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The current status of the elasmobranchs’ populations and distribution in the Canary Islands is uncertain. Here we investigate the abundance and distribution patterns of coastal elasmobranchs of Tenerife, with a special attention on the Aetomylaeus bovinus population using a photo-identification methodology. A total of 35 visual surveys were conducted in 5 coastal areas, in which pictures were taken every time an elasmobranch individual was sighted to conduct further biometry analyses. For A. bovinus sightings, pictures were used to identify each individual. Relative abundances and sex-ratios of the elasmobranchs recorded were calculated. The Western Coastline of the island was more diverse, showing 8 species out of the 9-total observed. The photo-identification technique allowed a preliminary estimation of the population size of A. bovinus in El Palmar area with 10 individuals identified and 10 re-sightings over a period of 12 months. This study provides the first relevant information regarding broad distribution, abundance and diversity of coastal elasmobranchs around Tenerife. The study also provides information regarding site fidelity of 3 mature individuals of Aetomylaeus bovinus in the South West area.

Self-amplifying mRNA (saRNA) represents a promising platform for nucleic acid delivery of vaccine immunogens. Unlike plasmid DNA, saRNA does not require entry into the nucleus of target cells for expression, having the capacity to drive higher protein expression compared to mRNA as it replicates within the cytoplasm. In this study, we examined the potential of stabilized native-like HIV-1 Envelope glycoprotein (Env) trimers to elicit immune responses when delivered by saRNA polyplexes (PLXs), assembled with linear polyethylenimine. We showed that Venezuelan equine encephalitis virus (VEEV) saRNA induces a stronger humoral immune response to the encoded transgene compared to Semliki Forest virus saRNA. Moreover, we characterized the immunogenicity of the soluble and membrane-bound ConSOSL.UFO Env design in mice and showed a faster humoral kinetic and an immunoglobulin G (IgG)2a skew using a membrane-bound design. The immune response generated by PLX VEEV saRNA encoding the membrane-bound Env was then evaluated in larger animal models including macaques, in which low doses induced high IgG responses. Our data demonstrated that the VEEV saRNA PLX nanoparticle formulation represents a suitable platform for the delivery of stabilized HIV-1 Env and has the potential to be used in a variety of vaccine regimens., Graphical abstract, Aldon, McKay, and colleagues evaluated the potential of formulated VEEV and SFV self-amplifying mRNA (saRNA) for the delivery of HIV-1 Env stabilized trimers. The authors demonstrate great potential for PEI-polyplex formulated VEEV-saRNA platform, which elicits strong humoral responses at low doses in non-human primates.

Objective: To determine the effects of dry needling (DN) in active myofascial trigger points in the teres major muscle compared to an untreated control group in pain during throwing actions, shoulder range of motion (ROM), strength, and extensibility of the tissues in professional handball (HB) athletes. Methods: A randomised, single-blinded, controlled clinical trial was designed. Thirty HB athletes with shoulder pain were randomly assigned to the DN group (n = 15) or control group (n = 15). The DN group received a single session of ultrasound-guided DN technique in the teres major muscle. The control group received no intervention. Pain intensity during throwing actions (Numeric Pain Rating Score), shoulder ROM (inclinometer), isometric strength (hand-held dynamometer), and extensibility (inclinometer) were measured before and after treatment. Results: DN group showed statistically significant improvements with large effect sizes for pain intensity (p &lt, 0.001, E.S: 1.3), internal rotation ROM (p &lt, E.S: 3.0) and extensibility (p &lt, E.S: 2.9) compared to the control group. No statistically significant differences were found for isometric strength (p &gt, 0.05). Conclusion: A single session of DN in the teres major muscle was effective for improving pain intensity during throwing actions, internal rotation ROM and extensibility in HB athletes with shoulder pain.

Periodontal disease encompasses gingivitis and periodontitis and is one of the most common chronic infections in the adult population. This study aimed to evaluate the influence of Spanish propolis extract (EEP) on the effect of the clinical and microbiological parameters as an adjuvant to scaling and root planning in patients undergoing supportive periodontal therapy (SPT). Forty chronic periodontitis patients were randomly assigned into two groups for the treatment. In the control group (n = 20), the sites were treated by scaling and root planing followed by gingival irrigation with physiological saline and in the test group (n = 20), the sites were treated by scaling and root planing followed by subgingival placement of EEP. At baseline (BL), bleeding on probing positive (BOP+) sites with probing pocket (PPD) ≥ 4 mm were defined as study sites. Plaque index, PPD, BOP, clinical attachment level (CAL), and subgingival plaque were evaluated at BL and 1 month later. The results showed a significant clinical improvement (p &lt, 0.05) in the PPD, CAL and BOP+ comparing them with BL and one month after the periodontal treatment and a significant reduction (p &lt, 0.05) for Tannerella forsythensis, Porphyromonas gingivalis, Prevotella intermedia and Treponema denticola in both groups. In addition, the improvement of clinical parameters was observed with subgingival use of EEP and also statistically significant differences between groups were observed (p &lt, 0.05) such as reductions of BOP+ % and reduced counts of T. forsythensis and P. gingivalis, considered as the “key pathogens” for the periodontal diseases. Our results suggest prophylactic and therapeutic potential for EEP against periodontal diseases, improving clinical parameters, reducing gingival bleeding and decreasing bacterial counts of T. forsythensis and P. gingivalis. The subgingival use of EEP represents a promising modality as an adjuvant in periodontal therapy to avoid microbial resistance and other adverse effects.

This work was supported by the Regional Government of Andalusia, Excellence Research Project No. P11-AGR-7648. M Garcia Burgos was supported by a fellowship from the Spanish Ministry of Education, Culture and Sport (MECD). J Moreno-Fernandez was supported by a postdoctoral contract (Contrato Puente) from the University of Granada. Funding for open access charge: University of Granada / CBUA., BACKGROUND: Iron deficiency and iron overload can affect the normal functioning of the innate and adaptive immune responses. Fermented milk products may enhance immune functions, but little is known about the effect of fermented milks on modulation of the immune response during iron deficiency anemia and recovery with normal or high dietary iron intake. Eighty male Wistar rats were randomly assigned to a control group fed a standard diet or to an anemic group fed a diet deficit in iron. Control and anemic groups were fed for 30 days with diets based on a fermented goat's or cow's milk product, with normal iron content or iron overload. RESULTS: In general, during anemia recovery lectin and alternative complement pathway activity and lactoferrin decreased, because it improves iron homeostasis, which is critically important in immune system functions. Fermented goat's milk diet enhanced immune function during iron deficiency recovery, suppressed oxidant-induced eotaxin and fractalkine expression due to the concurrent reduction of free radical production and pro-inflammatory cytokines, and decreased monocyte chemoattractant protein-1 and monocyte migration and adhesion. The increase in interferon-γ can confer immunological colonization of gut microbiota and downregulate inflammation. CONCLUSION: Fermented goat's milk consumption enhances immune function, modifying complement pathway activity and decreasing pro-inflammatory cytokines as well as lactoferrin concentration, due to the improvement of iron homeostasis, which is critically important in the normal function of the immune system., Regional Government of Andalusia, Excellence Research Project P11-AGR-7648, Spanish Ministry of Education, Culture and Sport (MECD), University of Granada, University of Granada / CBUA

