Molecular detection of Babesia spp. and Rickettsia spp. in coatis (Nasua nasua) and associated ticks from midwestern Brazil.

Procyonids are reservoirs of many zoonotic infectious diseases, including tick-borne pathogens. The role of coatis (Nasua nasua) in the epidemiology of piroplasmids and Rickettsia has not been fully addressed in Brazil. To molecularly study these agents in coatis and associated ticks, animals were sampled in two urban areas in Midwestern Brazil. Blood (n = 163) and tick (n = 248) DNA samples were screened by PCR assays targeting the 18S rRNA and gltA genes of piroplasmids and Rickettsia spp., respectively. Positive samples were further molecularly tested targeting cox-1, cox-3, ß-tubulin, cytB, and hsp70 (piroplasmid) and ompA, ompB, and htrA 17-kDa (Rickettsia spp.) genes, sequenced and phylogenetically analyzed. All coatis' blood samples were negative for piroplasmids, whereas five pools of ticks (2%) were positive for two different sequences of Babesia spp.. The first from Amblyomma sculptum nymphs was close (i.e., ≥ 99% nucleotide identity) to a Babesia sp. previously found in capybaras (Hydrochoerus hydrochaeris); the second from Amblyomma dubitatum nymphs and Amblyomma spp. larvae was identical (100% nucleotide identity) to a Babesia sp. detected in opossums (Didelphis albiventris) and associated ticks. Four samples (0.8%) were positive by PCR to two different Rickettsia spp. sequences, being the first from Amblyomma sp. larva identical to Rickettsia belli and the second from A. dubitatum nymph identical to Rickettsia species from Spotted Fever Group (SFG). The detection of piroplasmids and SFG Rickettsia sp. highlights the importance of Amblyomma spp. in the maintenance of tick-borne agents in urban parks where humans and wild and domestic animals are living in sympatry.

Perceptions of change in the environment caused by the COVID-19 pandemic: Implications for environmental policy.

COVID-19 lockdown measures have impacted the environment with both positive and negative effects. However, how human populations have perceived such changes in the natural environment and how they may have changed their daily habits have not been yet thoroughly evaluated. The objectives of this work were to investigate (1) the social perception of the environmental changes produced by the COVID-19 pandemic lockdown and the derived change in habits in relation to i) waste management, energy saving, and sustainable consumption, ii) mobility, iii) social inequalities, iv) generation of noise, v) utilization of natural spaces, and, vi) human population perception towards the future, and (2) the associations of these potential new habits with various socio-demographic variables. First, a SWOT analysis identified strengths (S), weaknesses (W), opportunities (O), and threats (T) generated by the pandemic lockdown measures. Second, a survey based on the aspects of the SWOT was administered among 2370 adults from 37 countries during the period from February to September 2021. We found that the short-term positive impacts on the natural environment were generally well recognized. In contrast, longer-term negative effects arise, but they were often not reported by the survey participants, such as greater production of plastic waste derived from health safety measures, and the increase in e-commerce use, which can displace small storefront businesses. We were able to capture a mismatch between perceptions and the reported data related to visits to natural areas, and generation of waste. We found that age and country of residence were major contributors in shaping the survey participants ´answers, which highlights the importance of government management strategies to address current and future environmental problems. Enhanced positive perceptions of the environment and ecosystems, combined with the understanding that livelihood sustainability, needs to be prioritized and would reinforce environmental protection policies to create greener cities. Moreover, new sustainable jobs in combination with more sustainable human habits represent an opportunity to reinforce environmental policy.

Molecular detection of zoonotic filarioids in Culex spp. from Portugal.

