Low-dose CT for lung cancer screening: position paper from the Italian college of thoracic radiology.

Smoking is the main risk factor for lung cancer (LC), which is the leading cause of cancer-related death worldwide. Independent randomized controlled trials, governmental and inter-governmental task forces, and meta-analyses established that LC screening (LCS) with chest low dose computed tomography (LDCT) decreases the mortality of LC in smokers and former smokers, compared to no-screening, especially in women. Accordingly, several Italian initiatives are offering LCS by LDCT and smoking cessation to about 10,000 high-risk subjects, supported by Private or Public Health Institutions, envisaging a possible population-based screening program. Because LDCT is the backbone of LCS, Italian radiologists with LCS expertise are presenting this position paper that encompasses recommendations for LDCT scan protocol and its reading. Moreover, fundamentals for classification of lung nodules and other findings at LDCT test are detailed along with international guidelines, from the European Society of Thoracic Imaging, the British Thoracic Society, and the American College of Radiology, for their reporting and management in LCS. The Italian College of Thoracic Radiologists produced this document to provide the basics for radiologists who plan to set up or to be involved in LCS, thus fostering homogenous evidence-based approach to the LDCT test over the Italian territory and warrant comparison and analyses throughout National and International practices.

Pioneering women of microbiology in Spain.

Presented herein are the trajectories of four women who can be considered pioneers of microbiology in Spain. Three of them have been studied before, but never presented as pioneers of microbiology, and their lives are briefly reviewed: Zoe Rosinach Pedrol, a pioneering microbiologist in the health care field; Isabel Torán del Carré, in the agri-food sector; and Luz Zalduegui Gabilondo in the veterinary sciences. Nevertheless, Trinidad del Pan Arana is presented from the first time as pioneering microbiologist in the natural sciences area. All of these women developed their professional activity during the first third of the twentieth century, contributing to the establishment of microbiology as a new scientific discipline in Spain.

Biotechnological applications of occlusion bodies of Baculoviruses.

The ability of Baculoviruses to hyper-express very late genes as polyhedrin, the major component of occlusion bodies (OBs) or polyhedra, has allowed the evolution of a system of great utility for biotechnology. The main function of polyhedra in nature is to protect Baculovirus in the environment. The possibility of incorporating foreign proteins into the crystal by fusing them to polyhedrin (POLH) opened novel potential biotechnological uses. In this review, we summarize different applications of Baculovirus chimeric OBs. Basically, the improvement of protein expression and purification with POLH as a fusion partner; the use of recombinant polyhedra as immunogens and antigens, and the incorporation of proteins into polyhedra to improve Baculoviruses as bioinsecticides. The results obtained in each area and the future trends in these topics are also discussed.

Novel biotechnological platform based on baculovirus occlusion bodies carrying Babesia bovis small antigenic peptides for the design of a diagnostic enzyme-linked immunosorbent assay (ELISA).

Baculoviruses are large DNA virus of insects principally employed in recombinant protein expression. Its ability to form occlusion bodies (OBs), which are composed mainly of polyhedrin protein (POLH), makes them biotechnologically attractive, as these crystals (polyhedra) can incorporate foreign peptides and can be easily isolated. On the other hand, peptide microarrays allow rapid and inexpensive high-throughput serological screening of new candidates to be incorporated to OBs. To integrate these 2 biotechnological approaches, we worked on Babesia bovis, one of the causative agents of bovine babesiosis. Current molecular diagnosis of infection with B. bovis includes enzyme-linked immunosorbent assay (ELISA) techniques, which use merozoite lysate obtained from infected bovine erythrocytes. However, it is important to produce recombinant antigens that replace the use of crude antigens. Here, we describe a new biotechnological platform for the design of indirect ELISAs based on 5 antigenic peptides of 15 amino acid residues of B. bovis (ApBb), selected from a peptide microarray and expressed as a fusion to POLH. An Sf9POLHE44G packaging cell line infected with recombinant baculoviruses carrying POLH-ApBb fusions yielded higher levels of chimeric polyhedra, highlighting the advantage of a trans-contribution of a mutant copy of polyhedrin. Finally, the use of dissolved recombinant polyhedra as antigens was successful in an ELISA assay, as B. bovis-positive sera recognized the fusion POLH-ApBb. Thus, the use of this platform resulted in a promising alternative for molecular diagnosis of relevant infectious diseases.

