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Objectives: A decline in mitochondrial function and increased susceptibility to oxidative stress is a hallmark of ageing. Exercise endogenously generates reactive oxygen species (ROS) in skeletal muscle and promotes mitochondrial remodelling resulting in improved mitochondrial function. It is unclear how exercise induced redox signalling results in alterations in mitochondrial dynamics and morphology. Methods: In this study, a Caenorhabditis elegans model of exercise and ageing was used to determine the mechanistic role of Peroxiredoxin 2 (PRDX-2) in regulating mitochondrial morphology. Mitochondrial morphology was analysed using transgenic reporter strains and transmission electron microscopy, complimented with the analysis of the effects of ageing and exercise on physiological activity. Results: The redox state of PRDX-2 was altered with exercise and ageing, hyperoxidised peroxiredoxins were detected in old worms along with basally elevated intracellular ROS. Exercise generated intracellular ROS and rapid mitochondrial remodelling, which was disrupted with age. The exercise intervention promoted mitochondrial ER contact sites (MERCS) assembly and increased DAF-16/FOXO nuclear localisation. The prdx-2 mutant strain had a disrupted mitochondrial network as evidenced by increased mitochondrial fragmentation. In the prdx-2 mutant strain, exercise did not activate DAF-16/FOXO, mitophagy or increase MERCS assembly. The results demonstrate that exercise generated ROS increased DAF-16/FOXO transcription factor nuclear localisation required for activation of mitochondrial fusion events that were blunted with age. Conclusions: The data demonstrate the critical role of PRDX-2 in orchestrating mitochondrial remodelling in response to a physiological stress by regulating redox dependent DAF-16/FOXO nuclear localisation.

Abstract Malaria can have severe long-term effects. Even after treatment with antimalarial drugs eliminates the parasite, survivors of cerebral malaria may suffer from irreversible brain damage, leading to cognitive deficits. Angiotensin II, a natural human peptide hormone that regulates blood pressure, has been shown to be active against Plasmodium spp., the etiologic agent of malaria. Here, we tested two Ang II derivatives that do not elicit vasoconstriction in mice: VIPF, a linear tetrapeptide, which constitutes part of the hydrophobic portion of Ang II; and Ang II-SS, a disulfide-bridged derivative. The antiplasmodial potential of both peptides was evaluated with two mouse models: an experimental cerebral malaria model and a mouse model of non-cerebral malaria. The latter consisted of BALB/c mice infected with Plasmodium berghei ANKA. The peptides had no effect on mean blood pressure and significantly reduced parasitemia in both mouse models. Both peptides reduced the SHIRPA score, an assay used to assess murine health and behavior. However, only the constrained derivative (Ang II-SS), which was also resistant to proteolytic degradation, significantly increased mouse survival. Here, we show that synthetic peptides derived from Ang II are capable of conferring protection against severe manifestations of malaria in mouse models while overcoming the vasoconstrictive side effects of the parent peptide.

Several reports have provided evidence that there are genetic variants of genes such as MSTN, BDRKB2, ACTN3 and ADRB2 that are involved in a better response to adaptation during resistance or strength training, while other genes such as GRB14, AGT and END1 are reported to be associated with the risk of suffering from some diseases such as diabetes, hypertension or obesity. A cross-sectional study from a Mexican Mestizo population was performed to estimate the frequency of 10 gene variants in 8 genes involved in athletic performance or chronic degenerative diseases, MSTN (rs1805085, rs1805086), BDKRB2 (rs1799722), FST (rs1423560), ACTN3 (rs1815739), ADRB2 (rs1042713, rs1042714), GRB14 (rs8192673), AGT (rs699) and EDN1 (rs5370), and to compare frequencies from 26 populations reported in the Database of 1000 Genomes project. Genotype frequencies fitted the Hardy–Weinberg equilibrium, except for MST rs1805086 and FST rs1423560, and our study revealed significant differences in the distribution of frequencies of some of these gene variants among populations reported in the 1000 Genomes Project. Our findings provide insights regarding the genetic background of our population, and future case–control studies can be carried out with more accurate sample sizes for genetic association studies. Our results may be also useful in recognizing the roles and mechanisms contributing to athletic performance and/or chronic degenerative diseases in Mexicans.

Muscular dystrophies (MDs) are a heterogeneous group of diseases of genetic origin characterized by progressive skeletal muscle degeneration and weakness. There are several types of MDs, varying in terms of age of onset, severity, and pattern of the affected muscles. However, all of them worsen over time, and many patients will eventually lose their ability to walk. In addition to skeletal muscle effects, patients with MDs may present cardiac and respiratory disorders, generating complications that could lead to death. Interdisciplinary management is required to improve the surveillance and quality of life of patients with an MD. At present, pharmacological therapy is only available for Duchene muscular dystrophy (DMD)—the most common type of MD—and is mainly based on the use of corticosteroids. Other MDs caused by alterations in dystrophin-associated proteins (DAPs) are less frequent but represent an important group within these diseases. Pharmacological alternatives with clinical potential in patients with MDs and other proteins associated with dystrophin have been scarcely explored. This review focuses on drugs and molecules that have shown beneficial effects, mainly in experimental models involving alterations in DAPs. The mechanisms associated with the effects leading to promising results regarding the recovery or maintenance of muscle strength and reduction in fibrosis in the less-common MDs (i.e., with respect to DMD) are explored, and other therapeutic targets that could contribute to maintaining the homeostasis of muscle fibers, involving different pathways, such as calcium regulation, hypertrophy, and maintenance of satellite cell function, are also examined. It is possible that some of the drugs explored here could be used to affordably improve the muscular function of patients until a definitive treatment for MDs is developed.

Diagnosis of developmental dysplasia of the hip (DDH) mostly relies on physical examination and ultrasound, and both methods are operator-dependent. Late detection can lead to complications in young adults. Current evidence supports the involvement of environmental and genetic factors, such as single nucleotide variants (SNVs). Incorporating genetic factors into diagnostic methods would be useful for implementing early detection and management of affected individuals. Our aim was to analyze environmental factors and SNVs in DDH patients. We included 287 DDH cases and 284 controls. Logistic regression demonstrated an association for sex (OR 9.85, 95% CI 5.55–17.46, p = 0.0001), family history (OR 2.4, 95% CI 1.2–4.5, p = 0.006), fetal presentation (OR 3.19, 95% CI 1.55–6.54, p = 0.002), and oligohydramnios (OR 2.74, 95%CI 1.12–6.70, p = 0.026). A model predicting the risk of DDH including these variables showed sensitivity, specificity, PPV, and NPV of 0.91, 0.53, 0.74, and 0.80 respectively. The SNV rs1800470 in TGFB1 showed an association when adjusted for covariables, OR 0.49 (95% CI 0.27–0.90), p = 0.02. When rs1800470 was included in the equation, sensitivity, specificity, PPV and NPV were 0.90, 0.61, 0.84, and 0.73, respectively. Incorporating no-operator dependent variables and SNVs in detection methods could be useful for establishing uniform clinical guidelines and optimizing health resources.

