The role of microRNAs in NBS-LRR gene expression and its implications for plant immunity and crop development.

Plants evolved, over millions of years, complex defense systems against pathogens. Once infected, the interaction between pathogen effector molecules and host receptors triggers plant immune responses, which include apoptosis, systemic immune response, among others. An important protein family responsible for pathogen effector recognition is the nucleotide binding site-leucine repeat rich (NBS-LRR) proteins. The NBS-LRR gene family is the largest disease resistance gene class in plants. These proteins are widely distributed in vascular plants and have a complex multigenic cluster distribution in plant genomes. To counteract the genetic load of such a large gene family on fitness cost, plants evolved a mechanism using post transcriptional gene silencing induced by small RNAs, particularly microRNAs. For the NBS-LRR gene family, the small RNAs involved in this silencing mechanism are mainly the microRNA482/2118 superfamily. This suppression mechanism is relieved upon pathogen infection, thus allowing increased NBS-LRR expression and triggering plant immunity. In this review, we will discuss the biogenesis of microRNAs and secondary RNAs involved in this silencing mechanism, biochemical and structural features of NBS-LRR proteins in response to pathogen effectors and the evolution of microRNA-based silencing mechanism with a focus on the miR482/2118 family. Furthermore, the biotechnological manipulation of microRNA expression, using both transgenic or genome editing approaches to improve cultivated plants will be discussed, with a focus on the miR482/2118 family in soybean.

Test station to characterize the emission of a LiDAR.

A test station setup devised to measure the emission characteristics and beam propagation parameters of a light detection and ranging (LiDAR) system is presented. The main blocks of the station to measure the accessible emission, wavelength peak and FWHM, pulse duration, pulse repetition rate, horizontal and vertical angular resolution, field of view, beam propagation factor M 2, beam waist size, waist location, and divergence are described. The performance of this test station was demonstrated using a commercial spinning LiDAR, a Velodyne VLP-16, which successfully enables these measurements for a laser beam with a wavelength of 913 nm.

High frequency of non-susceptibility to ciprofloxacin in nontyphoid Salmonella recovered from Brazilian broiler chicken.

1. This study evaluated the minimal inhibitory concentration (MIC) of ciprofloxacin and the presence of plasmid-mediated quinolone resistance (PMQR) mechanisms in 97 nontyphoidal Salmonella spp. isolated from broilers and carcases from three different regions in Brazil. The presence of mutations in quinolone resistance determination regions (QRDRS) was investigated in the ciprofloxacin-resistant strain by DNA sequencing.2. Most of the Salmonella spp. (85.6%) had intermediate resistance to ciprofloxacin and only one isolate was resistant. MIC breakpoints ranged from ≤0.03 to 1 µg/ml and 67.0% of the strains had a MIC of 0.25 µg/ml (n=65). Thirteen strains (13.4%) were susceptible to ciprofloxacin with MIC ≤0.06 µg/ml. The qnrB gene was detected in eight isolates with intermediate resistance and in two susceptible strains. The other PMQR genes, qnrA, qnrC, qnrD, qnrS, qnrVC, aac(6')-Ib, qepA, oqxAB and mutations in QRDR were not detected in any strain.3. There was a high frequency of ciprofloxacin intermediate resistant Salmonella from broiler and broiler carcases from Brazil. The presence of these strains in poultry and derived products poses a risk to public health.

Impetigo: A need for new therapies in a world of increasing antimicrobial resistance.

WHAT IS KNOWN AND OBJECTIVE: Impetigo is a highly contagious bacterial skin infection and is one of the most common skin infections in children. Antibiotics are the first-line treatment when multiple lesions exist, but with an increasing prevalence of antibiotic-resistant bacteria the successful management of impetigo in the future is an area of concern. COMMENT: Current treatment options that favour the use of oral antibiotic therapy are increasingly problematic. Widespread use of these agents contributes to antimicrobial resistance and has adverse consequences for individuals and communities. There is a need for new topical antimicrobials and antiseptics as an alternative treatment strategy. WHAT IS NEW AND CONCLUSION: To successfully treat impetigo into the future and ensure that therapy does not contribute to bacterial resistance, additional research is required to ascertain the usefulness of alternative agents, including new topical antimicrobials and antiseptics.