Presence of Oil Mineral Aggregates (OMAs) in Surface Sediments from Mexico's Exclusive Economic Zone, NW Gulf of Mexico.

We assessed the presence and distribution of oil mineral aggregates (OMAs) in surficial sediments of Mexican waters in the NW Gulf of Mexico, their potential sources and their correlation with polycyclic aromatic hydrocarbons (PAH). In summer of 2010, OMAs were detected in three shallow sites. In winter of 2011, OMAs were observed in ten sites, two of them in the northernmost area at > 1500 m depth. These particles were possibly advected from the north Gulf and Mississippi area following the deep-water currents of the zone. The OMAs from shallower sites may reflect local pollution sources. PAHs displayed low concentrations in both surveys (from 0.01 to 0.7 µg g-1 in summer, and from 0.01 to 0.51 µg g-1 in winter), and showed rather a local origin. The expansion of the oil and port industry in the region is accountable for most of the OMAs detected.

Periocular Involvement in Trigeminal Trophic Syndrome.

Trigeminal trophic syndrome (TTS) is an uncommon condition resulting from trigeminal nerve damage, characterized by persistent facial ulceration, loss of sensation, and paresthesia within the trigeminal dermatome, with ala nasi involvement being a key feature. Lesions develop from repeated self-inflicted manipulation and trauma of the dysesthetic skin. This report details three cases of TTS, highlighting periocular changes, with etiologies varying from cerebrovascular accidents to herpes zoster ophthalmicus.

Perioperative assessment of platelet function by Thromboelastograph Platelet Mapping in cardiovascular patients undergoing non-cardiac surgery.

UNLABELLED: Five percent of patients on dual antiplatelet therapy after coronary artery stent implantation will need non-cardiac surgery within the first year of therapy, and many more will need surgery later on. A function assay that evaluates platelet reactivity and inhibition by drug therapy is beneficial for such patients. Platelet Mapping assay (PM) using the TEG analyzer was tested in surgical patients. After IRB approval, 60 patients on combined aspirin and clopidogrel therapy were consented and enrolled. The TEG maximal amplitude (MA) and the percentage (%) platelet inhibition were recorded and analyzed. Fifty-seven patients (mean age 65.7 ± 10.9 years) had preoperative data only. Distribution of preoperative ADP (43.6 ± 24.4%) and AA inhibition (52.8 ± 30.2%) was determined, as well as for the preoperative MA ADP (43.1 ± 15.9 mm) and MA AA (37.2 ± 19.6 mm), showing an offset of the effect of both medications starting from day 3. Patients with complete pre- and postoperative data were stratified depending on duration off antiplatelet therapy (≤3 days, 3-7 days and >7 days): n = 27, ADP % preop inhibition (43.2 ± 21.6%), ADP % postop inhibition (32.3 ± 18.3%), p = 0.048. Distribution of immediate pre- and post- ADP and AA % inhibitions, showing a possible reduction in Δ of inhibition for clopidogrel at 3 days, were also assessed. CONCLUSION: According to the findings, the TEG PM assay might be a feasible approach to objectively evaluate the effects of aspirin and clopidogrel during the perioperative period and potentially guide drug management.

Organochlorine contaminants in sediments and factors influencing their distribution in the natural marine protected area in the gulf of Mexico.