Exercise generates a site-specific increase in Reactive Oxygen Species (ROS) within muscle that promotes changes in gene transcription and mitochondrial biogenesis, required for the beneficial adaptive response. We demonstrate that Peroxiredoxin 2 (Prdx2), an abundant cytoplasmic 2-Cys peroxiredoxin, is required for the adaptive hormesis response to physiological levels of H2O2 in myoblasts and following exercise in C. elegans. A short bolus addition of H2O2 increases mitochondrial capacity and improves myogenesis of cultured myoblasts, this beneficial adaptive response was suppressed in myoblasts with decreased expression of cytoplasmic Prdxs. Moreover, a swimming exercise protocol in C. elegans increased mitochondrial content, fitness, survival and longevity in wild type (N2) worms. In contrast, prdx-2 mutant worms had decreased fitness, disrupted mitochondria, reduced survival and lifespan following exercise. Global proteomics following exercise identified distinct changes in the proteome of N2 and prdx-2 mutants. Furthermore, a redox proteomic approach to quantify reversible oxidation of specific Cysteine residues revealed a more reduced redox state in the non-exercised prdx-2 mutant strain that become oxidized following exercise. In contrast, specific Cys residues from regulatory proteins become more reduced in the N2 strain following exercise, establishing the key regulatory role of PRDX-2 in a redox signalling cascade following endogenous ROS generation. Our results demonstrate that conserved cytoplasmic 2-Cys Peroxiredoxins are required for the beneficial adaptive response to a physiological redox stress.

Background: Plant-plant facilitation provides benefits to the protected species since the host species reduces the intensity of stressful physical conditions. However, it has been hypothesized that the outcome of the interaction could be regulated by the characteristics of the interacting species and that herbivores could counteract the benefits of protective plants. Question: Can cactus size and identity of neighboring plants influence the herbivory of the columnar cactus P. leucocephalus? Studied species / Mathematical model: Pilosocereus leucocephalus (Poselg.) Byles & G.D. Rowley /One-way ANOVA, Repeated measures ANOVA, Analysis of covariance and linear regression. Study site and dates: San Ignacio, Veracruz, Mexico. April to May 2019. Methods: We randomly selected 60 P. leucocephalus plants growing underneath the canopy of seven species of shrubs and P. leucocephalus plants growing ≥ 3 m away from other species and exposed to full sunlight conditions. We obtained the size of the P. leucocephalus plants (size of main stem from ground to apex) and determined the intensity (number of bites on the stem) of vertebrate herbivory. Results: The number of bite scars by small vertebrate species was significantly higher on the stems of cacti established in open spaces than on those underneath the canopy of seven shrub species and increased significantly with the stem size of the protected species. Conclusions: We conclude that herbivory damage by rodents on a giant cactus in a dry tropical forest depends on cactus size and the identity, but not the shade, of the nearest neighbor.

Rare diseases (RDs) are serious and often fatal uncommon conditions with a high subjacent genetic origin. Since RD are not prevalent, knowledge regarding these diseases among health care providers and health students is scarce. The management of RD is often inadequate and precarious. Therefore, the aim of the present study is to assess the degree of knowledge, perceptions and attitudes of healthcare practitioners and students in Mexico. Herein we present a reliable questionnaire to assess the degree of knowledge, perceptions, awareness and attitudes of healthcare practitioners and students in Mexico with regard to RDs. The pilot study showed satisfactory levels of knowledge and significant differences in perceptions, awareness and attitudes among the study groups.

Muscular dystrophies are a group of well-defined genetic disorders characterized by the variable distribution of muscle wasting and progressive weakness. The diagnosis and treatment of these diseases remain challenging due to genetic heterogeneity and clinical overlapping. Herein, we describe our 10 years’ experience with the diagnosis and management of muscular dystrophy patients. In total, 169 patients were screened for pathogenic variants in eleven genes linked to frequent muscular dystrophies using MLPA and NGS sequencing panels. Most frequent muscular dystrophies found in the Mexican population were dystrophinopathies, dysferlinopathies and calpainopathies. Novel variants were found in genes: DMD, CAPN3, DYSF, and FKRP. For Duchenne muscular dystrophy, improvements in early diagnosis and prolonged ambulation were achieved, on the contrary, for limb-girdle muscular dystrophies and congenital muscular dystrophies, uncomplimentary follow-up and lack of detection strategies were observed. For most common muscular dystrophies, improvements in diagnosis and management have been achieved in the last 10 years, due to a collaborative effort done nationwide.

Abstract The anti-inflammatory effects of the plant protease inhibitor BbCI (Bauhinia bauhinioides cruzipain inhibitor), which blocks elastase, cathepsin G, and L, and proteinase 3 has been demonstrated. Here, we investigated the recombinant rBbCI-His(6) (containing a histidine tail) in an experimental venous thrombosis model of vena cava (VC) ligature in rats, comparing to heparin. We evaluate the effects of the inhibitors (native or recombinant) or heparin on the activated partial thromboplastin time (aPTT) and prothrombin time (PT) in human and rat plasmas. The rats undergoing treatment received a saline solution or increasing concentrations of rBbCI-His(6), heparin, or a mixture of both. After 4 h of ligature VC, thrombus, if present was removed and weighed. aPTT, PT, and cytokines were measured in blood collected by cardiac puncture. aPTT, PT, and bleeding time (BT) were also measured at the time of VC (vena cava) ligature. rBbCI-His(6) (0.45 or 1.40 mg/kg) does not alter aPTT, PT or BT. No differences in coagulation parameters were detected in rBbCI-His(6) treated rats at the time of VC ligature or when the thrombus was removed. There was a significant decrease in the weight of thrombus in the animals of the groups treated with the rBbCI-His(6) (1.40 mg/kg), with the rBbCI-His(6) mixture (1.40 mg/kg) + heparin (50 IU/kg) and heparin (100 IU/kg) in relation to control group (saline). The growth-related oncogene/keratinocyte chemoattractant (GRO/KC) serum levels in rats treated with rBbCI-His(6) (1.40 mg/kg) or heparin (200 IU/kg) were reduced. In the experimental model used, rBbCI-His(6) alone had an antithrombotic effect, not altering blood clotting or bleeding time.

OBJECTIVES/GOALS: Non-alcoholic fatty liver disease increases with aging and may be associated with MCI. Thus, adults >65 years with NAFLD have a greater risk of MCI. Our objective is to associate NAFLD with MCI and, therefore, optimize the management of MCI through prevention strategies and early interventions. METHODS/STUDY POPULATION: We will recruit 100 patients >65 of all sexes from the GI and Geriatrics clinics of the University of Puerto Rico (UPR). NAFLD prevalence in aging adults will be estimated via ICD-10 codes and definition will be liver fat accumulation or steatosis (AASLD) without secondary causes. We will exclude patients RESULTS/ANTICIPATED RESULTS: We expect to find a higher NAFLD prevalence in aging adults >65 years old compared to younger ones. Furthermore, we aim to elucidate an association of NAFLD with MCI in aging adults. Besides, as the literature has shown, we anticipate that the main cognitive domains affected by NAFLD will be the visuospatial and executive functions. As the population ages, this study will help identify future targets for early interventions in people diagnosed with NAFLD that could have a greater risk for cognitive impairment. DISCUSSION/SIGNIFICANCE: To our knowledge, no study has determined the association of NAFLD with MCI in Hispanics >65 in PR. Raising awareness of NAFLD as a possible treatable or preventable risk factor for MCI by screening NAFLD patients for MCI may improve not only their global health but their quality of life as well.

Background: The association of leptin (LEP) and vascular endothelial growth factor A (VEGFA) genes with the susceptibility to knee osteoarthritis (OA) has been analyzed; however, the epistasis between them has not been investigated. Objective: The objective of the study was to analyze the association of LEP and VEGFA variants and their interaction with primary knee OA in a Mexican Mestizo population. Methods: A case-control study was developed. Cases were ≥40 years, BMI ≤27 kg/m2, with primary knee OA and radiologic Grade ≥2. Controls were participants with no knee OA and a radiologic Grade

Mutations in the human MYH7 gene, encoding a slow skeletal muscle/β-cardiac myosin heavy chain, cause different types of myopathies. The nematode model Caenorhabditis elegans has frequently been employed to study the molecular and physiological consequences of MYH7 mutations in muscle function by introducing mutations into the unc-54 gene, the worm MYH7 ortholog. We report here that the C. elegans model is not appropriate for such studies if they involve expression of the UNC-54 protein (wild-type or fused to green fluorescent protein) above endogenous levels.

