Epidemiology of Neospora caninum and Toxoplasma gondii infection in venison from Aguascalientes, Mexico.

The objective of the study was to estimate the seroprevalence of Neospora caninum and Toxoplasma gondii in venison from Aguascalientes, Mexico, their possible association with some risk factors, and to identify the presence of parasite DNA in blood and tissues. For this study, 5 farms and four species of venison were included, where 43 blood serum samples were obtained and in 37 of these animals a peripheral blood sample was also obtained; from hunted deer, 6 liver and 2 heart samples were obtained. The samples were analyzed by ELISA and PCR tests, respectively. The association between the serological status and the possible risk factors was estimated. The overall seroprevalence in N. caninum was 47% (20/43; CI 95% 31-62), with positive animals in all farms in a range of 18 to 100%, while for T. gondii it was 49% (21/43; CI 95% 33-64), with positive animals in 80% of farms in a range of 18 to 100%. The prevalence of N. caninum DNA detection in blood was 59% (22/37; CI 95% 42-74), with positive animals in all farms, in a range of 45 to 100%, while in T. gondii it was 76% (28/37; CI 95% 58-87), with positive animals in all farms, in a range of 56 to 100%. Age (> 4 years) was identified as associated with seroprevalence in N. caninum (OR 5.2) and in T. gondii (OR 12.7). DNA from both parasites was detected in the liver and heart samples. The results shown that venison populations included in the study are living in an environment highly contaminated with oocysts excreted by the definitive host.

EPIDEMIOLOGY OF TOXOPLASMA GONDII AND NEOSPORA CANINUM INFECTION IN GOATS FROM AGUASCALIENTES, MEXICO.

The objective of this study was to describe the epidemiology of Toxoplasma gondii and Neospora caninum infection by estimating seroprevalence and its association with certain risk factors in goats from Aguascalientes, Mexico. A total of 150 blood samples was taken from 10 farms and serum samples were subjected to enzyme-linked immunosorbent assay indirect test to detect T. gondii and N. caninum antibodies; the association between seroprevalence and some potential risk factors was estimated through logistic regression analysis. The general seroprevalence for T. gondii was 12.6%, observed in the farms in a range of 6.6 to 60%, finding seropositive animals in 80% of them; for N. caninum the seroprevalence was 3.3% and in farms a range of 6.6 to 13.3% was identified and 30% of them had at least 1 seropositive animal. The coinfection was 0.66%. The risk analysis for T. gondii identified a history of abortions (odds ratio 9.25) as a factor associated with seroprevalence; for N. caninum, no risk factor was identified.

Determination of the discriminant doses to identify resistance to fipronil, flumethrin and coumaphos, in Rhipicephalus sanguineus s.l. (Acari: Ixodidae) from Mexico.

Rhipicephalus sanguineus s.l. is an intra- and peridomiciliary tick of medical and veterinary importance, whose control is carried out through the application of various ixodicides, to which it can develop resistance. The objective of this work was to determine the discriminant doses (d.d.) to identify resistance against three ixodicides - fipronil, flumethrin and coumaphos - in R. sanguineus s.l. larvae originating from Mexico. Engorged ticks were collected from naturally infested dogs residing in rural communities in five Mexican states; the dogs had no history of recent treatment with ixodicides. All ticks were identified as R. sanguineus s.l. They were incubated for 25 days and their progeny was used in the larval package test (LPT) to be exposed to d.d. as determined in Rhipicephalus microplus, and those that were susceptible were analyzed using the LPT with six concentrations, for fipronil from 0.1 to 0.003125%, for flumethrin from 0.02 to 0.000625% and for coumaphos from 0.4 to 0.0125%. Mortality was analyzed with Probit methodology, to calculate the 50 and 99% lethal concentrations (LC50 and LC99). The d.d. was determined as a consensus value by multiplying the LC99 value × 2. Nine double susceptible samples (fipronil + flumethrin) and one triple susceptible sample (fipronil + flumethrin + coumaphos) were found; the d.d. determined were fipronil 0.05%, flumethrin 0.01% and coumaphos 0.43%. These doses can be used to rapidly and inexpensively identify resistant populations in samples collected in the field.