Pablo Mira-Rufino and Jorge Moreno-Fernandez are grateful to the Excellence Ph.D. Program "Nutricion y Ciencias de los Alimentos" from the University of Granada. Jorge Moreno-Fernandez was supported by the fellowship from the Ministry of Education, Culture and Sport (Spain). The authors thank the Fire Department of Granada for their participation in the current study., Bone and energy metabolism are profoundly influenced by exercise. The objective of this study was to determine for the first time whether a short-term supplementation with ubiquinol could have a modulating effect on bone turnover and energy metabolism associated with strenuous exercise. The participants (n= 100 healthy and well-trained firemen) were randomly divided into two groups: ubiquinol group (ubiquinol (200 mg day(-1))) and control group (placebo) for two weeks. The protocol consisted of conducting two identical strenuous exercise tests with a rest period between tests of 24 h. Blood samples were collected before supplementation (basal value) (T1), after supplementation (T2), after the first physical exercise test (T3), after 24 h of rest (T4), and after the second physical exercise test (T5). Parathyroid hormone (PTH), osteocalcin (OC), osteoprotegerin (OPG), osteopontin (OPN), sclerotin (SOST), alkaline phosphatase (AP), adrenocorticotropin (ACTH), insulin, leptin, adrenaline, noradrenaline and peroxisome proliferator activated receptor-gamma coactivator-1 alpha (PGC-1 alpha) were determined. Our protocol increased ACTH, SOST, PTH and OC levels, while it decreased OPN. This protocol also increased adrenaline, noradrenaline and PCG-1 alpha, and decreased insulin. After ubiquinol supplementation, PTH, OC, OPG, alkaline phosphatase, leptin, insulin, noradrenaline and PGC-1 alpha levels increased in the supplemented group compared to the control group after the exercise protocol. Strenuous exercise has a clear effect on energy metabolism and bone turnover. These effects are modulated by ubiquinol supplementation, which especially increases the biomarkers of bone formation during strenuous exercise. In addition, ubiquinol has a beneficial effect on the mobilization of energy sources, fact that it could represent an ergogenic and physiological advantage for skeletal muscles., Ministry of Education, Culture and Sport (Spain)

J.M.-F. was supported by two fellowships from the Ministry of Education, Culture and Sport (Spain), FPU and Traslados Temporales FPU (University of King´s College of London). J.M.-F. is grateful to the Excellence Program “Nutrición y Ciencias de los Alimentos” from the University of Granada. The authors are grateful to Susan Stevenson for her efficient support in the revision with the English language., Despite the crucial role of the liver as the central regulator of iron homeostasis, no studies have directly tested the modulation of liver gene and protein expression patterns during iron deficiency instauration and recovery with fermented milks. Fermented goat milk consumption improves the key proteins of intestinal iron metabolism during iron deficiency recovery, enhancing the digestive and metabolic utilization of iron. The aim of this study was to assess the influence of fermented goat or cow milk consumption on liver iron homeostasis during iron-deficiency anemia recovery with normal or iron-overload diets. Analysis included iron status biomarkers, gene and protein expression in hepatocytes. In general, fermented goat milk consumption either with normal or high iron content up-regulated liver DMT1, FPN1 and FTL1 gene expression and DMT1 and FPN1 protein expression. However, HAMP mRNA expression was lower in all groups of animals fed fermented goat milk. Additionally, hepcidin protein expression decreased in control and anemic animals fed fermented goat milk with normal iron content. In conclusion, fermented goat milk potentiates the up-regulation of key genes coding for proteins involved in iron metabolism, such as DMT1, and FPN1, FTL1 and down-regulation of HAMP, playing a key role in enhanced iron repletion during anemia recovery, inducing a physiological adaptation of the liver key genes and proteins coordinated with the fluctuation of the cellular iron levels, favoring whole-body iron homeostasis., Ministry of Education, Culture and Sport (Spain), Spanish Government, Traslados Temporales FPU (University of Kings College of London)

Salting is one of the oldest methods of preserving food. The main limitation of salting is its extended processing time due to slow salt diffusion. A moderate electric field (MEF) can improve the mass transfer rate through electroporation. Regularly, mass transfer processes are modeled with Fick’s second law. However, due to the anisotropic nature of food microstructures, it might be more appropriate to use an anomalous model. The main objective of this study was to search for a phenomenological explanation for salt and water diffusion in the salmon brining process coupled with MEF. Salmon fillets were cut into finite cylinders (0.025 × 0.025 m) and brined in two salt concentrations (6 and 24% w/w NaCl) at 6 °C for 20 h. MEFs were applied in the range of 0 to 2 V/cm. The salt and water contents of the salmon were measured during the process. Fick’s second law and anomalous model based on fractional calculus were used to describe the diffusion phenomena. The results showed that an MEF tended to reduce the brining processing time and increase the salt content of salmon. This effect is predominantly due to an increase in the equilibrium salt concentration in the salmon tissue. Mathematical analysis shows that the anomalous diffusion model is more suitable for representing the brining process, exhibiting superdiffusion behavior (α > 1). An MEF accelerates the salt mass transfer into salmon tissue even at lower temperatures, significantly reducing the processing time. In addition, the diffusion process can be characterized with an anomalous model.

Objective: to provide detailed information on iron status, body changes, food intake, hepatosomatic index, and body composition during recovery from severe iron deficiency anemia with fermented cow or goat milk. Material and methods: sixty male Wistar albino rats were divided into two experimental groups (control and anemic) and fed ad libitum an AIN-93G diet for 40 days, receiving a normal Fe diet (45 mg/kg of diet) or a low-Fe diet (5 mg/kg of diet), respectively. After induction of anemia, both the control and anemic groups were additionally fed for 30 days either a fermented cow milk-based or fermented goat milk-based diet with normal Fe content. Hematological and iron-related biochemical parameters, weight changes, food intake, hepatosomatic index, and body composition were assessed. Results: The hepatosomatic index was higher in the anemic group versus the control group. In both groups fed a fermented goat milk-based diet the hepatosomatic index was higher due to lower body weight and a higher liver weight. Food intake, weight gain, and total body fat were lower, whereas lean mass and free and total water were higher in the anemic group as compared to the control group. In both animal groups fed a fermented goat milk-based diet body weight and body fat were lower and the percentages of lean mass and free water and total water were higher. Conclusion: Iron deficiency decreased weight gain, lean mass, and body fat, indicating lower energy stores. Fermented goat milk-based diet recovers more efficiently iron status, decreased adiposity, and increased energy expenditure.Objetivo: determinar la relación entre estatus férrico, cambios ponderales, ingesta, índice hepatosomático y composición corporal durante la recuperación de la anemia ferropénica con dietas basadas en leche fermentada de cabra o vaca. Material y métodos: sesenta ratas macho Wistar albinas se han distribuido en dos grupos experimentales (control y anémico) y se han alimentado durante 40 días con dieta AIN-93G con contenido férrico normal (45 mg/kg de dieta) o bajo (5 mg/kg de dieta). Posteriormente, ambos grupos experimentales se alimentaron durante 30 días con dietas basadas en leche de cabra o vaca fermentada. Se han determinado los parámetros hematológicos y bioquímicos relacionados con el estatus de hierro, así como los cambios ponderales y la ingesta de alimento, relación hepatosomática y composición corporal. Resultados: el índice hepatosomático fue mayor en el grupo anémico . En ambos grupos de animales alimentados con dieta de leche fermentada de cabra, el índice hepatosomático fue mayor debido a menor peso corporal y mayor peso del hígado. La ingesta de alimento, ganancia de peso y grasa corporal fueron menores en el grupo anémico, mientras que la masa magra, el agua libre y el agua total fueron mayores. En ambos grupos de animales alimentados con dieta de leche de cabra fermentada, el peso y la grasa corporal fueron menores, mientras que los porcentajes de masa magra, agua libre y agua total fueron mayores . Conclusión: la ferrodeficiencia disminuyó la ganancia de peso, masa magra y grasa corporal, indicando menores almacenes de energía. La dieta basada en leche de cabra fermentada recupera más eficientemente el nivel de hierro durante la recuperación de la ferrodeficiencia, disminuye la adiposidad y aumenta el gasto energético.