To investigate the role of dipterans in the transmission of Onchocerca lupi and other zoonotic filarioids, samples were collected from different sites in Algarve, southern Portugal, morphologically identified and molecularly tested for filarioids. Culex sp. (72.8%) represented the predominant genus followed by Culicoides sp. (11.8%), Ochlerotatus sp. (9.7%), Culiseta sp. (4.5%), Aedes sp. (0.9%) and Anopheles sp. (0.3%). Nineteen (2.8%) specimens scored positive for filarioids, with Culex pipiens quinquefasciatus (2%) positive for Dirofilaria immitis (1.4%), Dirofilaria repens, Acanthocheilonema reconditum, Onchocerca lupi, unidentified species of Filarioidea (0.2%, each) and Onchocercidae (0.6%). Additionally, Culiseta longiareolata (6.5%), Ochlerotatus caspius (3%) and Culex laticinctus (0.2%) scored positive for unidentified Onchocercidae, A. reconditum and for O. lupi, respectively. This is the first report of the occurrence of DNA of O. lupi, D. repens and A. reconditum in Culex spp. in Portugal. Information regarding the vectors and the pathogens they transmit may help to adopt proper prophylactic and control measures.

Unipolar resistive switching behavior in Al2O3/HfO2 multilayer dielectric stacks: fabrication, characterization and simulation.

In this work, the impact of different HfO2/Al2O3-based multilayer dielectric stack (DS) configurations on the electrical characteristics and on the resistive switching (RS) performance of Ni/Insulator/Silicon devices has been systematically investigated. Significant differences are observed in the electrical characteristics of the fabricated bilayer, trilayer and pentalayer stacks compared to a single HfO2 layer of the same physical thickness. The RS analysis has shown similar low resistance state currents and set voltages for all the DS combinations whereas currents at the high resistance state and reset voltages depend on the DS. The shift of the reset voltage to lower values for HfO2 and Al2O3/HfO2/Al2O3 cases is explained by the results from thermal simulations that reveal that these differences could be associated to the different temperature distributions at the narrowest part of the conductive filament immediately before the thermally triggered reset process occurs.

Molecular detection of Leishmania (Sauroleishmania) tarentolae in human blood and Leishmania (Leishmania) infantum in Sergentomyia minuta: unexpected host-parasite contacts.

The detection of atypical Kinetoplastida in vertebrate hosts and vectors might suggest unexpected host-parasite contacts. Aside to major vectors of Leishmania (Leishmania) infantum in Italy (e.g. Phlebotomus perniciosus and Phlebotomus perfiliewi), the sand fly fauna also includes Sergentomyia minuta, herpetophilic and proven vector of Leishmania (Sauroleishmania) tarentolae, in which records of blood meal on mammals and detection of L. infantum DNA are increasing. This study was conducted in Central Italy aiming to molecularly detect potential atypical Leishmania host-vector contacts. Detection of Leishmania spp. DNA was performed by polymerase chain reaction (SSU rRNA, ITS1 targets) on field-collected sand fly females (N = 344), blood samples from humans (N = 185) and dogs (N = 125). Blood meal identification was also performed on engorged sand flies. Leishmania spp. DNA was found in 13.1% sand flies, 3.7% humans and 14.4% dogs. Sequence analysis identified L. infantum in S. minuta (4.4%), P. perniciosus (9.1%), humans (2.2%) and dogs (14.4%). Leishmania tarentolae was detected in S. minuta (12.6%), P. perfiliewi (6.6%) and human (1.6%) samples. Of 28 S. minuta examined for blood meal, 3.6 and 21.4% scored positive for human and lizard DNA, respectively. These results indicate the importance of one-health approach to explore new potential routes of transmission of leishmaniasis involving S. minuta.

Application of embryo biopsy and sex determination via polymerase chain reaction in a commercial equine embryo transfer program in Argentina.