Seroprevalence of Toxoplasma gondii Infection and Associated Risk Factors in Different Populations of Dogs from Aguascalientes, Mexico.

CONTEXT AND PURPOSE: This work aimed to study the seroprevalence of natural infection by T. gondii in different population segments of dogs from Aguascalientes, Mexico as well as its possible association with some risk factors. METHODS: The study included 210 clinically healthy dogs from three population segments: rural, stray and pet. A blood serum sample was collected on a single occasion and processed by indirect immunofluorescence considering a dilution of 1:16 as a positive reaction; the association between the presence of antibodies and the potential risk factors was estimated with logistic regression. RESULTS: The overall prevalence of anti-T. gondii antibodies in the studied population was 59% (123/210; CI 95% 51-65) being different between population segments, in rural dogs 74% (59/80; CI 95% 62-82), in stray dogs 60% (48/80; CI 95% 48-70) and in pet dogs 32% (16/50; CI 95% 19-46), the titers of the positive sera were in a range of 1:16-1:128. In rural dogs the seroprevalence according to the municipality was in a range of 55% to 100% finding positive dogs in all the studied municipalities. The zootechnical function of guarding the property in rural dogs was identified as a risk factor (OR 2.4), while in pet dogs it was living with cats (OR 7) as well as sharing the drinking water container (OR 4); in stray dogs it was not possible to identify any risk factor. CONCLUSIONS: This study documents the wide presence of anti-T. gondii antibodies in the population groups examined, this being more important in rural dogs.

Role of miR-15a-5p and miR-199a-3p in the inflammatory pathway regulated by NF-κB in experimental and human atherosclerosis.

BACKGROUND: Cardiovascular diseases (CVDs) prevalence has significantly increased in the last decade and atherosclerosis development is the main trigger. MicroRNAs (miRNAs) are non-coding RNAs that negatively regulate gene expression of their target and their levels are frequently altered in CVDs. METHODS: By RT-qPCR, we analysed miR-9-5p, miR-15a-5p, miR-16-5p and miR-199a-3p levels in aorta from apolipoprotein knockout (ApoE-/- ) mice, an experimental model of hyperlipidemia-induced atherosclerosis, and in human aortic and carotid atherosclerotic samples. By in silico studies, Western blot analysis and immunofluorescence studies, we detected the targets of the altered miRNAs. RESULTS: Our results show that miR-15a-5p and miR-199a-3p are significantly decreased in carotid and aortic samples from patients and mice with atherosclerosis. In addition, we found an increased expression in targets of both miRNAs that participate in the inflammatory pathway of nuclear factor kappa B (NF-κB), such as IKKα, IKKß and p65. In human vein endothelial cells (HUVECs) and vascular smooth muscle cells (VSMCs), the overexpression of miR-15a-5p or miR-199a-3p decreased IKKα, IKKß and p65 protein levels as well as NF-κB activation. On the other hand, miR-15a-5p and miR-199a-3p overexpression reduced ox-LDL uptake and the inflammation regulated by NF-κB in VSMCs. Moreover, although miR-15a-5p and miR-199a-3p were significantly increased in exosomes from patients with advanced carotid atherosclerosis, only in the ROC analyses for miR-15a-5p, the area under the curve was 0.8951 with a p value of .0028. CONCLUSIONS: Our results suggest that the decrease of miR-199a-3p and miR-15a-5p in vascular samples from human and experimental atherosclerosis could be involved in the NF-κB activation pathway, as well as in ox-LDL uptake by VSMCs, contributing to inflammation and progression atherosclerosis. Finally, miR-15a-5p could be used as a novel diagnostic biomarker for advanced atherosclerosis.

