AEDV Expert Consensus for the Management of Syphilis. / Documento de expertos de la AEDV para el manejo de la sífilis.

Syphilis -the "great simulator" for classical venereologists-is re-emerging in Western countries despite adequate treatment; several contributing factors have been identified, including changes in sexual behaviour, which won't be the topic of this article though. In 2021, a total of 6613 new cases of syphilis were reported in Spain, representing an incidence of 13.9×100 000 inhabitants (90.5%, men). Rates have increased progressively since 2000. The clinical presentation of syphilis is heterogeneous. Although chancroid, syphilitic roseola and syphilitic nails are typical lesions, other forms of the disease can be present such as non-ulcerative primary lesions like Follmann balanitis, chancres in the oral cavity, patchy secondary lingual lesions, or enanthema on the palate and uvula, among many others. Regarding diagnosis, molecular assays such as PCR have been replacing dark-field microscopy in ulcerative lesions while automated treponemal tests (EIA, CLIA) are being used in serological tests, along with classical tests (such as RPR and HAART) for confirmation and follow-up purposes. The interpretation of these tests should be assessed in the epidemiological and clinical context of the patient. HIV serology and STI screening should be requested for anyone with syphilis. Follow-up of patients under treatment is important to ensure healing and detect reinfection. Serological response to treatment should be assessed with the same non-treponemal test (RPR/VDRL); 3-, 6-, 12-, and 24-month follow-up is a common practice in people living with HIV (PLHIV). Sexual contacts should be assessed and treated as appropriate. Screening is advised for pregnant women within the first trimester of pregnancy. Pregnant women with an abortion after week 20 should all be tested for syphilis. The treatment of choice for all forms of syphilis, including pregnant women and PLHIV, is penicillin. Macrolides are ill-advised because of potential resistance.

[Translated article] AEDV Expert Consensus for the Management of Syphilis.

Syphilis -the "great simulator" for classical venereologists-is re-emerging in Western countries despite adequate treatment; several contributing factors have been identified, including changes in sexual behaviour, which won't be the topic of this article though. In 2021, a total of 6613 new cases of syphilis were reported in Spain, representing an incidence of 13.9×100 000 inhabitants (90.5%, men). Rates have increased progressively since 2000. The clinical presentation of syphilis is heterogeneous. Although chancroid, syphilitic roseola and syphilitic nails are typical lesions, other forms of the disease can be present such as non-ulcerative primary lesions like Follmann balanitis, chancres in the oral cavity, patchy secondary lingual lesions, or enanthema on the palate and uvula, among many others. Regarding diagnosis, molecular assays such as PCR have been replacing dark-field microscopy in ulcerative lesions while automated treponemal tests (EIA, CLIA) are being used in serological tests, along with classical tests (such as RPR and HAART) for confirmation and follow-up purposes. The interpretation of these tests should be assessed in the epidemiological and clinical context of the patient. HIV serology and STI screening should be requested for anyone with syphilis. Follow-up of patients under treatment is important to ensure healing and detect reinfection. Serological response to treatment should be assessed with the same non-treponemal test (RPR/VDRL); 3-, 6-, 12-, and 24-month follow-up is a common practice in people living with HIV (PLHIV). Sexual contacts should be assessed and treated as appropriate. Screening is advised for pregnant women within the first trimester of pregnancy. Pregnant women with an abortion after week 20 should all be tested for syphilis. The treatment of choice for all forms of syphilis, including pregnant women and PLHIV, is penicillin. Macrolides are ill-advised because of potential resistance.

Modulation of host plant immunity by Tobamovirus proteins.