The aim of this study was to determine the concentration of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in marine sediments of a natural marine protected area (NMPA), Veracruz Reef System (VRS), in the Gulf of Mexico (GoM). This NMPA represents one of the first coastal sites with port activity in Mexico, which causes a range of human activities that require adequate planning so as not to impact and destroy this marine protected area; In this sense, this work represents the first effort aimed at diagnosing the presence of organochlorine contaminants in reef sediments from the mainland, both from the adjacent port area and from more distant regions with dominant agricultural activities that ultimately affect this NMPA. Content of organic matter (OM), carbonates and grain-size composition were determined to investigate their influence on compounds distribution in the sediment through multivariate analysis. The potential risk represented by organochlorines concentrations was evaluated taking as reference Sediment Quality Guidelines (SQGs). Levels of ∑PCBs, ∑DDTs, ∑HCHs and ∑Cyclodienes ranged from 11.59 to 29.79, < 0.01 to 34.11, 22.10 to 102.80, and 17.21-104.45 ng g-1 dry weight (DW), respectively. The statistical analysis revealed that the content of mud and organic matter were the parameters that might influence PCBs distribution in the sediment while OCPs could be influenced by sand and carbonate. The theoretical risk assessment estimated with the results obtained showed that concentrations of lindane, endrin, dieldrin and p,p'-DDT could have an adverse effect on the reef benthic community and continued monitoring is required. The data obtained on the organochlorine contaminants (OCs) considered in this work will contribute to support the strategies proposed by the Mexican federal government entity to continue with the conservation and surveillance of this NMPA and thereby guarantee the least impact derived mainly from port and related agricultural activities.

Automated Coronary Artery Calcium and Quantitative Emphysema in Lung Cancer Screening: Association With Mortality, Lung Cancer Incidence, and Airflow Obstruction.

PURPOSE: To assess automated coronary artery calcium (CAC) and quantitative emphysema (percentage of low attenuation areas [%LAA]) for predicting mortality and lung cancer (LC) incidence in LC screening. To explore correlations between %LAA, CAC, and forced expiratory value in 1 second (FEV 1 ) and the discriminative ability of %LAA for airflow obstruction. MATERIALS AND METHODS: Baseline low-dose computed tomography scans of the BioMILD trial were analyzed using an artificial intelligence software. Univariate and multivariate analyses were performed to estimate the predictive value of %LAA and CAC. Harrell C -statistic and time-dependent area under the curve (AUC) were reported for 3 nested models (Model survey : age, sex, pack-years; Model survey-LDCT : Model survey plus %LAA plus CAC; Model final : Model survey-LDCT plus selected confounders). The correlations between %LAA, CAC, and FEV 1 and the discriminative ability of %LAA for airflow obstruction were tested using the Pearson correlation coefficient and AUC-receiver operating characteristic curve, respectively. RESULTS: A total of 4098 volunteers were enrolled. %LAA and CAC independently predicted 6-year all-cause (Model final hazard ratio [HR], 1.14 per %LAA interquartile range [IQR] increase [95% CI, 1.05-1.23], 2.13 for CAC ≥400 [95% CI, 1.36-3.28]), noncancer (Model final HR, 1.25 per %LAA IQR increase [95% CI, 1.11-1.37], 3.22 for CAC ≥400 [95%CI, 1.62-6.39]), and cardiovascular (Model final HR, 1.25 per %LAA IQR increase [95% CI, 1.00-1.46], 4.66 for CAC ≥400, [95% CI, 1.80-12.58]) mortality, with an increase in concordance probability in Model survey-LDCT compared with Model survey ( P <0.05). No significant association with LC incidence was found after adjustments. Both biomarkers negatively correlated with FEV 1 ( P <0.01). %LAA identified airflow obstruction with a moderate discriminative ability (AUC, 0.738). CONCLUSIONS: Automated CAC and %LAA added prognostic information to age, sex, and pack-years for predicting mortality but not LC incidence in an LC screening setting. Both biomarkers negatively correlated with FEV 1 , with %LAA enabling the identification of airflow obstruction with moderate discriminative ability.