Determination of a Discriminant Dose to Identify Resistance to Amitraz in Rhipicephalus sanguineus s.l. (Acari: Ixodidae) from Mexico.

The brown dog tick, Rhipicephalus sanguineus s.l., is considered the most widely distributed three-host tick in the world and has medical and veterinary importance; the control of infestation is carried out with acaricides, towards which it can develop resistance. This study aimed to determine the discriminant dose (d.d) of amitraz to identify resistance in R. sanguineus s.l. larvae natives from Mexico and to evaluate its application in field-collected ticks. Engorged ticks were collected from naturally infested dogs residing in rural communities and were incubated for 25 days, and their progeny was used in a larval immersion test (LIT) to be exposed to the d.d. determined in Rhipicephalus microplus, and those that were susceptible were analyzed using the LIT in six concentrations. Mortality was analyzed through probit methodology to calculate the lethal concentration (LC) 50 and 99. The d.d. was determined as a consensus value by multiplying the LC99 × 2, and then, we proceeded to evaluate it in in-field samples by using the LIT technique. The d.d. calculated was 4 ppm. The in-field evaluation found 64% of the resistant samples to amitraz with mortality percentages between 98.3% and 0.35%. This dose can be used to rapidly and inexpensively identify resistant populations in samples collected in the field.

Presence of the toxigenic fungi Aspergillus spp. and Fusarium spp. in Musca domestica L. (Diptera: Muscidae) collected from dairy farms.

The objective of the study was to identify the presence of toxigenic fungi Aspergillus spp. and Fusarium spp. in domestic flies collected from dairy farms. We selected 10 dairy farms distributed in the central valley of the state of Aguascalientes, México. The flies were trapped using entomological traps with an olfactory attractant in 7 sites of the farm (silo-cutting surface, feed store, milking parlor, 3 feeders, and the rearing room). The fungi were cultivated in Sabouraud agar through direct sowing by serial dilutions to obtain the isolates, and a taxonomical identification was carried out under the microscope. The aflatoxins and zearalenone production capacity of the pure isolates were quantified using the ELISA test. The flies were present in all of the capture sites (45.3 flies, 567 mg, trap per day). We obtained 50 isolates of Aspergillus spp. genus, 12 of which produced aflatoxins (327 ± 143 µg/kg), whereas from 56 of the Fusarium spp. isolates, 10 produced large quantities of zearalenone (3,132 ± 665 µg/kg). These results suggest that the presence of domestic flies on dairy farms can constitute a source of dissemination for toxigenic fungi that can eventually contaminate grains and forage that are part of the daily cattle diet.

Seroepidemiology of Toxoplasma gondii infection and parasite DNA in free-range chickens in Aguascalientes, Mexico.

The objectives of the study were to estimate the prevalence of anti-Toxoplasma gondii antibodies in free-range chickens in Aguascalientes, Mexico, its association with certain risk factors, and the frequency of parasite DNA in the heart. Eighty-one small rural family farms were included, and blood and heart samples were taken from 150 clinically healthy, adult, free-range chickens. Serum samples were processed by indirect immunofluorescence antibody test considering a dilution of 1:16 as a positive reaction, while the DNA detection was done by PCR. The correlation between the presence of antibodies and the potential risk factors was estimated with logistic regression. The overall seroprevalence in the examined populations was 67%, with a range from 33% to 100% among municipalities, and 78% of the farms having at least one seropositive bird. The PCR test identified the presence of parasite DNA for one case only. Among the variables, the presence of cats and the presence of other animal species on the farm were the ones that had the highest values, while the presence of feral cats and the presence of noxious fauna were also identified as potential risk factors. The results indicate high soil contamination by T. gondii oocysts, which implies the active role of cats living on the farms.

Implication of miR-155-5p and miR-143-3p in the Vascular Insulin Resistance and Instability of Human and Experimental Atherosclerotic Plaque.

