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Biomedical subjects

J Sanchez

Publications and source records attributed to J Sanchez.

At least 19 recordsLinked to original sources

Characterization of a murine model of intranasal infection suitable for testing vaccines against C. abortus.

Mouse models have been widely used to test candidate vaccines against Chlamydophila abortus infection in mice. Although the induction of a systemic infection by endogenous or intraperitoneal inoculation is a useful tool for understanding the immune mechanism involved in the protection conferred by the vaccination, a different approach is necessary to understand other factors of the infection, such as mucosal immunity or the colonization of target organs. To test whether C. abortus intranasal model of infection in mice is a useful tool for testing vaccines in a first group of experiments mice, were infected intranasally with C. abortus to characterize the model of infection. When this model was used to test vaccines, two inactivated experimental vaccines, one of them adjuvated with QS-21 and another with aluminium hydroxide, and a live attenuated vaccine (strain 1B) were used. Non-vaccinated control mice died within the first 8 days, after displaying substantial loss of weight. Histologically, the mice showed lobar fibrinopurulent bronchointerstitial pneumonia. Prior immunization with QS-21 adjuvated vaccine or 1B vaccine presented mortality and the recipients showed a greater number of T cells in the lesions, especially CD8(+) T cells, than the control mice and mice immunized with vaccine adjuvated with aluminium hydroxide. The results confirm that the C. abortus intranasal model of infection in mice is a useful tool for testing vaccines.

Abortion, Veterinary↗

Effect of eprinomectin pour-on treatment around calving on reproduction parameters in adult dairy cows with limited outdoor exposure.

The objective of this study was to investigate if treatment of cows with eprinomectin around calving had any beneficial effects on the calving to first artificial insemination interval, calving to conception interval, and number of services per conception in totally- and semi-confined dairy herds. In totally-confined herds lactating- and dry-cows were housed throughout the summer and had no access to pasture. In semi-confined herds lactating- and dry-cows had limited outdoor exposure to a small pasture or paddock but were still fed a ration that met all their nutritional requirements. The study was carried out between February 2002 and February 2003 in 35 herds (2381 cows) located in Quebec, Ontario and Minnesota (USA) participating in a larger clinical trial. The herds kept electronic reproduction records. Cows were randomly allocated to receive eprinomectin or a placebo, with treatment being administered on or close to the day of calving. Monthly bulk tank milk samples from each farm were tested with an indirect ELISA using a crude Ostertagia ostertagi antigen and these data were averaged over the study year. The optical density ratio (ODR) values were then dichotomized into high and low using a cut-point of 0.50. Treatment effects were analyzed using Cox proportional hazards survival models with herd frailty effects for calving to conception and calving to first service intervals. Aalen's linear hazards model was used to investigate time-varying effects in the Cox models. A random effects poisson regression model was used to model the number of services per conception. Other predictor variables tested in the models were lactation number, calving season, study site, peak milk production, ODR and the lactating- and dry-cow housing variables. Overall, there was no significant effect of treatment on the three indices of reproductive performance. The effect of season of calving depended on how much time had passed since calving. Presumably this effect reflected a seasonal effect at the time of breeding. Hazard of conception in younger cows was higher than in older cows. Early bred cows tended to have a higher number of inseminations per conception than those bred late. The results of the study suggested that eprinomectin treatment at calving was not beneficial to reproduction.

Animal Husbandry↗

Test characteristics from latent-class models of the California Mastitis Test.

We evaluated (using latent-class models) the ability of the California Mastitis Test (CMT) to identify cows with intramammary infections on the day of dry-off. The positive and negative predictive values of this test to identify cows requiring dry-cow antibiotics (i.e. infected) was also assessed. We used 752 Holstein-Friesian cows from 11 herds for this investigation. Milk samples were collected for bacteriology, and the CMT was performed cow-side, prior to milking on the day of dry-off. At the cow-level, the sensitivity and specificity of the CMT (using the four quarter results interpreted in parallel) for identifying all pathogens were estimated at 70 and 48%, respectively. If only major pathogens were considered the sensitivity of the CMT increased to 86%. The negative predictive value of the CMT was >95% for herds with major-pathogen intramammary-infection prevalence <15%, so that selective dry-cow therapy might be reasonable for such herds if cows were screened with the CMT.