Fermented dairy products are obtained from fermentation of milk, through the action of suitable and harmless microorganisms. In addition to lactic acid bacteria, fermented dairy products have bioactive compounds as well as bacteria derived metabolites produced during fermentation. Fermented dairy products, due to their special characteristics, are an excellent matrix for the incorporation of ingredients and/or nutrients that give the final product properties beyond purely nutritional, making them true functional foods, interesting in a healthy diet. This review is focused on existing scientific evidence regarding the consumption of fermented dairy products and their health benefits, principally hypocholesterolemic, antioxidant, bone, hypotensive effects and immunological benefits, such as effects on the hosts intestine and microbiome, anti-carcinogenic, immunomodulation and anti- ́ allergenic effects. This review has extensively assessed the fermented dairy products which could be used as possible nutraceutical agents in food and health industries., Ministry of Education, Culture and Sport (Spain), University of Granada

Background Iron (Fe) plays a crucial role in several fundamental processes, including erythropoiesis, cellular metabolism, and in cardiovascular disease. The aim of this work was to contribute to a better understanding of the physiology of and recovery from Fe deficiency by studying how fermented milk consumption affects vascular biomarkers during Fe repletion. Results The deleterious cardiovascular biomarkers cytokine-induced neutrophil chemoattractant 1, connective tissue growth factor (CTGF), interleukin-6, monocyte chemoattractant protein-1 (MCP-1), inhibitor of tissue plasminogen activator 1 total, metallopeptidase inhibitor 1 (TIMP-1), tumor necrosis factor alpha, vascular endothelial growth factor (VEGF), sE-selectin, and soluble intercellular adhesion molecule 1 (sICAM-1) decreased after fermented goat milk consumption in groups of fed animals either with normal Fe or Fe overload with respect to rats fed with fermented cow milk. The beneficial cardiovascular biomarkers caveolin-1 and adiponectin were higher in both control and anemic rats fed fermented goat milk either with normal Fe or Fe overload with respect to fermented cow milk. Anemia decreased TIMP-1 in rats fed fermented goat milk with Fe overload, whereas there was increased CTGF and MCP-1 in animals fed fermented cow milk with either normal or Fe overload. In addition, Fe overload increased VEGF. Conclusion Fermented goat milk consumption improves hematological status and promotes beneficial metabolic responses, which may attenuate cardiovascular risk factors during anemia recovery and iron overload to lessen the inflammatory response, macrophages activation and atherosclerosis development. © 2018 Society of Chemical Industry.

To date, no studies are available about adipose tissue modifications during anemia recovery; therefore, the aim of this study is to provide detailed information about adipose tissue homeostasis during anemia recovery with fermented milks. Forty male Wistar rats were placed on a pre-experimental period of 40 days, divided in two groups (normal-Fe diet and Fe-deficient diet). Then rats were fed fermented goat or cow milk-based diets with normal-Fe content during 30 days. Ghrelin and adiponectin decreased in both groups of animals fed fermented goat milk, whereas leptin and NEFA increased. UCP-1 decreased in anemic rats fed either fermented milk, and irisin greatly increased in both groups of animals fed fermented goat milk. Fermented goat milk reduces adiposity, inducing leptin elevation and ghrelin reduction. Conversely, plasma adiponectin concentrations decreased in animals fed fermented goat milk, showing an inverse correlation with NEFA, an important marker of lipid mobilization, indicating increased lipolysis. Irisin up-regulation in animals fed fermented goat milk contributes to a favorable metabolic profile and the browning of adipose tissue during anemia recovery.

The aim of the present study was to analyze the influence of a supervised concurrent exercise-training program, from the 17th gestational week until delivery, on cytokines in maternal (at 17th and 35th gestational week, and at delivery) and arterial and venous cord serum. Fifty-eight Caucasian pregnant women (age: 33.5 &plusmn, 4.7 years old, body mass index: 23.6 &plusmn, 4.1kg/m2) from the GESTAFIT Project (exercise (n = 37) and control (n = 21) groups) participated in this quasi-experimental study (per-protocol basis). The exercise group followed a 60-min 3 days/week concurrent (aerobic-resistance) exercise-training from the 17th gestational week to delivery. Maternal and arterial and venous cord serum cytokines (fractalkine, interleukin (IL)&ndash, 1&beta, IL-6, IL-8, IL-10, interferon (IFN)&ndash, &gamma, and tumor necrosis factor (TNF)&ndash, &alpha, ) were assessed using Luminex xMAP technology. In maternal serum (after adjusting for the baseline values of cytokines), the exercise group decreased TNF-&alpha, (from baseline to 35th week, p = 0.02), and increased less IL-1&beta, (from baseline to delivery, p = 0.03) concentrations than controls. When adjusting for other potential confounders, these differences became non-significant. In cord blood, the exercise group showed reduced arterial IL-6 and venous TNF-&alpha, (p = 0.03 and p = 0.001, respectively) and higher concentrations of arterial IL-1&beta, (p = 0.03) compared to controls. The application of concurrent exercise-training programs could be a strategy to modulate immune responses in pregnant women and their fetuses. However, future research is needed to better understand the origin and clearance of these cytokines, their role in the maternal-placental-fetus crosstalk, and the influence of exercise interventions on them.

Oxidative stress is a harmful factor for male reproductive function, and a major cause of infertility. On the other hand, fermented goat milk has positive effects on anemia recovery and mineral metabolism. This study evaluated the effect of feeding rats with fermented milks during anaemia recovery on molecular mechanisms linked to oxidative stress and inflammatory signalling in rats reproductive system. Forty male Wistar rats were placed on a pre-experimental period of 40 days (control group, receiving normal-Fe diet and Fe-deficient group, receiving low-Fe diet). Lately, rats were fed with fermented goat or cow milk-based diets during 30 days. After feeding the fermented milks, Total antioxidant status (TAS) and non-esterified fatty acids (NEFA) increased and 8-hydroxy-2'-deoxyguanosine (8-OHdG), 15-F2t-isoprostanes and thiobarbituric acid reactive substances (TBARS) decreased in testis. DNA oxidative damage in testis germ cells was lower with fermented goat milk. Fermented goat milk reduced IL-6 and TNF-α in control animals, increasing INF-γ in control and anaemic rats. NRF2 and PGC-1α protein levels increased in testis after fermented goat milk consumption in control and anaemic rats. Fermented goat milk also increased TAS and decreased oxidative damage, protecting the main testis cell bioconstituents (lipids, proteins, DNA, prostaglandins) from oxidative damage and reduced inflammatory activity, preventing injuries to testis germinal epithelium. Fermented goat milk enhanced lipolysis, fatty acids degradation and immune response, attenuating inflammatory signalling, representing a positive growth advantage for testicular cells.

Oxidative stress is a harmful factor for male reproductive function, and a major cause of infertility. On the other hand, fermented goat milk has positive effects on anemia recovery and mineral metabolism. This study evaluated the effect of feeding rats with fermented milks during anaemia recovery on molecular mechanisms linked to oxidative stress and inflammatory signalling in rats reproductive system. Forty male Wistar rats were placed on a pre-experimental period of 40 days (control group, receiving normal-Fe diet and Fe-deficient group, receiving low-Fe diet). Lately, rats were fed with fermented goat or cow milk-based diets during 30 days. After feeding the fermented milks, Total antioxidant status (TAS) and non-esterified fatty acids (NEFA) increased and 8-hydroxy-2’-deoxyguanosine (8-OHdG), 15-F2t-isoprostanes and thiobarbituric acid reactive substances (TBARS) decreased in testis. DNA oxidative damage in testis germ cells was lower with fermented goat milk. Fermented goat milk reduced IL-6 and TNF-α in control animals, increasing INF-γ in control and anaemic rats. NRF2 and PGC-1α protein levels increased in testis after fermented goat milk consumption in control and anaemic rats. Fermented goat milk also increased TAS and decreased oxidative damage, protecting the main testis cell bioconstituents (lipids, proteins, DNA, prostaglandins) from oxidative damage and reduced inflammatory activity, preventing injuries to testis germinal epithelium. Fermented goat milk enhanced lipolysis, fatty acids degradation and immune response, attenuating inflammatory signalling, representing a positive growth advantage for testicular cells., Jorge Moreno-Fernandez was supported by fellowship from the Ministry of Education, Culture and Sport (Spain) and is grateful to the Excellence Ph.D. Program “Nutrición y Ciencias de los Alimentos” from the University of Granada. This study was supported by Andalusian Government, Excellence Research Project No. P11-AGR-7648.