EDEM-1, EDEM-2 and EDEM-3 are key players for the quality control of newly synthesized proteins in the endoplasmic reticulum (ER) by accelerating disposal and degradation of misfolded proteins through ER Associated Degradation (ERAD). Although many previous studies reported the role of individual ERAD components especially in cell-based systems, still little is known about the consequences of ERAD dysfunction under physiological and ER stress conditions in the context of a multicellular organism. Here we report the first individual and combined characterization and functional interplay of EDEM proteins in Caenorhabditis elegans using single, double, and triple mutant combinations. We found that EDEM-2 has a major role in the clearance of misfolded proteins from ER under physiological conditions, whereas EDEM-1 and EDEM-3 roles become prominent under acute ER stress. In contrast to SEL-1 loss, the loss of EDEMs in an intact organism induces only a modest ER stress under physiological conditions. In addition, chronic impairment of EDEM functioning attenuated both XBP-1 activation and up-regulation of the stress chaperone GRP78/BiP, in response to acute ER stress. We also show that pre-conditioning to EDEM loss in acute ER stress restores ER homeostasis and promotes survival by activating ER hormesis. We propose a novel role for EDEM in fine-tuning the ER stress responsiveness that affects ER homeostasis and survival.

The TBX20 gene has a key role during cardiogenesis, and it has been related to epigenetic mechanisms in congenital heart disease (CHD). The purpose of this study was to assess the association between DNA methylation status and congenital septal defects. The DNA methylation of seven CpG sites in the TBX20 gene promoter was analyzed through pyrosequencing as a quantitative method in 48 patients with congenital septal defects and 104 individuals with patent ductus arteriosus (PDA). The average methylation was higher in patients than in PDA (p < 0.001). High methylation levels were associated with a higher risk of congenital septal defects (OR = 4.59, 95% CI = 1.57–13.44, p = 0.005). The ROC curve analysis indicated that methylation of the TBX20 gene could be considered a risk marker for congenital septal defects (AUC = 0.682; 95% CI = 0.58–0.77; p < 0.001). The analysis of environmental risk factors in patients with septal defects and PDA showed an association between the consumption of vitamins (OR = 0.10; 95% CI = 0.01–0.98; p = 0.048) and maternal infections (OR = 3.10; 95% CI = 1.26–7.60; p = 0.013). These results suggest that differences in DNA methylation of the TBX20 gene can be associated with septal defects.

Abstract In the frugivory networks of many arid and semi‐arid Mesoamerican ecosystems, columnar cacti act as keystone species that produce fruits with a high content of water and nutrients attractive to numerous vertebrates. The aim of this investigation was to assess the fruit removal patterns of two guilds of frugivores on the fruits of the woolly torch Pilosocereus leucocephalus. We assessed fruit pulp removal in two ways: by estimating the consumption of seeds given the amount of pulp removed per visit and by estimating the percentage of pulp removal over time. We put exclosures on unripe, intact fruits to keep frugivores from removing material. Once ripe, we removed the exclosures and tracked animal visitation of 69 fruits using camera traps. We obtained a total of 2,162 hr of footage (14:47 hours of them with effective pulp removal). The highest number of visitors is that of diurnal species (n = 12, all birds) versus only four nocturnal (three bats, one rodent). The most effective species in pulp removal are birds. Bats play a modest role in frugivory of this cactus. The significance of this work is twofold: (a) birds and bats consume the fruit pulp of this cactus and likely disperse its seeds, and (b) although bats rank high in pulp removal effectiveness, birds as a guild far outweigh their importance in this system, as they are not only more frequent but also remove more pulp and seeds. Both groups are known to be important in cacti seed dispersal, and our findings are essential in understanding the population dynamics of the woolly torch and in elucidating its seed dispersal ecology.

Abstract Background In several Apicomplexa, the formation of moving junctions (MJs) at the interface between the external membranes of the invading parasite and the host cell is essential for the process of parasite invasion. In Plasmodium falciparum and Toxoplasma gondii, the MJ is composed of the Apical Membrane Antigen 1 (AMA1) and Rhoptry Neck Proteins (RONs) complex; specifically, AMA1 interacts with RON2 during host cell invasion. Methods Recombinant proteins based on Plasmodium vivax RON2 (A2033-P2100) and its synthetic peptide fragments, one cyclic and one linear, based on PvRON2 (D2035-T2074) were generated and used to evaluate the interaction with P. vivax AMA1 (PvAMA1) by the far western blot, surface plasmon resonance (SPR), and isothermal titration microcalorimetry (ITC) methods. The structural studies of peptides were performed by circular dichroism, and the structural analysis of the complex of PvAMA1 with peptides based on PvRON2 (D2035-T2074) was conducted with small-angle X-ray scattering (SAXS). Results Surface plasmon resonance (KD = 23.91 ± 2.078 μmol/L) and ITC (K = 3 × 105 mol/L) studies conclusively showed an interaction between the cyclic peptide based on PvRON2 and PvAMA1-His6. In contrast, the linear peptide and recombinant PvRON2 (GST fusion protein) did not show an interaction with PvAMA1. However, the interaction among recombinant proteins PvRON2.2 and PvAMA1-His6 was possible to show by far western blot. Conclusions The results show that the PvRON2 structure, particularly the S–S bond between C2051 and C2063, is determinant for the existence of the interaction between PvAMA1 and PvRON2.

Plant protection product (PPP) applications to isolated olive trees are commonly performed with regular air-assisted sprayers, which are not adapted to their particular characteristics. Whilst strong efforts have been undertaken over the last years to improve technical aspects like canopy detection and automated proportional dosing, nearly no efforts have been made regarding the sprayer adaption to the crop. For this reason, three prototype sprayers were developed for traditional and intensive olive cultivations systems (P1: centrifugal fan; P2: six small side axial fans; P3: two axial fans in tower structure) with the purpose to improve the application efficiency. The main goal of the present study was to check spray quality and efficiency in comparison with the conventional sprayer in both cultivation systems. The sprayers were tested in two different olive groves and properly calibrated according to the tree dimensions. The spray deposition, coverage, drift, and losses to the ground were measured in five trees per cultivation system by placing the appropriate collectors. The sprayers performed very differently in both cultivation systems. In the intensive system, the spray deposition did not present significant differences (p = 0.105). However, it did in the traditional system (p = 0.003), with P3 obtaining the best results. The spray coverage followed the same trend, with significant differences only in the traditional orchard (p = 0.011), with the prototypes leading. The conventional equipment generated the highest spray losses in both cultivation systems. Crop adapted spraying can significantly improve the spray quality and efficiency in difficult crops like olive. This topic may have a key importance to match the environmentally sustainable use of PPP.

BackgroundExclusive breastfeeding promotes beneficial modifications on the microbiota of cesarean born infants, but little is known about the role of specific breast milk components in this modulation. Women with an active FUT2 gene (called secretors) secrete α1-2 fucosylated human milk oligosaccharides (HMOs), which promote Bifidobacterium in the infant's gut and may modulate the microbiota of cesarean born infants.ObjectiveTo compare the microbiota composition of cesarean and vaginally born infants breastfed by secretor mothers.MethodsMaternal secretor status was determined by the occurrence of 4 different α1-2 fucosylated HMOs in breast milk by LC-MS. The fecal microbiota composition from cesarean and vaginally born infants was analyzed by 16S rRNA gene sequencing and qPCR, stratified by the maternal secretor status, and compared.ResultsAlpha and beta diversity were not significantly different in cesarean born, secretor-fed infants (CSe+) compared to vaginally born, secretor-fed infants (VSe+). There were no significant differences in the fecal relative abundance of Bifidobacterium between CSe+ and VSe+ infants, but the prevalence of the species B. longum was lower in CSe+. The fecal relative abundance of Bacteroides was also lower, while Akkermansia and Kluyvera were higher in CSe+ infants.ConclusionCesarean and vaginally born infants fed with breast milk containing the α1-2 fucosylated HMOs fraction present similar amounts of Bifidobacterium in the feces, but differences are observed in other members of the microbiota.