(1) Background: Cardiovascular diseases (CVDs) are the main cause of death in developed countries, being atherosclerosis, a recurring process underlying their apparition. MicroRNAs (miRNAs) modulate the expression of their targets and have emerged as key players in CVDs; (2) Methods: 18 miRNAs were selected (Pubmed and GEO database) for their possible role in promoting atherosclerosis and were analysed by RT-qPCR in the aorta from apolipoprotein E-deficient (ApoE-/-) mice. Afterwards, the altered miRNAs in the aorta from 18 weeks-ApoE-/- mice were studied in human aortic and carotid samples; (3) Results: miR-155-5p was overexpressed and miR-143-3p was downregulated in mouse and human atherosclerotic lesions. In addition, a significant decrease in protein kinase B (AKT), target of miR-155-5p, and an increase in insulin-like growth factor type II receptor (IGF-IIR), target of miR-143-3p, were noted in aortic roots from ApoE-/- mice and in carotid plaques from patients with advanced carotid atherosclerosis (ACA). Finally, the overexpression of miR-155-5p reduced AKT levels and its phosphorylation in vascular smooth muscle cells, while miR-143-3p overexpression decreased IGF-IIR reducing apoptosis in vascular cells; (4) Conclusions: Our results suggest that miR-155-5p and miR-143-3p may be implicated in insulin resistance and plaque instability by the modulation of their targets AKT and IGF-IIR, contributing to the progression of atherosclerosis.

Expression, Purification, and Structure Determination of Human PTCH1-HH-N Complexes.

Patched-1 (PTCH1), a tumor suppressor, serves as the receptor of Hedgehog (HH) ligand and negatively regulates the HH signaling pathway. Mutations of PTCH1 are implicated in many human cancers. Structural investigation revealed the mechanism of PTCH1-mediated HH signal regulation, further facilitating the therapeutic development of cancers. Here, we describe the expression and purification of a nearly full-length functional PTCH1 variant, PTCH1*. With purified PTCH1* protein, two forms of PTCH1*-Sonic Hedgehog (SHH) complexes were assembled, and their structures subsequently determined by cryo-electron microscope (cryo-EM).

Concerted regulation of non-alcoholic fatty liver disease progression by microRNAs in apolipoprotein E-deficient mice.

The prevalence of non-alcoholic fatty liver disease (NAFLD) is constantly increasing, and altered expression of microRNAs (miRNAs) fosters the development and progression of many pathologies, including NAFLD. Therefore, we explored the role of new miRNAs involved in the molecular mechanisms that trigger NAFLD progression and evaluated them as biomarkers for diagnosis. As a NAFLD model, we used apolipoprotein E-deficient mice administered a high-fat diet for 8 or 18 weeks. We demonstrated that insulin resistance and decreased lipogenesis and autophagy observed after 18 weeks on the diet are related to a concerted regulation carried out by miR-26b-5p, miR-34a-5p, miR-149-5p and miR-375-3p. We also propose circulating let-7d-5p and miR-146b-5p as potential biomarkers of early stages of NAFLD. Finally, we confirmed that circulating miR-34a-5p and miR-375-3p are elevated in the late stages of NAFLD and that miR-27b-3p and miR-122-5p are increased with disease progression. Our results reveal a synergistic regulation of key processes in NAFLD development and progression by miRNAs. Further investigation is needed to unravel the roles of these miRNAs for developing new strategies for NAFLD treatment. This article has an associated First Person interview with the joint first authors of the paper.

Epidemiology of Neospora caninum infection in free-range chickens (Gallus gallus domesticus) in north central Mexico.