Extracellular vesicles (EVs) are small lipid vesicles released by either any prokaryotic or eukaryotic cell, or both, with a biological role in cell-to-cell communication. In this work, we characterize the proteomes and nanomechanical properties of EVs released by tissue-culture cell-derived trypomastigotes (mammalian infective stage; (TCT)) and epimastigotes (insect stage; (E)) of Trypanosoma cruzi, the etiologic agent of Chagas disease. EVs of each stage were isolated by differential centrifugation and analyzed using liquid chromatography with tandem mass spectrometry (LC-MS/MS), dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), electron microscopy and atomic force microscopy (AFM). Measurements of zeta-potential were also included. Results show marked differences in the surface molecular cargos of EVs between both stages, with a noteworthy expansion of all groups of trans-sialidase proteins in trypomastigote’s EVs. In contrast, chromosomal locations of trans-sialidases of EVs of epimastigotes were dramatically reduced and restricted to subtelomeric regions, indicating a possible regulatable expression of these proteins between both stages of the parasite. Regarding mechanical properties, EVs of trypomastigotes showed higher adhesion compared to the EVs of epimastigotes. These findings demonstrate the remarkable surface remodeling throughout the life cycle of T. cruzi, which shapes the physicochemical composition of the extracellular vesicles and could have an impact in the ability of these vesicles to participate in cell communication in completely different niches of infection., ERANet program, Instituto Carlos III, Ministerio de Sanidad, Gobierno de España, Fundación Ramón Areces funded the projects: “Research in prevention of congenital Chagas disease: parasitological, placental and immunological markers” (ERANet17/ HLH-0142 (Cochaco), ERANE-LAC HD 328/2014) and “Interactoma de las exovesículas de T. cruzi y de los inmunocomplejos que forman con las células del hospedador: implicaciones en la patología de la enfermedad de Chagas (2019)”, Ministerio de Ciencia y Tecnología of the government of Spain funded the project PGC2018-099424-B-I00

Background: Iron deficiency anemia (IDA) is one of the most common nutritional problems in the world, and it is accepted that reactive oxygen species (ROS) production is altered during IDA. The aim of this study was to assess the influence of fermented goat and cow milks on enzymatic antioxidant activities and gene expression, and their role in protecting from oxidative damage during anemia recovery.; Results: After feeding the fermented milks-based diets (cow or goat), a significant elevation of some antioxidant endogenous enzymes was found, together with an increase in total antioxidant status (TAS), and a decrease in 8-hydroxy-2'-deoxyguanosine (8-OHdG) was recorded in animals consuming fermented goat milk-based diet. In contrast, DNA strand breaks, hydroperoxides, 15-F2t-isoprostanes and protein carbonyl groups were lower in some tissues in animals fed fermented goat milk-based diet, revealing an improvement in both systemic and cellular antioxidant activity of plasma and tissues due to fermented goat milk consumption.; Conclusion: Fermented goat milk consumption induces a protective increase in TAS together with lower oxidative damage biomarkers, revealing that the milk protects main cell bioconstituents (lipids, protein, DNA, prostaglandins) from evoked oxidative damage during anemia recovery. © 2016 Society of Chemical Industry.; © 2016 Society of Chemical Industry.

Congenital erythropoietic porphyria is a rare autosomal recessive disease produced by deficient activity of uroporphyrinogen III synthase, the fourth enzyme in the heme biosynthetic pathway. The disease impacts many organs, can be life-threatening and currently lacks curative treatments. Inherited mutations most commonly reduce the enzyme's stability, altering its homeostasis and ultimately blunting intracellular heme production. This results in uroporphyrin by-product accumulation in the body, aggravating associated pathological symptoms such as skin photosensitivity and disfiguring phototoxic cutaneous lesions. Here we demonstrated that the synthetic marketed antifungal ciclopirox binds to the enzyme, stabilizing it. Ciclopirox targeted the enzyme at an allosteric site distant from the active center and did not affect the enzyme's catalytic role. The drug restored enzymatic activity in vitro and ex vivo, and was able to alleviate most clinical symptoms of congenital erythropoietic porphyria in a genetic mouse model of the disease at sub-toxic concentrations. Our findings establish a possible line of therapeutic intervention against congenital erythropoietic porphyria which is potentially applicable to the majority of deleterious missense mutations causing this devastating disease.

The aim of this research was to investigate the acceleration of brine-salting process of salmon when coupling it with moderate electric field (MEF). Salmon samples were cut into cylinder shape (0.025 +/- 0.001 m diameter and 0.025 +/- 0.001 m height), an

Display Omitted 10 SNPs in DEFB1 and CAMP modify the TFBS for 24 TFs related to HIV/AIDS and TB.We introduce the homotypic redundancy weight factor (HWF) to assess rSNP impacts.Our algorithm has 53% sensitivity and 84.6% specificity to detect functional rSNPs.Future ENCODE and 1000 genomes project releases will improve the proposed algorithm.This algorithm could be useful to probe design and personalized medicine. Single nucleotide polymorphisms (SNPs) in transcription factor binding sites (TFBSs) within gene promoter region or enhancers can modify the transcription rate of genes related to complex diseases. These SNPs can be called regulatory SNPs (rSNPs). Data compiled from recent projects, such as the 1000 Genomes Project and ENCODE, has revealed essential information used to perform in silico prediction of the molecular and biological repercussions of SNPs within TFBS. However, most of these studies are very limited, as they only analyze SNPs in coding regions or when applied to promoters, and do not integrate essential biological data like TFBSs, expression profiles, pathway analysis, homotypic redundancy (number of TFBSs for the same TF in a region), chromatin accessibility and others, which could lead to a more accurate prediction. Our aim was to integrate different data in a biologically coherent method to analyze the proximal promoter regions of two antimicrobial peptide genes, DEFB1 and CAMP, that are associated with tuberculosis (TB) and HIV/AIDS. We predicted SNPs within the promoter regions that are more likely to interact with transcription factors (TFs). We also assessed the impact of homotypic redundancy using a novel approach called the homotypic redundancy weight factor (HWF). Our results identified 10 SNPs, which putatively modify the binding affinity of 24 TFs previously identified as related to TB and HIV/AIDS expression profiles (e.g. KLF5, CEBPA and NFKB1 for TB; FOXP2, BRCA1, CEBPB, CREB1, EBF1 and ZNF354C for HIV/AIDS; and RUNX2, HIF1A, JUN/AP-1, NR4A2, EGR1 for both diseases). Validating with the OregAnno database and cell-specific functional/non functional SNPs from additional 13 genes, our algorithm performed 53% sensitivity and 84.6% specificity to detect functional rSNPs using the DNAseI-HUP database. We are proposing our algorithm as a novel in silico method to detect true functional rSNPs in antimicrobial peptide genes. With further improvement, this novel method could be applied to other promoters in order to design probes and to discover new drug targets for complex diseases.

