Ecogenomics and metabolic potential of the South Atlantic Ocean microbiome.

The unique combination of depth, salinity, and water masses make the South Atlantic Ocean an ecosystem of special relevance within the global ocean. Yet, the microbiome of this ecosystem has received less attention than other regions of the global Ocean. This has hampered our understanding of the diversity and metabolic potential of the microorganisms that dwell in this habitat. To fill this knowledge gap, we analyzed a collection of 31 metagenomes from the Atlantic Ocean that spanned the epipelagic, mesopelagic and bathypelagic zones (surface to 4000 m). Read-centric and gene-centric analysis revealed the unique taxonomic and functional composition of metagenomes from each depth zone, which was driven by differences in physical and chemical parameters. In parallel, a total of 40 metagenome-assembled genomes were obtained, which recovered one third of the total community. Phylogenomic reconstruction revealed that many of these genomes are derived from poorly characterized taxa of Bacteria and Archaea. Genomes derived from heterotrophic bacteria of the aphotic zone displayed a large apparatus of genes suited for the utilization of recalcitrant organic compounds such as cellulose, chitin and alkanes. In addition, we found genomic evidence suggesting that mixotrophic bacteria from the bathypelagic zone could perform carbon fixation through the Calvin-Benson-Bassham cycle, fueled by sulfur oxidation. Finally, we found that the viral communities shifted throughout the water column regarding their targeted hosts and virus-to-microbe ratio, in response to shifts in the composition and functioning their microbial counterparts. Our findings shed light on the microbial and viral drivers of important biogeochemical processes that take place in the South Atlantic Ocean.

New viral biogeochemical roles revealed through metagenomic analysis of Lake Baikal.

BACKGROUND: Lake Baikal is the largest body of liquid freshwater on Earth. Previous studies have described the microbial composition of this habitat, but the viral communities from this ecosystem have not been characterized in detail. RESULTS: Here, we describe the viral diversity of this habitat across depth and seasonal gradients. We discovered 19,475 bona fide viral sequences, which are derived from viruses predicted to infect abundant and ecologically important taxa that reside in Lake Baikal, such as Nitrospirota, Methylophilaceae, and Crenarchaeota. Diversity analysis revealed significant changes in viral community composition between epipelagic and bathypelagic zones. Analysis of the gene content of individual viral populations allowed us to describe one of the first bacteriophages that infect Nitrospirota, and their extensive repertoire of auxiliary metabolic genes that might enhance carbon fixation through the reductive TCA cycle. We also described bacteriophages of methylotrophic bacteria with the potential to enhance methanol oxidation and the S-adenosyl-L-methionine cycle. CONCLUSIONS: These findings unraveled new ways by which viruses influence the carbon cycle in freshwater ecosystems, namely, by using auxiliary metabolic genes that act upon metabolisms of dark carbon fixation and methylotrophy. Therefore, our results shed light on the processes through which viruses can impact biogeochemical cycles of major ecological relevance. Video Abstract.

[Methodology for Identification of Inverse Drug Distribution, Spain]. / Resultados de una metodología para la identificación de distribución inversa de medicamentos en oficinas de farmacia.

OBJECTIVE: The phenomenon of reverse drug trafficking in the legal supply chain is an unlawful practice to serious risks to public health. The aims was to identify proactively pharmacies that carry out these illegal activities. METHODS: An analysis was performed through the crossing billing data to SAS of 52 million packs of medicines for the 496 pharmacies in the province over a period of 29 months with the drug packaging data supplied by the distribution entities of the province with the implementation of specific indicator defined called 'percentage overbought' allows us to detect those pharmacies at high risk of being involved in this illicit trade. RESULTS: It was tested in two pharmacies one rural and other urban a detour of 5.130 medicine containers and an illicit profit obtained from € 9,591.78 for the first and 9.982 packaging and € 26,885.11 for the second; they had gone unnoticed in previous inspections. CONCLUSIONS: The methodology implemented to define a profile of infringing pharmacies high risk in these illicit practices, identify new ones that had not been sanctioned, weigh the drugs for illegal trade and to identify new drugs subject to diversion; also added as a challenge, it helps to adjust accurately and effectively calculate the illicit profit obtained.

