Recovery of active pathogenesis-related enzymes from the apoplast of Musa acuminata infected by Mycosphaerella fijiensis

The fungus Mycosphaerella fijiensis causes black Sigatoka (BS) disease, a major pathogen in the banana industry worldwide. Numerous molecular and biochemical studies have been done for the M. fijiensis, Musa acuminata interaction, but this is the first study describing the zymographic behavior of β-1,3-glucanase, chitinase and protease in the apoplast and symplast of healthy, BS-infected but asymptomatic and BS diseased banana leaves. In BS-infected tissues, β-1,3-glucanase enzymatic activity was associated with two polypeptides with retention index (Ri) values of 0.43 and 0.56. These were more notable in the apoplast than in the symplast. Chitinase activity in BS-infected tissue in both the apoplast and symplast was mainly associated with a single polypeptide (Ri = 0.89). Both β-1,3-glucanase and chitinase activities were apparently more intense in BS-infected leaves than in healthy leaves. Protease activity was associated with two polypeptides (Ri = 0.04 and 0.14). In both the apoplast and symplast, the Ri 0.04 polypeptide increased in intensity with disease progression, whereas Ri 0.14 polypeptide intensity decreased. Overall protease activity intensity was higher in the symplast. Maximum symplast contamination of the apoplast was 2% as estimated by glucose 6-phosphate dehydrogenase activity, a biochemical marker for symplast. Accumulation of pathogenesis-related enzymatic activities in the apoplast of M. acuminata leaf tissue was caused by hostcontrolled enzyme downloading in response to M. fijiensis infection. Clear differences were identified in the electrophoretic profiles of healthy and diseased banana plants. The results further support a putative role of these enzymes in the extracellular defense repertoire of banana and, more importantly, suggest that M. fijiensis possesses a mechanism for suppression and delay of defense response in M. acuminata.Key words: Black Sigatoka, glucose 6-phosphate dehydrogenase, pathogenesis-related (PR) proteins, polyacrylamide gel electrophoresis (PAGE), retention index (Ri), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)

Is it important to achieve physical activity recommendations at early stages of life to improve bone health?

Research in bone health during childhood is limited and important to prevent future diseases, particularly, osteoporosis. Bone parameters using DXA and pQCT in 295 Spanish children were evaluated and we found a benefit of meeting the World Health Organization physical activity recommendations in bone composition in childhood. Purpose To investigate the association between physical activity (PA) and bone health in a Spanish paediatric cohort, considering the influence of meeting/not meeting the current World Health Organization (WHO) PA recommendations and to elucidate if there are differences between boys and girls. Methods In a cohort of children born in the region of Aragon (Spain) in 2009, followed until the age of 7 years, bone parameters were assessed using dual-energy X-ray absorptiometry (DXA) (whole body scan) and peripheral quantitative computed tomography (pQCT) (tibia scanned at the 8% (distal) and 38% (diaphyseal) of the total tibia length) in 295 7-year-old children (154 boys) in the last evaluation performed between 2016 and 2017. PA was assessed using GT3X Actigraph accelerometers. Results Boys had significantly higher areal bone mineral density (aBMD), higher total bone mineral content (BMC) at the diaphyseal site and higher trabecular BMC and vBMD, and higher total bone area at the distal site than girls (p<0.01 for all of them). Both boys and girls complying with the WHO PA recommendations had significantly higher trabecular BMC than their inactive counterparts. Conclusions Meeting WHO PA recommendations has a beneficial effect in bone composition in childhood both in boys and in girls

A simplified courtship conditioning protocol to test learning and memory in Drosophila