Hepatocellular carcinoma (HCC) represents 80% of the primary hepatic neoplasms. It is the sixth most frequent neoplasm, the fourth cause of cancer-related death, and 7% of registered malignancies. Sorafenib is the first line molecular targeted therapy for patients in advanced stage of HCC. The present study shows that Sorafenib exerts free radical scavenging properties associated with the downregulation of nuclear factor E2-related factor 2 (Nrf2)-regulated thioredoxin 1 (Trx1) expression in liver cancer cells. The experimental downregulation and/or overexpression strategies showed that Trx1 induced activation of nitric oxide synthase (NOS) type 3 (NOS3) and S-nitrosation (SNO) of CD95 receptor leading to an increase of caspase-8 activity and cell proliferation, as well as reduction of caspase-3 activity in liver cancer cells. In addition, Sorafenib transiently increased mRNA expression and activity of S-nitrosoglutathione reductase (GSNOR) in HepG2 cells. Different experimental models of hepatocarcinogenesis based on the subcutaneous implantation of HepG2 cells in nude mice, as well as the induction of HCC by diethylnitrosamine (DEN) confirmed the relevance of Trx1 downregulation during the proapoptotic and antiproliferative properties induced by Sorafenib. In conclusion, the induction of apoptosis and antiproliferative properties by Sorafenib were related to Trx1 downregulation that appeared to play a relevant role on SNO of NOS3 and CD95 in HepG2 cells. The transient increase of GSNOR might also participate in the deactivation of CD95-dependent proliferative signaling in liver cancer cells.

Human selenium-binding protein 1 (SELENBP1) was originally identified as a protein binding selenium, most likely as selenite. SELENBP1 is associated with cellular redox and thiol homeostasis in several respects, including its established role as a methanethiol oxidase that is involved in degradation of methanethiol, a methionine catabolite, generating hydrogen sulfide (H2S) and hydrogen peroxide (H2O2). As both H2S and reactive oxygen species (such as H2O2) are major regulators of Caenorhabditis elegans lifespan and stress resistance, we hypothesized that a SELENBP1 ortholog in C. elegans would likely be involved in regulating these aspects.Here we characterize Y37A1B.5, a putative selenium-binding protein 1 ortholog in C. elegans with 52% primary structure identity to human SELENBP1. While conferring resistance to toxic concentrations of selenite, Y37A1B.5 also attenuates resistance to oxidative stress and lowers C. elegans lifespan: knockdown of Y37A1B.5 using RNA interference resulted in an approx. 10% increase of C. elegans lifespan and an enhanced resistance against the redox cycler paraquat, as well as enhanced motility. Analyses of transgenic reporter strains suggest hypodermal expression and cytoplasmic localization of Y37A1B.5, whose expression decreases with worm age. We identify the transcriptional coregulator MDT-15 and transcription factor EGL-27 as regulators of Y37A1B.5 levels and show that the lifespan extending effect elicited by downregulation of Y37A1B.5 is independent of known MDT-15 interacting factors, such as DAF-16 and NHR-49. In summary, Y37A1B.5 is an ortholog of SELENBP1 that shortens C. elegans lifespan and lowers resistance against oxidative stress, while allowing for a better survival under toxic selenite concentrations. Keywords: Selenium-binding protein, Stress signaling, Caenorhabditis elegans, Lifespan

The TBX5 gene regulates morphological changes during heart development, and it has been associated with epigenetic abnormalities observed in congenital heart defects (CHD). The aim of this research was to evaluate the association between DNA methylation levels of the TBX5 gene promoter and congenital septal defects. DNA methylation levels of six CpG sites in the TBX5 gene promoter were evaluated using pyrosequencing analysis in 35 patients with congenital septal defects and 48 controls. Average methylation levels were higher in individuals with congenital septal defects than in the controls (p < 0.004). In five CpG sites, we also found higher methylation levels in patients than in the controls (p < 0.05). High methylation levels were associated with congenital septal defects (OR = 3.91; 95% CI = 1.02–14.8; p = 0.045). The analysis of Receiver Operating Characteristic (ROC) showed that the methylation levels of the TBX5 gene could be used as a risk marker for congenital septal defects (AUC = 0.68, 95% CI = 0.56–0.80; p = 0.004). Finally, an analysis of environmental factors indicated that maternal infections increased the risk (OR = 2.90; 95% CI = 1.01–8.33; p = 0.048) of congenital septal defects. Our data suggest that a high DNA methylation of the TBX5 gene could be associated with congenital septal defects.

Abstract Background Primary Osteoarthritis (OA) of the knee is a multifactorial disease that has an important genetic component, and several genes have been associated with its development. The vitamin D receptor has a role in skeletal metabolism that suggests a relationship with OA. The aim of this study was to analyze the association of Vitamin D receptor gene (VDR) polymorphisms in Mexican Mestizo patients. Methods A case-control study was conducted in which 107 cases with primary OA of the knee and 114 controls were included. Cases were patients > 40 years of age with a Body mass index (BMI) of ≤27 and a radiological score for OA of the knee of ≥2. Controls were subjects > 40 years of age with a radiological score of

The European Cooperation in Science and Technology (COST) provides an ideal framework to establish multi-disciplinary research networks. COST Action BM1203 (EU-ROS) represents a consortium of researchers from different disciplines who are dedicated to providing new insights and tools for better understanding redox biology and medicine and, in the long run, to finding new therapeutic strategies to target dysregulated redox processes in various diseases. This report highlights the major achievements of EU-ROS as well as research updates and new perspectives arising from its members. The EU-ROS consortium comprised more than 140 active members who worked together for four years on the topics briefly described below. The formation of reactive oxygen and nitrogen species (RONS) is an established hallmark of our aerobic environment and metabolism but RONS also act as messengers via redox regulation of essential cellular processes. The fact that many diseases have been found to be associated with oxidative stress established the theory of oxidative stress as a trigger of diseases that can be corrected by antioxidant therapy. However, while experimental studies support this thesis, clinical studies still generate controversial results, due to complex pathophysiology of oxidative stress in humans. For future improvement of antioxidant therapy and better understanding of redox-associated disease progression detailed knowledge on the sources and targets of RONS formation and discrimination of their detrimental or beneficial roles is required. In order to advance this important area of biology and medicine, highly synergistic approaches combining a variety of diverse and contrasting disciplines are needed.

Objectives Chitosan has been widely investigated and used. However, the literature does not refer to the shelf life of this solution. This study evaluated, through the colorimetric titration technique and an analysis of dentin micro-hardness, the shelf life of 0.2% chitosan solution. Materials and Methods Thirty human canines were sectioned, and specimens were obtained from the second and third slices, from cemento-enamel junction to the apex. A 0.2% chitosan solution was prepared and distributed in 3 identical glass bottles (v1, v2, and v3) and 3 plastic bottles (p1, p2, and p3). At 0, 7, 15, 30, 45, 60, 90, 120, 150, and 180 days, the specimens were immersed in each solution for 5 minutes (n = 3 each). The chelating effect of the solution was assessed by micro-hardness and colorimetric analysis of the dentin specimens. 17% EDTA and distilled water were used as controls. Data were analyzed statistically by two-way and Tukey-Kramer multiple comparison (α = 0.05). Results There was no statistically significant difference among the solutions with respect to the study time (p = 0.113) and micro-hardness/time interaction (p = 0.329). Chitosan solutions and EDTA reduced the micro-hardness in a similar manner and differed significantly from the control group (p < 0.001). Chitosan solutions chelated calcium ions throughout the entire experiment. Conclusions Regardless of the storage form, chitosan demonstrates a chelating property for a minimum period of 6 months.