Animals↗

Geographical and temporal aspects of an outbreak of off-flavours in bulk-tank milk in Prince Edward Island, Canada.

The geographical and temporal dynamics of the outbreak of off-flavours in bulk-tank milk that occurred between September 2000 and June 2002 in Prince Edward Island (PEI) dairy herds were described using spatial, temporal and space-time scan statistics. Results suggested clustering both in time and space. We found one primary cluster (six case herds, feed off-flavour, October 2000 to January 2001) and two secondary clusters formed of feed (5 case herds, October 2000 to January 2001)) and rancid (five case herds, December 2000 to March 2001) off-flavours, respectively. The relative risk (RR) for producing off-flavoured milk was similar for these three clusters (RR=6.4).

Animal Feed↗

Seroprevalence of and risk factors for HIV-1 infection among female commercial sex workers in South America.

OBJECTIVE: Assessment of HIV prevalence and associated risk behaviours among female commercial sex workers (FCSW) across major cities in South America. METHODS: Seroepidemiological, cross sectional studies of 13 600 FCSW were conducted in nine countries of South America during the years 1999-2002. Participants were recruited in brothels, massage parlours, hotels, and streets where anonymous questionnaires and blood samples were collected. HIV infection was determined by enzyme linked immunosorbent assay (ELISA) screening and western blot confirmatory tests. RESULTS: The overall HIV seroprevalence was 1.2% (range 0.0%-4.5%). The highest HIV seroprevalences were reported in Argentina (4.5%) and Paraguay (2.6%); no HIV infected FCSW were detected in Venezuela and Chile. Consistent predictors of HIV seropositivity were: (1) a previous history of sexually transmitted infections (STI, AORs = 3.8-8.3), and (2) 10 years or more in commercial sex work (AORs = 2.2-24.8). In addition, multiple (> or =3) sexual contacts (AOR = 5.0), sex with foreigners (AOR = 6.9), use of illegal drugs (AOR = 3.2), and marijuana use (AOR = 8.2) were associated with HIV seropositivity in Southern Cone countries. CONCLUSIONS: Consistently low HIV seroprevalences were detected among FCSW in South America, particularly in the Andean region. Predictors of HIV infection across the continent were STI and length of commercial sex work; however, use of illegal drugs, especially marijuana, and sexual contacts with foreigners were also found to be associated risk factors in the Southern Cone region. Interventions for the control of HIV and other STI need to be region and country specific; drug use appears to have an ever increasing role in the spread of HIV among heterosexually active populations.

Adult↗

Temporal and spatial analysis of the 1999 outbreak of acute clinical infectious bursal disease in broiler flocks in Denmark.

The objective of this study was to describe the spatial and temporal dynamics of the 1999 outbreak of acute clinical infectious bursal disease (IBD) in broiler chicken farms in Denmark. The analysis was performed using data from all broiler farms located in the Jutland peninsula and the island of Funen (168 municipalities). The Moran's index I, K-functions and scan statistics were used to describe the dynamics of the epidemic. In addition, spatially correlated survival analyses were performed and the posterior frailties were mapped to identify areas with high or low survival times. From January to October 1999, a total of 43 farms (81 flocks) out of a total of 299 farms (2970 flocks) in 110 municipalities were infected with IBD. The outbreak developed more or less simultaneously in two regions, one in the north and the other in the center of the study area. The space-time descriptive methods suggested that the cases were more likely to occur during a short period of time and over relatively short distances, indicating that local factors facilitated the spread of the virus. For instance, the highest risk, as estimated by the K-function, was within 20 km and 20 days. The risk of transmission peaked in July and remained high until the end of the study. The spatially correlated survival analysis identified municipalities with low survival times that roughly corresponded to the clusters found using scan statistics. The analysis for the full hierarchical structure of the dataset identified variation (i.e. clustering) at the farm and municipality levels.

Animals↗

Assessing the agreement between Ostertagia ostertagi ELISA tests performed using the crude adult antigen and the adult and larval stage 4 excretory/secretory antigens.