Embryo biopsy for fetal sexing has clinical application, but few reports are available of its use within an active embryo transfer program. We evaluated results on biopsy of 459 embryos over one breeding season. There were no significant differences in pregnancy rate between biopsied and non-biopsied embryos (72% vs 73%) or for biopsied embryos recovered at the centre (73%) compared with those shipped overnight (72%). However, the pregnancy rate decreased significantly in shipped embryos biopsied ≥20h after collection. Overall, 86% of biopsies provided a sex diagnosis. The likelihood of a positive genomic (g) DNA result was significantly higher for biopsies from large blastocysts (96%) than from smaller embryos (70-85%). In total, 38% of biopsies were positive for Y chromosome DNA (Y-DNA) and were diagnosed as male. Subsequently, 95% of Y-DNA-positive embryos were confirmed as male and 78% of Y-DNA-negative embryos were confirmed as female. The accuracy of prediction of female (Y-DNA negative) was significantly higher when the biopsy sample was probed for Y-DNA only compared with probing for both gDNA and Y-DNA. We estimate that by transferring only Y-DNA-negative embryos, 3% of potential female pregnancies may have been lost, and production of male pregnancies was reduced by 72%.

Insecticide susceptibility status in Mexican populations of Stegomyia aegypti (= Aedes aegypti): a nationwide assessment.

In Mexico, mosquito vector-borne diseases are of public health concern as a result of their impact on human morbidity and mortality. The use of insecticides against adult mosquitoes is one of the most common ways of controlling mosquito population densities. However, the use of these compounds has resulted in the development of insecticide resistance. The aim of this study was to estimate susceptibility to six pyrethroids, two carbamates and two organophosphates in Mexican populations of Stegomyia aegypti (Linnaeus, 1762) (= Aedes aegypti) (Diptera: Culicidae) mosquitoes. Bottle insecticide susceptibility tests, with 1 h exposure, were performed on adult mosquitoes from 75 localities across 28 states. At 30 min of exposure, the proportion of fallen mosquitoes was recorded. After 60 min of exposure, mosquitoes were recovered in non-treated containers and mortality was determined at 24 h after the set-up of the experiment. In general, the carbamate insecticides represented the most effective group in terms of the proportion of mosquitoes fallen at 30 min (72-100%) and 24-h mortality (97-100%). High and widespread resistance to pyrethroids Types I and II and, to a lesser extent, to organophosphates was observed. Insecticide susceptibility among and within states was highly variable.

First report of Stegomyia aegypti (= Aedes aegypti) in Mexico City, Mexico.

Stegomyia aegypti (= Aedes aegypti) (Diptera: Culicidae) is a species of mosquito that is currently widespread in Mexico. Historically, the mosquito has been distributed across most tropical and subtropical areas lower than 1700 m a.s.l. Currently, populations that are found at higher altitudes in regions with cold and dry climates suggest that these conditions do not limit the colonization and population growth of S. aegypti. During a survey of mosquitoes in September 2015, larvae of S. aegypti mosquitoes were found in two different localities in Mexico City, which is located at about 2250 m a.s.l. Mexico City is the most populous city in Mexico and has inefficient drainage and water supply systems. These factors may result in the provision of numerous larval breeding sites. Mosquito monitoring and surveillance are now priorities for the city.

The flexibility and dynamics of protein disulfide isomerase.

We have studied the mobility of the multidomain folding catalyst, protein disulfide isomerase (PDI), by a coarse-graining approach based on flexibility. We analyze our simulations of yeast PDI (yPDI) using measures of backbone movement, relative positions and orientations of domains, and distances between functional sites. We find that there is interdomain flexibility at every interdomain junction but these show very different characteristics. The extent of interdomain flexibility is such that yPDI's two active sites can approach much more closely than is found in crystal structures-and indeed hinge motion to bring these sites into proximity is the lowest energy normal mode of motion of the protein. The flexibility predicted for yPDI (based on one structure) includes the other known conformation of yPDI and is consistent with (i) the mobility observed experimentally for mammalian PDI and (ii) molecular dynamics. We also observe intradomain flexibility and clear differences between the domains in their propensity for internal motion. Our results suggest that PDI flexibility enables it to interact with many different partner molecules of widely different sizes and shapes, and highlights considerable similarities of yPDI and mammalian PDI. Proteins 2016; 84:1776-1785. © 2016 Wiley Periodicals, Inc.