The objective of the present study was to investigate the epidemiology of natural infection by N. caninum in free-range chickens from Aguascalientes, Mexico, through identifying the presence of antibodies, its association with certain risk factors, and estimate the frequency of parasite DNA detection in the brain. The study included 150 clinically healthy chickens from 81 small family farms distributed throughout the state, from which blood and brain samples were taken. Serum samples were processed by indirect immunofluorescence considering a dilution of 1:50 as a positive reaction, while the DNA detection was done by PCR; the association between the presence of antibodies and the potential risk factors was estimated through logistic regression. The general anti-N. caninum presence of antibodies in the studied population was 46% (69/150, CI 95% 38-54), with titers of 1:50 in 38, 1:100 in 24, and 1:200 in 7, finding positive cases in 49% of the farms and in 91% of the municipalities that integrate the state. The presence of antibodies found in the municipalities was estimated in a range of 20 to 75%. The frequency of DNA detection was 6% (9/150, CI 95% 2-11). The grain supplementation was identified as the unique risk factor for infection (OR = 2.82; 95% CI, 1.18-6.73, p < 0.019).

Molecular basis of cholesterol efflux via ABCG subfamily transporters.

The ABCG1 homodimer (G1) and ABCG5-ABCG8 heterodimer (G5G8), two members of the adenosine triphosphate (ATP)-binding cassette (ABC) transporter G family, are required for maintenance of cellular cholesterol levels. G5G8 mediates secretion of neutral sterols into bile and the gut lumen, whereas G1 transports cholesterol from macrophages to high-density lipoproteins (HDLs). The mechanisms used by G5G8 and G1 to recognize and export sterols remain unclear. Here, we report cryoelectron microscopy (cryo-EM) structures of human G5G8 in sterol-bound and human G1 in cholesterol- and ATP-bound states. Both transporters have a sterol-binding site that is accessible from the cytosolic leaflet. A second site is present midway through the transmembrane domains of G5G8. The Walker A motif of G8 adopts a unique conformation that accounts for the marked asymmetry in ATPase activities between the two nucleotide-binding sites of G5G8. These structures, along with functional validation studies, provide a mechanistic framework for understanding cholesterol efflux via ABC transporters.

Control of Haematobia irritans and Stomoxys calcitrans with Metarhizium anisopliae in naturally infested cattle / Control de Haematobia irritans y Stomoxys calcitrans con Metarhizium anisopliae en ganado naturalmente infestado

ABSTRACT Objective. Assess the efficacy of Metarhizium anisopliae strain 135 (Ma135) applied by aspersion to reduce simultaneous infestation of Haematobia irritans and Stomoxys calcitrans in naturally infested cattle maintained in a production system which combine grazing and confinement in pens. Materials and methods. Was applied an aqueous formulation of Ma135 (1x108 conidia/ml) on a nine cows group in six occasions with seven days interval, while the control group received the same formulation without conidial content. The infestation index was estimated daily for each fly independently; the effectiveness of the formulation was calculated using the Abbott's formula. Results. The Ma135 formulation had a reduction in the fly population of 58% for H. irritans and 69% for S. calcitrans, taking into consideration the six study weeks, in addition to causing no negative effects on animal health. An Infestation index reduction was observed from the first week post-treatment (p<0.05) and maintained this trend throughout the study. Conclusions. The present study has demonstrated the potential of strain Ma135 to reduce the simultaneous infestation of both hematophagous flies in cattle under conditions of natural infestation.

RESUMEN Objetivo. Evaluar la eficacia de la cepa 135 de Metarhizium anisopliae (Ma135) aplicada por aspersión para reducir la infestación simultánea de Haematobia irritans y Stomoxys calcitrans en ganado naturalmente infestado mantenido en un sistema de producción de leche que combina el pastoreo y el confinamiento en corrales. Materiales y métodos. Se aplicó una formulación acuosa de Ma135 (1x108 conidios/ml) a un grupo de nueve vacas en seis ocasiones con un intervalo de siete días, mientras que el grupo control recibió una solución compuesta por agua, Monooleato de polioxietileno sorbitán en solución 0.01% y un adyuvante agrícola al 0.1%. Se estimó diariamente el índice de infestación para cada mosca de forma independiente; la efectividad de la formulación se calculó usando la fórmula de Abbott. Resultados. La formulación de Ma135 tuvo una eficacia en el control de la infestación de 58% para el caso de H. irritans y de 69% para S. calcitrans, tomando en cuenta las seis semanas de estudio, además de que no causó ningún efecto negativo en la salud de los animales. La reducción del índice de infestación se observó desde la primera semana post-tratamiento (p<0.05) y mantuvo esta tendencia durante todo el estudio. Conclusiones. El presente estudio ha demostrado el potencial de la cepa Ma135 para reducir la infestación simultánea de ambas moscas hematófagas en el ganado bajo condiciones de infestación natural.