The objective of this study was to determine the agreement between ELISA tests conducted using three O. ostertagia antigens: crude adult worm, larval stage 4 (L4) excretory/secretory (ES) and adult ES. This study was carried out on 289 Holstein cows from five herds in Prince Edward Island and one herd in Nova Scotia. Composite milk samples of these cows were collected (between May and September 2002) from the respective provincial laboratories and sent to the Atlantic Veterinary College where each sample was tested for antibodies to O. Ostertagi using an indirect microtitre ELISA test. Results were expressed as optical density ratio (ODR) values. Each milk sample was tested with three ELISA tests, with each test using a different O. ostertagi antigen. There was a slight rise in ODR values of both adult antigens, between May and August, with higher values obtained using the adult ES antigen. L4 ES ODR values were generally higher than those for both adult antigens during the study period, except for May. There was a more dramatic rise in L4 ES ODR values between May and August. Rises in ODR in May and end of July coincided with periods of mass maturation of L4 to adult worms. The results of the study showed that the concordance correlation coefficient (CCC) between tests performed using both ES and the crude antigens were low (crude adult versus adult ES=0.31, crude adult versus L4 ES=0.30). The highest CCC was observed between tests done using both ES antigens (CCC=0.56). Generally, the study results suggest that the antibody response (detectable by the ELISA) is mainly directed against ES antigens (especially L4) than the crude adult worm antigen.

Animals↗

The S-phase fraction of the aneuploid cell subpopulation is the biologically relevant one in aneuploid breast cancers.

BACKGROUND: In the case of DNA-aneuploid tumors there are no clear guidelines as to which S-phase fraction is the more relevant one: that corresponding to either the diploid or the aneuploid population, or rather an average of both. MATERIALS AND METHODS: We studied 280 breast cancer specimens from previously untreated patients. Histologically, 231 were ductal infiltrating carcinomas, 30 lobular infiltrating carcinomas and 19 corresponded to other, less frequent varieties. Postsurgically, 164 cases (58.6%) were classified as T1, 87 (31.1%) as T2 and 7 as T3. The remaining 22 cases were multifocal, diffuse tumors. Flow cytometry was performed on fresh tumor tissue, and immunohistochemistry for hormone receptors, Ki67, c-erb-B2 and p53 on paraffin-embedded material. RESULTS: In diploid tumors, a high S-phase (above the 75th percentile) correlated significantly with Ki67 expression > or =20% (p<0.0001). In aneuploid tumors, however, this was only the case for the aneuploid fraction of tumor cells (p< 0.0001). A high S-phase of diploid tumors correlated directly and significantly with a high histologic grade (p=0.04), a high nuclear grade (p=0.01), tumor size (p=0.0008), and inversely with estrogen (p<0.0001) and progesterone (p<0.0001) receptor expression. In aneuploid tumors, the aneuploid tumor fraction showed a direct and significant correlation with a high histologic grade (p=0.005), a high nuclear grade (p=0.001), mutant p53 expression (p=0.0009), and inversely with estrogen (p<0.0001) and progesterone (p=0.0001) receptor expression. A high S-phase of the diploid cell fraction of aneuploid tumors, on the other hand, just showed an inverse correlation with high nuclear grade of the tumors (p=0.02), and none whatsoever with all other tested parameters.

Aneuploidy↗

[Prevalence of impairments and social inequalities].

BACKGROUND: Population ageing and longer duration of the working career could increase the prevalence of impairments and disabilities whereas there have been few data to help prevention and care. This study aimed at describing the prevalence of various types of impairments and assessing their social inequalities in the Lorraine population (north-eastern France). METHODS: The sample included 6.214 subjects aged 15 or more from 8.000 households randomly selected in the Lorraine population. A mailed questionnaire including socio-demographical characteristics, job, and various types of impairments was used. The data were analysed with the chi2 independence test, the adjusted odds ratio (OR) and the Mantel-Haenszel test. RESULTS: The prevalence of at least one impairment was 30% in both sexes. It varied from about 20% for subjects aged less than 40 years to about 60% for the subjects aged 70 years or more. It was higher in men than in women among the individuals aged more than 50 years (OR adjusted on age: 1.22, 95% CI: 1.04-1.44). The impairments were generally multiple for the subjects aged over 50. The prevalence of at least one impairment was higher for workmen (OR adjusted on age: 2.11, 95% CI: 1.68-2.66), farmers, craftsmen, traders and heads of firms (OR adjusted on age: 1.60, 95% CI: 1.17-2.18) and for employees (OR adjusted on age: 1.62, 95% CI: 1.29-2.03) in comparison with executives, intellectual professionals and teachers. For workmen a higher prevalence was observed for all types of impairments: intellectual and psychological functions, language, hearing, vision, visceral functions, and skeleton and posture. Farmers, craftsmen, traders, heads of firms and employees showed an excess for most types of impairments. Social inequalities were significant for various age groups during their period of occupational activity, but not after retirement. Unemployed people also had an excess of impairments. CONCLUSION: The prevalence of impairments strongly increased with advancing age. Marked inequalities were observed between various socio-occupational categories. Men were more affected than the women over 50 years of age. Impairments represent a health index which is useful for prevention to reduce their causes and consequences.