Characterization of the autoimmune response against the nerve tissue S100ß in patients with type 1 diabetes.

Type 1 diabetes results from destruction of insulin-producing beta cells in pancreatic islets and is characterized by islet cell autoimmunity. Autoreactivity against non-beta cell-specific antigens has also been reported, including targeting of the calcium-binding protein S100ß. In preclinical models, reactivity of this type is a key component of the early development of insulitis. To examine the nature of this response in type 1 diabetes, we identified naturally processed and presented peptide epitopes derived from S100ß, determined their affinity for the human leucocyte antigen (HLA)-DRB1*04:01 molecule and studied T cell responses in patients, together with healthy donors. We found that S100ß reactivity, characterized by interferon (IFN)-γ secretion, is a characteristic of type 1 diabetes of varying duration. Our results confirm S100ß as a target of the cellular autoimmune response in type 1 diabetes with the identification of new peptide epitopes targeted during the development of the disease, and support the preclinical findings that autoreactivity against non-beta cell-specific autoantigens may have a role in type 1 diabetes pathogenesis.

Reliability of texture analysis using co-occurrence matrices (glcm) on photographic image in the assessment of cellulite in a Spanish population.

BACKGROUND: Despite the existence of diverse instruments to assess cellulite, its high prevalence and the continuous advances in its treatment makes the development of new, more objective methods for evaluation necessary. AIM: To study intraobserver validity and reliability (test-retest) of textural analysis using co-occurrence matrices on photographic images in the evaluation of cellulite in a Spanish population and its possible relationship to the degree of cellulite. METHODS: Twenty-seven women were selected for this reliability study (mean age 26.41 SD = 6.16). Digital photographs were taken under standardized conditions in contraction and relaxation of the femoral gluteus region. The areas of interest of the photographs were selected at two different times a month apart. Textural parameters studied were energy (ASM), entropy, contrast, Homogeneity (IDM) and textural correlation. Reliability was analysed by the intraclass correlation coefficient (ICC). Differences between laterality and between contraction and relaxation were performed by analysis of variance for repeated measurements. Correlation between Cellulite Severity Scale (CSS) and the textural parameters by means of the Pearson correlation coefficient was studied. CSS was re-coded to a binary variable, performing a differentiate analysis for each laterality with this variable and the textural parameters. RESULTS: In the intraobserver reliability analysis ICC was high (≥0.80) in seven parameters and excellent (≥0.90) in 35 parameters. In general, CSS and textural parameters showed more cellulite severity in right areas than in left ones. Correlation coefficients showed a moderate correlation between textural parameters and CSS score. The multivariate discriminant model obtained with textural parameters classified a high percentage of images (96% right side and 82% left side). CONCLUSION: Textural analysis used to assess cellulite on the backs of thighs and buttocks proved to be an instrument that has excellent reliability, moderate correlation with CSS and a satisfactory power classification model.

Variability in HfO2-based memristors described with a new bidimensional statistical technique.

A new statistical analysis is presented to assess cycle-to-cycle variability in resistive memories. This method employs two-dimensional (2D) distributions of parameters to analyse both set and reset voltages and currents, coupled with a 2D coefficient of variation (CV). This 2D methodology significantly enhances the analysis, providing a more thorough and comprehensive understanding of the data compared to conventional one-dimensional methods. Resistive switching (RS) data from two different technologies based on hafnium oxide are used in the variability study. The 2D CV allows a more compact assessment of technology suitability for applications such as non-volatile memories, neuromorphic computing and random number generation circuits.

3D simulation of conductive nanofilaments in multilayer h-BN memristors via a circuit breaker approach.