The effect of maternal DHA supplementation on expression of key genes and proteins involved in Fe metabolism and mineral placenta content was evaluated for the first time. One hundred and ten pregnant women were randomly assigned to one of the groups: control (n = 54, 400 ml/day of the control dairy drink; DHA-supplemented (n = 56, 400 ml/day of the DHA-enriched dairy drink). Placenta DMT1, FPN1, TfR1 and Hamp1 mRNA and protein expressions were analysed. Hepcidin concentration in the mother and umbilical cord and placenta mineral content were assessed. DMT1, FPN1, TfR1 and Hamp1 gene expressions together with serum hepcidin increased in umbilical cord and artery of the DHA-supplemented group. In this group, Fe concentration increased in plasma, umbilical cord vein and artery. Ca concentrations raised in umbilical cord vein and artery, Cu, V and Mn levels. Docosahexaenoic acid (DHA) supplementation improved transplacental Fe homeostasis, enhancing Fe transfer and boosting neonates Fe stores at delivery.

The effect of ohmic heating (OH) on the texture, microbial load, and cadmium (Cd) and lead (Pb) content was evaluated in Chilean blue mussel ( Mytilus chilensis ) at shucking and subsequent canning. Mussel samples were processed by OH and blanching (BLAN) at 50 ± 1 °C, 70 ± 1 °C and 90 ± 1 °C for shucking and then canning at 115 °C for 20 min. The Cd and Pb content in fresh mussels was 2.47 ± 0.18 μg g − 1 and 5.26 ± 0.55 μg g − 1 dry weight (dw). The greatest reduction in Cd content was reached at 90 ± 1 °C by OH with 1.67 ± 0.06 μg g − 1 (dw) and 1.69 ± 0.08 μg g − 1 (dw) by BLAN. At the same temperature, the Pb content was reduced to 4.18 ± 0.24 μg g − 1 dw for OH and 4.14 ± 0.30 μg g − 1 dw for BLAN. The cutting strength of fresh samples (21.35 ± 2.44 N) significantly decreased in samples processed by OH (15.65 ± 1.36 N) at 90 ± 1 °C compared with BLAN samples (20.41 ± 3.16 N) at the same temperature. The initial mean count for mesophilic aerobic microorganisms and Enterobacteriaceae was 3.8 log cfu/g and 2.5 log cfu/g, respectively. The mesophilic aerobic microorganism count was reduced by 1.7 logarithmical units, while Enterobacteriaceae counts were reduced to undetectable levels by OH at 90 ± 1 °C. Industrial relevance High temperature short time (HTST) processes rely on rapid convection heat transfer and are thus well suited to liquid foods. But, they are, however, limited in application to particulates since, for particles more than a couple of millimeters thick, the processing time is insufficient for heat to transfer to the center to give sterility. Ohmic heating is an emerging technology and very promising in food industries. It is a thermal process in which heat is generated internally in food and acts as a resistance to the flow of alternating electric current. This type of treatment prevents overheating by producing less deterioration in food components. It is a rapid procedure that allows food to conserve its natural properties; microorganisms and enzymes are also inactivated. The purpose of the present work was to evaluate the effect of Ohmic heating on mechanical properties, microbial load, Cd and Pb content in the opening and subsequent canning of Chilean blue mussel ( Mytilus chilensis ). This information could be used for the seafood industries in order to improve thermal heating processes in mussels.

Prion diseases are neurodegenerative conditions characterized by the conformational conversion of the cellular prion protein (PrPC), an endogenous membrane glycoprotein of uncertain function, into PrPSc, a pathological isoform that replicates by imposing its abnormal folding onto PrPC molecules. A great deal of evidence supports the notion that PrPC plays at least two roles in prion diseases, by acting as a substrate for PrPSc replication, and as a mediator of its toxicity. This conclusion was recently supported by data suggesting that PrPC may transduce neurotoxic signals elicited by other disease-associated protein aggregates. Thus, PrPC may represent a convenient pharmacological target for prion diseases, and possibly other neurodegenerative conditions. Here, we sought to characterize the activity of chlorpromazine (CPZ), an antipsychotic previously shown to inhibit prion replication by directly binding to PrPC. By employing biochemical and biophysical techniques, we provide direct experimental evidence indicating that CPZ does not bind PrPC at biologically relevant concentrations. Instead, the compound exerts anti-prion effects by inducing the relocalization of PrPC from the plasma membrane. Consistent with these findings, CPZ also inhibits the cytotoxic effects delivered by a PrP mutant. Interestingly, we found that the different pharmacological effects of CPZ could be mimicked by two inhibitors of the GTPase activity of dynamins, a class of proteins involved in the scission of newly formed membrane vesicles, and recently reported as potential pharmacological targets of CPZ. Collectively, our results redefine the mechanism by which CPZ exerts anti-prion effects, and support a primary role for dynamins in the membrane recycling of PrPC, as well as in the propagation of infectious prions.

Oxidative stress results from an imbalance between the formation and neutralization of reactive oxygen species (ROS), and it is imposed on cells because of one or more of the following factors: an increase in oxidant generation, a decrease in antioxidant protection, or a failure to repair oxidative damage. There is controversy about the susceptibility of cells to lipid peroxidation in iron deficiency anemia (IDA)—some investigators have claimed there is no difference in lipid peroxidation among patients with IDA compared with controls, but others have reported that among patients with IDA oxidants are increased and antioxidants decreased, so the oxidative/antioxidative balance is shifted toward the oxidative side. This apparent discrepancy may be due to different concentrations of ROS and antioxidant enzymes in the tissues studied, the subjects of the study, the severity of the iron deficiency, and the methods used for the assessment of the oxidative stress. However, in humans, where the degree of iron deficiency is not very high, it is accepted that iron deficiency increases oxidative stress, a fact that can also be attributed to the repletion process with several sources of iron. Several authors have reported increased lipid peroxidation products in patients with iron-deficiency anemia, which may be attributed to overproduction of ROS and a deficiency of antioxidant defense. Decreased superoxide dismutase (SOD) activity in iron-deficiency anemia may be linked to increased oxidative stress, because it is well known that ROS inhibits SOD activity. On the other hand, goat milk has beneficial effects, reducing oxidative stress and protecting cell bioconstituents from free radicals. Caprine milk alters the activities of enzymes that improve the total antioxidative defense capacity of the organism, a fact that limits the oxidative damage to the main biomolecules (lipids, protein DNA, and prostaglandins).

The beneficial effects of regular, nonexhaustive physical exercise are well-known, however, these beneficial effects are lost with strenuous exercise. High-intensity exercise induces inflammatory reactions together with high production of free radicals, muscle damage, and subsequent inflammation indicated by muscle soreness and swelling, prolonged loss of muscle function, and the leakage of muscle proteins, such as creatine kinase and myoglobin, into the circulation. The pineal hormone, melatonin (N-acetyl-5-methoxytryptamine), is a potent, endogenously produced and diet-derived free radical scavenger and broad-spectrum antioxidant, consequently, it might have positive effects on the recovery following the exercise session. Among other effects, several studies have shown that oral supplementation of melatonin during high-intensity exercise is efficient in reducing the degree of oxidative stress (decreasing lipid peroxidation, with a significant increase of antioxidant enzymes activity), which lead to the maintenance of the cell integrity and the reduction of secondary tissue damage. Data also indicate that melatonin has potent protective effects, preventing over-expression of pro-inflammatory mediators and inhibiting the effects of several pro-inflammatory cytokines. The aim of this chapter is to review the most relevant studies focused on the effect of melatonin on diverse negative aspects associated with strenuous exercise, such as oxidative stress or the inflammatory signaling, in human, animals, and in vitro assays. Melatonin supplementation can decrease muscle damage through modulation of oxidative stress, inflammatory signaling, improving recovery, and performance efficiency associated to the physical challenge.