Adolescent↗

Effect of eprinomectin treatment at calving on milk production in dairy herds with limited outdoor exposure.

The objective of this study was to determine the effect of anthelmintic treatment at calving in herds that were totally or semiconfined during the summer. In totally confined herds, lactating and dry cows were housed throughout the summer and had no access to pasture. In semiconfined herds, lactating and dry cows had limited outdoor exposure to a small pasture or paddock but were still fed a ration that met all their nutritional requirements. The study was carried out between February 2002 and February 2003 in 65 herds enrolled with DHI and distributed in 4 regions in Canada and 1 state in the United States. Cows were randomly allocated to receive eprinomectin or a placebo, with treatment being administered on or close to day of calving. In May and June 2002, 8 fecal samples were collected from each farm and fecal egg counts (FEC) were determined. Monthly bulk tank milk samples from each farm were tested with an indirect ELISA using a crude Ostertagia ostertagi antigen. Monthly test-day milk production data were recorded for 200 d after calving. In general, FEC were very low (mean = 1 egg per gram, range = 0 to 27). Mean herd bulk milk ELISA optical density ratio (ODR) values for the whole year ranged between 0.22 and 0.80. The ODR values were dichotomized into high and low using a threshold of 0.5. Treatment effects were analyzed using a linear mixed model with herd and cow as random effects. The analysis was restricted to 4789 cows (23,956 test-day records) treated between 21 d before and 7 d after calving. Overall, there was no significant effect of treatment. However, there was a tendency for an interaction between treatment and ODR, as illustrated by a larger numerical difference in treated vs. untreated cows in high-ODR herds than in low-ODR herds. However, the confidence intervals for the treatment effects (kg/d of milk per cow) in high-ODR herds (-0.33 to 1.10) and in low-ODR herds (-0.53 to 0.14) were wide and included zero. Therefore, this study failed to show a beneficial effect of eprinomectin treatment in these totally or semiconfined herds.

Animal Husbandry↗

De novo hemolytic-uremic syndrome/thrombotic microangiopathy in renal transplant patients receiving calcineurin inhibitors: role of sirolimus.

The aim of this study was to evaluate the outcome of ten renal transplant recipients who developed de novo hemolytic uremic syndrome/thrombotic microangiopathy (DnHUS) after treatment with calcineurin inhibitors among 3,862 patients transplanted during the period 2000-2001 in Spain, and the results of switching to sirolimus for resolution of this pathologic condition. No patient had end-stage disease due to primary HUS. The criteria of diagnosis were decreased renal function, biopsy-proven thrombotic microangiopathy, and no signs of acute rejection. Calcineurin inhibitors were completely removed and immediate treatment with sirolimus started after diagnosis. The follow-up period was 19.0+/-4.3 months, at least 12 months after diagnosis. One patient died of sepsis shortly after starting sirolimus therapy. The serum creatinine level in the series decreased from 5.2+/-2.6 mg/dL at the time of biopsy to 2.15+/-1.9 mg/dL 1 month later (P=.011). All but one of the nine recipients, who lost his graft 3 months later (80% success) maintained function, with a serum creatinine of 2.1+/-1.4 mg/dL and Cockroft index of 61.3+/-34 mL/min at the end of follow up. During this time, none of the patients experienced an acute rejection episode and sirolimus was maintained without any remarkable secondary effect. Sirolimus seems to be a promising alternative for the treatment of renal transplant patients who develop calcineurin inhibitor-induced DnHUS.

Adult↗

The effect of eprinomectin treatment at calving on reproduction parameters in adult dairy cows in Canada.