ROS (reactive oxygen species) are potentially damaging by-products of aerobic metabolism which, unchecked, can have detrimental effects on cell function. However, it is now widely accepted that, at physiological levels, certain ROS play important roles in cell signaling, acting as second messengers to regulate cell choices that contribute to the development, adaptation and survival of plants and animals. Despite important recent advances in the biochemical tools available to study redox-signaling, the molecular mechanisms underlying most of these responses remain poorly understood, particularly in multicellular organisms. As we will review here, C. elegans has emerged as a powerful animal model to elucidate these and other aspects of redox biology. Keywords: Aging, Caenorhabditis elegans, Cuticle, Innate immunity, Pathogen infection, Reactive oxygen species, Redox homeostasis, ROS detection, Wound healing

Abstract Chitosan is a natural, biocompatible chelating substance with potential for dental use. This study compared the effects of final canal irrigation with chitosan and EDTA on dentin microhardness, sealer dentin tubules penetration capacity, and push-out strength. Methodology: Fifty canine roots were distributed according to the final irrigation protocol (n=10): G1- 15% EDTA with conventional irrigation; G2- 15% EDTA with Endovac; G3- 0.2% chitosan with conventional irrigation; G4- 0.2% chitosan with Endovac; and G5- without irrigation. Specimens were obturated (AH Plus) and sectioned in 3 slices per root third. The first slice was used for microhardness and sealer penetration assessments under a laser confocal microscope. The second was utilized in a push-out strength test. The third slice was discarded. Data were analyzed using 2-way ANOVA and Tukey's post hoc test (α0.05). In general, all experimental groups presented similar bond resistance (p>0.05) that significantly differed from the control (p

Animal development requires the execution of specific transcriptional programs in different sets of cells to build tissues and functional organs. Transcripts are exported from the nucleus to the cytoplasm where they are translated into proteins that, ultimately, carry out the cellular functions. Here we show that in Caenorhabditis elegans, reduction of mRNA export strongly affects epithelial morphogenesis and germline proliferation while other tissues remain relatively unaffected. Epithelialization and gamete formation demand a large number of transcripts in the cytoplasm for the duration of these processes. In addition, our findings highlight the existence of a regulatory feedback mechanism that activates gene expression in response to low levels of cytoplasmic mRNA. We expand the genetic characterization of nuclear export factor NXF-1 to other members of the mRNA export pathway to model mRNA export and recycling of NXF-1 back to the nucleus. Our model explains how mutations in genes involved in general processes, such as mRNA export, may result in tissue-specific developmental phenotypes.

Introducción: La toxoplasmosis es una zoonosis ampliamente distribuida, un tercio de la población mundial tiene anticuerpos contra Toxoplasma gondii. El hombre se infecta principalmente por vía oral, por la ingesta de alimentos o aguas contaminadas, a través de la placenta al feto, por vía hematógena, por trasplantes de órganos o transfusiones de sangre contaminada. La circulación de Toxoplasma gondii se ha demostrado en la parte occidental de Cuba. Objetivo: Determinar la seroprevalencia en la región oriental de Cuba en donantes de sangre. Métodos: Las muestras de suero de 3365 donantes de sangre se evaluaron con previo consentimiento informado. La técnica de inmunofluorescencia indirecta se utilizó para detectar anticuerpos contra este parásito. Resultados: La positividad al Toxoplasma gondii fue de un (38,4 %) 1292 muestras positivas en los donantes estudiados, siendo la seropositividad del área rural mayor que en el área urbana, y no se observó relación entre seroprevalencia edad y sexo. Conclusiones: Se demuestra la exposición y la circulación del parásito en esta región. Se recomienda la certificación de donaciones de sangre para Toxoplasma gondii antes de su uso.

Thioredoxins (TRX) are traditionally considered as enzymes catalyzing redox reactions. However, redox-independent functions of thioredoxins have been described in different organisms, although the underlying molecular mechanisms are yet unknown. We report here the characterization of the first generated endogenous redox-inactive thioredoxin in an animal model, the TRX-1 in the nematode Caenorhabditis elegans. We find that TRX-1 dually regulates the formation of an endurance larval stage (dauer) by interacting with the insulin pathway in a redox-independent manner and the cGMP pathway in a redox-dependent manner. Moreover, the requirement of TRX-1 for the extended longevity of worms with compromised insulin signalling or under calorie restriction relies on TRX-1 redox activity. In contrast, the nuclear translocation of the SKN-1 transcription factor and increased LIPS-6 protein levels in the intestine upon trx-1 deficiency are strictly redox-independent. Finally, we identify a novel function of C. elegans TRX-1 in male food-leaving behaviour that is redox-dependent. Taken together, our results position C. elegans as an ideal model to gain mechanistic insight into the redox-independent functions of metazoan thioredoxins, overcoming the limitations imposed by the embryonic lethal phenotypes of thioredoxin mutants in higher organisms. Keywords: Caenorhabditis elegans, Dauer, Food-leaving, Lips-6, Longevity, Male, Redox, Skn-1, Thioredoxin

Part-through surface or corner 2D cracks are commonly found in structural components, even because practically all fatigue cracks tend to start this way. It is a reasonable hypothesis to model them assuming the shape of their fronts is approximately an elliptic arc, as supported by many fractographic observations. But their transition to a 1D through-crack, an important issue in many practical applications, is normally not properly addressed in fatigue life predictions. Although experimental results reveal that the frontier of surface cracks essentially retain their elliptical shape as they gradually grow into a through-crack, it is usual to assume they are immediately transformed into a through-crack when their depth reaches the cracked component thickness. This crude approximation may create a large jump in stress intensity values, leading to excessively conservative fatigue crack growth predictions, or else the crude shape jump hypothesis may induce false overload events that can much affect fatigue crack growth retardation models, leading to inadmissible non-conservative life predictions. To minimize such problems, an improved model to describe the transition of 2D surface cracks to 1D through-cracks is proposed and verified by crack propagation tests in two different materials, 4340 steel and polycarbonate (PC). Moreover, fatigue life predictions based on this improved model are compared with experimental results.

The nematode Caenorhabditis elegans has been used to examine the influence of epicatechin (EC), an abundant flavonoid in the human diet, in some stress biomarkers (ROS production, lipid peroxidation and protein carbonylation). Furthermore, the ability of EC to modulate the expression of some key genes in the insulin/IGF-1 signaling pathway (IIS), involved in longevity and oxidative or heat shock stress response, has also been explored. The final aim was to contribute to the elucidation of the mechanisms involved in the biological effects of flavonoids. The results showed that EC-treated wild-type C. elegans exhibited increased survival and reduced oxidative damage of biomolecules when submitted to thermal stress. EC treatment led to a moderate elevation in ROS levels, which might activate endogenous mechanisms of defense protecting against oxidative insult. The enhanced stress resistance induced by EC was found to be mediated through the IIS pathway, since assays in daf-2, age-1, akt-1, akt-2, sgk-1, daf-16, skn-1 and hsf-1 loss of function mutant strains failed to show any heat-resistant phenotype against thermal stress when treated with EC. Consistently, EC treatment upregulated the expression of some stress resistance associated genes, such as gst-4, hsp-16.2 and hsp-70, which are downstream regulated by the IIS pathway.