Iron deficiency anemia (IDA) is one of the most prevalent nutritional deficiencies worldwide. Iron plays critical roles in nervous system development and cognition. Despite the known detrimental consequences of IDA on cognition, available studies do not provide molecular mechanisms elucidating the role of iron in brain functions during iron deficiency and recovery with dairy components. In this study, 100 male Wistar rats were placed on a pre-experimental period of 40 days and randomly divided in two groups: a control group receiving a normal-Fe diet, (45 mg/kg), and an Fe-deficient group receiving a low-Fe diet (5 mg/kg). At day 40, 10 rats per group were sacrificed to anemia control, and 80 rats were divided into eight experimental groups fed with fermented goat or cow milk-based diets, with normal Fe content or Fe overload (450 mg/kg) for 30 days. IDA decreased most of the parameters related to brain molecular functions, namely dopamine, irisin, MAO-A, oxytocin, &beta, endorphin, and &alpha, MSH, while it increased synaptophysin. These alterations result in an impairment of brain molecular functions. In general, during anemia recovery, fermented goat milk diet consumption increased dopamine, oxytocin, serotonin, synaptophysin, and &alpha, MSH, and decreased MAO-A and MAO-B, suggesting a potential neuroprotective effect in brain functions, which could enhance brain molecular functions.

Prion diseases or Transmissible Spongiform Encephalopathies (TSEs) are a group of fatal neurodegenerative disorders affecting several mammalian species. Its causative agent, disease-associated prion protein (PrPd), is a self-propagating β-sheet rich aberrant conformation of the cellular prion protein (PrPC) with neurotoxic and aggregation-prone properties, capable of inducing misfolding of PrPC molecules. PrPd is the major constituent of prions and, most importantly, is the first known example of a protein with infectious attributes. It has been suggested that similar molecular mechanisms could be shared by other proteins implicated in diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis or systemic amyloidoses. Accordingly, several terms have been proposed to collectively group all these disorders. Through the stringent evaluation of those aspects that characterise TSE-causing prions, in particular propagation and spread, strain variability or transmissibility, we will discuss whether terms such as “prion”, “prion-like”, “prionoid” or “propagon” can be used when referring to the aetiological agents of the above other disorders. Moreover, it will also be discussed whether the term “infectious”, which defines a prion essential trait, is currently misused when referring to the other misfolded proteins.

Despite the crucial roles of duodenal cytochrome b (Dcytb), divalent metal transporter 1 (DMT1), ferritin light chain (Ftl1), ferroportin 1 (FPN1), transferrin receptor 1 (TfR1), and hepcidin antimicrobial peptide (Hamp) in Fe metabolism, no studies have investigated the modulations of these genes during Fe repletion with fermented milks. Analysis included Fe status markers and gene and protein expression in enterocytes of control and anemic animals fed fermented milks. Fermented goat's milk up-regulated enterocyte Dcytb, DMT1, FPN1, and Ftl1 and down-regulated TfR1 and Hamp gene expression in control and anemic animals. Anemia decreased Dcytb, DMT1, and Ftl1 in animals fed fermented cow's milk and up-regulated TfR1 and Hamp expression. Fe overload down-regulated Dcytb and TfR1 in animals fed fermented cow's milk and up-regulated DMT1 and FPN1 gene expression. Fermented goat's milk increased expression of duodenal Dcytb, DMT1, and FPN1 and decreased Hamp and TfR1, improving Fe metabolism during anemia recovery.

Prion diseases are rare neurodegenerative conditions associated with the conformational conversion of the cellular prion protein (PrPC) into PrPSc, a self-replicating isoform (prion) that accumulates in the central nervous system of affected individuals. The structure of PrPSc is poorly defined and likely to be heterogeneous, as suggested by the existence of different prion strains. The latter represents a relevant problem for therapy in prion diseases, as some potent anti-prion compounds have shown strain-specificity. Designing therapeutics that target PrPC may provide an opportunity to overcome these problems. PrPC ligands may theoretically inhibit the replication of multiple prion strains, by acting on the common substrate of any prion replication reaction. Here, we characterized the properties of a cationic tetrapyrrole [Fe(III)-TMPyP], which was previously shown to bind PrPC and inhibit the replication of a mouse prion strain. We report that the compound is active against multiple prion strains in vitro and in cells. Interestingly, we also find that Fe(III)-TMPyP inhibits several PrPC-related toxic activities, including the channel-forming ability of a PrP mutant and the PrPC-dependent synaptotoxicity of amyloid-β (Aβ) oligomers, which are associated with Alzheimer’s Disease. These results demonstrate that molecules binding to PrPC may produce a dual effect of blocking prion replication and inhibiting PrPC-mediated toxicity.

Gender is a crucial determinant of life span, but little is known about gender differences in free radical homeostasis and inflammatory signaling. The aim of the study was to determine gender-related differences concerning oxidative stress and inflammatory signaling of healthy neonates and mothers. Fifty-six mothers with normal gestational course and spontaneous delivery were selected. Blood samples were collected from the mother (at the beginning of delivery and start of expulsive period) and from neonate (from umbilical cord vein and artery). The mothers of girls featured a higher total antioxidant status and lower plasma hydroperoxides than the mother of boys. Regarding the neonates, the girls featured a higher total antioxidant status and lower plasma membrane hydroperoxides in umbilical cord artery together with higher catalase, glutathione peroxidase, and superoxide dismutase activities. Lower levels of interleukin 6, tumor necrosis factor alpha, and prostaglandin E2 were observed in the mothers of girls and higher level of soluble tumor necrosis factor receptor II. In the neonates, lower levels of interleukin 6 and tumor necrosis factor alpha were observed in umbilical artery and higher soluble tumor necrosis factor receptor II in umbilical cord vein and artery of girls. An association between gender, oxidative stress, and inflammation signaling exists, leading to a renewed interest in the neonate’s sex as a potential risk factor to several alterations.

Anaemia is associated with fatigue and diminished muscular oxygenation, which may affect skeletal muscle (SM). No studies are available about the SM modifications during anaemia recovery; therefore, the aim of this study is to study SM homeostasis during anaemia recovery with fermented milks. Forty male Wistar rats were placed on a pre-experimental period of 40 days, divided in two groups (control group receiving normal-Fe diet and Fe-deficient group receiving low-Fe diet). Lately, rats were fed with fermented goat or cow milk-based diets, with normal-Fe content during 30 days. After feeding the fermented milks, leptin, adiponectin, non-esterified fatty acids (NEFA) and protein expression (UCP1, PepT1 and irisin) within the SM were assessed. Adiponectin decreased in both groups of animals fed fermented goat milk, while leptin and NEFA increased. UCP1 protein expression increased in control and anaemic animals fed fermented goat milk. UCP1 also increased in both group of anaemic animals fed either fermented cow or goat milk in comparison with their controls. Irisin increased in both group of animals fed fermented goat milk. Finally, PepT1 also showed an increased expression in control and anaemic rats fed fermented goat milk and the anaemia also induced an over-expression of this transporter in animals fed either fermented cow or goat milk. Fermented goat milk consumption during anaemia recovery diminishes adiposity depots and enhances lipolysis, increasing UCP1, PepT1 and irisin protein expression, featuring an ergogenic effect in the SM which is an important endocrine regulator of body metabolism.

The effect of ohmic heating (OH) and vacuum impregnation (VI) on the osmotic dehydration kinetics and microstructure of pears was studied. Three different dehydration levels (30, 40 and 50 °C) were used, by applying VI or not (OD) and OH (100 V). Dehydrated samples showed that the application of OH during the osmotic treatments had significant effects on the kinetics of water loss and sugar gain as well as on the microstructure of samples. The greatest water loss was observed with the OD–OH. The largest amount of solute gain and the smallest firmness loss were obtained in the VI–OH. In some treatments, the process time was reduced by as much as 40%. The SEM observations showed that cell deformation and cell rupture were significant in the OD–OH than on the VI–OH samples. The increases in the permeability of cell by OH explain the acceleration of mass transference and process time reduction.