OBJETIVO: El fenómeno del tráfico inverso en la cadena legal de suministro de medicamentos es una práctica ilícita con graves riesgos para la Salud Pública. El objetivo de este trabajo fue identificar de forma proactiva las oficinas de farmacia que realizan estas conductas ilegales. METODOS: Se cruzaron los datos de facturación al SAS de 52 millones de envases de medicamentos con los de las 496 farmacias de la provincia en un periodo de 29 meses (abril de 2012 a agosto de 2014). Junto con la aplicación del indicador específico definido denominado "porcentaje de sobrecompra", permitió identificar los establecimientos farmacéuticos con alto riesgo de estar implicados en el comercio ilícito. RESULTADOS: Se comprobó que hubo desvío en dos oficinas de farmacia, una rural (de 5.130 envases de medicamentos y beneficio ilícito obtenido de 9.591,78 €) y otra urbana (9.982 envases y 26.885,11 €), los cuales habían pasado desapercibidos en anteriores actuaciones inspectoras. CONCLUSIONES: La metodología implantada permite definir un perfil de oficina de farmacia infractora de alto riesgo en estas prácticas ilícitas, identificarlas, ponderar los fármacos destinados a este comercio ilegal y determinar nuevos medicamentos objeto de desvío. Ayuda a ajustar de forma precisa el cálculo del beneficio ilícito obtenido.

Production of conidia of Beauveria bassiana in solid-state culture: current status and future perspectives.

Beauveria bassiana is an important entomopathogenic fungus widely commercialized in the world. Recent progress and achievements on conidia production have focused on a yield goal of 10(9) to 10(10) conidia per gram of dry substrate. Due to cost-competitive perspectives, these yields should be associated with better production rates or productivities. This study presents a review of relevant studies of B. bassiana conidia production on solid-state cultures and the parameters that should be taken into account to maintain constant quality in the product to be commercialized. Conditions for maximizing production and infectivity of B. bassiana conidia are also analysed.

Arrival of radionuclides released by the Fukushima accident to Tenerife (Canary Islands).

Two weeks after the accident at the Fukushima-Daichi nuclear power plant, 131I, 137Cs and 134Cs activities were measured in two different stations located in Tenerife (Canary Islands), situated at 300 (FIMERALL) and 2400 (IZAÑA) m.a.s.l, respectively. Peak measured activity concentrations were: 1.851 mBq/m3 (131I); 0.408 mBq/m3 (137Cs) and 0.382 mBq/m3 (134Cs). The activities measured at the FIMERALL station were always higher than at IZAÑA station, suggesting that the radioactive plume arrived to the island associated with low altitude air masses. Simulations of potential dispersion of the radioactive cloud (137Cs) after the nuclear accident in reactor Fukushima I show that radioactive pollution reached remote regions such as the Canary Islands in the Eastern subtropical North Atlantic. The corresponding effective dose to the local population was 1.17 nSv, a value less than one millionth of the annual limit for the general public. Therefore, there was no risk to public health.

Drug-eluting stents for off-label indications in real clinical world: evidence based or 'intuition' based clinical practice?

BACKGROUND: The use of drug-eluting stents (DES) is an example of the disparity between recommendations given by regulatory agencies and the real clinical world. Such disparity might lead cardiologists to adopt different routines in the use of DES. We aimed to assess variability of off-label DES use between hospitals and to what extent it can be explained by differences in patient or hospital characteristics. METHODS: Characteristics of consecutive patients receiving DES in 29 hospitals were recorded. Individual and hospital determinants of receiving DES for off-label indications were assessed by multilevel logistic regression. RESULTS: 1903 patients were recruited and 1188 (62.4%) received DES for off-label indications. Individual variables associated with off-label use were age (OR 1.01 (1-1.02)), previous percutaneous (OR 2.24 (1.68-2.97)) or surgical (2.41 (1.52-3.84)) revascularization, repeated procedure at the same admission (OR 4.66 (2.7-8.05)), receiving two (OR 4.17 (3.24-5.37)) or three or more DES (OR 14.12 (9.08-21.96)) vs one. Adjusting for individual variables, the Odds of receiving DES for off-label indication was higher in public funding hospitals with surgery availability vs private hospitals: 1.49 (0.86-2.6), and in public hospitals without surgery vs public with surgery availability: OR 1.76 (1.02-3.03). Interhospital variability reminded significant after adjustment for individual and contextual variables. CONCLUSION: Off-label DES use is highly variable between centers. Although this variability is partially determined by hospital type of funding and cardiac surgery availability, the substantial interhospital variability after multilevel adjustment suggests heterogeneity in the process of care.