BACKGROUND: To establish successful infection, plant viruses produce profound alterations of host physiology, disturbing unrelated endogenous processes and contributing to the development of disease. In tobamoviruses, emerging evidence suggests that viral-encoded proteins display a great variety of functions beyond the canonical roles required for virus structure and replication. Among these, their modulation of host immunity appears to be relevant in infection progression. SCOPE: In this review, some recently described effects on host plant physiology of Tobacco mosaic virus (TMV)-encoded proteins, namely replicase, movement protein (MP) and coat protein (CP), are summarized. The discussion is focused on the effects of each viral component on the modulation of host defense responses, through mechanisms involving hormonal imbalance, innate immunity modulation and antiviral RNA silencing. These effects are described taking into consideration the differential spatial distribution and temporality of viral proteins during the dynamic process of replication and spread of the virus. CONCLUSION: In discussion of these mechanisms, it is shown that both individual and combined effects of viral-encoded proteins contribute to the development of the pathogenesis process, with the host plant's ability to control infection to some extent potentially advantageous to the invading virus.

Direct and maternal additive effects are not the main determinants of Iberian piglet perinatal mortality.

Data of 127,800 Iberian piglets were used to study genetic parameters of mortality at birth at the piglet level. These records proceed from three data sets: 4,987 litter of 2,156 sows of a dam line, 2,768 litter of 817 sows of a complete diallel cross between four Iberian strains and 7,153 litter of 2,113 sows of the Torbiscal composite line. Perinatal mortality was considered as a binary trait, and Bayesian threshold animal models were fitted to separately analyse the three data sets. The posterior means of direct heritability were 0.010, 0.004 and 0.003, and those of maternal heritability were 0.034, 0.011 and 0.014 for dam line, diallel cross and Torbiscal line, respectively. Important effects of litter size and parity order were inferred in the three data sets, of within-breed cross-breeding parameters in the diallel cross and of sex and sow handling in the Torbiscal line Therefore, the inclusion of perinatal mortality in the objective of selection is questionable in this breed and strategies for reducing piglet mortality successful in other breeds should be considered.

Novel yeast isolated from broilers' feedstuff, gut and faeces as aflatoxin B1 adsorbents.

AIMS: To isolate and characterize native yeast strains from broilers' environment as feedstuff, faeces and gut, and to evaluate their binding capacity for aflatoxin B1 (AFB1 ). METHODS AND RESULTS: A total of nine yeast strains were isolated: three from feedstuff identified as Pichia kudriavzevii (2) and Clavispora lusitaniae (1), two from gut identified as Candida tropicalis and four from faeces identified as Cl. lusitaniae (3) and Cyberlindnera fabianii (1). AFB1 binding percentages varied among yeast strains and with AFB1 concentrations. To carry out adsorption studies, one strain from each genus and each origin was selected as follows: Cl. lusitaniae and P. kudriavzevii from feedstuff, Cl. lusitaniae and Cy. fabianii from faeces and Ca. tropicalis from gut. The most appropriate concentrations for cells and toxin were 107 cells per ml and 100 ng ml-1 of AFB1 respectively. All the tested yeast strains showed similar adsorption capacities independently of the origin. The adsorption isotherm studies in all yeasts assayed showed behaviour of L type or Langmuir and a varied affinity for the toxin. The stability of the AFB1 -yeast complex demonstrated the irreversibility of the binding process. CONCLUSION: Yeast strains tested in this study constitute potential AFB1 adsorbents and they possess the advantage to be native from the avian environment. SIGNIFICANCE AND IMPACT OF THE STUDY: This study makes a contribution to using native yeasts from broilers' environment for controlling chronic aflatoxicosis in avian production.

Subaerial biofilms on granitic historic buildings: microbial diversity and development of phototrophic multi-species cultures.

Microbial communities of natural subaerial biofilms developed on granitic historic buildings of a World Heritage Site (Santiago de Compostela, NW Spain) were characterized and cultured in liquid BG11 medium. Environmental barcoding through next-generation sequencing (Pacific Biosciences) revealed that the biofilms were mainly composed of species of Chlorophyta (green algae) and Ascomycota (fungi) commonly associated with rock substrata. Richness and diversity were higher for the fungal than for the algal assemblages and fungi showed higher heterogeneity among samples. Cultures derived from natural biofilms showed the establishment of stable microbial communities mainly composed of Chlorophyta and Cyanobacteria. Although most taxa found in these cultures were not common in the original biofilms, they are likely common pioneer colonizers of building stone surfaces, including granite. Stable phototrophic multi-species cultures of known microbial diversity were thus obtained and their reliability to emulate natural colonization on granite should be confirmed in further experiments.