A 3D simulation of conductive nanofilaments (CNFs) in multilayer hexagonal-BN memristors is performed. To do so, a simulation tool based on circuit breakers is developed including for the first time a 3D resistive network. The circuit breakers employed can be modeled with two, three and four resistance states; in addition, a series resistance and a module to account for quantum effects, by means of the quantum point contact model, are also included. Finally, to describe real dielectric situations, regions with a high defect density are modeled with a great variety of geometrical shapes to consider their influence in the resistive switching (RS) process. The simulator has been tuned with measurements of h-BN memristive devices, fabricated with chemical-vapour-deposition grown h-BN layers, which were electrically and physically characterized. We show the formation of CNFs that produce filamentary charge conduction in our devices. Moreover, the simulation tool is employed to describe partial filament rupture in reset processes and show the low dependence of the set voltage on the device area, which is seen experimentally.

Inhibition of HIV-1 protease: the rigidity perspective.

MOTIVATION: HIV-1 protease is a key drug target due to its role in the life cycle of the HIV-1 virus. Rigidity analysis using the software First is a computationally inexpensive method for inferring functional information from protein crystal structures. We evaluate the rigidity of 206 high-resolution (2 Å or better) X-ray crystal structures of HIV-1 protease and compare the effects of different inhibitors binding to the enzyme. RESULTS: Inhibitor binding has little effect on the overall rigidity of the protein homodimer, including the rigidity of the active site. The principal effect of inhibitor binding on rigidity is to constrain the flexibility of the ß-hairpin flaps, which move to allow access to the active site of the enzyme. We show that commercially available antiviral drugs which target HIV-1 protease can be divided into two classes, those which significantly affect flap rigidity and those which do not. The non-peptidic inhibitor tipranavir is distinctive in its consistently strong effect on flap rigidity. CONTACT: jack.heal@warwick.ac.uk; r.roemer@warwick.ac.uk SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Cellulite's aetiology: a review.

Cellulite, highly prevalent among women, represents a serious problem for many of them, and one of their main aesthetic concerns. It is difficult to pinpoint its aetiology and physiology/pathophysiology, as there are many factors that are involved in it, affect it, and many processes that are taking place simultaneously and sequentially. Our objective is therefore, to review the scientific scholarship on cellulite to explore the causes of its origin. We carried out a preliminary search of the Medline, Cochrane, and Web of Knowledge databases covering the period from 1978 to April 2011. As there is no specific key word for the phenomenon at hand, we used the following descriptors: adipose tissue, subcutaneous fat, subcutaneous tissue, connective tissue, skin, skin disease and dermis. This resulted in a retrieval of 26 articles contributing to relevant information on the aetiology of cellulite. As a result of our first research, we concluded that cellulite is a physiological phenomenon or at least, that it has a physiological origin, which is characteristic of women, and multi-causal, with the coexistence of a number of factors that trigger, perpetuate, or exacerbate it. The outstanding factors include, among others, connective tissue architecture, oestrogen action, microvascular alterations and certain genetic and hormonal characteristics. All of them provide us with future and novel clues to cellulite treatment, and is necessary to take some or all of these factors into account in developing an effective therapy. However, we are aware of the necessity of further investigation in this field.

Intra- and inter-observer reliability of the application of the cellulite severity scale to a Spanish female population.

