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OBJECTIVES/SPECIFIC AIMS: The Title V Cooperative Project of the UPR-MSC and UCC has demonstrated that educational interventions in CTR are very effective in fulfilling the objective of promoting awareness, stimulate interest and increase the knowledge, skills and opportunities, to US, GS and F (participants) in CTR. METHODS/STUDY POPULATION: The training sessions (TS) offered through the Title V initiative have become an engine for the involvement in CTR for participants from higher education institutions island-wide. TS consisted of cycles –level 1 and 2–: Research Education Towards Opportunities (RETO,I,II) and Mentorship Offering Training Opportunities for Research (MOTOR,I,II), ending in the formation of the Clinical and Translational Mentoring Teams (CTMT)s, in which participants, paired by their research interests, were mentored by a well-established CT researchers in their research project, to be developed in the Intensive Development and Experiences in Advancement of Research and Increased Opportunities (IDEARIO). RESULTS/ANTICIPATED RESULTS: Up to date, 4 TS-level 1 and 2 TS-level 2 were offered. Eighty (80) participants completed level 1, distributed: 42 (52.5%) US in RETO, 21 (26.25%) GS and 17 (21.25%) F in MOTOR and 17 participants completed level 2, distributed: 4 (23.52%) US in RETO, 6 (25.29%) GS and 7 (41.17%) F in MOTOR. From which, 15, with 8 CT researchers, formed 5 CTMTs in different research areas – cardio, neuro, liver, renal, Zika–. DISCUSSION/SIGNIFICANCE OF IMPACT: US, GS and F were integrated in the active process throughout educational levels for their development in CTR.
This is the first report on the development and characterization of eight monoclonal antibodies (MABs) generated against whole- and membrane-enriched tachyzoite extracts of the apicomplexan parasite Besnoitia besnoiti. Confocal laser scanning immunofluorescence microscopy was used to localize respective epitopes in B. besnoiti tachyzoites along the lytic cycle. A pattern compatible with dense granule staining was observed with MABs 2.A.12, 2.F.3 and 2.G.4, which could be confirmed by immunogold electron microscopy for MABs 2.A.12 and 2.F.3. In particular, MABs 2.F.3 and 2.G.4 were secreted during early invasion, proliferation and egress phases. MABs 3.10.8 and 5.5.11 labelled the tachyzoite surface, whilst MABs 1.17.8, 8.9.2 and 2.G.A recognized the apical tip, which is reminiscent for microneme localization. Besides, the epitopes recognized by the latter two (MABs 8.9.2 and 2.G.A) exhibited a redistribution from the anterior part across the parasite surface towards the posterior end during invasion. Most MABs developed were genus-specific. Indeed, the MABs cross-reacted neither with T. gondii nor with N. caninum tachyzoites. In summary, we have generated MABs that will be useful to study the key processes in the lytic cycle of the parasite and with additional promising diagnostic value. However, the molecular identity of the antigens recognized remains to be elucidated.
This study aimed to molecularly survey Bartonella in dogs from Chile. Quantitative real-time PCR (qPCR) for Bartonella spp. based on nuoG gene was performed in 139 blood samples taken from dogs belonging to rural localities of the Valdivia Province, Los Ríos region, southern Chile. nuoG qPCR-positive samples were submitted to conventional PCR assays for ftsZ, gltA, rpoB and nuoG genes and sequencing for speciation and phylogenetic analysis. Based upon qPCR results, Bartonella spp. occurrence in dogs was 4.3% (6/139). Out of six nuoG qPCR-positive samples, six, three, two and none showed positive results in cPCR assays based on gltA, ftsZ, rpoB and nuoG genes, respectively. Consistent sequencing results were obtained only for the ftsZ gene from sample #1532 (GeneBank accession number: MG252491), and gltA gene from samples #1535 (MG252490) and #1532 (148 bp fragment that was not deposited in GenBank). Phylogenetic analysis of ftsZ and gltA genes allowed speciation of two nuoG-positive samples, one as Bartonella vinsonii subsp. berkhoffii and the other as B. henselae. Bartonella vinsonii subsp. berkhoffii and B. henselae are detected for the first time in dogs from Chile, highlighting the importance of the canine population as a source of zoonotic agents and potential infection risk to humans.