Validity of the Ages and Stages Questionnaire to detect risk of Developmental Coordination Disorder in preschoolers.

BACKGROUND: A reliable and valid screening tool for detecting children at risk for Developmental Coordination Disorder (DCD) is needed. The purpose of this study was to evaluate the ability of the Ages and Stages Questionnaire-Third Edition (ASQ-3) motor scales to detect children at risk for DCD in a community-based sample of children aged 3.5-5.5 years. METHODS: One hundred and sixty parent-child pairs were recruited from community-based organizations. Children were eligible if they spoke English and had no known physical impairments. Eligible parents were asked to fill out the ASQ-3, following which their child's fine motor and gross motor proficiency was assessed using the Movement Assessment Battery for Children-Second Edition (MABC-2). DCD risk was defined as those children scoring at or below the 16th percentile on the MABC-2. Sensitivity and specificity of the fine and gross motor areas of the ASQ-3 were examined and referenced against the DCD risk classification. RESULTS: The ASQ-3 total motor score correlated moderately with overall standard score on the MABC-2 (r = 0.41; p < 0.001). Regardless of the ASQ-3 cut-off used, sensitivities of the ASQ-3 fine motor or gross motor scale to detect DCD risk were low at 21-47%, whereas specificities were high at 89-96%. CONCLUSION: Early identification of motor skill delays is important in order to intervene and hopefully prevent the associated negative health consequences. However, because of the low sensitivity of the ASQ-3 motor scales, these results suggest that the ASQ-3 is not an appropriate screening tool to identify children at risk for DCD in the preschool population.

Assessing effective population size, coancestry and inbreeding effects on litter size using the pedigree and SNP data in closed lines of the Iberian pig breed.

The complete pedigree of two closed Iberian pig lines (Gamito and Torbiscal), with 798 and 4077 reproducers, has been used to measure the evolution of coancestry (f) and inbreeding (F) for autosomal and X-linked genes along 16 and 28 respective equivalent discrete generations. At the last generation, the mean values of each line were f = 0.41 and 0.22, F = 0.35 and 0.18, fX  = 0.46 and 0.22 and FX  = 0.47 and 0.19, respectively. Other calculated parameters were the effective number of founders (final values, 6.8 and 35.2) and non-founders (1.5 and 2.4), founder genome equivalents (1.2 and 2.3) and effective population size (16.0 and 57.7). Measures of Torbiscal effective size based on rates of coancestry (66.1), inbreeding (65.0) and linkage disequilibrium (71.0) were estimated from whole-genome SNP genotyping data. Values of new and old inbreeding and their respective rates by generation were computed to detect purging effects of natural selection. The analysis of 6854 Torbiscal litters showed significant negative impacts of new and fast inbreeding on litter size, as expected from the purging hypothesis: -0.20 born piglets per litter by a 10% of new inbreeding, and -0.03 and -0.02 piglets by 1% of total and new inbreeding rates, respectively. The analysis performed on 1274 litters of the Gamito line failed to show purging effects. The only significant results were reductions in -0.91 and -0.17 piglets by a 10% of old and X-linked genes inbreeding, respectively. These results may be useful for some practical issues in conservation programs of farm or captive wild animals.

Cardiac arrhythmias in stroke unit patients. Evaluation of the cardiac monitoring data. / Arritmias cardiacas en la unidad de ictus: análisis de los datos de la monitorización cardiaca.