Prevalence and risk factors of Neospora caninum infection in ovine flocks of central-western Mexico.

This study, carried out in 13 sheep flocks of central-western Mexico, aimed to identify the prevalence of anti-N. caninum antibodies, to develop a risk analysis of the infection and to estimate the prevalence of parasite DNA in blood. A total of 368 serum and blood samples were subjected to ELISA and PCR tests, respectively, and the association between the prevalence of antibodies and some management factors was estimated. The overall prevalence of anti-N. caninum antibodies was 13.5% [50/368; 95% confidence interval (CI) 10-17], ranging from 2.7 to 90% per flock, and 92% of the farms had seropositive animals. In ewes the prevalence was 14% (48/348, 95% CI 10-17) and in rams 10% (2/20; 95% CI 1- 33). The mean prevalence of Neospora DNA in blood was 27% (99/368, 95% CI 22-31), implying a range between 16 and 82%. In rams, the prevalence was 5% (1/20 95% CI 2-26), while in ewes it was 28% (98/348, 95% CI 23-33). The agreement between the tests was k = 0.19. The presence of other domestic animal species in the farms [odds ratio (OR) 4.4] and the consumption of placental debris, fetuses and stillborn lambs by dogs living in the farms (OR 5.8) were demonstrated to be risk factors.

Implementación de círculos de calidad para la mejora del registro de Enfermería / Implementation of quality control circles for the improvement of nursing records

El registro de Enfermería es una herramienta que facilita la recolección y consulta de datos para el desarrollo de planes de cuidados, el intercambio de información y como medio de referencia. Sin embargo, por factores asociados a la labor de Enfermería no siempre cumplen con todos los estándares, incluido los de índole legal. El propósito de este trabajo es dar a conocer cómo se llevó a cabo un proceso de mejora de la calidad del registro de Enfermería de un hospital público, sobre todo, en los aspectos relacionados con el cumplimiento de la normativa institucional. Para ello, se definieron enfermeros líderes del método que se iba a usar mediante el establecimiento de círculos de calidad, quienes recibieron capacitación, asesoría y seguimiento durante todo el proceso. Paralelamente se incluyeron otros profesionales enfermeros que formarían parte de estos círculos de calidad. Por otro lado, cada líder pudo identificar áreas de mejora, que conllevaron actividades orientadas a la mejora del registro de Enfermería. Al final del proceso, participaron 31 enfermeras y enfermeros (siete líderes y 24 integrantes del círculo), de los 51 que eran responsables de la cumplimentación de los registros (participación del 61%), distribuidos en siete círculos de calidad, consiguiendo una mejora del 75% respecto a la cumplimentación de calidad de los registros. Sin duda, para favorecer el cumplimiento de este objetivo se requirió, no solo la capacitación de los profesionales en la mejora de la ejecución de los registros, sino también su participación constante y un alto grado de compromiso y responsabilidad. En esta ocasión el uso de los círculos de calidad fue una metodología pertinente y factible en la detección de áreas de mejora en beneficio de la organización