Philometra saltatrix (Nematoda: Philometridae) in the ovary of the bluefish, Pomatomus saltatrix (Linnaeus, 1766), off the coast of the state of Rio de Janeiro, Brazil.

The aims of the present study were to identify and describe the occurrence of nematode parasites in the gonads of bluefish Pomatomus saltatrix from off the coast of the state of Rio de Janeiro, Brazil. Only females were found to be parasitized by the nematodes, which were identified as P. saltatrix using morphological, morphometric and molecular data. Infection of female bluefish by this nematode had the following values: prevalence, 48.7%; mean intensity, 2.6; mean abundance, 1.3; and range of infection, 1-10 specimens. Histopathological examination of transverse and longitudinal sections of the parasitized ovaries showed nematodes at different stages of development among oocytes, but no indication of any associated inflammatory reaction. The presence of nematodes in the ovaries of bluefish is an important indication of fish hygiene, and parasitized fish are usually rejected by consumers because of their repugnant appearance.

The land morphology approach to flood risk mapping: An application to Portugal.

In the last decades, the increasing vulnerability of floodplains is linked to societal changes such as population density growth, land use changes, water use patterns, among other factors. Land morphology directly influences surface water flow, transport of sediments, soil genesis, local climate and vegetation distribution. Therefore, the land morphology, the land used and management directly influences flood risks genesis. However, attention is not always given to the underlying geomorphological and ecological processes that influence the dynamic of rivers and their floodplains. Floodplains are considered a part of a larger system called Wet System (WS). The WS includes permanent and temporary streams, water bodies, wetlands and valley bottoms. Valley bottom is a broad concept which comprehends not only floodplains but also flat and concave areas, contiguous to streams, in which slope is less than 5%. This will be addressed through a consistent method based on a land morphology approach that classifies landforms according to their hydrological position in the watershed. This method is based on flat areas (slopes less than 5%), surface curvature and hydrological features. The comparison between WS and flood risk data from the Portuguese Environmental Agency for the main rivers of mainland Portugal showed that in downstream areas of watersheds, valley bottoms are coincident with floodplains modelled by hydrological methods. Mapping WS has a particular interest in analysing river ecosystems position and function in the landscape, from upstream to downstream areas in the watershed. This morphological approach is less demanding data and time-consuming than hydrological methods and can be used as the preliminary delimitation of floodplains and potential flood risk areas in situations where there is no hydrological data available. The results were also compared with the land use/cover map at a national level and detailed in Trancão river basin, located in Lisbon metropolitan area, an urbanized basin that suffered heavy flooding in the last decades. This study also contributes to a better understanding of the basin morphology at a local-scale and the effects of soil sealing in downstream flood risks. This work will contribute to the understanding of the morphology, ecology and land use of watersheds that could be used to reduce runoff and downstream flood risk. This can be accomplished by using natural water retention and infiltration methods or higher-level based planning instead of a reaction to local decisions on flood hazards. This morphological approach to map landforms, including wet system, is a valuable tool to assist policy makers and planners in flood risk and land use management, floodplain restoration, agricultural land management practices, and location of human activities according to ecological suitability.

Soybean seed protein storage vacuoles for expression of recombinant molecules.

Soybean is one of the most important protein sources for human consumption and livestock feed. Soy production also allows the biosynthesis of edible oils, biodiesel, and biofertilizers. With the advent of modern agricultural biotechnology, soybean plants have also converted into bioreactors of therapeutic proteins and industrial enzymes. Soybean's characteristics, such as protein storage vacuoles (PSVs) and other unique organelles, allow the plant to be exploited as an accumulator of heterologous proteins under high stability and scalability conditions, and that maintains its basic functions. This review reports the main aspects of heterologous protein accumulation in soybean PSVs.