A clinical trial was carried out in two provinces of Canada to investigate the effect of treatment with eprinomectin at calving on production parameters in adult dairy cattle in 1999-2000. One of the objectives of this study was to evaluate the impact of treatment on reproductive performance as measured by: calving-to-conception interval, calving-to-first service interval and number of services per conception. The ability of an indirect ELISA using a crude adult Ostertagia ostertagi antigen to predict response to treatment also was evaluated. All lactating cows in 20 dairy herds were allocated randomly to receive either eprinomectin pour-on or placebo at calving. Information on reproductive parameters was obtained from computerised cow records. Survival models were used to evaluate the effect of treatment on the two intervals and a Poisson model was used to evaluate the number of services to conception. A total of 549 cows were included in these analyses. A marginally significant treatment effect on calving-to-conception interval was observed (hazard ratio=1.24, P=0.06) but not on calving-to-first service interval. A significant reduction in the number of breedings to conception for treated animals also was observed with a longer effect in cows with short interval to first service. Milk samples from a subset of 109 late-lactation cows were tested for antibodies against O. ostertagi. The ELISA optical-density ratio (ODR) values obtained between 120 days before calving and drying off were categorised as high ODR (>or=0.5) and low ODR (<0.5). Among untreated animals, the hazard of conception was lower (hazard ratio=0.38, 95% CI=[0.19,0.75]) for high-ODR cows compared to low ODR cows suggesting that higher parasite burdens had an adverse effect on reproductive performance. Treated high-ODR cows had a hazard of conception equivalent to the hazard for all cows in the low-ODR group (indicating that treatment prevented the negative effect associated with these higher parasite burdens).

Animals↗

Evaluation of the repeatability of a crude adult indirect Ostertagia ostertagi ELISA and methods of expressing test results.

An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against Ostertagia ostertagi using a crude adult worm antigen was evaluated using serum and milk samples from adult cows, as well as from bulk tank milk. Within and between plate repeatabilities were determined. In addition, the effects of factors such as antigen batch, freezing, preserving of the samples and somatic cell counts (SCCs) of the samples were evaluated. Raw optical densities (ODs) and normalized values were compared using the concordance correlation coefficient (CCC), the coefficient of variation (CV), Bland-Altman plots (BA). Based on raw OD values, there was a high repeatability within a plate (CCC approximately 0.96 and CV<10%). Repeatability between plates was evaluated following normalization of OD values by four methods. Computing normalized values as (OD-Nt)/(Pst-Nt), gave the most repeatable results, with the CCC being approximately 0.95 and the CV approximately 11%. When the OD values were higher than 1.2 and 0.3 for the positive and the negative controls, respectively, none of the normalization methods evaluated provided highly repeatable results and it was necessary to repeat the test. Two batches of the crude antigen preparation were evaluated for repeatability, and no difference was found (CCC=0.96). The use of preservative (bronopol) did not affect test results, nor did freezing the samples for up to 8 months. A significant positive relationship between ELISA OD for milk samples and SCC score was found. Therefore, the use of composite milk samples, which have less variable SCC than samples taken from each quarter, would be more suitable when the udder health status is unknown. The analytical methods used to evaluate repeatability provided a practical way to select among normalization procedures.

Animals↗

Identification of a novel cathepsin C mutation (p.W185X) in a Brazilian kindred with Papillon-Lefèvre syndrome.

Papillon-Lefèvre syndrome (PLS) is an autosomal recessive palmoplantar keratoderma caused by cathepsin C (CTSC) gene mutations. This study reports CTSC mutational and enzyme analyses in a consanguineous Brazilian family with PLS, representing the first enzymatic analysis of a Brazilian kinship with PLS. This family segregates a novel PLS-related mutation, p.W185X, that is associated with a complete loss of enzymatic activity.

Amino Acid Sequence↗

Penicillin acylase purification with the aid of pseudo-affinity chromatography.

The introduction of affinity chromatography has opened a new dimension in protein purification. This article reviews the current techniques used in the penicillin acylase purification process, especially pseudo-affinity chromatography. A profile for a suitable ligand is established. An aromatic ring and the presence of one or several amino groups seem essential for proper interaction. Immobilized metal affinity chromatography now seems to be a good competitor.

Chromatography, Affinity↗