210Pb geochronology and metal concentrations in sediment cores recovered in the Alvarado Lagoon system, Veracruz, Mexico.

Three sediment cores recovered from the Alvarado Lagoon System (ALS) in the Gulf of Mexico were used to reconstruct the history of metals and metalloids and their environmental importance. The sedimentary profiles were dated with 210Pb and verified with 137Cs. Maximum ages of 77 and 86 years were estimated. Sediment provenance was described by sedimentological and geochemical proxies. The chemical alteration index (CIA) and weathering index (CIW) revealed moderate to high intensity of weathering in the source area that is controlled tropical climatic conditions, runoff, and precipitation in the basin that feeds sediments to this coastal lagoon. The Al2O3/TiO2 ratios indicated that the sediments were derived from intermediate igneous rocks. The enrichment factor values revealed the lithogenic and anthropic contribution of metals and metalloids. Cd is classified under the category extremely severe enrichment; agricultural activities, fertilizers, herbicides, and pesticides containing Cd are expected to supply this metal to the ecosystem. Factor Analysis and Principal Components provided two main factors, terrigenous and biological origins; ANOVA indicated that there are significant differences between the cores for the parameters analyzed and revealed that there are differences in depositional environments between the recovery zones of the cores. The ALS presented natural variations associated with the climatic conditions, terrigenous input, and its relationship with the hydrological variations of the main rivers. The contribution of this work was to identify the magnitude of the natural component versus the human contribution, mainly of risk metals such as Cd, to support better management of the hydrological basin that affects the ALS.

Pollution by organochlorine pesticides in Navachiste-Macapule, Sinaloa, Mexico.

The lagoon system of Navachiste-Macapule is located in northern Mexico, in the state of Sinaloa, with an area of 24,000 ha. The main economic activity in the area is agriculture, and the lagoon lies next to the irrigation district ID-063 which covers 116,615 ha. The purpose of this study is to evaluate the levels of pollution generated by organochlorine pesticides (OC) in the surface sediments of the lagoon and in the agricultural drains of the ID-063 that are supposedly transported into the system as a result of agricultural activities and runoff from adjacent land. For this, between 2006 and 2007, 45 surface sediment samples were collected (warm dry, rainy and cold dry) from 15 sampling sites, during the three climatic seasons. Of these, eight were located inside the lagoon in marine conditions (salinity >31 PSU) and seven in the agricultural drains of the ID-063 in freshwater conditions (salinity <5 PSU). The average concentration of the OC in the sediments was 44.75 ng g(-1), among which the group of the alicyclic compounds presented the greatest concentrations. The average value of the total organic carbon (TOC) in the sediments of the system was 0.90%. The sediments collected inside the lagoon had an average OC concentration of 18.97 ng g(-1), and the predominant type of sediment was fine to very fine sand. The average OC concentration in the sediments collected in the agricultural drains was 75.69 ng g(-1), where fine sediments (silt) were predominant. The presence of methoxychlor, endrin and heptachlor suggested that these compounds were continuously used in the system, even though their use is forbidden in Mexico.

A 3D method to estimate the effect of derotational osteotomies over femoral maltorsion. / Un método 3D para estimar el efecto de osteotomías desrotadoras sobre alteraciones torsionales de fémur.