BACKGROUND: 'Hexsel, dal'Forno and Hexsel Cellulite Severity Scale' (CSS) was developed to evaluate cellulite with an objective and easy to apply tool. Objective Study CSS intra- and inter-observer reliability in a Spanish female population by evaluating patients' cellulite through photographs of their overall gluteofemoral zone as opposed to its creators who distinguished between buttocks and thigh. METHODS: Cellulite Severity Scale was applied to 27 women, evaluating gluteofemoral cellulite, differentiating between left and right. Evaluations were made by three expert examiners each at three times with a 1-week separation. Variables were the five CSS dimensions (number of evident depressions; depth of depressions; morphological appearance of skin surface alterations; grade of laxity, flaccidity, or sagging skin; and the Nürnberger and Müller classification scale), and the overall CSS score. Cronbach's alpha, intra-class correlation and item total correlation were analysed. RESULTS: Cronbach's alpha values were 0.951 (right) and 0.944 (left). In the intra-observer reliability analysis, intra-class correlation coefficient ranged from 0.993 to 0.999 (P < 0.001) and in the inter-observer analysis were 0.937 (right) and 0.947 (left) (P < 0.001). Item total correlation showed all dimensions to be needed except grade of laxity, flaccidity or sagging skin (0.959 right; 0.955 left). CONCLUSION: Cellulite Severity Scale has excellent reliability and internal consistency when used to evaluate cellulite on the buttocks and back of the thighs considered together. Nevertheless, the dimension grade of laxity, flaccidity or sagging skin does not contribute positively to the final consistency of the scale. This dimension needs to be analysed in greater depth in future studies.

Organic law regulating euthanasia: knowledge and involvement of doctors in Spain one year after its implementation.

BACKGROUND AND AIMS: The Organic Law Regulating Euthanasia (LORE, for its initials in Spanish) came into force in June 2021. This study aims to examine knowledge of the LORE among physicians licensed in Spain as well as their involvement with and the impact of the law. METHODS: This work is a descriptive, cross-sectional study conducted by means of a survey. Information was gathered through a self-administered questionnaire. RESULTS: The survey was answered by 1446 physicians licensed in Spain. The samples' demographic characteristics were as follows: 54.7% were women, the mean age was 52 ±â€¯14 years, and 66.0% worked in a hospital. Catalonia was the autonomous community with the most participants (44.6%). Regarding specialties, anesthesiology and resuscitation had the highest number of participants (21.9%), followed by family and community medicine (18.5%). The LORE was known in detail by 24.3% of physicians, 58.0% had a positive opinion of it, and 31.1% had direct experience with the euthanasia procedure. Practitioners working in the hospital setting perceived the law more favorably compared to those in the primary care setting (62.3% vs. 47.3%, p < 0.01). CONCLUSIONS: Most doctors did not have in-depth knowledge of the LORE, although a majority supported its existence, particularly those in hospital medicine. Most physicians who viewed the LORE negatively were male, older, and worked in primary care. A minority of physicians considered registering as conscientious objectors.

Mansonella sp. and associated Wolbachia endosymbionts in ring-tailed coatis (Nasua nasua) in periurban areas from Midwestern Brazil.

Coatis (Nasua nasua) are wild carnivorous well adapted to anthropized environments especially important because they act as reservoirs hosts for many arthropod-borne zoonotic pathogens. Information about filarioids from coatis and associated Wolbachia spp. in Brazil is scant. To investigate the diversity of filarial nematodes, blood samples (n = 100 animals) were obtained from two urban areas in midwestern Brazil and analyzed using blood smears and buffy coats and cPCR assays based on the cox1, 12S rRNA, 18S rRNA, hsp70 and myoHC genes for nematodes and 16S rRNA for Wolbachia. When analyzing coati blood smears and buffy coats, 30% and 80% of the samples presented at least one microfilaria, respectively. Twenty-five cox1 sequences were obtained showing 89% nucleotide identity with Mansonella ozzardi. Phylogenetic analyses clustered cox1 sequences herein obtained within the Mansonella spp. clade. Sequences of both myoHC and two hsp70 genes showed 99.8% nucleotide identity with Mansonella sp. and clustered into a clade within Mansonella sp., previously detected in coatis from Brazil. Two blood samples were positive for Wolbachia, with a 99% nucleotide identity with Wolbachia previously found in Mansonella perstans, Mansonella ozzardi and Mansonella atelensis and in ectoparasites of the genus Pseudolynchia, Melophagus and Cimex. The study showed a high prevalence of Mansonella sp. in the coati population examined, suggesting that this animal species play a role as reservoirs of a novel, yet to be described, species within the Onchocercidae family.