Recent droughts in Puerto Rico and throughout the Caribbean have emphasized the region's agricultural vulnerability to this hazard and the increasing need for adaptation mechanisms to support sustainable production. In this study, we assessed the geographic extent of agricultural conservation practices incentivized by US Department of Agriculture Natural Resources Conservation Service (NRCS) and evaluated their large-scale contribution to drought adaptability. We identified concentrations of drought-related practices (e.g. cover crops, ponds) applied between 2000 and 2016. Using information from spatial databases and interviews with experts, we assessed the spatial correlation between these practices and areas exposed to drought as identified by the US Drought Monitor. Between 2000 and 2016, Puerto Rico experienced seven drought episodes concentrated around the south, east and southeastern regions. The most profound drought occurred between 2014 and 2016 when the island experienced 80 consecutive weeks of moderate drought, 48 of severe drought and 33 of extreme drought conditions. A total of 44 drought-related conservation practices were applied at 6984 locations throughout 860 km2 of farmlands between 2000 and 2016 through the NRCS-Environmental Quality Incentives Program (EQIP). Practices related to water availability were statistically clustered along the coasts, whereas soil and plant health practices were clustered in the mountainous region. While these concentrations strongly correlated with areas exposed to moderate drought conditions, >80% did not coincide with areas that experienced severe or extreme drought conditions, suggesting that areas highly exposed to drought conditions generally lacked drought preparedness assisted by EQIP. Climate projections indicate an increase in the frequency and intensity of drought events, particularly in the eastern region of Puerto Rico. Our analysis highlighted the need to implement more conservation practices in these areas subject to drought intensification and exposure. Government programs intended to address vulnerabilities and enhance capacity and resilience may not be reaching areas of highest exposure. Recommendations include raising producer awareness of past and future exposure and making programs more accessible to a broader audience.
The temporal correlation between neutron and hard X-ray (HXR) emissions from a hundred joules plasma focus device (PF-400J) was studied. A method, time history analysis, to estimate the time of origin of neutrons with respect to HXRs is applied. In most of the discharges, it was found that neutrons are originated before HXRs in the axial direction and after HXRs in the radial direction. In some discharges, the time difference between HXRs and neutrons origin was found large enough, so that it can be interpreted that those neutrons would have been originated before the pinch. A qualitative discussion is conjectured to explain the experimental observations.
Protected areas have been established historically in residual places where the potential for extractive uses is low, implying that places at risk are usually under-protected. Argentina is no exception, with few protected areas established in productive regions that are prone to conversion. Here, using reptiles as a study group and considering the most important human threats in north-western Argentina, we estimated priority conservation areas where we expect species to persist in the face of climate change and land conversion. Protected areas cover no more than 9% of the study region, but represent less than 15% of reptile distributions. There are great opportunities for improving the conservation status in the region by protecting only 8% more of north-western Argentina, with the level of species protection inside the protected area network increasing almost four-fold, reaching 43% of species distributions on average and 59% of the distributions of threatened reptiles. Fortunately, the highest diversity of reptiles in the region does not match the places targeted for agriculture expansion. Our findings suggest that future prioritization schemes should embrace other groups that are especially diverse in the Chaco ecoregion, which overlaps with our study area.
Salmonella is a leading cause of bacterial foodborne illness. We report the collaborative investigative efforts of US and Canadian public health officials during the 2013–2014 international outbreak of multiple Salmonella serotype infections linked to sprouted chia seed powder. The investigation included open-ended interviews of ill persons, traceback, product testing, facility inspections, and trace forward. Ninety-four persons infected with outbreak strains from 16 states and four provinces were identified; 21% were hospitalized and none died. Fifty-four (96%) of 56 persons who consumed chia seed powder, reported 13 different brands that traced back to a single Canadian firm, distributed by four US and eight Canadian companies. Laboratory testing yielded outbreak strains from leftover and intact product. Contaminated product was recalled. Although chia seed powder is a novel outbreak vehicle, sprouted seeds are recognized as an important cause of foodborne illness; firms should follow available guidance to reduce the risk of bacterial contamination during sprouting.
Benthic hydrozoans are one of the most speciose and characteristic taxa from the Antarctic region, with a high number of endemic species, but diversity at the genus level is low and some families with world wide distribution are unrepresented. This is the case of the family Aglaopheniidae. A new species to science of the genus Aglaophenia Lamouroux has been found in the eastern end of the Weddell Sea, at depths of 65–116 m, within the material obtained by the German Antarctic expedition ANT XV/3. This finding constitutes a new record for the Weddell Sea fauna, the first evidence of the genus for the Polar Regions, and even the family Aglaopheniidae from Antarctic waters. The material has been accurately examined and described. Literature concerning the species of Aglaophenia from the sub-Antarctic and other close areas has been reviewed and, as a result, a checklist of 20 species, with their corresponding distribution, is given. The material examined does not agree with any of the species and therefore it is described as a new species. Some possible scenarios for the presence of an aglaopheniid in Antarctic waters are discussed (e.g. alien species, relict, global climate change, microhabitat).