Resumo: o artigo objetiva apresentar o mundo rural do estado do Tocantins a partir do olhar de autores clássicos da literatura goiana e tocantinense: Hugo de Carvalho Ramos, Bernardo Élis, Carmo Bernardes, Bariani Ortêncio, Eli Brasiliense e Moura Lima. Não há pretensão de fazer uma hermenêutica de cada autor e de suas obras, mas apresentar elementos chaves da constituição da realidade específica do camponês tocantinense a partir da produção literária. Incialmente o texto define e caracteriza o que vem a ser o sistema de poder do coronelismo, na sequência são trabalhados aspectos do mundo rural goiano e tocantinense tal como retratado pelos autores da literatura regional. Nas considerações finais, aponta-se que o avanço do capitalismo no campo não eliminou as características seculares da exploração do camponês, apenas agregou novos contornos. Palavras-chave: Campesinato. literatura regional. coronelismo.

Pneumatic spraying is especially sensitive to spray drift due to the production of small droplets that can be easily blown away from the treated field by the wind. Two prototypes of environmentally friendly pneumatic spouts were developed. The present work aims to check the effect of the spout modifications on the spray quality, to test the convenience of setting the liquid hose out of the spout in cannon-type and hand-type pneumatic nozzles and its effect on the droplet size, homogeneity and driftability in laboratory conditions. Laboratory trials simulating a real sprayer were conducted to test the influence of the hose insertion position (HP), including conventional (CP), alternative (AP), outer (OP) and extreme (XP), as well as the liquid flow rate (LFR) and the airflow speed (AS) on the droplet size (D50, D10 and D90), homogeneity and driftability (V100). Concurrently, the droplet size spectra obtained by the combination of aforementioned parameters (HP × LFR × AS) in both nozzles were also classified according to the ASABE S572.1. Results showed a marked reduction of AS outside the air spout, which led to droplet size increase. This hypothesis was confirmed by the droplet size spectra measured (D50, D10, D90 and V100). A clear influence of HP was found on every dependent variable, including those related with the droplet size. In both nozzles, the longer the distance to CP, the coarser the sprayed drops. Moreover, LFR and AS significantly increased and reduced droplet size, respectively. A higher heterogeneity in the generated drops was obtained in XP. This position yielded V100 values similar to those of the hydraulic low-drift nozzles, showing an effective drift reduction potential. The classification underlines that the variation of HP, alongside AS and LFR, allowed varying the spray quality from very fine to coarse/very coarse, providing farmers with a wide range of options to match the drift-reducing environmental requirements and the treatment specifications for every spray application.

The importance of antimicrobial peptides (AMPs) in relation to the survival of invertebrates is well known. The source and the mode of action on the insects' immune system of these molecules have been described from different perspectives. Insects produce their own AMPs as well as obtain these molecules from various sources, for example by absorption through the intestinal tract, as previously described for Boophilus microplus. Blood-sucking barber bug Triatoma infestans attracts social, economic and medical interest owing to its role in the transmission of Chagas disease. Despite new studies, descriptions of AMPs from this insect have remained elusive. Thus, the aims of this work were to characterize the antimicrobial potential of human fibrinopeptide A (FbPA) obtained from the T. infestans haemolymph and identify its natural source. Therefore, FbPA was isolated from the T. infestans haemolymph through liquid chromatography and identified by mass spectrometry. This peptide exhibited antimicrobial activity against Micrococcus luteus. Native FbPA from human blood and the synthetic FbPA also exhibited antimicrobial activity. The synthetic FbPA was conjugated with fluorescein isothiocyanate and offered to the insects. The haemolymph collected after 72 h exhibited fluorescence at the same wavelength as fluorescein isothiocyanate. Our experiments show that beyond intrinsic AMP production, T. infestans is able to co-opt molecules via internalization and may use them as AMPs for protection.

Cisplatin and derivatives are commonly used as chemotherapeutic agents. Although the cytotoxic action of cisplatin on cancer cells is very efficient, clinical oncologists need to deal with two major difficulties, namely the onset of resistance to the drug and the cytotoxic effect in patients. Here, we used Caenorhabditis elegans to investigate factors influencing the response to cisplatin in multicellular organisms. In this hermaphroditic model organism, we observed that sperm failure is a major cause of cisplatin-induced infertility. RNA sequencing data indicate that cisplatin triggers a systemic stress response, in which DAF-16/FOXO and SKN-1/NRF2, two conserved transcription factors, are key regulators. We determined that inhibition of the DNA damage-induced apoptotic pathway does not confer cisplatin protection to the animal. However, mutants for the pro-apoptotic BH3-only gene ced-13 are sensitive to cisplatin, suggesting a protective role of the intrinsic apoptotic pathway. Finally, we demonstrated that our system can also be used to identify mutations providing resistance to cisplatin and therefore potential biomarkers of innate cisplatin-refractory patients. We show that mutants for the redox regulator trxr-1, ortholog of the mammalian thioredoxin reductase 1 TRXR1, display cisplatin resistance. By CRISPR/Cas9, we determined that such resistance relies on the presence of the single selenocysteine residue in TRXR-1. This article has an associated First Person interview with the first author of the paper.

ABSTRACT Acinetobacter baumannii is a significant human pathogen associated with hospital-acquired infections. While adhesion, an initial and important step in A. baumannii infection, is well characterized, the intracellular trafficking of this pathogen inside host cells remains poorly studied. Here, we demonstrate that transcription factor EB (TFEB) is activated after A. baumannii infection of human lung epithelial cells (A549). We also show that TFEB is required for the invasion and persistence inside A549 cells. Consequently, lysosomal biogenesis and autophagy activation were observed after TFEB activation which could increase the death of A549 cells. In addition, using the Caenorhabditis elegans infection model by A. baumannii, the TFEB orthologue HLH-30 was required for survival of the nematode to infection, although nuclear translocation of HLH-30 was not required. These results identify TFEB as a conserved key factor in the pathogenesis of A. baumannii. IMPORTANCE Adhesion is an initial and important step in Acinetobacter baumannii infections. However, the mechanism of entrance and persistence inside host cells is unclear and remains to be understood. In this study, we report that, in addition to its known role in host defense against Gram-positive bacterial infection, TFEB also plays an important role in the intracellular trafficking of A. baumannii in host cells. TFEB was activated shortly after A. baumannii infection and is required for its persistence within host cells. Additionally, using the C. elegans infection model by A. baumannii, the TFEB orthologue HLH-30 was required for survival of the nematode to infection, although nuclear translocation of HLH-30 was not required.

DNA fragment assembly represents an important challenge to the development of efficient and practical algorithms due to the large number of elements to be assembled. In this study, we present some graph theoretical linear time algorithms to solve the problem. To achieve linear time complexity, a heap with constant time operations was developed, for the special case where the edge weights are integers and do not depend on the problem size. The experiments presented show that modified classical graph theoretical algorithms can solve the DNA fragment assembly problem efficiently.

Accurate crown characterization of large isolated olive trees is vital for adjusting spray doses in three-dimensional crop agriculture. Among the many methodologies available, laser sensors have proved to be the most reliable and accurate. However, their operation is time consuming and requires specialist knowledge and so a simpler crown characterization method is required. To this end, three methods were evaluated and compared with LiDAR measurements to determine their accuracy: Vertical Crown Projected Area method (VCPA), Ellipsoid Volume method (VE) and Tree Silhouette Volume method (VTS). Trials were performed in three different kinds of olive tree plantations: intensive, adapted one-trunked traditional and traditional. In total, 55 trees were characterized. Results show that all three methods are appropriate to estimate the crown volume, reaching high coefficients of determination: R2 = 0.783, 0.843 and 0.824 for VCPA, VE and VTS, respectively. However, discrepancies arise when evaluating tree plantations separately, especially for traditional trees. Here, correlations between LiDAR volume and other parameters showed that the Mean Vector calculated for VCPA method showed the highest correlation for traditional trees, thus its use in traditional plantations is highly recommended.