Nursing record is a tool which facilitates data collection and search for the development of care plans, information exchange, and as information and means of reference. However, due to factors associated with the nursing job, these records won’t always meet all standards, including those of legal nature. The objective of this article is to provide information about the manner in which a process for improving the quality of nursing records was conducted in a public hospital, mostly regarding those aspects associated with meeting the institutional rules. To this aim, there was a determination of nurses leading the method that would be used, through the implementation of quality control circles; these nurses received training, counselling, and follow-up throughout the process. At the same time, other nursing professionals were included, who would form part of these quality control circles. On the other hand, each leader could identify areas for improvement, which entailed activities orientated to an improvement in nursing records. At the end of the process, 31 male and female nurses were involved (seven leaders and 24 circle members), of the 51 nurses who were responsible for record completion (61% participation), distributed into seven quality control circles, and achieving a 75% improvement regarding the quality of record completion. Undoubtedly, meeting this objective not only required the training of professionals on the improvement of record execution, but also their constant involvement, and a high level of commitment and responsibility. In this occasion, the use of quality control circles was a relevant and feasible methodology for detecting areas for improvement that would benefit the organization

Neospora caninum Infection during the First Gestation of Holstein Heifers That Consume Food Contaminated Naturally with Zearalenone under Field Conditions.

BACKGROUND: This work studied the natural infection of Neospora caninum during the first gestation of heifers in a dairy farm in animals consuming a ration co ntaminated naturally with Zearalenone (ZEA), and to find out effect of mycotoxin in the levels of estrogen (E) and progesterone (P4) and that's relation to the infection to N. caninum and in the abortions. METHODS: The study was conducted in a dairy farm located in El Llano municipality, in Aguascalientes, Mexico, in 2013. Two groups were formed, the group "A" with 20 seronegative animals to N. caninum, and group "B" with 20 seropositive. Once a month was determined the levels of total IgG to N. caninum, the serum concentration of E and P4, and the level of ZEA in the ration; in cases of abortion, fetal brain samples were taken to identify the presence of N. caninum DNA. RESULTS: In group "A", was observed two subgroups: seronegative (60%) and seroconverted (40%), and three abortions. In group "B", all animals maintain their serostatus, and three animals aborted. All abortions were positive for N. caninum DNA. The level of ZEA in the ration has an average of 426 µg/kg; during the gestation did not identify that animals suffer any alteration in the levels of E or P4. No statistical differences among the studied variables (levels of E and P) in time (nine months of gestation) were detected. It was not identified any interaction with the natural exposure to ZEA intake in any of the groups under study. CONCLUSION: The chronic ingestion of ZEA does not affect serum concentrations of E and P4 during gestation of heifers under study and cannot be related to the infection for N. caninum and the abortion.

Genetic characterization of Neospora caninum from aborted bovine foetuses in Aguascalientes, Mexico.

The cyst-forming protozoan parasite Neospora caninum is one of the main causes of bovine abortion worldwide and is of great economic importance in the cattle industry. Recent studies have revealed extensive genetic variation among N. caninum isolates based on multilocus microsatellite genotyping. Currently, the most extensive study reported is based on the N. caninum genotyping of 96 samples from four countries on two continents (Spain, Argentina, Germany and Scotland) that demonstrate different clusters of multilocus genotypes (MLGs) implicated in cattle abortions as well as the population sub-structuring of N. caninum, which is partially associated with the geographical origin. The aim of this study was to genotype N. caninum from aborted bovine foetuses that originated from Mexico within the region of Aguascalientes and to investigate their genetic diversity. Parasite DNA was detected in 27 out of the 63 analysed foetuses recovered from 10 different herds. Complete or nearly complete profiles based on 9 microsatellite markers were obtained from 11 samples. Diverse N. caninum MLGs were implicated in the occurrence of abortion in each herd. All of the Mexican MLGs differed from the MLGs previously determined for the Argentinean, Spanish, German and Scottish N. caninum populations. The Mexican MLGs failed to cluster by eBURST analyses. The MLG relationships using PCoA showed a close genetic relationship between the Spanish population and a portion of the Mexican population, but a more distant genetic relationship with the Argentinean genotypes. These results demonstrate the genetic diversity of N. caninum in the studied areas that differed from other populations of N. caninum around the world.

Effect of Bosentan on Claudication Distance and Endothelium-Dependent Vasodilation in Hispanic Patients With Peripheral Arterial Disease.