Notes on the conservation threats to the western lesser spot-nosed monkey (Cercopithecus petaurista buettikoferi) in the Bijagós Archipelago (Guinea-Bissau, West Africa).

The lesser spot-nosed monkey (Cercopithecus petaurista) is a widely distributed West African guenon, which is generally considered less vulnerable to local extinctions than many sympatric primate species. Guinea-Bissau harbours the westernmost populations of the species, which is thought to be very rare or even extinct on the mainland, but to have putative populations on some islands of the Bijagós Archipelago. However, due to a lack of regional studies, baseline information on these insular populations is missing. We collected baseline data on the anthropogenic activities that possibly threaten the long-term conservation of this primate by using non-systematic ethnographic methodologies. The species was reported to be decreasing in number or rare by locals on two of the islands, and we identified two main conservation threats to it: generalised habitat loss/degradation, and hunting. While subsistence hunting has been recorded before in these areas, we report, to the best of our knowledge for the first time for these islands, the presence of a semi-organised commercial wild meat trade. The carcasses of western lesser spot-nosed monkeys were observed being stored and shipped from seaports to be sold at urban hubs (Bissau and Bubaque Island). The effect of commercial trade on the species could be severe, considering the small, naturally occurring, carrying capacities typical of insular ecosystems. The results of this study highlight the importance of understanding the leading social drivers of wild meat hunting of lesser spot-nosed monkeys on the Bijagós Archipelago, and the need to conduct baseline research on these insular populations, for which qualitative and quantitative methods could be combined.

Soybeans as bioreactors for biopharmaceuticals and industrial proteins.

Plants present various advantages for the production of biomolecules, including low risk of contamination with prions, viruses and other pathogens, scalability, low production costs, and available agronomical systems. Plants are also versatile vehicles for the production of recombinant molecules because they allow protein expression in various organs, such as tubers and seeds, which naturally accumulate large amounts of protein. Among crop plants, soybean is an excellent protein producer. Soybean plants are also a good source of abundant and cheap biomass and can be cultivated under controlled greenhouse conditions. Under containment, the plant cycle can be manipulated and the final seed yield can be maximized for large-scale protein production within a small and controlled area. Exploitation of specific regulatory sequences capable of directing and accumulating recombinant proteins in protein storage vacuoles in soybean seeds, associated with recently developed biological research tools and purification systems, has great potential to accelerate preliminary characterization of plant-derived biopharmaceuticals and industrial macromolecules. This is an important step in the development of genetically engineered products that are inexpensive and safe for medicinal, food and other uses.

Correct targeting of proinsulin in protein storage vacuoles of transgenic soybean seeds.

Soybean plants are promising bioreactors for the expression of biochemically complex proteins that cannot be produced in a safe and/or economically viable way in microorganisms, eukaryotic culture cells or secreted by transgenic animal glands. Soybeans present many desirable agronomic characteristics for high scale protein production, such as high productivity, short reproductive cycle, photoperiod sensitivity, and natural organs destined for protein accumulation in the seeds. The significant similarities between plant and human cells in terms of protein synthesis processes, folding, assembly, and post-translational processing are important for efficient accumulation of recombinant proteins. We obtained two transgenic lines using biolystics, incorporating the human proinsulin gene under control of the monocot tissue-specific promoter from sorghum gamma-kafirin seed storage protein gene and the alpha-coixin cotyledonary vacuolar signal peptide from Coix lacryma-jobi (Poaceae). Transgenic plants expressed the proinsulin gene and accumulated the polypeptide in mature seeds. Protein targeting to cotyledonary protein storage vacuoles was successfully achieved and confirmed with immunocytochemistry assays. The combination of different regulatory sequences was apparently responsible for high stability in protein accumulation, since human proinsulin was detected after seven years under room temperature storage conditions.

Ultrasound-guided cytology of spleen and liver: a prognostic tool in canine cutaneous mast cell tumor.