Use of GFP-tagged strains of Penicillium digitatum and Penicillium expansum to study host-pathogen interactions in oranges and apples.

Penicillium digitatum and Penicillium expansum are responsible for green and blue molds in citrus and pome fruits, respectively, which result in major monetary losses worldwide. In order to study their infection process in fruits, we successfully introduced a green fluorescent protein (GFP) encoding gene into wild type P. digitatum and P. expansum isolates, using Agrobacterium tumefaciens-mediated transformation (ATMT), with hygromycin B resistance as the selectable marker. To our knowledge, this is the first report describing the transformation of these two important postharvest pathogens with GFP and the use of transformed strains to study compatible and non-host pathogen interactions. Transformation did not affect the pathogenicity or the ecophysiology of either species compared to their respective wild type strains. The GFP-tagged strains were used for in situ analysis of compatible and non-host pathogen interactions on oranges and apples. Knowledge of the infection process of apples and oranges by these pathogens will facilitate the design of novel strategies to control these postharvest diseases and the use of the GFP-tagged strains will help to determine the response of P. digitatum and P. expansum on/in plant surface and tissues to different postharvest treatments.

Development of a green fluorescent tagged strain of Aspergillus carbonarius to monitor fungal colonization in grapes.

An enhanced green fluorescent protein has been used to tag an OTA-producing strain of Aspergillus carbonarius (W04-40) isolated from naturally infected grape berries. Transformation of the fungus was mediated by Agrobacterium tumefaciens. The most efficient transformation occurred when the co-cultivation was done with 10(4) conidia due to higher frequency of resistance colonies (894 per 10(4) conidia) and lower background obtained. To confirm the presence of the hph gene in hygromycin resistant colonies, 20 putative transformants were screened by PCR analysis. The hph gene was identified in all the transformants. Variation on the expression levels of the eGFP was detected among the transformants and 50% of them appeared bright green fluorescent under the microscope. Microscopic analysis of all the bright fluorescent transformants revealed homogeneity of the fluorescent signal, which was clearly visible in the hyphae as well as in the conidia. eGFP expression in A. carbonarius was shown to be stable in all transformants. Confocal Laser scanning microscopy images of grape berries infected with the eGFP transformant demonstrated fungal penetration into the berry tissues. OTA production was importantly increased in the eGFP transformant in comparison with the wild type strain and pathogenicity on grape berries was slightly decreased after four days of inoculation. However, no differences in virulence were found after seven days of inoculation, thus allowing utilization of this eGFP mutant for in situ analysis of A. carbonarius infection of grape berries. To our knowledge, this is the first report describing the construction of a GFP-tagged strain belonging to Aspergillus section Nigri for monitoring Aspergillus rot on grape berries.

Radiometric analysis of farmed fish (sea bass, gilthead bream, and rainbow trout) from Tenerife Island, Spain.

This study analyzed the content of gamma-emitting radionuclides in fish farmed on the island of Tenerife (Canary Islands, Spain). The fish species included in this study were sea bass, gilthead bream, and rainbow trout. The first two species are produced in offshore enclosures, while the third is produced in a freshwater fish farm. All measurements were performed using two high-purity germanium gamma-ray detectors. The content of gamma-emitting radionuclides in the fodder used to feed the different species of farmed fish studied was also determined. The following nuclides were often detected in the analyzed samples: 137Cs, 40K, 235U, 228Ac, 214Bi, 208Tl, 212Pb, and 214Pb. As a complement to this analysis, 210Po concentrations in two fish samples were determined by alpha spectrometry. The nuclide presenting the highest concentration was, as expected, the naturally occurring 40K, with an average concentration of 0.13 +/- 0.01 Bq/g (wet weight) (Bq/gww) in gilthead bream and sea bass and 0.12 +/- 0.01 Bq/gww in rainbow trout. The 235U concentrations determined in the same fish species were 0.6 +/- 0.5, 0.8 +/- 0.7, and 1.6 +/- 1.0 mBq/gww, respectively. This nuclide is seldom reported in fish samples. The concentrations of 137Cs (the only artificial nuclide determined in this study) in gilthead bream and sea bass were 0.026 +/- 0.006 and 0.044 +/- 0.01 mBq/gww, respectively. In addition to the radiometric analysis, the contribution of the analyzed nuclides to the effective dose from the mean daily intake of the fish was calculated. The calculated contribution, in terms of dose per person, produced by intake of the analyzed fish was 0.8 microSv/year. This value does not represent a significant risk to the local population.