INTRODUCTION AND OBJECTIVES: Accurate measurement of femoral torsion is essential to diagnosing, correct surgical decision-making, and the preoperative planning of derotational osteotomies in a subgroup of anterior knee pain patients and patellofemoral instability. Several measurement techniques have been described with great variability in the magnitude of the values. Moreover, none of them can predict the effect of rotational osteotomy on the femoral version value. The purpose of the present work is to develop a method to reliably measure femoral torsion as well as to evaluate the effects of osteotomy on the femoral version angle. MATERIAL AND METHODS: 30 patients without previous signs neither symptoms of patellofemoral pain or patellofemoral instability and with a lower limbs CT were selected. Their femoral torsion was measured using the classic Jeanmart and the Murphy's method by two independent radiologists. Independently, two orthopedic surgeons measured femoral torsion with the 3D method described in this study. The intraclass coefficient correlation and the Bland-Altman tests were used to analyze qualitatively and quantitatively the intraobserver and interobserver variability of the method regarding the Jeanmart's and Murphy's method. The confidence interval of the Normal values of the new method were defined and a lineal regression analysis between the 3D-Jeanmart's and 3D-Muphy's methods was performed. RESULTS: The mean femoral version with the 3D method was 12.74° (SD 5.96°, 95% CI: 11.67° to 13.82°). With Jeanmart's method, it was 12.84° (SD 8.60°, 95% CI: 11.28° to 14.39°) and 15.87° (SD 10.68°, 95% CI: 13.94° to 17.80°) with Murphy's method, being these differences not statistically significant. Both interobserver and intraobserver agreement of the 3D method were high. CONCLUSION: The method presented in this study allows for the reliable quantification of femoral torsion in a reproducible way. This method can be applied in several open access 3D image programs what makes its implementation in the healthcare centers possible from now on.

[Translated article] A 3D method to estimate the effect of derotational osteotomies over femoral maltorsion.

INTRODUCTION AND OBJECTIVES: Accurate measurement of femoral torsion is essential to diagnosing, correct surgical decision-making, and the preoperative planning of derotational osteotomies in a subgroup of anterior knee pain patients and patellofemoral instability. Several measurement techniques have been described with great variability in the magnitude of the values. Moreover, none of them can predict the effect of rotational osteotomy on the femoral version value. The purpose of the present work is to develop a method to reliably measure femoral torsion as well as to evaluate the effects of osteotomy on the femoral version angle. MATERIAL AND METHODS: 30 patients without previous signs neither symptoms of patellofemoral pain or patellofemoral instability and with a lower limbs CT were selected. Their femoral torsion was measured using the classic Jeanmart and the Murphy's method by two independent radiologists. Independently, two orthopedic surgeons measured femoral torsion with the 3D method described in this study. The intraclass coefficient correlation and the Bland-Altman tests were used to analyze qualitatively and quantitatively the intraobserver and interobserver variability of the method regarding the Jeanmart's and Murphy's method. The confidence interval of the Normal values of the new method were defined and a lineal regression analysis between the 3D-Jeanmart's and 3D-Muphy's methods was performed. RESULTS: The mean femoral version with the 3D method was 12.74° (SD 5.96°, 95% CI: 11.67°-13.82°). With Jeanmart's method, it was 12.84° (SD 8.60°, 95% CI: 11.28°-14.39°) and 15.87° (SD 10.68°, 95% CI: 13.94°-17.80°) with Murphy's method, being these differences not statistically significant. Both interobserver and intraobserver agreement of the 3D method were high. CONCLUSION: The method presented in this study allows for the reliable quantification of femoral torsion in a reproducible way. This method can be applied in several open access 3D image programmes what makes its implementation in the healthcare centres possible from now on.

Development of a novel set of Gateway-compatible vectors for live imaging in insect cells.

Insect genomics is a growing area of research. To exploit fully the genomic data that are being generated, high-throughput systems for the functional characterization of insect proteins and their interactomes are required. In this work, a Gateway-compatible vector set for expression of fluorescent fusion proteins in insect cells was developed. The vector set was designed to express a protein of interest fused to any of four different fluorescent proteins [green fluorescent protein (GFP), cyan fluorescent protein (CFP), yellow fluorescent protein (YFP) and mCherry] by either the C-terminal or the N-terminal ends. Additionally, a collection of organelle-specific fluorescent markers was assembled for colocalization with fluorescent recombinant proteins of interest. Moreover, the vector set was proven to be suitable for simultaneously detecting up to three proteins by multiple labelling. The use of the vector set was exemplified by defining the subcellular distribution of Mal de Río Cuarto virus (MRCV) outer coat protein P10 and by analysing the in vivo self-interaction of the MRCV viroplasm matrix protein P9-1 in Förster resonance energy transfer (FRET) experiments. In conclusion, we have developed a valuable tool for high-throughput studies of protein subcellular localization that will aid in the elucidation of the function of newly described insect and virus proteins.