INTRODUCTION: Cardiac arrhythmias are frequent in acute stroke. Stroke units are widely equipped with cardiac monitoring systems. Pre-existing heart diseases and heart-brain interactions may be implicated in causing cardiac arrhythmias in acute stroke. This article analyses cardiac arrhythmias detected in patients hospitalised in a stroke unit. METHOD: Prospective observational study of consecutive patients admitted to a stroke unit with cardiac monitoring. We collected clinical data from patients and the characteristics of their cardiac arrhythmias over a 1-year period (2013). Time of arrhythmia onset, associated predisposing factors, and the therapeutic decisions made after detection of arrhythmia were examined. All patients underwent continuous cardiac monitoring during no less than 48hours. RESULTS: Of a total of 332 patients admitted, significant cardiac arrhythmias occurred in 98 patients (29.5%) during their stay in the stroke unit. Tachyarrhythmia (ventricular tachyarrhythmias, supraventricular tachyarrhythmias, complex ventricular ectopy) was present in 90 patients (27.1%); bradyarrhythmia was present in 13 patients (3.91%). Arrhythmias were independently associated with larger size of brain lesion and older age. In 10% of the patient total, therapeutic actions were taken after detection of significant cardiac arrhythmias. Most events occurred within the first 48hours after stroke unit admission. CONCLUSIONS: Systematic cardiac monitoring in patients with acute stroke is useful for detecting clinically relevant cardiac arrhythmias. Incidence of arrhythmia is higher in the first 48hours after stroke unit admission. Age and lesion size were predicted appearance of arrhythmias. Detection of cardiac arrhythmias in a stroke unit has important implications for treatment.

Routine invasive strategy in acute coronary syndrome patients with renal dysfunction. Results of the ARIAM-SEMICYUC registry. / Estrategia invasiva de rutina en el síndrome coronario agudo sin elevación del segmento ST con disfunción renal. Resultados del registro ARIAM-SEMICYUC.

OBJECTIVE: To evaluate the use and effectiveness of a routine invasive strategy (RIS) in patients with acute coronary syndrome without persistent ST-segment elevation with renal dysfunction in the real world scenario. METHODS: A retrospective cohort study based on the ARIAM-SEMICYUC Registry (2011-2014) was carried out. Renal dysfunction was defined as GFR (Cockroft-Gault)<60ml/min (moderate dysfunction) or<30ml/min (severe dysfunction). Patients in which early angiography (<72h) was performed due to cardiogenic shock or recurrent myocardial ischemia were excluded. The primary endpoint was hospital mortality. Confounding factors were controlled using propensity score analysis. RESULTS: A total of 4,279 patients were analyzed, of which 26% had moderate renal dysfunction and 5% severe dysfunction. Patients with renal dysfunction had greater severity and comorbidity, higher hospital mortality (8.6 vs. 1.8%), and lesser use of the RIS (40 vs. 52%). The adjusted OR for mortality in patients without/with renal dysfunction were 0.38 (95% confidence interval [95%CI] 0.17 to 0.81) and 0.52 (95%CI 0.32 to 0.87), respectively (interaction P-value=.4779). The impact (adjusted risk difference) of RIS was higher in the group with renal dysfunction (-5.1%, 95%CI -8.1 to -2.1 vs. -1.6%, 95%CI -2.6 to -0.6; interaction P-value=.0335). No significant interaction was detected for the other endpoints considered (ICU mortality, 30-day mortality, myocardial infarction, acute renal failure or moderate/severe bleeding). CONCLUSIONS: The results suggest that the effectiveness of IRS is similar in patients with normal or abnormal renal function, and alert to the under-utilization of this strategy in such patients.

Could a patient with SMC1A duplication be classified as a human cohesinopathy?

The disorders caused by mutations in genes encoding subunits and accessory proteins of cohesin complex are collectively termed as cohesinopathies. The best known cohesinopathy is Cornelia de Lange Syndrome (CdLS), which is a multisystem developmental disorder characterized by facial dysmorphism, limb malformations, growth and cognitive impairment. Mutations in five genes, encoding subunits of the cohesin complex (SMC1A, SMC3, RAD21) and its regulators (NIPBL, HDAC8), are responsible for ∼ 70% of CdLS cases. We describe a 16-year-old boy with facial dysmorphism, growth retardation, intellectual disability, hirsutism and small hands, who has a small Supernumerary Marker Chromosome (sSMC) present in mosaic form. sSMC is composed of two duplicated segments encompassing 17 genes including SMC1A gene, at the regions Xp11.22 and Xp11.21q11.1. Clinical comparison between our patient with a previously reported individual with a SMC1A duplication and four male carriers of similar sSMC reported in databases, suggest that they all share clinical features related to cohesinopathies. Although our patient does not have the classical CdLS craniofacial phenotype, he has pre and postnatal growth retardation, intellectual disability and mild musculoskeletal anomalies, features commonly seen in patients with cohesinopathies.