[Leigh syndrome caused by the mitochondrial DNA G14459A mutation in a Mexican family]. / Síndrome de Leigh causado por la mutación G14459A del ADN mitocondrial en una familia mexicana.

INTRODUCTION: Leigh syndrome is a neurodegenerative and progressive disease that appears usually in childhood due to defects in nuclear or mitochondrial genome. The mutation G14459A in mitochondrial DNA has been associated previously to Leber hereditary optic neuropathy and recently to Leigh syndrome. CASE REPORT: A 10 months-old Mexican girl diagnosed of Leigh syndrome. Molecular-genetic studies detected the mutation G14459A in a percentage close to homoplasmy and in low heteroplasmy in her mother. The rest of the maternally related family members analyzed were negative. CONCLUSION: The G14459A mutation, although not very frequently associated to Leigh syndrome, should be analyzed in patients that do not present the most common point mutations.

NARP syndrome in a patient harbouring an insertion in the MT-ATP6 gene that results in a truncated protein.

BACKGROUND: Neurogenic muscle weakness, ataxia and retinitis pigmentosa (NARP) syndrome have been associated to m.8993T>G/C mutations in the subunit 6 of the ATP synthase (p.MT-ATP6). METHODS: We have performed a mutational screening of the mitochondrial DNA gene encoding for this protein in 62 patients with the disease, that do not carry any of the common mutations described to date. RESULTS: We report clinical and molecular data in one patient who harbours a de novo insertion in the MT-ATP6 gene that results in a truncated subunit. The mutation was heteroplasmic (85%) in muscle DNA and the BN-PAGE analysis showed a clear decrease in the amount of ATP synthase. CONCLUSION: Molecular analysis of NARP patients cannot be limited to the search of the m.8993T>G/C and either the ATP6 or the whole mtDNA should be sequenced.

[Mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS) with the A3243G mutation of the tRNALeu(UUR) gene of mtDNA in native American haplogroup B2]. / Encefalomiopatía mitocondrial, acidosis láctica y accidentes cerebrovasculares (MELAS) con la mutación A3243G en el gen ARNtLeu(UUR) del ADNmt en el haplogrupo B2 nativo americano.

INTRODUCTION: Mitochondrial encephalopathy, lactic acidosis and cerebrovascular accident syndrome (MELAS) is, from the clinical point of view, one of the best studied mitochondrial multisystemic disorders. This disease has mainly been associated to the mitochondrial desoxyribonucleic acid (mtDNA) mutation A3243G located in the tRNALeu(UUR) gene. Although a relation between European haplogroups and the presence of the 3243 mutation has not been described, nothing is known about the presence of this mutation in native American haplogroups. CASE REPORT: A 12 year-old female Mexican patient diagnosed with MELAS is reported. Besides neurological, biochemical and cytological examination, we also analyzed the particular mtDNA mutations related to MELAS and the whole genome was sequenced to determine the mitochondrial haplogroup. The A3243G mutation was detected in the patient and maternal relatives (mother and siblings, all of them asymptomatic). The genotype corresponds to the native American haplogroup B2 and contains two private non-synonymous polymorfisms. CONCLUSION: All the members of the family studied present different percentage of the A3243G mutation, being the patient who presented the highest value. The mtDNA genotype corresponds to the native American haplogroup B2 and the private polymorphisms do not confer any phenotypic modification in MELAS syndrome.

[Diseases of the human mitochondrial oxidative phosphorylation system]. / Enfermedades del sistema de fosforilación oxidativa mitocondrial humano.