In Drosophila, a male that has previously been sexually rejected reduces its courtship behavior when confronted again with an unreceptive female. This reduced courting time reflects a memory formation process. Here, we describe a simplified protocol to perform the courtship conditioning assay for assessing the reduced courting time, using regular lab equipment and handmade tools. Every step of the procedure, from raising flies and training to testing and quantification of this memory-related behavior, can be implemented in any practice laboratory.We would like to thank Javier Gil Castillo for its invaluable help and advices in 3D printing. We also thank the flies from Bloomington Stock Center. We would like to thank BioRender (www.biorender.com) for the open-access platform used to create the graphical abstract. This work was supported by the Spanish Research Agency (Ministerio de Innovacion y Ciencia [MICINN]) under the grant PGC2018-094630-B-100 to F.A.M., cofinanced by the European Regional Development Fund (ERDF) to F.A.M. F.A.M. is a recipient of a RyC-2014-14961 contract. B.G.-M. is a recipient of a FPI-UAM predoctoral fellowship, grant number SFPI/2020/00878. C.G.B. is a recipient of a FPU predoctoral fellowship, grant number FPU19/04449 (MEFP). S.P.-F. is a recipient of a JAE intro fellowship, grant number JAEINT_21_02520 (CSIC)

Fructooligosaccharides production by solid-state fermentation with Aspergillus niger PSH

Fructoligosacharides (FOS) belong to prebiotics group that are non- digestible oligosacharides but fermentable by the bacteria in the gut microbiota1 . FOS can be produced by inulin degradation, which results in products with long fructoligomer chains2 . Alternatively, FOS can be obtained by conversion of the sucrose using fructosyltransferase (FTase) or β- fructofuranosidase (FFase) enzymes from bacterial and fungal sources2 . The curent proces for industrial production of FOS uses FTase produced by Aspergilus niger, and reaches an yield value of aprox. 60%3 . The present study evaluated the FOS production by solid-state fermentation (SSF) with the objective of finding a technology able to produce FOS with higher yield

ANTIMICROBIAL EFFECT OF VANCOMYCIN ELECTRO-TRANSFERRED WATER AGAINST METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS VARIANT

Background: There is a number of alternative and complementary therapeutics that are unproven or have not been properly tested. For past twenty years, the transfer of bio-energetic information has been recognized as a novel scientific approach capable of contributing to improved therapy in the management of several diseases through the so-called bio-resonance therapy (BRT). Although BRT was discovered in the late 1980s, it is still poorly studied. The aim of this study was to evaluate the antibacterial effect of water samples transferred with electronic information of vancomycin, a well known drug against methicillin-resistant Staphylococcus aureus (MRSA), by using a BRT device on bacterial cultures. Material and Methods: MRSA cultures were treated with vancomycin electro-transferred water samples, vancomycin (4.0 and 8.0 µg/mL), sham electro-transferred (water to water) and non-transferred water samples (medium alone). Growth inhibition was evaluated in liquid and solid culture medium, spectrophotometrically and by CFU determination respectively. Results: The obtained data showed that by transferring vancomycin (4.0 and 8.0 µg/mL) information to water samples, the growth of cultured MRSA was significantly (p< 0.05) inhibited (up to 35%), compared with those cultures treated with electro-transferred water to water or cultured in medium alone (0% growth inhibition). Conclusion: This in vitro study suggests that water samples that are electronically transferred with vibration sustained information of vancomycin are capable of inhibiting growth of axenically cultured methicillin resistant S. aureus

Biological Activity and Implications of the Metalloproteinases in Diabetic Foot Ulcers

Inadequate metabolic control predisposes diabetic patient to a series of complications on account of diabetes mellitus (DM). Among the most common complications of DM is neuropathy, which causes microvascular damage by hyperglycemia in the lower extremities which arrives characterized by a delayed closing. The global prevalence of diabetic neuropathy (DN) was 66% of people with diabetes in 2015, representing the principal cause of total or partial lower extremities amputation, with 22.6% of the patients with DN. Matrix metalloproteinases (MMPs) are involved in healing. The function that these mainly play is the degradation during inflammation that has as consequence the elimination of the extracellular matrix (ECM), the disintegration of the capillary membrane to give way to angiogenesis and cellular migration for the remodeling of damaged tissue. The imbalance in MMPs may increase the chronicity of a wound, what leads to chronic foot ulcers and amputation. This chapter focuses on the role of MMPs in diabetic wound healing