Investigating whether high-risk human papillomavirus (HR-HPV) types tend to become grouped in a particular way and whether factors are associated with such grouping is important for measuring the real impact of vaccination. In total, 219 women proving positive for HPV as detected by real-time PCR were included in the study. Each sample was analysed for detecting and quantifying six viral types and the hydroxymethylbilane synthase gene. Multiple correspondence analysis led to determining grouping patterns for six HR-HPV types and simultaneous association with multiple variables and whether viral load was related to the coexistence of other viral types. Two grouping profiles were identified: the first included HPV-16 and HPV-45 and the second profile was represented by HPV-31, HPV-33 and HPV-58. Variables such as origin, contraceptive method, births and pregnancies, educational level, healthcare affiliation regime, atypical squamous cells of undetermined significance and viral load were associated with these grouping profiles. Different socio-demographic characteristics were found when coinfection occurred by phylogenetically related HPV types and when coinfection was due to non-related types. Biological characteristics, the number of viral copies, temporality regarding acquiring infection and competition between viral types could influence the configuration of grouping patterns. Characteristics related to women and HPV, influence such interactions between coexisting HPV types reflecting the importance of their evaluation.
A major challenge in nanofabrication is the manipulation and exact placement of nano-objects on a specific template. Artificial DNA nanostructures such as DNA origami have garnered significant interest as templates for incorporating nanomaterials at precise sites while the structures are self-assembled. This work shows the usage of the DNA origami technique in the design and fabrication of nanostructures with the shapes of a circle and a triangle using the third part of the M13 virus genome, named mini-M13, as a scaffold. These DNA origami templates were modified to have DNA binding sites with a uniquely coded sequence. This method is used to attach 5 nm gold nanoparticles functionalized with the complementary DNA sequence. Two new metallic nanostructures with different nanoparticle arrays having minimum size but recognizable morphology are provided. The formation and dimensions of the nanostructures were verified using AFM and agarose gel electrophoresis.
To design and develop a questionnaire that can account for an individual’s adherence to a Mediterranean lifestyle including the assessment of diet and physical activity patterns, as well as social interaction.
The Mediterranean Lifestyle (MEDLIFE) index was created based on the current Spanish Mediterranean food guide pyramid. MEDLIFE is a twenty-eight-item derived index consisting of questions about food consumption (fifteen items), traditional Mediterranean dietary habits (seven items) and physical activity, rest and social interaction habits (six items). Linear regression models and Spearman rank correlation were fitted to assess content validity and internal consistency.
A subset of participants in the Aragon Workers’ Health Study cohort (Zaragoza, Spain) provided the data for development of MEDLIFE.
Participants (n 988) of the Aragon Workers’ Health Study cohort in Spain.
Mean MEDLIFE score was 11·3 (sd 2·6; range: 0–28), and the quintile distribution of MEDLIFE score showed a significant association with each of the individual items as well as with specific nutrients and lifestyle indicators (intra-validity). We also quantified MEDLIFE correspondence with previously reported diet quality indices and found significant correlations (ρ range: 0·44–0·53; P<0·001) for the Alternate Healthy Eating Index, the Alternate Mediterranean Diet Index and Mediterranean Diet Adherence Screener.
MEDLIFE is the first index to include an overall assessment of lifestyle habits. It is expected to be a more holistic tool to measure adherence to the Mediterranean lifestyle in epidemiological studies.
Understanding the dietary needs of syntopic species is essential for examining species coexistence and resource partitioning. We analysed stable isotopes of carbon (δ13C) and nitrogen (δ15N) to estimate the diet of two putative nectarivorous bats on Puerto Rico, the brown flower bat (Erophylla bombifrons) and the Greater Antillean long-tongued bat (Monophyllus redmani). Isotopic ratios of δ13C and δ15N were obtained from whole blood of both species of bat and tissues of available plant foods and insect prey over 15 wk at the same locality. We used a concentration-dependent Bayesian mixing model to determine probability distributions of feasible dietary contributions for major potential foods used by each species of bat. Additionally, separate dietary estimates were conducted for males and non-reproductive, pregnant and lactating females to determine differences due to reproductive condition. Insects were an important source of protein for M. redmani, whereas E. bombifrons obtained most of its protein from plants. In both species of bat, lactating females had lower assimilated nitrogen than males, suggesting more reliance on plants for protein. We observed no intraspecific differences in assimilated carbon among reproductive conditions. Flight and lactation are energetically expensive and may explain the increased consumption of high-energy foods, such as fruit or nectar, in lactating female bats. Comparison of isotopes between E. bombifrons and M. redmani illustrate the differential use of food resources by these insular syntopic species of bat.