BACKGROUND: In the clinical staging of cutaneous mast cell tumors (cMCT), the diagnosis of metastasis is controversial based on cytological examination of lymph nodes, spleen, liver, bone marrow, and blood. OBJECTIVES: To define the prognostic role of ultrasound-guided cytology of spleen and liver in cMCT. The results of cytological evaluation were compared in relation with survival time. ANIMALS: Fifty-two client-owned dogs with a diagnosis of cMCT. METHODS: Selection of cases was based on cytological evaluation of liver and spleen to detect infiltration at distant sites. The Kaplan Meier method was used to compare survival in dogs with and without infiltration of spleen and liver (log-rank test P < .05). RESULTS: Ten dogs with cMCT had mast cell infiltration of spleen, liver, or both and 4 of these dogs had involvement of the regional lymph nodes. The majority of dogs had 2 or more ultrasonographically abnormal findings simultaneously in spleen and liver. Nine dogs had grade II cMCT, and 1 had grade III cMCT. Dogs with positive evidence of mast cell infiltration to spleen, liver, or both had shorter survival times (34 versus 733 days) compared with dogs negative for mast cell infiltration at distant sites. CONCLUSION AND CLINICAL IMPORTANCE: Dogs with evidence of mast cell infiltration at distant sites have a shorter survival times than dogs without evidence of infiltration at distant sites. This study suggests that cytology of spleen and liver is indicated either for ultrasonographically normal or for ultrasonographically abnormal spleen and liver in dogs with cMCT.

Mucoid degeneration of the anterior cruciate ligament: magnetic resonance imaging findings of an underdiagnosed entity.

Although imaging and clinical criteria are already established for the diagnosis of mucoid degeneration of the anterior cruciate ligament (ACL), many radiologists remain unaware of their existence. Once regarded as a rare occurrence, it has been recently suggested that its incidence is in fact very much higher than previously thought, and that it is probably underdiagnosed or misdiagnosed as partial ruptures of the ACL or as cystic ganglia. In this review, the authors revisit the literature concerning this subject and present their personal experience with it, stressing the paramount importance of magnetic resonance imaging and correlation with clinical data for a correct diagnosis.

Growth parameters estimates for a small fish of the Pantanal, Brazil: Moenkhausia dichroura (Characiformes; Characidae).

Growth parameters were estimated for Moenkhausia dichroura (Kner, 1858) (Characiformes, Characidae), a small-sized and very abundant fish of the Pantanal lentic habitats commonly known as "pequira ". A method based on the length frequencies distribution and the ELEFAN I routine from the FISAT program were used. The fish were collected in the Baia da Onça, an oxbowlake of the sub-region Pantanal of Aquidauana, Mato Grosso do Sul, Brazil, from June to December 1988. The standard length of the captured fishes ranged from 29 to 76 mm with an average of 53 mm. The estimated growth parameters were L(infinity) = 81 mm (standard length), k = 0.85 year(-1), C = 0.89, WP = 0.6 (Rn = 0.285). The WP indicated that growth reduction occurred in July, when the lowest temperature of the year was registered. The growth curve showed that captured individuals belonged to three cohorts. The obtained results seem robust and quite compatible with the biology of the fish and its adjustment to the environment. M. dichroura, in spite of not being a direct fishing interest, is an important species in terms of its ecological aspects, due to its abundance and high growth rate, and as a great food source for aquatic organisms and specially for larger fish of economic value. Considering the information gap about small fish, the parameters estimated for pequira constitute a comparison base for other growth studies of small-sized fish species of tropical environments.

Low serum levels of vitamin B12 in older adults with normal nutritional status by mini nutritional assessment.