This study was conducted in order to improve our understanding of how phenolics and aroma compounds change in wine grapes during postharvest dehydration. Pinot noir grapes grown in the Willamette Valley of Oregon were harvested at 22.0 and 24.0 °Brix. Grapes harvested at 22.0 °Brix were divided into three equal lots with one lot immediately used for wine production, and the remaining two lots placed inside an air tunnel with an air speed of 1.0–1.8 m s−1, 38% relative humidity and a temperature of 22 °C. The soluble solids content and weight loss were measured daily and wines were made from grapes when they reached 24.8 and 26.7 °Brix. The soluble solids of grapes increased about 1 °Brix per day; therefore, on the third and fourth day the berries reached the desired concentration; weight loss was 14 and 16%, respectively. Results from berry phenolic analysis indicated that per berry anthocyanin amount remained unchanged during dehydration. The composition of proanthocyanidins isolated from berries changed during dehydration. Volatile compounds in wines made from dehydrated grapes contained more terpenes and norisoprenoids (β-ionone, β-damascenone) when compared to wine made from the original fruit. Wines made from increasingly dehydrated grapes tended to resemble the composition and flavour profile of wines made from grapes left on the vine (i.e. with extended ripening). The results of this study suggest that postharvest flavour changes consistent with changes during fruit ripening can occur in grapes when harvested early and allowed to dehydrate under controlled conditions prior to fermentation.

The aim of the current study was to assess the influence of fermented goat or cow milk on melatonin levels and antioxidant status and during anemia recovery. Eighty male Wistar rats were placed on a pre-experimental period of 40 days and randomly divided into two groups, a control group receiving normal-Fe diet (45 mg kg(-1)) and the Fe-deficient group receiving low-Fe diet (5 mg kg(-1)). Then, the rats were fed with fermented goat or cow milk-based diets with a normal-Fe content or Fe-overload (450 mg kg(-1)) for 30 days. After 30 days of feeding the fermented milks, the total antioxidant status (TAS) was higher in both groups of animals fed fermented goat milk with the normal-Fe content. Plasma and urine 8-OHdG were lower in control and anemic rats fed fermented goat milk. Melatonin and corticosterone increased in the anemic groups during Fe replenishment with both fermented milks. Urine isoprostanes were lower in both groups fed fermented goat milk. Lipid and protein oxidative damage were higher in all tissues with fermented cow milk. During anemia instauration, an increase in melatonin was observed, a fact that would improve the energy metabolism and impaired inflammatory signaling, however, during anemia recovery, fermented goat milk had positive effects on melatonin and TAS, even in the case of Fe-overload, limiting the evoked oxidative damage.

The effect of osmotic dehydration (OD) at atmospheric pressure and vacuum impregnation (VI) treatments on some physiochemical parameters of papaya (a x ,ph, color, firmness, and microstructure) was analysed. Osmotic treatments were carried ont on papapaya with 55°Bris and 65°Brix sucrose solutions at 30 °C. VI with 65°Brix osinatic sohition was the most effectise in redin log a due to the highest senrsre gain during osmotic treatment. Color differences were auseciated to loss of clarity in line with transparency gain. Scanning electron microscopy observations show that osmotic dehydration caused strape changes and the reduction of papaya cells; also differences in microstructural features were observed batween OD treated and VI treated samples. Moreover, the largest flrmness observed in VI samples sempared with OD treatments was associated with the thlcknress of the middle lamella between cells, whith was greater in VI than OD treatments. Improvement in rexture and palatability of papaya was obtained with VI rather than OD treatment compared with fresh papaya.

Bioassay-guided purification of the organic crude extract of Alternaria solani resulted in the isolation of three metabolites responsible for causing necrosis on potato leaves. These phytotoxins were identified as 2-(2",3"-dimethyl-but-1-enyl)-zinniol (1), 8-zinniol methyl ether (2). and 8-zinniol methyl ether based on their spectroscopic data (IR, MS, 1H and 13CNMR). Metabolites 1 and 2 have been identified as new phytotoxins structurally related to zinniol (4). Additionally, 5-(3',3'-dimethylallyloxy)-7-methoxy-6-methyl-phthalide and 8-zinniol-2-(phenyl)-ethyl ether (3) were also isolated during the purification process.

In a first experiment, five pigs were inoculated intranasally with porcine rubulavirus (PoRV) at 5 days of age and killed 7 days post-infection (pi). In a second experiment, four pigs were infected with the same virus at 17 days of age and killed at 9 or 15 days pi. Control piglets in each experiment received uninfected cell culture supernate. All PoRV-infected pigs developed respiratory and nervous signs, and histological lesions of non-suppurative encephalitis and interstitial pneumonia. All control pigs remained clinically normal and did not have histological lesions. Significantly increased numbers of apoptotic cells were detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL) in tonsil and lymph nodes of the pigs infected at 7 days of age and killed at 7 days pi. Significantly increased percentages of CD2 + and CD8 + T lymphocytes were also found in peripheral blood of these animals at this time, while the percentages of CD4 + and MHC class II lymphocytes were significantly reduced. Significantly increased numbers of apoptotic cells were detected in lymphoid tissues of the pigs infected at 17 days of age and killed at 9 days pi. The percentages of CD2 + , CD8 + and MHC class II lymphocytes in peripheral blood were also significantly increased at this time; the percentage of MHC class II lymphocytes remained elevated at 15 days pi. These results indicate that induction of apoptosis is an important mechanism in the pathogenesis of PoRV infection in young pigs, and that this virus induces changes in lymphocyte subpopulations in peripheral blood.

A blocking enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to porcine rubulavirus (La Piedad Michoacan Virus [LPMV]) in serum samples from pigs. The test, based on a monoclonal antibody against the LPMV hemagglutinin-neuraminidase glycoprotein, had a sensitivity of 99% and a specificity of 97%. The results of this test were in agreement with those obtained by an indirect ELISA and hemagglutination inhibition, indirect immunofluorescence, and virus neutralization tests. The blocking ELISA is considered the most suitable test for routine screening for antibodies against LPMV.

The complete nucleotide sequence of the porcine rubulavirus LPMV (La Piedad Michoacan virus) large (L) protein gene was determined and analysed. The L mRNA was found to span 6786 nucleotides, containing one single large open reading frame (ORF), putatively encoding a polypeptide of 2551 amino acids. By aligning the amino acid sequence of the LPMV L-protein with L-proteins of a number of viruses belonging to the order mononegavirale, a high degree of similarity between the LPMV L-protein and other rubula virus L-proteins was demonstrated, extending through almost the whole protein. Additionally we could identify several regions as being highly conserved among all studied viruses of the order mononegavirale. The significance of these regions are discussed.

Complementary DNA clones representing the fusion (F) protein gene of the porcine rubulavirus LPMV were isolated and sequenced. The F gene was found to be 1,845 nucleotides long containing one long open reading frame capable of encoding a protein of 541 amino acids. The cleavage motif for F0 into F1 and F2 is His-Arg-Lys-Lys-Arg. A sequence comparison and a phylogenetic analysis was performed in order to identify possible functional domains of paramyxovirus fusion proteins and also to classify the porcine rubulavirus. The F gene of LPMV is most closely related to the human mumps virus and simian virus type 5 F genes, and is therefore classified into the rubulavirus genus. A coding region for a small hydrophobic protein was however not found between the F and hemagglutinin-neuraminidase (HN) genes as previously found in both SV5 and mumps.

