still unexplored. Herein, we report on the geochemistry and prokaryotic diversity in the subsurface (down to a depth of 166 m) of the Iberian Pyritic belt using an array of geochemical and complementary molecular ecology techniques. Using an antibody microarray, we detected polymeric biomarkers (lipoteichoic acids and peptidoglycan) from Gram-positive bacteria throughout the borehole. DNA microarray hybridization confirmed the presence of members of methane oxidizers, sulfate-reducers, metal and sulfur oxidizers, and methanogenic Euryarchaeota. DNA sequences from denitrifying and hydrogenotrophic bacteria were also identified. FISH hybridization revealed live bacterial clusters associated with microniches on mineral surfaces. These results, together with measures of the geochemical parameters in the borehole, allowed us to create a preliminary scheme of the biogeochemical processes that could be operating in the deep subsurface of the Iberian Pyrite Belt, including microbial metabolisms such as sulfate reduction, methanogenesis and anaerobic methane oxidation.

Acute paralysis after seafood ingestion.

The first European case of tetrodotoxin intoxication is reported in a patient who ingested a trumpet shellfish from the Atlantic Ocean in Southern Europe. He suffered general acute paralysis with respiratory failure necessitating ventilation. Early neurophysiologic studies showed complete peripheral nerve inexcitability, with no recordable sensory or motor responses, and normal electroencephalography. Tetrodotoxin was detected in high quantities in the patient's blood and urine through high performance liquid chromatography-mass spectrometry analysis. Seventy-two hours after admission the patient recovered normal strength, reflexes and sensation.

Effect of apolipoprotein C3 and apolipoprotein A1 polymorphisms on postprandial response to a fat overload in metabolic syndrome patients.

OBJECTIVES: Apolipoprotein C-III (APOC3) is a component of triglyceride rich lipoproteins, and SstI polymorphism has been associated with hypertriglyceridemia. Apolipoprotein A-I (APOA1) is the major component of HDL and MspI polymorphism has been associated with APOA1 and HDL-C levels. Thus, we study the influence of these polymorphisms in the postprandial response in metabolic syndrome (MS). DESIGN AND METHODS: 73 MS patients and 21 healthy subjects underwent a fat overload, with measurements of their fasting and postprandial lipid profile. The APOC3 SstI and the APOA1MspI polymorphisms were genotyped. RESULTS: No significant differences were found in the lipid profile with respect to the MspI genotype. Patients with the S2S2 APOC3 genotype had significantly higher fasting and postprandial triglyceride levels and postprandial APOC3 and chylomicron-triglyceride levels compared with the other SstI APOC3 genotypes. CONCLUSIONS: Homozygosity for the minor allele of the APOC3 SstI polymorphism was associated to a worse postprandial response in MS patients.

Pseudoaneurisma de arteria segmentaria lumbar tras introducción de tornillo transpedicular L5. Una rara complicación vascular / Lumbar segmental artery pseudoaneurysm after L5 pedicle screw placement. A rare vascular complication

La fusión instrumentada lumbar por vía posterior se realiza de manera habitual hoy en día, aunque en algunas ocasiones puede inducir complicaciones que pueden llegar a ser devastadoras. Una de las causas, aunque poco frecuentes, de complicación mayor es la malposición de los tornillos pediculares, de ahí la importancia de ser metódicos a la hora de su colocación, comprobando el correcto labrado del trayecto y su introducción. Presentamos un caso de sangrado masivo tras la introducción de un tornillo pedicular lumbar durante una cirugía por estenosis de canal. La malposición del tornillo conllevó la inestabilidad hemodinámica intraoperatoria de la paciente tras el fracaso de los métodos habituales de control de sangrado en el campo quirúrgico. La realización de una TAC con contraste evidenció lesión de la arteria intersegmentaria lumbar que fue finalmente controlada mediante embolización e implantación de coil vascular (AU)

Posterior lumbar screw fixation is a common surgical procedure nowadays. However, it can sometimes produce complications that can be devastating. One of the less common causes of major complication is the misplacement of a pedicle screw. This highlights the importance of being methodical when placing pedicle screws, and checking that the pathway has been created correctly and their placement. We present a case of a massive bleed after a pedicular screw placement during lumbar canal stenosis surgery. Screw malposition led to intraoperative haemodynamic instability after failed attempts to control bleeding in the surgical site. Contrast enhanced CT imaging revealed a lumbar intersegmentary artery injury that was eventually controlled by means of a coil embolisation (AU)

Molecular detection of Ehrlichia ruminantium infection in Amblyomma variegatum ticks in The Gambia.

In West Africa, losses due to heartwater disease are not known because the incidence/prevalence has not been well studied or documented. To develop a diagnostic tool for molecular epidemiology, three PCR-based diagnostic assays, a nested pCS20 PCR, a nested map1 PCR and a nested reverse line blot (RLB) hybridization assay, were evaluated to determine their ability to detect infection in vector ticks, by applying them simultaneously to A. variegatum field ticks to detect Ehrlichia ruminantium, the causative agent of heartwater. The nested pCS20 PCR assay which amplified the pCS20 gene fragment showed the highest detection performance with a detection rate of 16.6%; the nested map1 PCR, which amplified the gene encoding the major antigenic protein1 (map1 gene) showed a detection rate of 11% and the RLB, based on the 16S rDNA sequence of anaplasma and ehrlichial species, detected 6.2%. The RLB, in addition, demonstrated molecular evidence of Ehrlichia ovina, Anaplasma marginale and Anaplasma ovis infections in The Gambia. Subsequently, the pCS20 assay was applied to study the prevalence and distribution of E. ruminantium tick infection rates at different sites in five divisions of The Gambia. The rates of infection in the country ranged from 1.6% to 15.1% with higher prevalences detected at sites in the westerly divisions (Western, Lower River and North Bank; range 8.3-15.1%) than in the easterly divisions (Central River and Upper River; range 1.6-7.5%). This study demonstrated a gradient in the distribution of heartwater disease risk for susceptible livestock in The Gambia which factor must be considered in the overall design of future upgrading programmes.