Use of recA gene sequence analysis for the identification of Staphylococcus equorum strains predominant on dry-cured hams.

Spanish dry-cured ham is an uncooked meat product highly appreciated due to its characteristics flavour. In this study, we examined the accuracy of biochemical tests and 16S rDNA sequencing in the identification of 56 staphylococcal strains isolated during industrial Spanish dry-cured ham processes. Important differences were observed comparing genotypic and phenotypic data. Staphylococcus xylosus was the prevalent species identified by biochemical methods (87.5%), however, sequencing of the 16S rDNA resulted in an unambiguous identification of Staphylococcus equorum (73.2%) and Staphylococcus vitulinus (8.9%) strains. Reliable identification of meat staphylococci, mainly among S. xylosus and S. equorum strains could be also achieved by means of recA gene sequence comparison. Two degenerate primers previously described for lactic acid bacteria were used to amplify an internal fragment of the recA gene. This fragment was amplified from twelve staphylococcal type strains representing frequent meat species. The results indicated that recA sequencing is an adequate method to discriminate among meat staphylococci. In addition, S. xylosus and S. equorum strains could be more accurately discriminated by recA sequencing than 16S rDNA or sodA sequencing. The S. equorum sequence diversity showed at the intra-species level by recA gene sequencing confirmed the high heterogeneity described among S. equorum strains.

Synthesis of propyl gallate by transesterification of tannic acid in aqueous media catalysed by immobilised derivatives of tannase from Lactobacillus plantarum.

Immobilised derivatives of tannase from Lactobacillus plantarum were able to catalyse the transesterification of tannic acid by using moderate concentrations of 1-propanol in aqueous media. Transesterification of tannic acid was very similar to transesterification of methyl gallate. The synthetic yield depended on the pH and concentration of 1-propanol, although it did not vary much when using 30% or 50% 1-propanol. Synthetic yields of 45% were obtained with 30% of 1-propanol at pH 5.0. The product was chromatographically pure, and the reaction by-product was 55% pure gallic acid. On the other hand, immobilised tannase was fairly stable under optimal reaction conditions.

Chronic Coronary Syndrome: Overcoming Clinical Practice Guidelines. The role of the COMPASS Strategy.

Current European guidelines on chronic coronary syndromes recommend the use of low-dose aspirin (or clopidogrel if intolerance or contraindication occurs) throughout life. However, as the risk of recurrent vascular events is high, particularly in some patients (i.e. diffuse multivessel coronary artery disease, diabetes, recurrent myocardial infarction, peripheral artery disease, or chronic kidney disease,…), these guidelines also consider that in those patients at moderate or high risk of ischemic events, but without a high bleeding risk, dual antithrombotic therapy should be considered. According to these guidelines, treatment options for dual antithrombotic therapy in combination with aspirin may include clopidogrel 75 mg/daily, prasugrel 10 mg/daily, ticagrelor 60 mg bid or rivaroxaban 2.5 mg bid. Remarkably, despite the results of the clinical trials that sustain these recommendations clearly diverge, guidelines do not differentiate between them. However, although all these drugs have demonstrated a significant reduction in major cardiovascular events in patients with stable atherosclerotic disease, only the addition of rivaroxaban has been associated with a reduction in cardiovascular and overall mortality in the secondary analysis. This may be related to the fact that the activation of platelets and factor X plays a key role in the development of atherothrombosis, and, consequently, both targets should be considered for the appropriate management of these patients.

Current treatments of Alzheimer disease: are main caregivers satisfied with the drug treatments received by their patients?