The persistence of porcine rubulavirus (PorPV-LPMV) in five pigs that had survived an outbreak of a natural infection was determined. After the resolution of the outbreak, each animal was housed in an isolation pen together with one sentinel pig. Approximately every 2 months thereafter one group of animals was euthanized and tissue samples taken for virological and serological analysis. Infectious virus was not isolated from any samples; antibodies to PorPV-LPMV were detected in convalescent pigs by virus neutralisation test and blocking ELISA but not in sentinel pigs. PorPV-LPMV mRNA of the nucleoprotein (NP) and phosphoprotein (P) genes was detected by a nested polymerase chain reaction (nPCR) in samples of trigeminal and optic nerves, cervical spinal cord, tonsils, salivary gland, lung and pancreas from convalescent pigs. mRNA was also detected in the midbrain, corpus callosum, or olfactory bulb in four out of five pigs by nRT-PCR, this result was confirmed by the sequencing of a 260bp PCR product of P gene region. The highest average viral copies/μg of total RNA occurred in the olfactory bulb and pancreas tissues of convalescent pigs and midbrain, tonsil and pancreas of sentinel pigs housed with the convalescent pigs. Satellitosis and gliosis of the midbrain, olfactory bulb, corpus callosum, medulla oblongata or choroid plexus were microscopically observed in four convalescent pigs. The control pig remained negative in all tests. The results indicate that PorPV-LPMV mRNA persists and induces a durable humoral immune response in pigs that have recovered from a natural infection. After a possible reactivation of the virus, it was transmitted to sentinel pigs in contact with the convalescent pigs.

La Piedad Michoacan Paramyxovirus (LPMV) is a recently recognized paramyxovirus infecting pigs throughout Mexico. Disease syndromes observed in field cases associated with LPMV infection include neurologic, respiratory, and reproductive disorders. Clinical signs and the distribution of LPMV virus and antigen in tissue samples from pigs experimentally infected with LPMV by natural routes were studied. Severe neurologic disease and death occurred following experimental inoculation of 3- and 17-day-old pigs. All of the pigs inoculated at 3 days of age were either dead or moribund by 8 days after inoculation, whereas 30% of the pigs inoculated at 17 days of age were affected. Virus was consistently recovered from or demonstrated in tissues from the respiratory tract of both groups of pigs. LPMV and antigen were also demonstrated in central nervous system (CNS) tissues from these pigs; however, differences in virus distribution within the CNS were demon- strated in the 2 groups. In the pigs inoculated at 17 days of age, isolation of LPMV was restricted to the olfactory bulb and midbrain. In contrast, in the pigs inoculated at 3 days of age, isolation of LPMV was more widespread throughout the CNS tissue examined. Virus excretion studies indicated that nasal spread of LPMV was more important than fecal spread. Comparatively large quantities of infectious LPMV were consistently recovered from urine samples of experimentally infected pigs. An outbreak of a fatal disease of pigs in Mexico (El Sindrome del Ojo Azul), associated with an unrecog- nized infectious agent, was first reported in 1981. 10 This outbreak of disease was characterized by enceph- alomyelitis and corneal opacity (blue eye) in young pigs and reproductive disorders in adult pigs. The clinical signs observed in this outbreak of disease were variable and dependant on the age of the pigs infected. Typi- cally, the disease in piglets 2-15 days of age usually started with fever, a rough hair coat, and an arched back. These signs were quickly followed by progressive nervous signs of ataxia, weakness, rigidity, muscle tremor, abnormal posture, and death within 48 hours. In older pigs, clinical signs were much less severe, con- sisting of transient fever, sneezing, and coughing. Ner

We report that the genomes of reindeer papillomavirus (RPV), European elk papillomavirus (EEPV), and deer papillomavirus (DPV) contain a short conserved translational open reading frame (ORF), E9, which is located between the E5 ORF and the early polyadenylation site. In RPV, DPV, and EEPV, E9 ORFs have the potential to encode extremely hydrophobic polypeptides of approximately 40 amino acids. In mouse C127 cells transformed by EEPV and RPV, there exists a unique, abundant mRNA species of approximately 700 nucleotides which has the capacity to encode an E9 polypeptide. This mRNA is transcribed from a previously unrecognized promoter at position 4030 in the EEPV genome. The EEPV E9 ORF exhibits weak transforming activity in C127 cells and primary rat embryo fibroblasts. We also show that EEPV E5 is the major oncogene in the EEPV genome when assayed in C127 cells, although it is less efficient in transformation than the E5 genes of bovine papillomavirus type 1, DPV, and RPV.

La neosporosis es una parasitosis de importancia en el sector ganadero por ser causante de abortos y mortalidad neonatal. El objetivo del estudio fue determinar la presencia de anticuerpos contra Neospora caninum en búfalos de agua procedentes de cinco centros de crianza ubicados en el distrito Jenaro Herrera, departamento de Loreto, Perú, en el año 2008. Se evaluaron 83 muestras de suero, obtenidas de búfalas en edad productiva, criadas para la producción de leche. Las muestras se evaluaron por ELISA indirecto e inmunofluorescencia indirecta (IFI). No se halló evidencia de anticuerpos contra N. caninum a las diluciones 1:25 y 1:100 utilizando ELISA indirecto, así como a la dilución 1:100 con IFI. Los resultados sugieren una baja o nula exposición de los búfalos de agua al parásito en la zona estudiada., Neosporosis is an important parasitic disease that causes reproductive problems as abortion and neonatal mortality in diary cattle. The aim of the study was to determine the presence of antibodies against Neospora caninum in water buffaloes reared in five farms located in Jenaro Herrera district, Loreto, Peru in 2008. Blood samples were collected from 83 female dairy buffalo older than 2 years. Serum samples were tested by indirect ELISA and immunnefluorescent antibody test (IFAT). None of sera presented antibodies against N. caninum to the dilution 1:25 and 1:100 using indirect ELISA or to the dilution 1:100 by IFAT. The results suggest a low exposition of water buffaloes to N. caninum in the studied area.

Protein and genomic studies of a previously uncharacterized porcine paramyxovirus (designated LPMV) confirmed that it was a member of the paramyxovirus genus. The nucleotide sequences and deduced amino acids of the complete P-gene, M-gene, F-gene and HN-gene as well as the intergenic sequences have been determined. Comparative sequence analysis of the M-gene of LPMV revealed the closest relationship of LPMV was to human mumps virus with a homology of 46% and 55% at the amino acid and nucleotide levels respectively. The P-gene of LPMV is transcribed to V protein mRNA and by editing of the gene to the P protein mRNA. The LPMV P-gene has the coding capacity for an additional protein of 126 amino acids, a C protein.

The nucleotide sequence of the P gene of the porcine paramyxovirus La-Piedad-Michoacan-Mexico virus (LPMV) was analysed. Three long open reading frames (ORFs) were found in the mRNA sense. Insertion of two G residues is necessary to obtain an ORF encoding the P protein, which gives a P protein of 404 amino acids with a calculated M r of 42475. This form of editing was demonstrated, two non-templated G residues being added in a portion of the mRNA transcripts. The LPMV V protein, which has a conserved cysteine-rich C-terminal region, is encoded by an exact copy of the P gene. The third ORF has the capacity to encode a protein of 126 amino acids, which may resemble the C proteins found in some paramyxoviruses. The ORF starts from an AUG codon down-stream of the first AUG codon of the P/V ORF.

1. 1. Seasonal variations in the biochemical composition, lipid classes and fatty acids in the digestive gland of the shrimp Pleoticus muelleri from San Jorge Gulf, Argentina, were studied during 1984–1985. 2. 2. The relative size of the digestive gland decreased significantly in fully mature shrimps of both sexes during the summer. 3. 3. Immature females and mature males gradually increased the level of total lipids from winter toward the autumn as a reflex of the annual cycle of feeding. 4. 4. Total lipids fell markedly during the ovary maturation in summer whilst protein increased. Free fatty acids, triacylglycerols and phospholipids were apparently transferred to the developing ovary; however, the immediate dietary supply of lipids and protein appeared to be preponderant from incipient maturation until the completion of ovary development. 5. 5. Total lipids decreased with the maturation of the male reproductive system in spring with the suggestion of their utilization during spermatogenesis. 6. 6. Maturing females appeared to use selectively fatty acids 16:0, 16:1 and 18:1 of neutral lipids during oogenesis in summer whereas males used the mentioned acids of polar lipids apparently due to metabolic stress. 7. 7. The results are discussed in terms of concentration and absolute amounts in relation to physiological and ecological factors.