Endothelin (ET) is involved in the etiopathogenesis of peripheral arterial disease (PAD). We hypothesized that ET antagonism might improve the endothelial function, inflammatory status, and symptoms in PAD. This pilot randomized clinical trial was designed to determine the clinical efficacy, pleiotropic effects, and safety of dual ET-receptor antagonist bosentan in Hispanic patients with PAD presenting intermittent claudication. The Bosentan Population-Based Randomized Trial for Clinical and Endothelial Function Assessment on Endothelin Antagonism Therapy was a 12-month, randomized, controlled, parallel-group, double-blind, proof-of-concept pilot study evaluating the effect of bosentan on absolute claudication distance (primary efficacy end point), flow-mediated arterial dilation, and C-reactive protein levels (primary pleiotropic end points) in patients with PAD with Rutherford category 1 to 2 of recent diagnosis. Secondary end points included ankle-brachial index, subjective claudication distance, and safety. Of the 629 screened subjects, 56 patients were randomized 1:1 to receive bosentan for 12 weeks (n = 27) or placebo (n = 29). Six months after the initiation, a significant treatment effect in flow-mediated arterial dilation of 2.43 ± 0.3% (95% CI 1.75 to 3.12; p = 0.001), absolute claudication distance of 283 ± 23 m (95% CI 202 to 366; p = 0.01), ankle-brachial index of 0.16 ± 0.03 (95% CI 0.09 to 0.23; p = 0.001), and a decrease in C-reactive protein levels of -2.0 ± 0.5 mg/L (95% CI -2.8 to -1.1; p = 0.02) were observed in the bosentan-treated group compared to the control group. No severe adverse effects were found in the bosentan group. In conclusion, in Hispanic patients with intermittent claudication, bosentan was well tolerated and improved endothelial function and claudication distance as well as inflammatory and hemodynamic states.

Attenuation of early atherosclerotic lesions by immunotolerance with ß2 glycoprotein I and the immunomodulatory effectors interleukin 2 and 10 in a murine model.

OBJECTIVE: This study assessed the effect of cellular and humoral autoimmune response inhibition after immunization with ß2-glycoprotein I (ß2-GPI) and the effect of immunomodulation with interleukin (IL)-2 and IL-10 in the development of early atherosclerotic vascular lesion in a murine model. Atherosclerosis is increasingly considered a chronic inflammatory disease with pathogenic autoimmune processes. Regulatory T cells, and their cytokines, have been implicated in the inhibition of the development of atherosclerotic lesions and involved in the immunologic tolerance induction. METHODS: Eight-week-old male C57BL6 LDL-receptor deficient (LDLR(-/-)) mice were fed a cholesterol-rich (2.8%), high-saturated-fat (82%) diet for a week and divided in five groups. The groups received the following intravenous immunizations: group I (control group): one dose of 5 µg ß2-GPI; group II: 5 µg ß2-GPI I and 1 µg IL-2; group III: 5 µg ß2-GPI and 0.75 µg of IL-10; and group IV: 5 µg ß2-GPI, 1 µg IL-2, and 0.75 µg IL-10. The aortas of the mice were assessed 8 weeks after inoculation to determine the aortic lesion size and composition in all groups. RESULTS: ß2-GPI immunization attenuated the early atherosclerotic lesions development compared with the control group (P = .001). Macroscopic and histologic aortic atherosclerotic lesions were significantly decreased in the IL-2 and IL-10-treated groups in ß2-GPI-tolerant mice compared with the ß2-GPI-tolerant group without cytokine injection (P = .001). The association of both cytokines did not provoke a major inhibition in the atherosclerosis development when compared with groups injected with the two cytokines separately. CONCLUSIONS: The immunotolerance induction against ß2-GPI attenuates the development of atherosclerosis lesions in an animal model, enhanced by downregulation of the cellular and humoral autoimmune response provoked by IL-2 and IL-10.