BACKGROUND/AIMS: A full comparison of the satisfaction with treatment using the current Alzheimer's disease (AD) therapies from the perspective of caregivers has not yet been done. The aim of this study was thus to find out the degree of satisfaction with the main available drug treatments in monotherapy for AD from this point of view. METHODS: A cross-sectional, multicentre study of patients with possible/probable AD according to DSM-IV/NINCDS-ADRDA criteria, on monotherapy with donepezil, galantamine, rivastigmine or memantine, was carried out. Treatment satisfaction was measured by a caregiver proxy-administration of the generic SATMED-Q questionnaire [range: 0 (not satisfied at all) to 100 (totally satisfied)], overall and in 6 domains: tolerability, efficacy, medical care, ease and convenience, impact on daily activities and overall satisfaction. RESULTS: A total of 829 patients were included: 63.3% women, aged 78.2 +/- 6.8 years; 546 (67.3%) on donepezil, 106 (13.1%) on rivastigmine, 99 (12.2%) on galantamine and 60 (7.4%) on memantine. SATMED-Q scores p values were adjusted by MMSE and treatment duration. Caregivers of patients on donepezil showed significantly higher SATMED-Q total (71.8 +/- 12.3; p < 0.05) and overall satisfaction domain scores (81.6 +/- 18.4; p < 0.01) than those of patients on any other drugs, as well as significantly higher ease and convenience of use domain (81.5 +/- 17.4; p < 0.01) and undesirable effects domain (96.0 +/- 12.9; p < 0.05) scores than those of rivastigmine- and galantamine-treated patients. Of the caregivers of donepezil-treated patients, 76.7% were satisfied with treatment versus 68.7, 61.4 and 46.7% of those caregivers whose patients were treated with galantamine, rivastigmine and memantine, respectively (p = 0.0002). CONCLUSION: Caregivers of AD patients undergoing donepezil monotherapy seem to be more satisfied with treatment than those of patients receiving the other usual AD treatments in this study, particularly due to the ease and convenience of use of this drug. The higher level of satisfaction of these caregivers could be explained by the fact that, within the donepezil group, a high percentage of patients were treated with orally disintegrating tablets, which are easier for the patient to swallow.

Multilocus sequence typing of oenological Saccharomyces cerevisiae strains.

This study describes the application of a multilocus sequence typing (MLST) analysis for molecular discrimination at the strain level of Spanish wine yeast strains. The discrimination power of MLST is compared to mitochondrial RFLP analysis. Fragments of the ADP1, ACC1, RPN2, GLN4, and ALA1 genes were amplified by PCR from chromosomal DNA of 18 wine Saccharomyces cerevisiae strains. Ten polymorphic sites were found in the five loci analyzed showing 13 different genotypes, with 11 of them represented by only one strain. RFLP analysis of the same 18 wine yeast strains showed seventeen different mitochondrial patterns. Phylogenetic relationships among the strains analyzed, inferred by MLST data, showed wine isolates of S. cerevisiae as a rather homogeneous group. The discrimination potential of mitochondrial RFLP analysis was superior to the MLST scheme used in this work. However, MLST analysis allowed an easy construction of reliable phylogenetic trees. MLST analysis offers the possibility of typing wine S. cerevisiae strains simultaneously to the study of the genetic relationship among them.

Peripheral neurostimulation in supraorbital neuralgia refractory to conventional therapy.

Supraorbital neuralgia has been identified as an infrequent cause of headache that may prove very difficult to control pharmacologically. Peripheral nerve stimulation using electrodes to stimulate the nerve segmentally responsible for the zone of pain may constitute a management alternative in such cases. We present the case of a patient with headache because of posttraumatic supraorbital neuralgia, refractory to medical treatment, with good analgesic control after peripheral nerve stimulation. Peripheral nerve stimulation may be considered a safe, reversible treatment for patients with headache secondary to supraorbital neuralgia who respond poorly to pharmacological treatment, thus avoiding irreversible alternatives such as surgery.