Differential expression pattern of microRNAs in CD4+ and CD19+ cells from asymptomatic patients with systemic lupus erythematosus.

OBJECTIVE: The aim of this study was to investigate the pattern of microRNA (miRNA) expression in CD19+ and CD4+ cells from asymptomatic patients with systemic lupus erythematosus (SLE). METHODS: A screening of the expression of 377 miRNAs was performed in human CD4+ and CD19+ cells isolated from the peripheral blood by using a TaqMan Human MicroRNA Array. Validation of differential expression pattern of those was performed using TaqMan assays in these cell populations obtained from a larger cohort of patients and controls. RESULTS: According to the screening assays, three miRNAs were differentially expressed (p value <0.1) in cell populations from both patients and controls: hsa-miR-143, hsa-miR-224 and hsa-miR-576-5p for CD4+ cells, and hsa-miR-10a, hsa-miR-31 and hsa-miR-345 for CD19+ cells. After validation, significant differences (p value <0.05) were confirmed only for hsa-miR-143 and hsa-miR-224 in CD4+ cells and for hsa-miR-10a and hsa-miR-345 in CD19+ cells. In all cases, the miRNAs were over expressed in SLE patients compared with healthy donors. CONCLUSIONS: Our results support a different pattern of miRNA expression in SLE patients.

Copy number variants in a highly inbred Iberian porcine strain.

We carried out a comprehensive genomic analysis of porcine copy number variants (CNVs) based on whole-genome SNP genotyping data and provided new measures of genomic diversity (number, length and distribution of CNV events) for a highly inbred strain (the Guadyerbas strain). This strain represents one of the most ancient surviving populations of the Iberian breed, and it is currently in serious danger of extinction. CNV detection was conducted on the complete Guadyerbas population, adjusted for genomic waves, and used strict quality criteria, pedigree information and the latest porcine genome annotation. The analysis led to the detection of 65 CNV regions (CNVRs). These regions cover 0.33% of the autosomal genome of this particular strain. Twenty-nine of these CNVRs were identified here for the first time. The relatively low number of detected CNVRs is in line with the low variability and high inbreeding estimated previously for this Iberian strain using pedigree, microsatellite or SNP data. A comparison across different porcine studies has revealed that more than half of these regions overlap with previously identified CNVRs or multicopy regions. Also, a preliminary analysis of CNV detection using whole-genome sequence data for four Guadyerbas pigs showed overlapping for 16 of the CNVRs, supporting their reliability. Some of the identified CNVRs contain relevant functional genes (e.g., the SCD and USP15 genes), which are worth being further investigated because of their importance in determining the quality of Iberian pig products. The CNVR data generated could be useful for improving the porcine genome annotation.

Skin infections caused by community-acquired methicillin-resistant Staphylococcus aureus: clinical and microbiological characteristics of 11 cases.

INTRODUCTION: Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emerging pathogen that causes skin and soft-tissue infections. OBJECTIVE: To describe the clinical characteristics of skin infections caused by CA-MRSA and correlations with the available demographic and microbiological data. MATERIAL AND METHODS: This was a descriptive study of patients with a microbiologically confirmed diagnosis of CA-MRSA infection treated in a dermatology department between June 2009 and December 2011. We recorded demographic details, the clinical characteristics of lesions, and the treatments used. RESULTS: We studied 11 patients (5 men and 6 women); 91% were under 40 years of age and had no relevant past medical history. The most common presentation was a skin abscess (with or without cellulitis). In all such cases, marked tissue necrosis and little or no purulent exudate was observed when the abscess was drained. Fifty percent of these abscesses had been treated previously with ß-lactam antibiotics, and in all cases the lesions resolved after surgical drainage, which was combined in 63% of cases with quinolones or cotrimoxazole. CONCLUSIONS: Today, skin infections due to CA-MRSA affect healthy young athletes who have no contact with healthcare settings. The most common presentation is a skin abscess characterized by marked tissue necrosis and little or no purulent exudate. In cases with these characteristics in susceptible patients, the involvement of CA-MRSA as the causative agent should be suspected. The abscesses should be drained whenever possible and, if necessary, antibiotic treatment should be prescribed; empirical use of ß-lactam antibiotics should be avoided.