AIM: The diseases of the oxidative phosphorylation system consist of a group of disorders originated by a deficient synthesis of adenosine triphosphate (ATP). These diseases are increasingly being diagnosed among patients with multisystemic disorders. Mitochondrial deoxyribonucleic acid (mtDNA) mutations are usually maternally inherited, but they also can be sporadic or secondary to nuclear mutations, that are inherited in a Mendelian mode, or due to environmental hazards. In this review we will update, from a genetic point of view, the knowledge on human mitochondrial diseases, remarking the difficulties to study these pathologies. DEVELOPMENT: To mirror these difficulties, we will use selected examples of mutations in the mitochondrial genome, and review recent advances on mitochondrial pathology due to mutations in the nuclear genes codifying for mitochondrial proteins that participate in a good performance of the oxidative phosphorylation system. CONCLUSIONS: Sequencing of the complete human mtDNA should be part of the basic profile in the study of mitochondrial diseases. Due to the increasing number of nuclear genes involved in the oxidative phosphorylation system performance, their analysis should be based on solid biochemical clues.

Skeletal muscle mitochondrial DNA content in exercising humans.

Several weeks of intense endurance training enhances mitochondrial biogenesis in humans. Whether a single bout of exercise alters skeletal muscle mitochondrial DNA (mtDNA) content remains unexplored. Double-stranded mtDNA, estimated by slot-blot hybridization and real time PCR and expressed as mtDNA-to-nuclear DNA ratio (mtDNA/nDNA) was obtained from the vastus lateralis muscle of healthy human subjects to investigate whether skeletal muscle mtDNA changes during fatiguing and nonfatiguing prolonged moderate intensity [2.0-2.5 h; approximately 60% maximal oxygen consumption (Vo(2 max))] and short repeated high-intensity exercise (5-8 min; approximately 110% Vo(2 max)). In control resting and light exercise (2 h; approximately 25% Vo(2 max)) studies, mtDNA/nDNA did not change. Conversely, mtDNA/nDNA declined after prolonged fatiguing exercise (0.863 +/- 0.061 vs. 1.101 +/- 0.067 at baseline; n = 14; P = 0.005), remained lower after 24 h of recovery, and was restored after 1 wk. After nonfatiguing prolonged exercise, mtDNA/nDNA tended to decline (n = 10; P = 0.083) but was reduced after three repeated high-intensity exercise bouts (0.900 +/- 0.049 vs. 1.067 +/- 0.071 at baseline; n = 7; P = 0.013). Our findings indicate that prolonged and short repeated intense exercise can lead to significant reductions in human skeletal muscle mtDNA content, which might function as a signal stimulating mitochondrial biogenesis with exercise training.

[Five-year follow-up of urinary incontinence in older people in a Spanish rural population]. / Seguimiento durante 5 años de la incontinencia urinaria en los ancianos de una población rural española.

OBJECTIVE: To determine prevalence, incidence and remission rates and change patterns of urinary incontinence (UI) over a 5-year time span. DESIGN: Longitudinal study. LOCATION: Basic Health Zone of Cabra (Cordoba). PATIENTS: A random age-stratified sample of 827 subjects selected from a total of 5139 persons > or = 65 years of age and followed up between 1996 and 2001. INTERVENTIONS: Home interview. MEASUREMENTS: Questions about detection and level of severity of UI, sociodemographic data, and level of functioning. Two logistic regression models were created to identify risk factors for the probability to be alive and to develop UI, respectively. RESULTS: UI prevalence reached 36% at baseline and 46% five years later. Mortality rates were similar in continent and incontinent subjects. 54.5% of continent subjects in 1996 remained so in 2001. Five-year incidence and remission rates were 29% and 15%, respectively. In both sexes, slight and moderate incontinence mainly progressed to moderate and severe degrees, whereas severe UI remained unchanged in 40.1%. The main risk factors for increased survival and for UI incidence were self-rated health and level of functioning. CONCLUSIONS: Prevalence of UI is high, incidence is moderate and remission is low. Urinary incontinence is a dynamic problem and does not affect mortality. Impaired mobility has a strong influence on UI incidence.