To assess the agreement between self-reported and parent-reported dietary and physical activity habits in children; and to evaluate the socio-economic determinants of healthier habits (Mediterranean diet and physical activity) among children.
Cross-sectional analysis of children recruited to a cluster-randomized controlled trial (Program SI!). Information about children’s and parents’ dietary and physical activity habits was obtained through validated questionnaires (Program SI! questionnaires, Kidmed, Krece Plus and Predimed scores).
Twenty-four schools in Madrid, Spain.
Children (n 2062) aged 3–5 years and their parents (n 1949).
There was positive agreement between parental- and self-reporting for three of the six children’s habits examined. Parents’ dietary and physical activity patterns were associated with those of their children. The main determinants of higher scores in children were higher parental age, the mother’s scores, Spanish origin and higher awareness of human health (P<0·005). Children from parents with a low educational level had lower odds for scoring positively on items such as using olive oil (OR=0·23; 95 % CI 0·13, 0·41) and not skipping breakfast (OR=0·36; 95 % CI 0·23, 0·55), but higher odds for meeting the recommendations for consuming pulses (OR=1·71; 95 % CI 1·14, 2·55). Other habits being influenced by parental socio-economic status included the consumption of vegetables, fish, nuts, avoidance of fast food, and consumption of bakery products for breakfast.
Children’s habits may be influenced by their parents’ health awareness and other socio-economic characteristics. These findings suggest that intervention strategies, even in very young children, should also target parents in order to achieve maximum success.
Free-living amoebae belonging to the genus Acanthamoeba are the causative agents of infections such as amoebic keratitis (AK), granulomatous amoebic encephalitis (GAE) and cutaneous lesions. The mechanisms involved in the establishment of infection are unknown. However, it is accepted that the initial phase of pathogenesis involves adherence to the host tissue. In this work, we analysed surface molecules with an affinity for epithelial and neuronal cells from the trophozoites of Acanthamoeba castellanii. We also investigated the cellular mechanisms that govern the process of trophozoite adhesion to the host cells. We first used confocal and epifluorescence microscopy to examine the distribution of the A. castellanii actin cytoskeleton during interaction with the host cells. The use of drugs, as cytochalasin B (CB) and latrunculin B (LB), revealed the participation of cytoskeletal filaments in the adhesion process. In addition, to identify the proteins and glycoproteins on the surface of A. castellanii, the trophozoites were labelled with biotin and biotinylated lectins. The results revealed bands of surface proteins, some of which were glycoproteins with mannose and N-acetylglucosamine residues. Interaction assays of biotinylated amoebae proteins with epithelial and neuronal cells showed that some surface proteins had affinity for both cell types. The results of this study provide insight into the biochemical and cellular mechanisms of the Acanthamoeba infection process.
The induced diffusion of tracers in a bacterial suspension is studied theoretically and experimentally at low bacterial concentrations. Considering the swimmer–tracer hydrodynamic interactions at low Reynolds number and using a kinetic theory approach, it is shown that the induced diffusion coefficient is proportional to the swimmer concentration, their mean velocity and a coefficient
, as observed experimentally. This paper shows that
increases as a result of the interaction with solid surfaces. The coefficient
scales as the tracer–swimmer cross-section times the mean square displacement produced by single scattering events, which depends on the swimmer propulsion forces. Considering simple swimmer models (acting on the fluid as two monopoles or as a force dipole), it is shown that
increases for decreasing swimming efficiencies. Close to solid surfaces, the swimming efficiency degrades and, consequently, the induced diffusion increases. Experiments on wild-type Escherichia coli in a Hele-Shaw cell, under buoyant conditions, are performed to measure the induced diffusion on tracers near surfaces. The modification of the suspension pH varies the swimmers’ velocity over a wide range, allowing the
coefficient to be extracted with precision. It is found that solid surfaces modify the induced diffusion: decreasing the confinement height of the cell,
increases by a factor of 4. The theoretical model reproduces this increase, although there are quantitative differences, probably attributed to the simplicity of the swimmer models and to the estimates for the parameters that model E. coli.