Porcine colonization of the Americas: a 60k SNP story.

The pig, Sus scrofa, is a foreign species to the American continent. Although pigs originally introduced in the Americas should be related to those from the Iberian Peninsula and Canary islands, the phylogeny of current creole pigs that now populate the continent is likely to be very complex. Because of the extreme climates that America harbors, these populations also provide a unique example of a fast evolutionary phenomenon of adaptation. Here, we provide a genome wide study of these issues by genotyping, with a 60k SNP chip, 206 village pigs sampled across 14 countries and 183 pigs from outgroup breeds that are potential founders of the American populations, including wild boar, Iberian, international and Chinese breeds. Results show that American village pigs are primarily of European ancestry, although the observed genetic landscape is that of a complex conglomerate. There was no correlation between genetic and geographical distances, neither continent wide nor when analyzing specific areas. Most populations showed a clear admixed structure where the Iberian pig was not necessarily the main component, illustrating how international breeds, but also Chinese pigs, have contributed to extant genetic composition of American village pigs. We also observe that many genes related to the cardiovascular system show an increased differentiation between altiplano and genetically related pigs living near sea level.

Measuring inbreeding and inbreeding depression on pig growth from pedigree or SNP-derived metrics.

Multilocus homozygosity, measured as the proportion of the autosomal genome in homozygous genotypes or in runs of homozygosity, was compared with the respective pedigree inbreeding coefficients in 64 Iberian pigs genotyped using the Porcine SNP60 Beadchip. Pigs were sampled from a set of experimental animals with a large inbreeding variation born in a closed strain with a completely recorded multi-generation genealogy. Individual inbreeding coefficients calculated from pedigree were strongly correlated with the different SNP-derived metrics of homozygosity (r = 0.814-0.919). However, unequal correlations between molecular and pedigree inbreeding were observed at chromosomal level being mainly dependent on the number of SNPs and on the correlation between heterozygosities measured across different loci. A panel of 192 SNPs of intermediate frequencies was selected for genotyping 322 piglets to test inbreeding depression on postweaning growth performance (daily gain and weight at 90 days). The negative effects on these traits of homozygosities calculated from the genotypes of 168 quality-checked SNPs were similar to those of inbreeding coefficients. The results support that few hundreds of SNPs may be useful for measuring inbreeding and inbreeding depression, when the population structure or the mating system causes a large variance of inbreeding.

Causes of low vision in children: A systematic review.

OBJECTIVE: To identify the ocular pathologies that are reported as causes of low vision in children. MATERIAL AND METHODS: The systematic search was carried out in Medline (PubMed), Embase and Lilacs. Observational studies with populations between 0-18 years of age, reporting visual acuity data between 20/60-20/400 and reporting the frequency of ocular pathologies were selected. Studies in which the diagnosis of the condition had not been verified by a professional, or which covered only cases of blindness, uncorrected refractive errors, or amblyopia, were excluded. The methodological quality of the articles was evaluated using the Joanna Briggs Institute instrument for prevalence studies. RESULTS: 27 studies conducted in Asia (13 publications), Africa (6 studies), Oceania (4 studies), Europe and South America (2 studies each) were included. The most reported causes of low vision were: cataract, with prevalence between 0.8% and 27.2%; albinism with from 1.1% to 47%; nystagmus, with prevalence between 1.3% and 22%; retinal dystrophies between 3.5% and 50%; retinopathy of prematurity (ROP) with prevalence between 1.1% and 65.8%, optic atrophy between 0.2% and 17.6%, and glaucoma from 2.4% to 18.1%. CONCLUSIONS: Cataract, albinism and nystagmus are the ocular pathologies most mentioned by studies as a cause of low vision in children, as well as retinal diseases such as ROP and optic nerve diseases such as atrophy. However, there are numerous eye conditions that can result in low vision in the pediatric population.