Challenges in Estimating Insecticide Selection Pressures from Mosquito Field Data

Insecticide resistance has the potential to compromise the enormous effort put into the control of dengue and malaria vector populations. It is therefore important to quantify the amount of selection acting on resistance alleles, their contributions to fitness in heterozygotes (dominance) and their initial frequencies, as a means to predict the rate of spread of resistance in natural populations. We investigate practical problems of obtaining such estimates, with particular emphasis on Mexican populations of the dengue vector Aedes aegypti. Selection and dominance coefficients can be estimated by fitting genetic models to field data using maximum likelihood (ML) methodology. This methodology, although widely used, makes many assumptions so we investigated how well such models perform when data are sparse or when spatial and temporal heterogeneity occur. As expected, ML methodologies reliably estimated selection and dominance coefficients under idealised conditions but it was difficult to recover the true values when datasets were sparse during the time that resistance alleles increased in frequency, or when spatial and temporal heterogeneity occurred. We analysed published data on pyrethroid resistance in Mexico that consists of the frequency of a Ile1,016 mutation. The estimates for selection coefficient and initial allele frequency on the field dataset were in the expected range, dominance coefficient points to incomplete dominance as observed in the laboratory, although these estimates are accompanied by strong caveats about possible impact of spatial and temporal heterogeneity in selection

Final Report of the Fifth Meeting of Scientific Experts on Fish Stocks in the Central Arctic Ocean

This report provides a summary of the 5th meeting of scientific experts on Fish Stocks in the Central Arctic Ocean (FiSCAO) on October 24‐26, 2017, in Ottawa, Canada. At the request of the 10 parties negotiating on an agreement to prevent unregulated commercial fishing in the High Seas portion of the Central Arctic Ocean (CAO), participants of the 5th FiSCAO meeting were tasked with addressing four Terms of Reference, summarized below: ToR 1. Design a 1‐3 year long mapping program. ToR 2. Design a monitoring program. ToR 3. Identify human, financial, vessel/equipment resources needed for mapping and monitoring. ToR 4. Develop data collection, sharing, and hosting protocols that outline the details of what and how data shall be collected, shared, and hosted for consideration by the Parties. The 5th FiSCAO meeting included scientific representatives from seven states including Canada, the People's Republic of China, the European Union, Iceland, the Republic of Korea, the Kingdom of Norway and the United States of America. The meeting also included representatives from the International Council for the Exploration of the Sea (ICES), the North Pacific Marine Science Organization (PICES) and the Arctic Council’s Protection of the Arctic Marine Environment (PAME) and Conservation of Arctic Flora and Fauna (CAFF) working groups. The report summarizes the elements for collecting baseline data (i.e., a mapping program) in the high seas CAO to achieve the goals of documenting species distributions, relative abundances and key ecosystem parameters (ToR 1). The mapping program describes the priority areas to sample, the types of data to collect and possible data collection approaches to employ. Participants emphasized that existing planned surveys are very limited, and that significant dedicated resources will be required to implement the mapping program. The report outlines a strategy for monitoring indicators of fish stocks and ecosystem components (ToR 2). The report includes a list of existing monitoring programs and a prioritized list of indicators to detect environmental change in the high seas CAO. Further refinement of a monitoring program will use information from the mapping program (ToR 1). Participants emphasized the need to begin monitoring as soon as possible and that additional research is required to operationalize monitoring indicators. The report summarizes the preliminary cost estimates (ToR 3) to implement a mapping program to collect data in the high seas portion of the CAO using a vessel of opportunity and in the Pacific Gateway region of the CAO using an independently‐organized survey. Cost implications for the monitoring program and other scientific activities are also listed (e.g., data analysis, data management). The report includes a draft data sharing policy as the foundation for a future data sharing protocol, including the technical specifications for data sharing (ToR 4). The development of the data sharing protocol will require negotiation and legal review among the participating states. A data management and data sharing pilot study on a CAO fish database is suggested to test a framework