Undernutrition as well as low levels of vitamin B12 and folic acid are common problems among older adults. However, recommended routine nutritional status assessment tools may result in inadequate vitamin serum levels to go unnoticed. Therefore, the aim of this study is to evaluate the inadequacy of serum levels of vitamin B12 and folic acid within Mini Nutritional Assessment (MNA) classification categories among older adults. A cross-sectional study was conducted with 97 older adults residing in care homes in Portugal. Undernutrition was identified through the MNA, and serum levels of vitamin B12 and folic acid were measured using chemiluminescence. Cognitive function, depressive symptoms and functional characteristics were also assessed using the Abbreviated Mental Test Score, the Epidemiologic Studies Depression Scale and the Barthel Index, respectively. The mean age of older adults was 82.2 (6.3) years; 3.1% were undernourished and 26.8% were at undernutrition risk. In the MNA normal nutritional status group, 11.8% presented vitamin B12 deficiency (<200 pg/ml), 32.4% had low serum levels (200-400 pg/ml) and 4.4% had folic acid deficiency (<3 ng/ml). A high proportion of older adults with low serum levels of vitamin B12 presenting normal nutritional status by MNA was identified. This finding emphasizes the need to evaluate serum vitamin B12 levels, independently of the MNA results.

Proposal of an iELISA for Mycoplasma bovis diagnosis in dairy cattle and associated risk factors / [Proposta de um iELISA para diagnóstico de Mycoplasma bovis em bovinos leiteiros e fatores de risco associados]

Mycoplasma bovis is a highly contagious agent associated with several pathologies in cattle. The detection of reactive antibodies to M. bovis by Indirect Enzyme-Linked Immunosorbent Assay (iELISA) identifies if there was an exposure to the microorganism. The current study aimed to optimize an iELISA from M. bovis total cell antigen, applying it to bovine serum samples, and to evaluate risk factors. Serum samples were obtained from 400 cows from 17 herds from Southeast Brazil. In the optimization of iELISA, the following was established: 2 µg/mL of antigen, sera dilution 1:300, and conjugate dilution 1:15000. The frequency was 62.3% (249/400) of reactive animals and 100% (17/17) of reactive herds. Risk factors were: herds with more than 100 animals (OR= 3.1; CI= 95%); Holstein breed (OR= 72.5; CI= 95%); cows (OR= 29.7; CI= 95%); intensive breeding system (OR= 3.3; CI= 95%); associated small ruminant production (OR= 4.4; CI= 95%); milk production above 500L (OR= 2.9; CI= 95%); no quarantine (OR= 1.5; CI= 95%); mechanical milking (OR= 5.5; CI= 95%) and cases of mastitis (OR= 5.5; CI= 95%). The proposed iELISA was able to detect antibodies reactive to M. bovis in bovine serum. Knowledge of these risk factors can assist in the implementation of prophylactic measures.(AU)

Mycoplasma bovis é um agente altamente contagioso relacionado a várias patologias em bovinos. A detecção de anticorpos reativos a M. bovis por Ensaio de Imunoadsorção Enzimática Indireto (iELISA) identifica se houve exposição ao microrganismo. O presente estudo teve como objetivo otimizar um iELISA de antígeno celular total de M. bovis, aplicando-o a amostras de soro bovino, bem como avaliar fatores de risco. Amostras de soro foram obtidas de 400 vacas de 17 rebanhos da Região Sudeste do Brasil. Na otimização do iELISA foram obtidos: 2µg/mL de antígeno, diluição dos soros 1:300 e do conjugado 1:15000. A frequência de animais reativos foi de 62,3% (249/400) e de 100% (17/17) para os rebanhos. Os fatores de risco foram: rebanhos com mais de 100 animais (OR= 3,1; IC= 95%); raça Holandesa (OR= 72,5;IC= 95%); vacas (OR= 29,7;IC= 95%); sistema intensivo (OR= 3,3; C= 95%); produção de pequenos ruminantes (OR= 4,4;IC=95%); produção de leite acima de 500L (OR= 2,9;IC= 95%); sem quarentena (OR= 1,5;IC= 95%); ordenha mecânica (OR= 5,5;IC= 95%) e casos de mastite (OR= 5,5;IC= 95%). O iELISA proposto foi capaz de detectar anticorpos reativos a M. bovis no soro bovino. O conhecimento desses fatores de risco pode auxiliar na implementação de medidas profiláticas.(AU)