Transgenic expression of Tobacco mosaic virus capsid and movement proteins modulate plant basal defense and biotic stress responses in Nicotiana tabacum.

Plant viruses cause metabolic and physiological changes associated with symptomatic disease phenotypes. Symptoms involve direct and indirect effects, which result in disruption of host physiology. We used transgenic tobacco expressing a variant of Tobacco mosaic virus (TMV) coat protein (CP(T42W)) or movement protein (MP), and a hybrid line (MP×CP(T42W)) that coexpresses both proteins, to study the plant response to individual viral proteins. Findings employing microarray analysis of MP×CP(T42W) plants and silenced mp×cp(T42W)* controls revealed that altered transcripts were mostly downregulated, suggesting a persistent shut-off due to MP×CP(T42W) expression. Next, we showed that MP triggered reactive oxygen species (ROS) accumulation, reduction of total ascorbate, and expression of ROS scavenging genes. These effects were enhanced when both proteins were coexpressed. MP and MP×CP(T42W) plants showed increased levels of salicylic acid (SA) and SA-responsive gene expression. Furthermore, these effects were partially reproduced in Nicotiana benthamiana when GMP1 transcript was silenced. CP(T42W) seems to be playing a negative role in the defense response by reducing the expression of PR-1 and RDR-1. MP and MP×CP(T42W) transgenic expression promoted a recovery-like phenotype in TMV RNA infections and enhanced susceptibility to Pseudomonas syringae and Sclerotinia sclerotiorum. The individual effects of viral proteins may reflect the ability of a virus to balance its own virulence.

Basic characterization and partial purification of ß-glucosidase from cell-free extracts of Oenococcus oeni ST81.

AIMS: This study was designed to characterize a ß-glucosidase of Oenococcus oeni ST81, a strain isolated from a Spanish wine of the origin appellation Ribeira Sacra. METHODS AND RESULTS: The ß-glucosidase of O. oeni ST81 seems to have a periplasmic localization into the cells. This activity was strongly inhibited by gluconic acid, partially inhibited by glucose and not inhibited by fructose, lactate, malate, mannitol or sorbitol. Ethanol increased the activity of this enzyme up to 147%. Among the several metal ions assayed, only Fe²âº (10 mmol l⁻¹) and Cu²âº (5 mmol l⁻¹) exhibited a partial inhibitory effect (40%). This enzyme was partially purified using a combination of ammonium sulfate precipitation and chromatographic methods. The single peak because of ß-glucosidase in all chromatographic columns indicates the presence of a single enzyme with an estimated molecular mass of 140 kDa. The calculated K(m) and V(max) values for 4-nitrophenyl-ß-D-glucopyranoside were 0·38 mmol l⁻¹ and 5·21 nmol min⁻¹, respectively. The enzyme was stable at pH 5·0 with a value of t(1/2) = 50 days for the crude extract. CONCLUSIONS: The ß-glucosidase of O. oeni ST81 is substantially different from those characterized from other wine-related lactic acid bacteria (LAB), such as Lactobacillus plantarum and Lactobacillus brevis; however, it appears to be closely related to a ß-glucosidase from O. oeni ATCC BAA-1163 cloned into Escherichia coli. The periplasmic localization of the enzyme together with its high tolerance to ethanol and fructose, the low inhibitory effect of some wine-related compounds on the enzymatic activity and long-term stability of the enzyme could be of interest for winemaking. SIGNIFICANCE AND IMPACT OF THE STUDY: Information regarding a ß-glucosidase from O. oeni ST81 is presented. Although the release of aroma compounds by LAB has been demonstrated, little information exists concerning the responsible enzymes. To our knowledge, this study contains the first characterization of a native ß-glucosidase purified from crude extracts of O. oeni ST81.