Biomarker-based sperm analysis elevates the treatment of human infertility and ameliorates reproductive performance in livestock. The negative biomarker-based approach focuses on proteins and ligands unique to defective spermatozoa, regardless of their morphological phenotype, lending itself to analysis by flow cytometry (FC). A prime example is the spermatid specific thioredoxin SPTRX3/TXNDC8, retained in the nuclear vacuoles and superfluous cytoplasm of defective human spermatozoa. Infertile couples with high semen SPTRX3 are less likely to conceive by assisted reproductive therapies (ART) and more prone to recurrent miscarriage while low SPTRX3 has been associated with multiple ART births. Ubiquitin, a small, proteolysis-promoting covalent posttranslational protein modifier is found on the surface of defective posttesticular spermatozoa and in the damaged protein aggregates, the aggresomes of spermiogenic origin. Semen ubiquitin content correlates negatively with fertility and conventional semen parameters, and with sperm binding of lectins LCA (Lens culinaris agglutinin; reveals altered sperm surface) and PNA (Arachis hypogaea/peanut agglutinin; reveals acrosomal malformation or damage). The Postacrosomal Sheath WWI Domain Binding Protein (PAWP), implicated in oocyte activation during fertilization, is ectopic or absent from defective human and animal spermatozoa. Consequently, FC-parameters of PAWP correlate with ART outcomes in infertile couples and with fertility in bulls. Assays based on the above biomarkers have been combined into multiplex FC semen screening protocols, and the surface expression of lectins and ubiquitin has been utilized to develop nanoparticle-based bull semen purification method validated by field artificial insemination trials. These advances go hand-in-hand with the innovation of FC-technology and genomics/proteomics-based biomarker discovery.

Ultrasonic sensors are often used to adjust spray volume by allowing the calculation of the crown volume of tree crops. The special conditions of the olive tree require the use of long-range sensors, which are less accurate and faster than the most commonly used sensors. The main objectives of the study were to determine the suitability of the sensor in terms of sound cone determination, angle errors, crosstalk errors and field measurements. Different laboratory tests were performed to check the suitability of a commercial long-range ultrasonic sensor, as were the experimental determination of the sound cone diameter at several distances for several target materials, the determination of the influence of the angle of incidence of the sound wave on the target and distance on the accuracy of measurements for several materials and the determination of the importance of the errors due to interference between sensors for different sensor spacings and distances for two different materials. Furthermore, sensor accuracy was tested under real field conditions. The results show that the studied sensor is appropriate for olive trees because the sound cone is narrower for an olive tree than for the other studied materials, the olive tree canopy does not have a large influence on the sensor accuracy with respect to distance and angle, the interference errors are insignificant for high sensor spacings and the sensor’s field distance measurements were deemed sufficiently accurate.

Abstract This study evaluates the influence of root dentin treatment with NaOCl alone and combined with EDTA, with and without ultrasound activation, on the push-out bond strength (BS) of fiber-reinforced posts in weakened roots, cemented with RelyX or Panavia. The root canals of 42 maxillary canines were instrumented with Reciproc and 2.5% NaOCl. In the coronal 12mm of all canals, experimental weakening of the roots was produced by reducing dentin thickness with 2.44mm diameter diamond burs. The roots were assigned to 3 groups (n = 14) according to root dentin treatment: 2.5% NaOCl; 2.5% NaOCl + 17% EDTA; and 2.5% NaOCl + 17% EDTA, with solutions agitated using passive ultrasonic irrigation. After cementation of the fiber-reinforced posts the roots were divided in thirds. The first slice of each third was used for the push-out BS test, the second slice for confocal laser scanning microscopy and dentin microhardness (Knoop) analysis. Data were analysed by a two-way ANOVA and Tukey test (a = 0.05). NaOCl + EDTA provided highest BS values than NaOCl (p < 0.0001). Specimens cemented with Panavia presented significantly higher BS than those with RelyX in the three root thirds (p < 0.0001). The highest BS values occurred in the cervical third (p < 0.001). Ultrasound-activated NaOCl + EDTA promoted the greatest reduction in dentin microhardness, followed by NaOCl/EDTA and NaOCl. Ultrasonic activation of NaOCl and EDTA reduced root dentin microhardness, but did not improve the push-out BS of resin-based cements. Panavia presented higher BS than RelyX. RelyX was not influenced by the root dentin treatment protocols.

Limb-Girdle Muscular Dystrophy type 2 (LGMD2) is a group of autosomally recessive inherited disorders defined by weakness and wasting of the shoulder and pelvic girdle muscles. In the past, several population isolates with high incidence of LGMD2 arising from founder mutation effects have been identified. The aim of this work is to describe the results of clinical, epidemiologic, and molecular studies performed in a Mexican village segregating numerous cases of LGMD2. A population census was conducted in the village to identify all LGMD affected patients. Molecular analysis included genome wide homozygosity mapping using a 250K SNP Affymetrix microarray followed by PCR amplification and direct nucleotide sequencing of the candidate gene. In addition, DNA from 401 randomly selected unaffected villagers was analyzed to establish the carrier frequency of the LGMD2 causal mutation. A total of 32 LGMD2 patients were identified in the village, rendering a disease prevalence of 4.3 (CI: 2.9-5.9) cases per 1,000 habitants (1 in 232). Genome wide homozygosity mapping revealed that affected individuals shared a 6.6 Mb region of homozygosity at chromosome 15q15. The identified homozygous interval contained CAPN3, the gene responsible for LGMD2 type A (LGMD2A). Direct sequencing of this gene revealed homozygosity for a novel c.348C>A mutation (p.Ala116Asp) in DNA from all 20 affected subjects available for genetic screening, except one which was heterozygous for the mutation. In such patient, a heterozygous c.2362AG>TCATCT deletion/insertion was recognized as the second CAPN3 mutation. Western blot and autocatalytic activity analyses in protein lysates from skeletal muscle biopsy obtained from a p.Ala116Asp homozygous patient suggested that this particular mutation increased the autocatalytic activity of CAPN3. Thirty eigth heterozygotes of the p.Ala116Asp mutation were identified among 401 genotyped unaffected villagers, yielding a population carrier frequency of 1 in 11. This study demonstrates that a cluster of patients with LGMD2A in a small Mexican village arises from a novel CAPN3 founder mutation. Evidence of allelic heterogeneity is demonstrated by the recognition of an additional CAPN3 mutation in a single affected. Our study provides an additional example of genetic isolation causing a high prevalence of LGMD and of successful molecular characterization of the disease by means of homozygosity mapping. The identification of a very high carrier frequency of the LGMD2-causing mutation has implications for more rational genetic counseling in this community.

La Asociación Iberoamericana de Educación Superior a Distancia (AIESAD) fue fundada en octubre de 1980 en Madrid, durante el I Simposio Iberoamericano de Rectores de Universidades Abiertas y a Distancia. Su objetivo fundacional fue el de aunar esfuerzos para lograr mejorar la educación superior a distancia y promover el intercambio entre las universidades abiertas y a distancia en Iberoamérica. Fue una de las primeras instituciones en nuestra región que tuvo una visión integradora y una proyección de trabajo cooperado en esta modalidad de estudios. Este año, la AIESAD arriba a su XXX aniversario, por lo que el objetivo de este trabajo es hacer un recuento en apretada síntesis de la significación histórica de esta Asociación en el desarrollo y calidad de la educación superior abierta y a distancia.

Antibiotic resistance is at dangerous levels and increasing worldwide. The search for new antimicrobial drugs to counteract this problem is a priority for health institutions and organizations, both globally and in individual countries. Sarconesiopsis magellanica blowfly larval excretions and secretions (ES) are an important source for isolating antimicrobial peptides (AMPs). This study aims to identify and characterize a new S. magellanica AMP. RP-HPLC was used to fractionate ES, using C18 columns, and their antimicrobial activity was evaluated. The peptide sequence of the fraction collected at 43.7 min was determined by mass spectrometry (MS). Fluorescence and electronic microscopy were used to evaluate the mechanism of action. Toxicity was tested on HeLa cells and human erythrocytes; physicochemical properties were evaluated. The molecule in the ES was characterized as sarconesin II and it showed activity against Gram-negative (Escherichia coli MG1655, Pseudomonas aeruginosa ATCC 27853, P. aeruginosa PA14) and Gram-positive (Staphylococcus aureus ATCC 29213, Micrococcus luteus A270) bacteria. The lowest minimum inhibitory concentration obtained was 1.9 μM for M. luteus A270; the AMP had no toxicity in any cells tested here and its action in bacterial membrane and DNA was confirmed. Sarconesin II was documented as a conserved domain of the ATP synthase protein belonging to the Fli-1 superfamily. The data reported here indicated that peptides could be alternative therapeutic candidates for use in infections against Gram-negative and Gram-positive bacteria and eventually as a new resource of compounds for combating multidrug-resistant bacteria.

Canopy characterization has become important when trying to optimize any kind of agricultural operation in high-growing crops, such as olive. Many sensors and techniques have reported satisfactory results in these approaches and in this work a 2D laser scanner was explored for measuring canopy trees in real-time conditions. The sensor was tested in both laboratory and field conditions to check its accuracy, its cone width, and its ability to characterize olive canopies in situ. The sensor was mounted on a mast and tested in laboratory conditions to check: (i) its accuracy at different measurement distances; (ii) its measurement cone width with different reflectivity targets; and (iii) the influence of the target’s density on its accuracy. The field tests involved both isolated and hedgerow orchards, in which the measurements were taken manually and with the sensor. The canopy volume was estimated with a methodology consisting of revolving or extruding the canopy contour. The sensor showed high accuracy in the laboratory test, except for the measurements performed at 1.0 m distance, with 60 mm error (6%). Otherwise, error remained below 20 mm (1% relative error). The cone width depended on the target reflectivity. The accuracy decreased with the target density.

This paper reports a case of gemination in a maxillary lateral incisor with two root canals and crown-root dilaceration. A 16-year-old male patient was referred for endodontic treatment of the maxillary left lateral incisor and evaluation of esthetic and functional complaints in the anterior region. The patient reported trauma to the anterior primary teeth. There was no spontaneous pain, but the tooth responded positively to the vertical percussion test and negatively to the pulp vitality test. Clinical examination showed esthetic and functional alterations and normal periodontal tissues. CBCT imaging confirmed the suspicion of gemination and crown-root dilaceration and also revealed the presence of two root canals and periapical bone rarefaction. The root canals were instrumented with Reciproc R40 and 1% NaOCl irrigation and were filled by lateral condensation of gutta-percha and AH Plus sealer. The tooth was definitely restored with composite resin to recover esthetics. Continued follow-up over 6 months has shown absence of pain or clinical alterations as well as radiographic image suggestive of apical repair.

Abstract The aim of this study was to evaluate the influence of drying protocols (DP) on the apical sealing (AS) and on the bond strength (BS) of teeth filled with different sealers. The root canals of one hundred and fifty-six roots of maxillary canines were prepared with Reciproc rotary files (R50). The teeth were randomly divided into four groups (n = 39), according to the DP: GI–paper points; GII–70% isopropyl alcohol + aspiration with NaviTip points; GIII–95% ethanol + paper points; GIV–EndoVac + paper points. Each group was divided into subgroups, according to the sealer used: AH Plus, Sealapex and MTA Fillapex, using a single-cone technique. Evaluation of AS and BS was performed with fluid filtration (FF) and push-out (PO) methods, respectively. The PO test consisted of sectioning the roots, and subjecting a single slice from each third to testing and analysis for failure type. The data was submitted to two-way and three-way variance analysis (ANOVA) and Tukey (α = 5%). The AS showed no drying protocol influence. The FF results revealed a statistically significant difference between MTA and Sealapex (p < 0.05) sealers. The BS test values showed that there was no statistical significant difference among the canal thirds (p > 0.05), but that there was such a difference among the sealers (p < 0.05), among the protocols (p < 0.05), and in the interaction between sealers and protocols (p < 0.05). AH Plus revealed the highest BS values among the sealers; the highest BS results for the sealers occurred with the specimens used with isopropyl alcohol, compared with ethanol and EndoVac.

Los riesgos de contaminación con Toxoplasma gondii a través de las transfusiones, están incrementados por la incidencia de este parásito en donantes y la tolerancia de este a los procesos de preparación y almacenamiento de la sangre, por lo que resulta peligroso para los grupos de riesgo. Esta enfermedad no es de declaración obligatoria en Cuba. El interés de conocer el comportamiento epidemiológico de esta parasitosis nos condujo a estudiar 562 muestras de donantes de sangre de la provincia de Guantánamo, donde resultaron positivos para IgG anti T. gondii el 47,0 %, con más del 62 % de positividad en los municipios de Baracoa y Maisí, con mayor prevalencia en áreas rurales. Los valores de seropositividad observados en los grupos etários estudiados y la relación entre sexo no fue significativa, aunque el contacto con el parásito en el grupo de edad entre 18 y 40 años fue superior. Estos datos seroepidemiológicos alertan sobre la necesidad de proponer acciones de control en el uso de hemoderivados en los grupos de riesgo.The risk of contamination with Toxoplasma gondii via blood transfusion is increased by the incidence of this parasite in donors and its tolerance to blood processing and storage processes, which turns it into a hazard for risk groups. This disease is not notifiable in Cuba. The interest in learning about the epidemiological behavior of this parasitosis led us to study 562 blood donor samples from the province of Guantánamo, 47.0 % of which tested positive for anti-T. gondii IgG, with over 62 % positivity in the municipalities of Baracoa and Maisí, and higher prevalence in rural areas. Neither the seropositivity values found in the age groups studied nor their sex distribution were significant, though contact with the parasite was higher in the 18-40 age group. These seroepidemiological data point to the need to propose control actions for the use of blood products in risk groups.

En el ensayo de microlinfocitotoxicidad, el suero de conejo como fuente de complemento desempeña un rol esencial en la clasificación del complejo mayor de histocompatibilidad humano, tanto para la tipificación antigénica como en el cross-match linfocitario de la pareja donante-receptor antes de realizar los trasplantes. En este trabajo, se obtuvieron 3 lotes experimentales de dicho hemoderivado con óptimos indicadores de rendimiento, actividad biológica y estabilidad. El exanguíneo de los animales se realizó por la técnica de yugulación. El suero fue separado por centrifugación en condiciones ambientales y de temperatura controladas. Posteriormente, el producto se sometió a filtración esterilizante y se tomaron muestras para los ensayos de calidad. El estudio mostró que el producto conserva la actividad biológica al menos durante 18 meses y que los valores de citotoxicidad se encuentran dentro de los límites de aceptación para su empleo en los ensayos de histocompatibilidad pretrasplante.Microlinfocitotoxicity assay is one of the serological methods used to classify human major histocompatibility complex. In this technique the rabbit´s serum as complement source, plays an essential role in antigens determination and in lymphocytes cross match assay as necessary stage before the selection of the best donor-receiver pair to realize the organ transplant. Three experimental lots of normal rabbit serum were developed with excellent indicators of efficiency, biological activity and stability. Rabbit's blood was obtained through jugulating technique and serum was separated by centrifugation in controlled environment and temperature´s conditions. The sterilization process was performed by filtration and the samples were taken for quality´s control assays. Stability studies showed that the product kept the biological activity at least during 18 months after it was processed and cytotoxicity's values were adequate for its employment in histocompatibility pre-transplant assays.